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Open access peer-reviewed chapter
Recovering Ancient Grapevine Varieties: From Genetic Variability to In Vitro Conservation, A Case Study
By Carmina Gisbert, Rosa Peiró, Tania San Pedro, Antonio Olmos, Carles Jiménez and Julio García
Submitted: March 27th 2017Reviewed: September 20th 2017Published: December 20th 2017
DOI: 10.5772/intechopen.71133
Home > Books > Grapes and Wines - Advances in Production, Processing, Analysis and Valorization
A great number of varieties have been described in grapevine; however, few of them are currently in use. The increasing concern on varietal diversity loss has encouraged actions for recovering and preserving grapevine germplasm, which represents valuable resources for breeding as well as for diversification in grapevine-derived products. On the other hand, it is expected that this important crop, which is distributed in warm areas worldwide, will suffer the climate changes. Therefore, it is also convenient the identification of intravarietal variability and the recovery of accessions well adapted to particular environments. In this chapter, we will contribute to highlight the importance of recovering ancient materials, the usefulness of SSR markers to determine their molecular profile, the importance to analyze their virus status, and the possibilities that offer biotechnological tools for virus sanitation and in vitro storage as a complement of field preservation. In this context, we have evaluated different grapevine accessions and developed in vitro culture protocols for micropropagation, sanitation, and storage grapevine cultivars. In this work, we report the results obtained for the historic variety "Valencí Blanc" (or "Beba") and the historic and endangered variety "Esclafagerres" ("Esclafacherres" or "Esclafacherris").
Valencí Blanc
Esclafagerres
Esclafacherres
varietal identification
in vitro culture
chapter and author info
Carmina Gisbert*
Polytechnic University of Valencia (Universitat Politècnica de València), Spain
Rosa Peiró
Tania San Pedro
Valencian Institute of Agrarian Investigations (IVIA), Spain
Antonio Olmos
Carles Jiménez
Plant Health Service of the Generalitat Valenciana, Spain
Julio García
Conselleria of Agriculture, Environment, Climate Change and Rural Development, Spain
*Address all correspondence to: cgisbert@btc.upv.es
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Grapes and WinesAdvances in Production, Processing, Analysis and ValorizationEdited by António M. Jordão
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Grapevine (Vitis viniferaL.) is a crop of major economic importance distributed in warm areas worldwide [1] with a wine production of 2910 million hectoliters in 2014 and 75,866 square kilometers dedicated to grapevine culture [2]. The majority of the world's wine-producing regions are found between the temperate latitudes of 30 and 50° in both hemispheres [3]. Grapes are mainly used for making wine but also can be eaten fresh as table grapes or used for making jam, juice, jelly, grape seed extract, raisins, vinegar, and grape seed oil. Approximately 71% of world grape production is used for wine, 27% as fresh fruit, and 2% as dried fruit. In the VitisInternational Variety Catalog (VIVC; http://www.vivc.de/), supported by Biodiversity and the International Organization of Vine and Wine (OIV), there are around 24,500 accessions which include cultivars, breeding lines, and different Vitisspecies. Around 50% (12,679) of the varieties correspond to Vitis viniferaLinné Subsp. vinifera(or sativa), and 30% (7714) correspond to Vitisinterspecific crossing. Around 25% of the cultivars were registered in France (5602), followed by the United States (2401) and Italy (2348) with approximately 10% each one. Spain has registered a total of 734 varieties, being most of them (631) V. vinifera. According to Lacombe [4], a total of 1902 grape varieties (both scions and rootstocks) are officially authorized for cultivation in at least one country of the European Union. Around 65% of these grape varieties are registered only in one country, meaning the responsibility to preserve these varieties is too focused. On the other hand, four varieties ("Cabernet Sauvignon," "Merlot," "Chardonnay Blanc," and "Sauvignon Blanc") were maintained in at least 60 different institutions. Nowadays, not only these cultivars but also "Syrah" (or "Shiraz") dominates vineyards worldwide [5]. Considering that most major wine-producing regions could become by 2050 unsuitable for currently grown cultivars [6, 7], the preservation of genetic variability and the selection and/or development of cultivars well adapted to upcoming climate changes are important. The long juvenile period of grapevine makes breeding a slow process; therefore, the knowledge of the raw material and their availability is very important to speed up breeding programs.
1.1. Grapevine: gain and loss of diversity
Vitis vinifera subsp. viniferawas domesticated in the Neolithic period (ca8500–4000 BC) [8] from wild grapevines (Vitis viniferaL. subsp. sylvestris(Gmelin) Hegi) [9, 10, 11, 12]. Grapevine domestication appears to have occurred between the seventh and the fourth millennia BC, in a geographical area between the Black Sea and Iran [8, 13, 14, 15, 16], and the earliest evidence for large-scale winemaking was found in the North of Zagros Mountains and in the Caucasian region around 6000–5000 BC [17]. Cuttings of cultivated grapevines would have been spread by humans in the Near East, Middle East, and Central Europe. As a result, these areas may have constituted secondary domestication centers [18, 19] where spontaneous hybridizations among cultivars or local wild plants generated the pattern of admixture that is observed in current cultivars [19, 20, 21, 22, 23]. In consequence, genetic variability of grapevine has increased due to the contribution of different genetic pools in the process of grapevine spreading. The appearance of spontaneous mutations [24] and the selective pressure by humans which depended on the different uses of grapevine (fresh consumption, raisin, or wine production) [25] were also contributed to increase the genetic variability of this crop.
Along the years, genetic erosion has occurred in both cultivated and wild grapevines. Anthropogenic pressure on the wild natural habitats greatly decreased the wild grapevine populations that were also affected by the phylloxera aphid (Daktulosphaira vitifoliaeFitch) introduced from North America during the second part of the nineteenth century [26]. The phylloxera pest devastated the vineyards in all Europe. Since that time, grapevines need to be grafted onto phylloxera-resistant rootstocks. This fact reduced the number of grapevine cultivars used as scions which provoke genetic erosion [12]. On the other hand, the creation of denominations of origin (DO), each one including a reduced number of authorized varieties, has also contributed to reduce the varieties cultured in a specific area. Therefore, the preservation of grapevine minor cultivar and that on risk of disappearance together with Vitis vinifera subsp. sylvestrisis a major stake in grapevine preservation.
1.2. Grapevine preservation
The importance of germplasm preservation focused on their putative use, in the present or in the future. It is the source of genes to face new pathogens or climate constraints. Genetic diversity of grapevine is maintained normally as living plants in the field [27, 28]. Several important ex situ grapevine collections exist like "The Domaine de Vassal" in Montpellier (France), the "Julius Kühn Institute" in Siebeldingen (Germany), and "La colección de vides de El Encín" in the IMIDRA (Instituto Madrileño de Investigación y Desarrollo Rural, Agrario y Alimentario) center of Alcalá de Henares (Spain). The French collection houses 7800 accessions ca50 countries, representing 2300 different grape varieties, including wild species, rootstocks, hybrids, and mutants. Its transfer to the INRA Pech Rouge Experimental Unit (Gruissan, Aude) is under progress (https://www6.montpellier.inra.fr/vassal_eng/). The German collection of the Institute for Grapevine Breeding Geilweilerhof holds more than 3000 accessions of cultivars and wild species as well as important breeding lines. Beyond many others old and neglected cultivars from Germany, Switzerland, and Austria and rare germplasm from Eastern Europe can be found (https://www.julius-kuehn.de/en/grapevine-breeding/fields-of-activity/genetic-resources-and-information-centre-vine-and-wine/). The Spanish collection consists of 3532 accessions that are grouped into 852 rootstocks; 69 interspecific hybrids; 111 Vitisspp.; 1852 V. viniferavarieties, of which 1178 are for wine use and 674 for table use; and 648 V. vinifera sylvestris(http://www.madrid.org/coleccionvidencin/index.php?option=com_content&view=article&id=9&Itemid=2). In order to avoid the loss of the stored materials which are exposed to environmental disasters and pest attacks, the duplication of accessions for storage in different collections is a common strategy, although limited by budget constraints.
Another complementary strategy very useful in vegetative propagated plants is the in vitro preservation that offers the possibility to maintain plants under controlled and slow growing conditions and their micropropagation and transference to the field when need it. Although the first attempts to store grapevine under in vitro culture conditions were reported in the 1980s [29, 30, 31], this strategy is not usual in grapevine, although it is commonly used in other vegetative propagated crops like banana (preserved both through standard in vitro conditions and cryopreservation in the International Network for the Improvement of Banana and Plantain germplasm bank, in Leuven, Belgium). Cryopreservation is the storage of viable tissues, generally meristems or embryos, at ultralow temperature [32]. The success of in vitro conservation is tightly related to the choice of an adequate conservation method with the development of the corresponding methodology [33].
In grapevine, it is convenient to check by molecular markers that the variety to be stored really corresponds to it because homonymies (similar name for different cultivars) and/or synonymies (different names for a same cultivar) are commonly found. The identification of homonymies is important to avoid the loss of variability (loss of genotypes). On the contrary, the detection of synonymies avoids the maintenance of duplicated materials that do not contribute to increase variability but increase the cost. It is also very important to check the sanitary status of the plants, sanitize them if necessary, and provide suboptimal culture conditions that limit and slow down plant development, without causing physiological damage to the plant material. Grapevine can be infected by numerous viruses [34], and a high incidence of virus infection is commonly found in autochthonous cultivars [35, 36]. The EU Directive 2002/11/EC rules require that the initial plant material for vegetative propagation is free of Grapevine fanleaf virus(GFLV), Arabis mosaic virus(ArMV), Grapevine fleck virus(GFkV), Grapevine leafroll–associated virus-1(GLRaV-1), and Grapevine leafroll–associated virus-3(GLRaV-3). When samples to be storage are virus infected, different approaches to regenerated virus-free plants can be used. Since the middle late of the twentieth century, meristem culture and thermotherapy were applied with this aim for grapevine sanitation [37, 38]. Other techniques such as chemotherapy, electrotherapy, cryotherapy, and somatic embryogenesis were also reported [39, 40, 41].
For in vitro preservation under standard or limiting conditions, the development of protocols adjusted to the variety to be preserved is necessary. For this kind of storage, it is important to choose the adequate culture medium and environmental conditions in order to reduce the number of subcultures and hence minimize the cost and the putative errors that could arise in each subculture. Protocols for storage grapevine under in vitro culture have been reported by several authors [42, 43]. Recently, we reported the effectiveness of the MW medium to store a broad spectrum of grapevine cultivars, including endangered varieties, as well as the modifications of this medium (reduction of sucrose or elimination of indole-3-butyric acid (IBA) in the medium) to reduce growth in the faster growing varieties [44]. Cryopreservation protocols have also been developed for grapevine [45, 46, 47]. When developing cryopreservation protocols, the evaluation of the cryopreservation solution toxicity in the varieties to be preserved is required. Pre-culture in culture media that facilitate the dehydration of tissues (i.e., in medium with high content of sugars) is also a common step.
Finally, studies of genetic variability among varieties under conservation are important because they allow the detection of mutations and the study of relationships among them. The determination of molecular profiles is also of great importance in the development of core collections which represent the higher variability present in a whole collection in a reduced number of accessions. The core collections are very useful for breeders because studies on the core collection provide an overall view of the properties to be found in the whole collection [48]. In the following schema, the different steps to be carried out for germplasm storage are shown (Figure 1).
Scheme including the main steps required for germplasm storage.
2. Recovering ancient germplasm at the Comunitat Valenciana: the historic varieties "Valencí Blanc" and "Esclafagerres"
2.1. Richness of grapevine germplasm in the Comunitat Valenciana
The Comunitat Valenciana which includes the provinces of Alicante, Valencia, and Catellón is located in the Mediterranean coast of Spain and has been an important viticulture area since historic times to nowadays. For instance, in the archeological place of "L'Alt de Benimaquia" (Denia, Alicante), dated back to the end of the seventh century BC, significant quantities of vinification residues (tartaric acid and seeds) were found [49]. Important vineyards must be also located in the Requena-Utiel plateau (Valencia) as evidenced by the big presses dated in the fifth century BC found at Las Pilillas site [50]. Nowadays there exist three DO for wine production (DO Alicante (http://www.vinosalicantedop.org/), DO Valencia (http://www.dovalencia.info/), and DO Utiel-Requena (http://utielrequena.org/)), one DO for table grape (DO Uva de mesa del Vinalopó (http://uva-vinalopo.org/wp/)) and one protected geographical indication in Castellón (http://www.igpcastello.com/).
The richness in grapevine cultivars before the arrival to the phylloxera pest in the provinces of Alicante and mainly, in that of Valencia, is well documented [51, 52]. In a report about grapevine varieties cultured in Spain in 1889, it is pointed that more than 150 varieties were cultured in different locations of the Valencia province. These varieties included varieties with berries of black, white, and red color. In comparison with other provinces that also appeared in this report, Valencia was one of the richest [52]. The phylloxera aphids that devastated European vineyards invaded Spain in 1878 from three areas (Girona, Málaga, and Portuguese border). Its arrival to the Comunitat Valenciana, with the consequent loss of grapevine variability, occurred in 1912 when the aphids spread to Sagunto, Líria, and Requena [53].
In the context of the research project CGL2015–70843-R, we initiated different approaches in order to contribute in the recovering of ancient varieties in the risk of disappearance from Alicante and Valencia provinces. The objectives of this project include the analysis of grapevine germplasm diversity and the development of protocols for virus sanitation and in vitro conservation. Different prospections have been performed in order to rescue ancient varieties. The determination of SSR profiles is being useful to confirm the identity and to detect synonymies and homonymies that are very common in grapevine. The analysis of the genetic variability will identify accessions which may carry useful mutations for adaptation to specific environments. As occurred in other areas, grapevine cultivars were found commonly infected by the viruses GFLV, GLRaV, and GFkV. Sanitation of cultivars to be preserved in vitro is being carried out through meristem culture, although other alternative sanitation procedures are being developed [42, 54]. In this work, we report the SSR profiles of different accessions of the historic variety "Valencí Blanc" or "Beba" and the historic and endangered variety "Esclafagerres" (or "Esclafacherres/is"). The methodologies used for their sanitation and in vitro conservation are also summarized.
2.2. The historic varieties "Valencí Blanc" and "Esclafagerres"
The "Valencí Blanc" variety also known as "Beba" is a minor cultivar usually used as white table grape (Figure 2A). Despite in the past it was used for wine and raisin production [52, 55], today it is cultured as table grape for minor consume, and it is authorized for wine production in DO Ribera del Guadiana (Spain), where it is also named as "Eva" (http://riberadelguadiana.eu/esp/). The origin of this variety is unknown although it is proposed an oriental or North African origin [56]. The name "Valencí" ("Valensi" and "Balansí" in older reports) remembers to the name of Valencia City [57]. Oliver-Fuster (1980) cited by [58] proposed that this variety maybe was introduced by Balearic people who emigrate to Argelia. The most antique synonymy assigned for this variety is "Calop" [59] although other synonymies like "Ain el Kelp," "Tebourbi," "Panse the Provence" and "Grumer" are reported [56, 58, 59, 60]. In a report about grapevines cultivated in 1889 [52], the culture of "Grumer" is noted, among other 18 grapevine varieties in the Alicante province and in Albaida and Onteniente (locations nearest to the Alicante province). In the same report, the culture of "Valensí" in Alberique and Enguera (nearest to Valencia City) appeared. Recently, with the name of "Grumer," we have identified some accessions from the Alicante province that do not correspond to "Valencí" but grouped with "Muscat Istambul" [61]. Lacombe et al. [62] proposed as the origin of "Muscat Istambul" the cross of "Muscat of Alexandria" × "Valencí Blanc" which was confirmed by Mena et al. [63]. Therefore, it could be easy to found this homonym. In the VIVC database, 71 synonymies for "Beba" appeared. However, some of these ("Chelva," "Hebén," "Mantúo," "Teta de Vaca," and "Uva de Planta") were rejected as they did not share the same SSR profile [64]. Probably other of those proposed are also false synonymies. On the other hand, in the report about grapevine varieties cultured in Spain in 1889 [52], the variety "Valencí" was included in the groups of cultivars with black and white berries. The comparison among the SSR profiles of some accessions of "Valencí" with white grapes ("Valencí Blanc") and with black grapes ("Valencí Tinto" or "Valencí Negre" in the Comunitat Valenciana) indicates that they are not mutant for berry color but resulting from different crosses. Comparing the SSR profile of the variety "Heben" (or "Gibi"), proposed as parent of "Valencí Blanc," with the SSR profile of the accession of "Valencí Tinto" hold in the VIVC database, no relationship between them was observed. However, one or two alleles were shared between "Valencí Blanc" and "Valencí Tinto." Therefore, the unknown parent from "Valencí Blanc" could be the parent of "Valencí Tinto." The name of "Valensi Chaselas" also appeared in the report of grapevines cultured in 1889, concretely as cultured in the Valencian location of Gandía.
Grapes of "Valencí Blanc" (A) and "Esclafagerres" (B) varieties.
"Esclafagerres" variety (Figure 2B), which name means that which bursts the jars, is also an ancient variety with white berries commonly grown on the Alicante and Valencia provinces. Some old references that mentioned the culture of this variety in the Alicante province are reported by several authors [65, 66, 67]. In DGAIC [52], the culture of "Esclafagerres" appeared in the Alicante and in the Valencian locations of Albaida, Onteniente, and Sagunto, where it was included among the varieties with white and also with black berries. This variety was usually mixed with other grapevine varieties like "Merseguera" for wine production. The "Esclafagerres" variety gives high yields (probably the meaning of the name is related to this) and has grapes with low sugar content despite it was commonly cultured under dry land.
In this work, we report the assays performed with both varieties in order to determine their genetic profiles and resume the strategies performed for virus sanitation and in vitro conservation.
2.3. Determining the SSR profiles
A total of 14 samples of "Valencí Blanc" from different locations (Table 1) and two samples of "Esclafagerres" from La Mata and Monforte del Cid were used for DNA extraction and SSR analysis. DNA was extracted from fully expanded leaves using the commercial DNeasy Plant Mini Kit (Qiagen) according to the manufacturers' instructions. DNA quality and quantity were assessed using gel electrophoresis and spectrophotometry. Fifteen SSR markers (VVS2, VVMD5, VVMD6, VVMD7, VVMD24, VVMD25, VVMD27, VVMD28, VVMD32, VrZAG62, VRzAG64, VrZAG79, VrZAG83, and VMC1b11) were analyzed using two sets of multiplex PCR reactions. Each multiplex was carefully assembled according to the compatibility of the SSRs during PCR and the molecular size of their amplicons. The forward primer of the SSR markers was labeled with one of the four fluorescent dyes: carboxyfluorescein (FAM), carboxytetramethylrhodamine (TAMRA), hexachloro-6-carboxyfluorescein (HEX), or 6-carboxytetramethyl rhodamine (ROX) [61]. Multiplex PCR was carried out in a total volume of 11.00 μL, using 1.25 μL of commercial Master Mix PCR Multiplex (Takara Multiplex Hot Short PCR, Takara), 20–40 ng of genomic DNA, 0.1 μL of Takara Taq Hot Start, and labeled multiplexed SSR primers (from 5.5 to 35.0 μmol). The amplification was performed in an ABI 9700 thermocycler, and the amplification conditions were 95°C for 14 min followed by 30 cycles of 95°C for 30 s, 55°C for 90 s, and 72°C for 60 s and a final extension of 72°C for 30 min. Previous to PCR fragment size determination, the multiplex PCR product was previsualized using gel electrophoresis. The electrophoresis was carried out on an ABI 3100 platform (Applied Biosystems, Foster City, CA, USA). For PCR fragment size determinations, 0.13 μL of an internal size standard (GeneScan™ 500 LIZ, Applied Biosystems) was mixed with 1.00 μL of PCR product and 10.87 μL of formamide. The mixture was heated at 94°C for 3 min and then cooled in icy water. The size of the SSR fragments was determined with the software package GeneScan 3.7 (Applied Biosystems).
VVMD27
VVMD5
VrZAG83
VMC1b11
Vb-Pe0 Penàguila (Alicante) 179 187 233 237 136 144 197 197 242 246 189 205 134 158 241 247 206 208 254 270 255 255 185 189 243 257 209 211 248 254
Vb-Pe10 Penàguila (Alicante) 179 187 233 237 136 144 197 197 242 246 189 205 134 158 241 247 206 208 254 270 255 255 185 189 243 257 209 211 248 254
Vb-Be1 Benirarres (Alicante) 179 187 233 237 136 144 197 197 242 246 189 205 134 158 241 247 206 208 254 270 255 255 185 189 243 257 209 211 248 254
Vb-Al1 Alicante 179 187 233 237 136 144 197 197 242 246 189 205 134 158 241 247 206 208 254 270 255 255 185 189 243 257 209 211 248 254
Vb-FF1 La Font de la Figuera (Valencia) 179 187 233 237 136 144 197 197 242 246 189 205 134 158 241 247 206 208 254 254 255 255 185 189 243 257 209 211 248 254
Vb-FA1 Fontanars dels Alforins (Valencia) 179 187 233 237 136 144 197 197 242 246 189 205 134 158 241 247 206 208 254 254 255 255 185 189 243 257 209 211 248 254
Vb-On1 Ontinyent (Valencia) 179 187 233 237 136 144 197 197 242 246 189 205 134 158 241 247 206 208 254 254 255 255 185 189 243 257 209 211 248 254
Vb-Ba1 Fuente del Maestre (Badajoz) 179 187 233 237 136 144 197 197 242 246 189 205 134 158 241 247 206 208 254 254 255 255 185 189 243 257 209 211 248 254
Vb-Cu1 Campillo de Altobuey (Cuenca) 179 187 233 237 136 144 197 197 242 246 189 205 134 158 241 247 206 208 254 254 255 255 185 189 243 257 209 211 248 254
Es-Ma1 La Mata (Alicante) 187 191 233 237 146 152 193 197 242 246 189 197 134 136 237 247 206 208 254 270 241 255 171 189 233 247 209 211 248 248
Es-Mo1 Monforte del Cid (Alicante) 187 191 233 237 146 152 193 197 242 246 189 197 134 136 237 247 206 208 254 270 241 255 171 189 233 247 209 211 248 248
SSR profiles for 14 accessions of "Valencí Blanc" (Vb-Pe0, Vb-Pe1, Vb-Pe2, Vb-Pe3, Vb-Pe4, Vb-Pe5, Vb-Pe10, Vb-Be1, Vb-Al1, Vb-FF1, Vb-FA1, Vb-On1, Vb-Ba1, Vb-Cu1) and two of "Esclafagerres" (Es-Ma1, Es-Mo1) varieties.
Allele sizes are expressed as base pairs.
The SSR profiles of the analyzed accessions are shown in Table 1. Whereas similar SSR profiles were found for both accessions of "Esclafacherre," some variability is found among "Valencí Blanc" accessions. Among the 15 SSRs analyzed, differences in the SSR VVMD32 were found: the accessions collected in the province of Alicante have two alleles in this loci VVMD32 (254; 270), whereas the rest of accessions from the provinces Badajoz, Cuenca, and Valencia have the allele 254 in homozygosity. We can consider that among the analyzed germplasm, there are two variants of "Valencí Blanc" as in other cases in which two plants showed identical SSR profiles for all the SSR markers studied except for one or two alleles. This could be attributable to slight clonal polymorphism [68]. They may have originated in a similar place and then spread to different areas. The accession of "Valencí Blanc" in the VIVC database (accession number 22710) and that reported by Lacombe et al. [62] had the same profile for the comparable SSRs (including the VVMD32) to the accessions from Alicante. Similarly, accessions from Alicante showed also identical SSR profile to two accessions of "Beba" from El Encín grapevine collection analyzed by Mena [64].
The comparison of the SSR profile of the "Esclafagerres" accessions to SSR profiles in the VIVC database (including 3265 accessions), those in the International Vitis database (including 3430 accessions), as well as those reported in several publications [62, 63, 69] did not match with any of the included varieties. No matches were found when the SSR profile of "Esclafagerres" was blasted to the Italian Vitis database. Therefore, this profile should correspond with that of the "Esclafagerres" variety which has not been reported before.
2.4. Virus analysis and sanitation
To analyze the putative virus infection in the original samples, the methodology described by López-Fabuel et al. [70] was used. Briefly, extracts were prepared from leaves 1/20 in phosphate buffered saline (PBS) buffer, pH 7.2, supplemented with 0.2% diethyldithiocarbamic acid (DIECA) and 2% polyvinylpyrrolidone-10 (PVP-10) in individual plastic bags with a heavy net (Plant Print Diagnostics). Total RNA was extracted from 200 μL of crude extract using an Ultraclean Plant RNA isolation kit (Mobio) following the manufacturer's instructions. The real-time multiplex reverse transcription polymerase chain reaction (RT-PCR) was performed for the simultaneous detection of ArMV, GFLV, GFkV, GLRaV-1, and GLRaV-3 using a StepOnePlus thermocycler (Applied Biosystems) and a reaction mixture containing 1× AgPath-ID One-step RT-PCR buffer (Ambion) and 1.5× AgPath-ID One-step RT-PCR enzyme mix (Ambion); 5 μL of sample; 400 nM of GFLV, ArMV, GFkV, and GLRaV-1 primers; 800 nM of GLRaV-3 primers; and 200 nM of each probe. The amplification protocol consisted of an RT step at 45°C for 10 min and a denaturation step at 95°C for 10 min, followed by 45 cycles of amplification (95°C, 15 s; 50°C, 15 s; and 60°C, 60 s). As positive controls viral isolates maintained at the Instituto Valenciano de Investigaciones Agrarias (IVIA) were included. When amplification was observed for a specific virus, it was confirmed by real-time uniplex RT-PCR using the corresponding primers.
Meristem culture was used in a previous project (RTA2011–00067-C04) to obtain virus-free plants of "Esclafagerres." In the context of the project CGL2015–70843-R, "Valencí Blanc" (sample Vb-Pe0), which resulted in infection with GFkV and GLRaV-3, was sanitized through both meristem culture and somatic embryogenesis. Meristems (n = 35) from plants of "Valencí Blanc" were extracted using a binocular lent and cultured in vitro on plates (90 × 15 mm) containing the medium MW, selected for "Monastrell" micropropagation [71] supplemented with 6-benzylaminopurine (BAP) at 1.8 μM. Low light conditions were used for the two first weeks of culture. Only 54.3% of meristems grew after 20 days of culture (Figure 3A), and two develop into plants after transferring to tubes with MW, 70 days after (Figure 3B). Damage of meristem during extraction and/or the composition of the culture medium that could need to be enriched with other nutrients is putatively the cause of the low and slow regeneration of meristems. The analysis for virus presence of these two plants was carried out as described before, and one of them (50%) resulted free for both viruses. Therefore, from this plant, clones were obtained for in vitro conservation. Despite the fact that meristem culture is an efficient technology for virus sanitation, it is needed to obtain an adequate size of the meristem in order to avoid virus transmission allowing meristem development.
Strategies for virus sanitation of "Valencí Blanc" (accession Vb-Pe0). (A) Meristems cultured on MW supplemented with 6-benzylaminopurine (BAP) at 1.8 μM and without indole butyric acid (IBA) after 20 days of culture. (B) Plants from meristems cultured in MW medium after 90 days of culture. (C) Cut seeds cultured in EIM2 medium at day 0. (D) Seeds with some somatic embryos after 45 days of culture. (E) Plants regenerated from somatic embryos.
The other methodology used for virus sanitation was the induction of somatic embryos as reported in Peiró et al. [41]. Briefly, seeds of "Valencí Blanc" were extracted from grapes and disinfected and cut previously to be cultured on the embryogenesis induction medium (EIM2) which contained TDZ (thidiazuron) at 0.9 μM. Thirteen percent of explants responded after 60 days of culture on this medium (Figure 3C–D). Despite in grapevine a high percentage of somatic embryos are not able to develop into normal plants [72, 73, 74], in our work, germination of somatic embryos occurred directly in the induction medium and grew correctly (Figure 3E). Ten developed plants were analyzed for evaluating the success of virus cleaning. All plants regenerated from somatic embryos resulted virus-free for both GFkV and GLRaV-3 viruses. Therefore, 100% of sanitation was obtained. The result obtained for GFkV was expected because this virus is not seed transmitted [75]. With respect to GLRaV-3, it is not clear if it is present in seeds [34, 75], but we have found this virus in some regenerated plants resulting from somatic embryos induced from seeds of other infected grapevines, which would indicate its presence in the seeds [54]. Induction of somatic embryos from stamens or pistils was also reported in grapevine to cure plants of GLRaV [76, 77]. We also analyzed the SSR profile of regenerated virus-free plants in order to select those regenerated from mother tissues of the seeds, which will show the mother genotype. The 15 SSRs used for determining the SSR profile of "Valencí Blanc" accessions were used. One of ten analyzed plants showed the same SSR profile as the mother plant, that is, a 10% of regenerated plants were obtained from mother tissue and not from the embryo.
2.5. In vitro storage
Sanitized plants of "Valencí Blanc" and "Esclafagerres" are maintained in tubes with MW medium in an in vitro culture growth chamber under standard conditions (25± 2°C; 16 h light). The MW medium is adequate to storage a broad spectrum of grapevine cultivars including "Valencí Blanc" and "Esclafagerres" [44]. Both cultivars grew less than 4 cm after 40 days of culture in this medium. We consider that this speed of growth is acceptable to maintain these cultivars under standard conditions with small number of subcultures. A reduction of sugar or the elimination of IBA in the culture medium is used for maintaining cultivars that grew faster [44]. The higher the number of subcultures, the higher the cost and the higher the possibility to make nomenclature errors [78].
Another strategy to germplasm ex situ storage using in vitro culture is cryopreservation. With this methodology, the metabolism is greatly reduced, and few requirements are needed for the maintenance of tissue samples. Meristems of "Esclafagerres" from micropropagated virus-free plants maintained in vitro were used to initiate the cryopreservation assays using the methodology described in Gisbert et al. [79]. The first results indicated that 50 min of incubation in the plant vitrification solution 2 (PVS2) is adequate for recuperating cryopreserved meristems of "Esclafagerres" (Figure 2). Recently, Pathirana et al. [47] have reported a positive effect on grapevine regeneration when a pretreatment with salicylic acid was performed prior to cryopreservation. In both works [47, 79], the droplet vitrification protocol was used.
As a result of different actions performed in the context of the projects CGL2015-70843-R and RTA2011-00067-C04, a broad spectrum of grapevine varieties are being evaluated in order to determine the varietal identification and their variability and also their capacity for in vitro culture, plant regeneration, and germplasm storage. Different strategies for virus cleaning have been developed and applied to rescue virus-free plants. Among the analyzed materials, the historic varieties "Valencí Blanc" and "Esclafagerres" were sanitized and currently are maintained under in vitro culture conditions. Differences for the microsatellite VVMD32 were found among "Valencí Blanc" accessions, clustering the accessions from Alicante and that of other origins. The SSR profile for the variety "Esclafagerres" was firstly reported in the present work.
The authors thank the support of the projects CGL2015-70843-R and RTA2011-0020067-C04 both co-funded by FEDER Funds.
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2.Recovering ancient germplasm at the Comunitat Valenciana: the historic varieties "Valencí Blanc" and "Esclafagerres"
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Carmina Gisbert, Rosa Peiró, Tania San Pedro, Antonio Olmos, Carles Jiménez and Julio García (December 20th 2017). Recovering Ancient Grapevine Varieties: From Genetic Variability to In Vitro Conservation, A Case Study, Grapes and Wines - Advances in Production, Processing, Analysis and Valorization, António Manuel Jordão and Fernanda Cosme, IntechOpen, DOI: 10.5772/intechopen.71133. Available from:
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Grapes and Wines
Edited by António M. Jordão
Fingerprints of Anthocyanins and Flavonols in Wild Grapes (Vitis vinifera L. ssp. sylvestris (Gmelin) Hegi)
By Eugenio Revilla, Rosa Arroyo-Garcia, Alberto Bellido, David Carrasco, Anna Puig and Leonor Ruiz-Garcia
Advances in Grape and Wine BiotechnologyEdited by Antonio Morata
Advances in Grape and Wine Biotechnology
Edited by Antonio Morata
First chapter
The Microvine: A Versatile Plant Model to Boost Grapevine Studies in Physiology and Genetics
By Anne Pellegrino, Charles Romieu, Markus Rienth and Laurent Torregrosa
We are IntechOpen, the world's leading publisher of Open Access books. Built by scientists, for scientists. Our readership spans scientists, professors, researchers, librarians, and students, as well as business professionals. We share our knowledge and peer-reveiwed research papers with libraries, scientific and engineering societies, and also work with corporate R&D departments and government entities.


This book examines the transformation of the state in Central and Eastern Europe since the end of communism and adoption of market oriented reform in the early 1990s, exploring the impact of globalization and economic liberalization on the region's states, societies and political economy. It compares the different policies and national strategies adopted by key Central and Eastern European states, including the Czech Republic, Poland, Hungary and Slovakia, showing how initial internally oriented strategies of market reform, privileging domestic sources of investment, had by the late 1990s given way to externally oriented strategies emphasising the promotion of competitiveness by attracting foreign investment. It explores the reasons behind this convergence, considering the influence of internal and external forces, and the roles of interests, institutions and ideas. It argues that internationalization of the state is forged in the processes through which domestic groups linked to transnational capital attain domestic influence necessary to shape state policy and strategy. These groups — the comprador service sector in particular — constitute and organize political, social and institutional support of the competition state in the region. Overall, this book not only provides a detailed account of the political economy of post-communist transformation in Central and Eastern Europe, but also the processes by which states adapt to the forces of globalization.
Jan Drahokoupil is a research fellow at the Max Planck Institute for the Study of Societies in Cologne. He obtained a PhD at the Central European University in Budapest. His recent publications include European Neoliberal Governance and Beyond (co-edited with Bastiaan van Apeldoorn and Laura Horn).


VLEs or virtual learning environments have been at the forefront of learning technology provision for the first part of the 21st century in the guise of Webct, Blackboard and Moodle just to name a few, and have formed the back bone of institutional 'Learning Management Systems' (LMS).
In more recent times however the dominance of the VLE and its possible future role or indeed "survival" as been put into question. This was best illustrated by the ALT-C Conference (2009) entitled "Is the VLE Dead?" during which some panel members argue that many VLEs are not fit for purpose and VLEs provide a negative experience and believing that the VLE is dead and that personal learning environments (PLE) is the solution and institutionalized VLEs are slow to respond to change.
At the same conference other members of the panel sort to defend the VLE so indirect contrast Wheeler's, James Clay argues that "the VLE is not a dead concept, but a starting point of the journey" and Clay consolidates this in his BlogSpot (Clay, 2009) by stating that he believes "most learners do not know how to use web 2.0 technology effectively" this being the basis of a PLE, and "it is not dead yet!".
In conclusion what I believe is needed is a structured and intuitive system, flexible in its use and useable by the students and personalisation is the key but embedded with in VLE structure. VLEs have the capacity within the formal structure to personalize learning by embedding social and collaborative learning with use of for example Twitter and Wikis to mention a few.
Bloxham & al, e., 2007. Summary Report on the UKCLE/BILETA VLE Project. Journal of Information Law and Technology, p. 8.
Dillenbourg, P., 2000. Virtual Learning Environments, Geneva: TEFCA.
Stiles, M., 2007. Death of the VLE?: A challenge to a new orthodoxy.. Serials, pp. 31-36.
Vogel, M. & Oliver, M., 2006. Designing for learning in Virtual Learning Environments – insider perspectives, London: Centre for Excellence in Learning Technology.


text:
July 29, 2017 February 19, 2020 historyradio
"Bruce Lee – An Ethnic Role Model" by Josef Tzegai Yohannes
Josef Tzegai Yohannes (b.1980) is the creator of the The Urban Legend comic books. Inspired by Bruce Lee and Nelson Mandela, the books are available in over 8 languages.
What does Bruce Lee mean to the world, or maybe I should ask myself what does he mean to me? Bruce Lee means a lot of things to a lot of people around the world and that is why he is an icon today across the globe. His legacy has grown bigger and bigger for each year and people are still mesmerized by his skills, charisma and determination that shined so bright throughout his short life.
When I was growing up as a kid in Denmark and Norway, I had very few role models I could relate to, and the moment I saw Bruce Lee in action on the big screen for the first time, I was just so captivated by his persona, what he stood for and how on earth he could beat 50 guys at the same time. But Bruce showed it was possible and how I could be done and led the way. He was the first real martial artist action star the world had ever seen, there was no one before him and there hasn't been anyone bigger than him after he passed away in 1973 at the age of 32. He put martial arts on the map and gave it a face and a purpose and in many ways made it universal. So many of todays action stars like Jean Claude Van Damme have said that after they saw Bruce Lee in Enter the Dragon, they wanted to become an action star. Bruce defined and changed a whole genre, came up with his own martial art style (Jeet Kune Do) and left a mark on the world that can never be replaced. He also helped change the way Asians was looked upon in movies.
I remember when I was 8 years old, living in Copenhagen being at an Eritrean party. I had helped a friend of mine earlier that day so he gave me two Bruce Lee postcards as a gift, it was the famous picture of Bruce Lee holding his nun chakus. Unfortunately that night I have had a lot of soft drinks, so I had to go to the bathroom, so I put the postcards underneath my seat and rushed to the bathroom and when I got back my postcards where gone. I was so angry, that I almost started to cry and I walked up to everyone at the party asking them if they had seen my Bruce Lee postcards, but no one has seen them. I was devastated and my parents tried to comfort me by saying don't worry, something bigger and better will come one day.
A few days later a friend of mine came to cheer me up with a Bruce Lee movie called "Game of Death," which I had never seen before. The moment I saw Bruce Lee in his famous yellow and black tracksuit in the movie, I feel in love with it right there and then. It looked so amazing on him. I begged my parents to go and buy that tracksuit, so I could practice martial arts and be just like him. So we went to almost every store that were in Copenhagen looking for that famous tracksuit to no avail. No store had it and this was way before E-bay.
When I got the idea to create my own superhero called The Urban Legend, I named him after Malcolm X, my father Tzegai and Nelson Mandela, but I also put some reference to Bruce Lee in it. When I came to the stage where I had to get my superhero his own suit, Bruce Lee´s famous yellow and black tracksuit instantly came to mind, so I choose that as a tribute to Bruce Lee (and for making up for a lost childhood). In one issue of The Urban Legend I have used a Bruce Lee quote and you see Bruce Lee posters hanging on the wall of the superhero.
To me Bruce Lee symbolizes power, love, strength, self-improvement, dignity and all the good values we want see in our fellow human beings.
Josef Tzegai Yohannes
historyradio
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BusCon offers budget-conscious bus operators extraordinary value, with a full conference pass ranging from $115 to $150, which includes two networking receptions, educational programming, the exhibit hall, and more.
With the idea of mobility evolving and impacting all facets of bus transportation, BusCon 2018 will feature all of the latest vehicles and technology, as well as host discussions on some of the hottest topics, including autonomous vehicles and electric bus usage, when it makes its return to Indianapolis' Indiana Convention Center Oct. 1 to 3. Registration at the event begins Monday, Oct. 1 from Noon to 5 p.m. at the convention center.
In addition to taking a look at end-user usage of alternative-fueled and battery-electric vehicles, the conference will also address autonomous vehicles, including how autonomous shuttles are being used and what the future for the technology may look like as it matures, as well as the positive impact it can have on paratransit users.
As the idea of partnering grows to better provide transportation options for increased mobility and so-called complete trips, the "Forging Partnerships to Improve Mobility and Boost Ridership Session" will host a discussion on microtransit, as well as take a look at how developing partnerships can help effectively grow an operation's paratransit coverage.
New for this year, the National Transit Institute will host two training sessions — "Overview of Transit Procurement for Senior Leadership" and "Transit Asset Management Implementation for Tier II Providers and Sponsors" — that are available for the first 50 attendees to RSVP for each. For those interested in Lithium-ion battery technology, EnerDel will host two training courses, which will provide an overview of electrochemistry and discussions on Li-ion characteristics, life expectancy, and cell components, as well as designs that make for effective and safe battery solutions.
The United Motorcoach Association will also return to BusCon with two sessions on Monday, Oct. 1, including a look at how to expand your bus operation through diversification, which is ideal for those entering the motorcoach industry or for operators looking to get into using smaller-sized vehicles.
Other topics include how to use social media to sell your company; preparing your operation for pandemics such as this past flu season; and how the evolution of bus safety technologies will impact the industry both now and in the future.
Technology, safety, electric bus charging, and more will be discussed at this year's Transit Maintenance Forum (TMF) for senior-level transit maintenance professionals, which returns to Indianapolis in conjunction with BusCon.
TMF offers the chance to learn about some of the latest trends faced in maintenance shops, affording transit professionals the unique opportunity to network and share challenges with peers and keep up-to-date with the newest vehicles, technologies, and products on the market today.
This year's TMF will focus on the latest in technology, including "How A.I. is Helping Manufacturers Build More Reliable, Efficient Buses" session, which will look at how manufacturers are using Big Data analytics and Artificial Intelligence algorithms to build better buses, while also allowing fleets to prevent breakdowns and minimize maintenance costs. Also on tap, John Hronich, from BAE Systems, will discuss the benefits that can be derived from having all-electric accessories on diesel and CNG buses, while Carl Harr, from Liquid Spring LLC, will talk about the company's revolutionary technology, which can drastically improve the smoothness of a bus's ride.
Meanwhile, Bus Maintenance Consultant and well-known Industry Expert Halsey King will review a number of fascinating facts about bus fires and give participants insight into the work of a fire investigator in hopes of preventing bus fire situations at their operation, during his "Bus Fires: Stop them Before they Start" session.
As more and more transit agencies move to all-electric fleets, TMF also includes the "Considerations for Creating a Long-Term Electric Bus Charging Plan" session, which will focus on best practices for upgrading infrastructure and take a look at how two transit agencies are tackling the challenge.
John Schiavone of the Transportation Learning Center will also examine a national effort by the center and the U.S. Department of Labor to establish a registered apprenticeship program specifically for transit bus technicians.
For an economical $115, attendees have the opportunity to not only attend TMF, but also all of BusCon's networking events and the Keynote Address, as well as visit the extensive show floor.
In addition to taking a look at end-user usage of alternative-fueled and battery-electric vehicles, the conference will also address autonomous vehicles.
"North America's Biggest Bus Show" offers the largest quantity and widest variety of vehicle and products on the show floor, giving attendees the chance to see more than 60 vehicles, from shuttles to motorcoaches and from public transit buses to vans, including electric, propane, and hybrid vehicles. The show floor will be open Tuesday, Oct. 2 from Noon to 5 p.m. and Wednesday, Oct. 3 from 11a.m. to 2 p.m., with attendees able to use BusCon's mobile app, which is available via iTunes or Google Play, to navigate the show.
Platinum Sponsor Forest River, a Berkshire Hathaway Co., and its six bus brands — Battisti Customs, Berkshire Coach, Elkhart Coach, Glaval Bus, Starcraft Bus, and StarTrans Bus — will mark its return to BusCon with a nearly 7,000-square-foot booth showcasing a range of vehicles suitable for a variety of public and private sector transportation providers.
Other manufacturers scheduled to display vehicles include Alexander Dennis, ARBOC Specialty Vehicles, Blue Bird Commercial, BYD, Complete Coach Works, Embassy by SVO Group, Ford Commercial Vehicles, Freightliner Custom Chassis, General Motors Fleet, Karsan/Morgan Olson, Mercedes-Benz Vans, MobilityTRANS, Motor Coach Industries, New England Wheels, New Flyer, Ram Trucks, Thomas Built Buses, XL, and more.
In addition to vehicles, many other suppliers and technology providers will be on hand to showcase all the latest solutions on the market, with more than 30 new companies taking part in this year's BusCon.
In addition to vehicles, many other suppliers and technology providers will be on hand to showcase all the latest solutions on the market, with more than 30 new companies taking part in this year's BusCon. Some of those companies include technology solutions provider ViriCiti LLC; bio-decontamination system provider CURIS System; and bus-wrap company Turbo Images.
Additional new suppliers, include AM Equipment, Creative Carriage Ltd., VIS Check, and Vehicle Technical Consultants.
Attendees will also not want to miss the BusCon Breakfast Awards Ceremony on Wednesday, featuring the return of METRO's Innovative Solutions Awards, honoring operations and their supplier partners who launched new programs that positively impacted their business. New to the program is the Transit Maintenance Director of the Year Award, which will honor two directors who have gone above and beyond to create a well-run maintenance shop. Also on hand to present awards will be the Propane Education & Research Council, who will honor operators out in front in the use of propane-fueled vehicles.
For more information and to register, visit www.BusConExpo.com or call (800) 576-8788.


text:
from __future__ import print_function
from time import localtime, mktime, strftime
import email.utils
import feedparser
from twython import Twython, TwythonError
from docopt import docopt
import conf
from setting import FILE, URL

__usage__="""
Usage:
    keibanews-bot.py tweet [--user=<account>] [--no-tweet]

Options:
    -h --help           Show this screen
    --version           Show version
    --user=<account>    tweet user [default: keiba_news]
    --no-tweet          tweet to stdout

"""

def file_open(f):
    try:
        with open(f) as fp:
            return fp.read()
    except:
        pass

def parse_feed(u):
    data = feedparser.parse(u)
    feed = []
    for entry in data.entries:
        feed.append([entry.published, entry.title, entry.link])
    feed.reverse()
    return feed

def tweet_news():
    # OAuth setting
    args = docopt(__usage__)
    config = conf.ParseConf()
    oauth_conf = config.get_items('oauth_conf')
    if args['--user']:
        access_conf = config.get_items(args['--user'])
    twitter = Twython(
    oauth_conf['consumer_key'],
    oauth_conf['consumer_secret'],
    access_conf['access_token'],
    access_conf['access_secret']
    )
    feed = parse_feed(URL)
    latest = file_open(FILE)

    with open(FILE, 'w+') as fp:
        for entry in feed:
            _date, title, url = entry[0], entry[1], entry[2]
            entry_date = strftime('%Y/%m/%d %H:%M',localtime(mktime(email.utils.\
                                                        parsedate(_date))+32400))
            if not latest or entry_date > latest:
                post = u'{0} {1} {2}'.format(entry_date, title, url)
                if args['--no-tweet']:
                    print(post)
                else:
                    try:
                        twitter.update_status(status = post)
                    except TwythonError as e:
                        print(e)
                latest = entry_date
        fp.write(latest)

if __name__ == '__main__':
    tweet_news()

redpajama_set_name: RedPajamaGithub


If you want to know whether the website is as per the recommendations of SEO documents then SEO Doctor an extension in Mozilla Firefox is the right tool to analyse the website based on Google webmaster, Image, SEO starter guidelines. It will analyse for any problems and provides the score for your webpage.
There are different flag indication in the status bar to indicate the SEO Score. If Green Flag everything is fine. Yellow flag represents noncritical warning & Red flag indicates some of SEO elements can be improved better.
It has other features like Links which shows the no of external links, Visits which displays the visitors count, the Backlinks which shows the backlinks counts for the domain.
SEO Doctor analyses for Meta Description which is used to describe the page still used by some of search engines to display in search engines, how Header Tags are used in the webpage,Analyse For ALt Image tag which is important for image search,get details on Inbound links & checks for SEO friendly URL's whether link is static or dynamic.It also analyses for Page loading time which is important for reader as well as page rank.
A really awesome extension to use. Maybe you'd like to try it out?
Thanks Abdul. Will be interesting to learn every new tools. Will try out SEOQuake.
Hi Vipin, There is always a new learning in every tool or plugins. Try out this. Thanks for your comments.
I use SEO doctor. It is a great addons.


text:
Q: Jquery find number of table rows with specific values in 2 columns I have a table with 2 columns "id" (unique value) and "name". Using jQuery I would like to return the number (.length) of rows where the ID matches an array of ids (e.g. SR1, B1, B2) AND the value in the "name" field for those rows is empty.
So far I have:
var empty = $('[tabulator-field="name"]:empty').length;
var sr1 = $('.tabulator-row:nth-of-type(29) [tabulator-field="name"]:empty').length;
var sr2 = $('.tabulator-row:nth-of-type(30) [tabulator-field="name"]:empty').length;
var b1 = $('.tabulator-row:nth-of-type(7) [tabulator-field="name"]:empty').length;
var b2 = $('.tabulator-row:nth-of-type(8) [tabulator-field="name"]:empty').length;
var c10 = $('.tabulator-row:nth-of-type(27) [tabulator-field="name"]:empty').length;
var c11 = $('.tabulator-row:nth-of-type(28) [tabulator-field="name"]:empty').length;
var emptySR = sr1 + sr2 + b1 + b2 + c10 + c11;
var emptyBeds = empty - emptySR;

However, this seems like an inefficient way to calculate this and furthermore, selecting the rows I am interested in by their position in the table (rather than by their ID values) using :nth-of-type returns incorrect results if the table is sorted by a different field as the order of rows will change.
Please can anyone suggest a robust function for achieving what I need?

A: Use a selector to match the empty field, then use a filter function to test if the ID is in the array.
var bedIds = ['SR1', 'SR2', 'B1', 'B2', 'C10', 'C11'];
var emptyBeds = $('.tabulator-row:has([tabulator-field="name"]:empty) [tabulator-field="id"]').filter(function() {
    return bedIds.indexOf(this.textContent) != -1;
}).length;

redpajama_set_name: RedPajamaStackExchange


Why is my dog aggressive towards other dogs? You may expect your dog to get along with other dogs as if they have some special bond. However, you may find out that your dog is aggressive towards other dogs. Does your dog simply not like some dogs just like not all people like each other? Is this type of aggression normal? This article can hopefully answer those questions and more.
Aggression between dogs is somewhat normal in the fact that it does happen. However, you don't want to allow your dog to be aggressive. It can actually cause problems for not only your dog, but you. If your dog is overly aggressive it may start fighting with other dogs. Dog fights can lead to injuries and possibly death. You don't want your dog to get injured and you don't want to be responsible another dog's injuries.
Aggression in dogs is usually growling, snarling lips, raised back or hair, biting, etc. What causes dogs to be so aggressive? Dogs such as pit bulls may have aggression in their genes since many are bred to fight. Some dogs are very happy and nice, but act aggressive when they feel like they need to protect their house or family. Dogs that are trying to establish their place on a type of dog hierarchy may be aggressive towards others to prove their dominance. Other cases of aggression in dogs are actually the result of pain, discomfort, and illness.
Take your dog to a veterinarian if they are behaving aggressively. You want to rule out any medical problems. If it is a medical issue catching it early often helps and treatment options may be available. If the aggression is a behavioral issue, you should look into getting a professional dog trainer or researching things that you can do to train your dog to stop its aggressive ways.


Now more than anytime in the past 100 years, coastal Georgia stands at a crossroads.As we see thousands of acres of land being developed every year in the eleven counties of this region, more and more people are questioning the accumulating consequences. Public opinions, both formally surveyed and casually observed, express deep concern that the landscape, environmental quality, and character of the coast are slipping away.
Many share the sense that our region is at the mercy of external forces that no authority can control, and which are perhaps inevitable. As coastal Georgia's economy has flourished over past decades, we now seem to be laboring under the momentum of obsolete goals, or at least negligent assumptions about the nature and benefits of growth, and only vague understanding about the choices available to us in moving toward a consciously determined future.
Despite noble intentions, after more than a decade of planning efforts under the Georgia Planning Act most cities and counties continue to passively accommodate, if not welcome, virtually anyone who proposes to build a new project. Decisions about land use and public facilities are shaped by no long-term plan, guided by no vision for our communities or region. Although some may claim that these concerns are unfounded, held by only a margin of environmental extremists, the record suggests otherwise. Consider that three-quarters of the DNR marsh and shore permits appealed over the past twelve years were filed by property owners and developers, not by environmental groups.
Disputes over the use of natural resources, delays caused by permitting decisions and their legal challenge, and conflicts between property-rights advocates and defenders of public interest are yet further indication that we are in a period of crucial change. The future of coastal Georgia could be irreversibly altered through critical decisions made (or not made) in the next 3 to 5 years. Can we afford to continue passively neglecting this challenge by yielding to change on a case-by-case basis, allowing others to determine our region's fate by default? Instead, we urge creating and implementing a strategy that galvanizes public policy, political resolve, and entrepreneurial spirit to work toward a shared regional vision served by a multitude of individuals devoted to complementary objectives.
If we are to formulate a successful regional strategy, it must be based not only on general public consensus, but also informed understanding of the scope and nature of the problems being faced by our region. This report is a first attempt to provide an overview of the trends of coastal Georgia's growth, environmental conditions, and implications for the future. Although imprecise and incomplete by virtue of the data and resources available, the report's findings and recommendations are instructive. The State of the Coast Report presents information and analysis to stimulate public dialogue and raise awareness about critically important issues affecting coastal Georgia and its five watersheds. These issues must be responsibly resolved if we are to build the consensus and commitment needed to shape our shared future, using means that are within our grasp. We urge all concerned citizens, decision-makers, and other stakeholders to review and comment on our report.
For nearly a century following the Civil War, coastal Georgia suffered economically and socially. Between Reconstruction and World War II, this region's income, employment, education, and general prosperity lagged behind most of rest of the nation by a large margin. By the early 1950's a number of factors converged to reverse the historical trend, but these took several more decades to achieve significant effect.
Beginning in the 1930s, pulp and paper manufacturing research had developed a process for using the more resinous but faster growing southern pine as the main source of fiber in making paper, which had been infeasible until then. This opened unprecedented opportunities for forest products in the southeastern United States, and along with that demand came thousands of jobs in forestry and manufacturing.
Land and labor were cheaper in the South and energy costs were lower for expanding industry.
Domestic military facilities at Hunter (Chatham County), Fort Stewart (Liberty, Long, and Bryan Counties), and Kings Bay (Camden County) provided great economic stimulus. From 1950 to 1990 military and civilian defense jobs in the region grew by more than 70%.
Federal civil rights legislation gradually improved living conditions and economic opportunities for African Americans from the South, who returned to their families' native state in vast numbers.
Extensive improvement of the federal highway system and federal subsidy of state roads resulted in far greater mobility, in combination with vastly expanded trucking transport. Both within the state and beyond, this brought together people and markets with advanced flexibility and speed.
As international trade grew, demand for ports sustained commerce in coastal areas. With the advent of containers used in shipping and thriving demand for foreign-made products, the two coastal Georgia ports at Savannah and Brunswick enjoyed unprecedented growth in both volume and value of commodities handled. World demand, U.S. trade and agricultural policy, and farming productivity also favored export of Georgia's rural products.
The Sunbelt phenomenon took hold as air-conditioning became more affordable while millions of Northerners reached retirement age, and as expanding businesses moved south. No other urban area epitomized the impacts of these factors more than Atlanta, where metropolitan population grew from less than a half million in 1970 to more than 3 million by 2000. And as Atlanta became the economic "engine" of the state, it had enormous trickle-down effect on the rest of Georgia - both favorable and adverse.
Coastal growth rates nationally have outstripped other geographic areas, and (with the exception of Atlanta's metropolitan area) Georgia is no exception. Fully 60% of the nation's population lives within 50 miles of an ocean or one of the Great Lakes, and that portion of total population is projected to grow as high as 80% within 20 years.
The national transition to a service-based economy has considerably supplemented the economy and growth prospects of coastal Georgia, which thrives on a tourism industry valued at more than $2 billion annually. Many service businesses also depend on labor markets that fit this region's profile, and these jobs are more than compensating for the loss of manufacturing employment.
All of these trends compounded and interacted to produce remarkable changes for coastal Georgians in the half century from 1950 to 2000. As beneficial as many of these changes have been, they are not without adversities. Among these are the loss of historic character and natural landscapes; a troubling sense that rampant growth and economic advancement have disrupted our communities and increased stress while imposing demands that weaken control over our own lives; and, underlying these losses, the reduced capacity and quality of certain natural resources, despite efforts to protect them under state and federal laws.
Surveys report that increasing numbers of Americans in general, and Georgians in particular, are concerned about the quality of the air they breathe, the water they drink, and the health and security of their families. These issues also heighten the importance of political decisions influencing business growth, income and job creation, education, public health, and other factors that provide incentives and penalties (or at least obstacles) for various segments of the population in their pursuit of individual opportunities. As population and urbanization expand, it is inevitable that conflicts result over the use of natural resources, trade-offs between advocates of private property rights and public interest, and the implications of choices available to voters, workers, and consumers. One important objective of this report is to stimulate and inform public dialogue that is essential to making responsible decisions representing the diverse interests of our citizens and the needs of future generations.
Rapidly expanding areas of disturbed landscape, including use of poorly drained and unsuitable areas.
Driven by rising affluence of residential markets and growing use of cars, the land area needed to support a given number of households has substantially increased in recent decades. Although regional studies have not been done here, nationally the acreage of land development has increased at more than double the rate of population growth over the past 30 years.* Compounding this problem, since many of the highest and best-drained areas of coastal Georgia were developed first, new growth tends to occur in areas prone to periodic flooding (or requiring more filling), with higher potential for pollution from non-point sources such as yard chemicals and septic systems.
*Note: Due to large areas of the region in use for commercial forest and extensive public lands, overall population density is quite low and increasing only moderately; however, area of newly developed land is expanding faster than population growth, resulting in disproportionate burden on natural resources.
Mounting threats to property from exposure to erosion, flooding, and storms, with related public costs.
Development of marginal lands and of scenic marshfront and shorefront areas that are often preferred for homesites introduces heightened risk to both property and natural resources. Attempts to control natural forces such as flooding and erosion (bulkheads, seawalls and jetties), can not only fail to work, but also may actually accelerate related problems in adjacent areas while substantially raising the public costs of new growth - such as extensive drainage systems, revetments, and other efforts to stabilize shorelines. Erosion and erosion-control structures can also harm aquatic and marine habitat. A national study indicated that the dollar value risk of property loss from storm surges is far greater in the Southeast than in any other region of the country, and rapidly increasing.
Critical limits on water supply for both urban and rural needs, raising disputes among various user groups.
As evidenced by the rapid rise in the number of news articles and other media coverage of water issues in recent years, aggravated by regional drought since 1998, this vital natural resource has become critical to development decisions. Though many Georgians are aware of the Œwater wars' between Georgia, Alabama, and Florida, too few realize that similar battles are raging within the state. Ironically, many internal Georgia conflicts parallel the external interstate ones. For example, water needed to sustain the growth of Atlanta and other metropolitan areas may be taken from rivers and aquifers historically available to rural and other downstream regions. This is of particular concern in coastal Georgia, which is at the downstream end of five watersheds that encompass more than 60% of Georgia's land area. If coastal Georgia loses significant water supply and/or water quality due to continuing growth elsewhere in the state, both our economic prospects and quality of life will suffer.
Lack of understanding about ecosystem functions and their value are adding risks to human health and business potential.
There is perhaps no other region of Georgia that is more dependent on the health, diversity, and productivity of natural resources than the coastal area. Our 250,000 acres of tidal wetlands, a considerable portion of the remaining salt marsh on the Atlantic seaboard, is highly important to the fisheries of the state and nation. Actions that reduce the flow of fresh water to our estuaries, or that otherwise impair water quality here, could bring disastrous consequences - including the loss of many jobs and much income, as well as future business growth. With recreational and commercial fishing alone supporting an estimated 20,000 jobs and over a half billion dollars a year in commerce here, it is abundantly clear that protecting natural resources is a major business interest. Beyond this is the troubling record of this region's exposure to toxic materials (such as mercury, PCBs, dioxins, and petrochemicals related to wood preserving, chemical manufacturing, and other industrial processes), each of which present serious health risks that are largely unrecognized by the affected populations.
Increasing discharge of contaminants into air and water, threatening human health and nature-based business.
Contrary to common belief, although more activities affecting the environment are regulated than ever before, because the number of permitted activities is far greater, the total amount of pollutant discharged into air and water is often higher than in the past. With the urbanization of Georgia, according to EPA's Toxic Waste Inventory, in the ten years from 1989 through 1998, the amount of toxic chemicals entering Georgia waters rose by more than 80%. This computation only includes permitted (regulated) releases, not non-point source pollution or any other form of broadcast pollution, such as atmospheric deposition released in other states - air pollution that contaminates Georgia's land and water, carried by wind, from industries up to hundreds of miles beyond our state line. Moreover, the ten-year comparison does not account for new toxins that were not formerly used or controlled.
Accelerating impacts of public facilities (water & sewer lines, roads, etc.) needed to support continuing growth.
Analysis of development trends often underestimates or overlooks the impacts of public facilities that are needed to support development. For example, a number of important environmental violations in Georgia have been caused by improperly maintained or operated municipal sewage treatment plants. Although no comprehensive study has been done, it is commonly believed that many water pollution problems are also caused by domestic septic systems that are inadequately designed, installed, or maintained. Similarly, ditching done to control flooding and/or mosquitoes can aggravate the adverse effects of development by reducing the natural functions of wetlands, including flood control, water filtration, and moderation of stream flow needed to support fish and wildlife.
Displacement of traditional neighborhoods and cultural groups due to increasing property taxes.
With the rise in market value of land, tax assessment adjustments result in higher taxes even if tax rates remain unchanged, and over time property tax increases can be substantial. The average value of land in many coastal communities has quadrupled in the past 20 - 30 years, rising at a much higher rate than many family incomes. For those with modest or fixed incomes, the only solution may be to sell land that has been in their family for generations. Not only does this trend deprive historically established groups of their rightful heritage, but it also reduces the racial and cultural diversity of our communities - producing social homogenization that diminishes the quality of life.
Issues of water quality and environmental contamination invariably include aspects of environmental justice, since low income citizens are more likely to live in areas of greater exposure risk. In coastal Georgia, many families of low to moderate income depend on locally caught seafood for nourishment and they eat proportionally more local fish and shellfish than families with higher incomes. In addition to many "fish consumption advisories" in coastal Georgia, there are several fishing prohibitions in local waters that have been contaminated by site-specific hazardous materials. Despite efforts to restrict or prohibit human consumption of such fish, lower income people continue to catch and consume them, adding significant threats to their health. Thus, environmental problems can have adverse distributional effects that impose a greater burden on some segments of the population than others.
Similarly, many lower income families use shallow wells for their drinking water. Such wells are far more susceptible to contamination from toxic materials and other harmful waste than deeper wells. Shallow wells in the vicinity of hazardous sites, leaking septic systems, illegal dumping, and a variety of commonly used chemicals (such as fertilizers, herbicides, and pesticides) are in danger of health-threatening contamination. In Glynn County, residents in the area of a now-abandoned wood treatment facility are still at risk despite extensive efforts over many years by local activists to induce state and federal regulators to improve protection of groundwater from known contamination.
Another factor in this issue is the proximity of low-income neighborhoods to industrial activities that are continuing sources of contamination, including air pollution. In Brunswick and Savannah, grassroots organizations have been confronting industrial polluters and state regulators for many years, gradually increasing attention to these issues. Respiratory illnesses and premature deaths are believed to be linked to excessive exposure to stack emissions in residential areas close to industrial sites, at least some of which are alleged to be in violation of federal standards under the Clean Air Act. Presently there is a legal action being taken against EPA claiming failure of the state's Environmental Protection Division to properly enforce these standards in issuing an air emissions permit to an industry in Brunswick.
As further documented in this report, there are ample reasons for concern about contaminants in our environment and other risks to natural resources that are likely to lead to further legal actions on behalf of the public. Such actions are often necessary because existing regulations are not properly interpreted and enforced, especially when the adversely affected population is politically weak and economically disadvantaged.
If appropriate action is delayed until incontrovertible evidence is available, damage to the environment and human health are likely to be greater, possibly much greater, than if a more proactive approach is taken.
Although the capacity of the environment to absorb and recover from contamination and other forms of abuse is often unknown, it is certain that there are limits. Once these limits are exceeded, not only will human health suffer, but the costs of environmental restoration will be enormous, assuming recovery is even possible. Engineering substitutions for natural functions like flood control, water storage and filtration, and waste assimilation are extremely expensive and not always feasible.
(3) nature-based business, now worth at least $ 1 billion annually and steadily increasing.
Instead of playing a passive role and waiting until scenic quality, human and environmental health, and/or nature-based business suffer the consequences, it is far more responsible to use preliminary analysis to guide public policy toward a more desirable future. This report is intended to assist decision-makers, community leaders, and concerned citizens by providing such guidance and urging attention to these vital issues.
Coastal Georgia's population will double to a million or more within 30 years.
Developed land is still a small portion of region's geographic area.
Services dominate economic development trend, including tourism and nature-based activities.
State growth patterns and intensity inland from the coast is a risk to the coastal environment.
Along with the current population growth there is increasing pressure by both the public and private sectors for more visitation to, and entrepreneurship on, state and federally managed islands.
Water quality is still a problem - due to both point and non-point source pollution.
Water demand throughout the state imposing unprecedented demand on resources.
Wetlands are being lost, largely due to questionable interpretation of Supreme Court ruling.
Though sampling and data are limited, toxic contamination of air, land and water remains a problem and the trends are risky.
Salinity increases in inter-tidal areas and water contamination present risks to fish and human health.
Coastal Georgia citizens are concerned about growth trends and implications for their environment.
Majority of survey respondents gave Georgia low rating for regulatory enforcement.
Environmental education is seen as important to resolving issues of growth and resource protection.
Environmental ethic is advanced by education, but far more education is needed.
Expressed support for environmental policy is not reflected in consumer and voting behavior.
A variety of important research is being done, but much more is needed.
There are major problems in structuring and translating research for application in public policy.
Promising initiatives are being taken to apply "sound science" in policy, but funding is in jeopardy.
Improved understanding of ecosystem capacity is critically needed to support human demands.
Data for analyzing trends is inconsistent and patchy - numerous sources, varying methods.
Environmental monitoring and assessment, both general and specific to permit enforcement, is seldom linked to research, despite great potential for improving both.
State budget for environmental protection and research is insufficient and declining relative to need.
State policy tends to be dominated by perceived economic interests, often based on misinformation.
Political influence tends to override science and rational precaution in local and state permitting and related policies.
Regulation and environmental analysis are overly fragmented, lack necessary comprehensiveness.
Environmental monitoring and assessment are not adequately used in refining environmental standards and procedures.
There is too little recognition of the economic value of environmental resources and functions.
Understanding about private and common (public) property rights is inadequate, distorting many decisions affecting the use and condition of natural resources.
More opportunities are needed for public dialogue about development, trends, implications, and options.
1. Improve use of information in environmental monitoring, enforcement, and response under current laws.
Expand existing sampling of water, sediments, and fish to achieve more comprehensive assessment.
Ensure environmental permitting review and ongoing assessment with adequate funding of DNR/EPD.
Report all results of resource sampling and permit monitoring in media in readily understandable language.
Alert potentially affected residents and property-owners about permit violations and substandard conditions.
Use results of monitoring and regulatory permit review as guidance for environmental research priorities.
2. Build stewardship into environmental regulations - protect resources using standards based on goals for desired conditions (i.e., fishable & swimmable waters), instead of merely minimum legally acceptable limits.
Use best possible science to determine instream-flow requirements for healthy ecosystem support in rivers and estuaries - under all conditions, including drought - and apply uniformly as standards for permit decisions.
Restore wetland functions on forestry and agricultural lands - using proven best management practices.
Implement a comprehensive water conservation program, including further reductions by major user groups.
When impact assessment information is inconclusive but conditions are risky, permits should not be issued.
3. Base future development on more complete assessment of soils, hydrological characteristics, habitat and other ecosystem features.
Base land use decisions on comprehensive planning and environmental factors, including greenspace needs.
Get serious about implementing soil erosion controls - with better local tech support, penalties for violators.
Adopt land-use ordinances that encourage use of landscape buffers, control of impervious surfaces.
Diversify and monitor economic development within the limits of environmental support capacity.
Promote compatible nature-based business based on assessment of markets, environment, and job needs.
5. The State should take measures to verify the accuracy of water use reporting that includes water consumption and water return figures while developing sound water policies for the energy industry.
6. Create and promote public programs in environmental education at all levels - preschool through college - and in a variety of public forums on a regular basis.
7. Establish a comprehensive program for surveying citizens about their use and valuation of natural resources, and report findings in area newspapers, broadcast media, and public forums.
8. Develop regional plan and strategy for identifying, acquiring, and managing conservation areas based on assessment of scenic quality, ecosystem support, and vulnerability to development trends.
9. Promote responsible diversification of the nature-based economy through research, tax incentives, technical assistance, and start-up assistance, based on a continuing, broad-based participatory process for analyzing trends, revising policies, and evaluating development alternatives.
10. Establish a regional board for improving review and transfer of environmental research into public policy and to evaluate and recommend needed changes in all related public policies.
11. Research and assess potential for ecosystem restoration, determine feasibility and priorities, and adopt program for restoring health of identified areas that have been compromised or degraded.
12. Identify, protect and support a network of high-quality state research reserves strategically located to provide scientists with ecosystem-level baseline information and experimental sites that are required to achieve the best possible scientific information to guide legislative and management actions in coastal Georgia.
Such indicators can be developed for particular ecosystems, such as estuaries, for a single human community or county, or for a metropolitan area or region. In each case, the indicators selected are based on goals for improving conditions in the chosen geographic area, combined with practical considerations about the availability of existing information, costs of acquiring new information, and the reliability of such information in measuring characteristics relevant to stated goals.
- relevant; they show you something about the system that you need to know.
- easy to understand, even by people who are not experts.
- reliable; you can trust the information that the indicator is providing.
- based on accessible data; information is available or can be gathered while there is still time to act.
- Number/percentage of adults with high school diploma; technical training; college degree other.
- Crime rate and number - violent, non-violent.
- Distribution of income and property ownership by race, national origin.
Based on the assessment conducted in this report, we recommend that a set of environmental indicators be developed for the coastal region. We further recommended that these indicators be established by a coalition of individuals representing the diverse interests of coastal Georgia, including property owners, entrepreneurs (especially those in nature-based business), educators, recreational fishing enthusiasts, local and state elected officials, conservationists, and many other groups. Once an initial set of indicators is agreed upon, a plan should be developed for implementing them on a trial basis through a plan adopted by collaborating participants, such as: cities and counties, schools and colleges, homeowner associations, and non-profit organizations. This work should be coordinated by an appropriate public agency or qualified environmental professional under funding allocated especially for this work.
Over time, indicator values should then be evaluated and compared with valid sampling of public opinions about the goals and resources in question. Based on this analysis, indicators should then be refined or replaced as justified. As funding allows and results justify it, the use of indicators should be expanded and integrated into a formal evaluation process and reported to the public at least once annually.


Name: Magic Food Banana Hand Milk.
How: 1. Use a small amount on your hands (1 squeeze will be enough for backs and palms).
2. Spread evenly over the area.
Recommended for: All skin types, especially the dry ones.
Pros: Quick absorbing. Non-greasy formula. Smooth feel. Intense hydration. Cool packaging.
My favorite because… '…put a smile on your face' a quote by Tonymoly to describe some of their products, and I totally agree with this statement, the funky packaging in some of their products are really cool, innovative and sometimes funny. A great example of this is the Banana Hand Milk, and literally, you can see the banana shape in this hand's moisturizer. Moreover the stylish container, on the inside you'll find an aromatic moisturizer -of course banana scent- that will keep your hands out of dryness and chapped.


Nestled in between the Sacramento River and downtown, Old Sacramento is a location pleasantly caught in time – a window to the way Sacramentans lived more than a century ago.
Today, the historic hub bustles with locals and visitors alike soaking in the soul of this once Wild West metropolis. The 28-acre district is a state historic park, national historic landmark, and thriving retail district with numerous restaurants, shops, and boutiques. History thrives with a plethora of museums, including the ever-popular California State Railroad Museum, Sacramento History Museum, Delta King Riverboat, and Old Sacramento Schoolhouse Museum.
While most visitor-driven neighborhoods offer uninspired dining, Old Sac's culinary offerings are plentiful. Touted eateries include Indio Café, Old Town Smokehouse, the Firehouse Restaurant, Ten 22, and Danny's Mini Donuts (not really a restaurant, but a must-stop if you're in the area).
The area is also a hot spot for happenings. The Sacramento Music Festival & Jubilee, St. Patrick's Parade, Mardi Gras Parade, and Pacific Rim Street Festival all call this historic hamlet home, and the holidays light up with a Theatre of Lights and New Year's Eve fireworks.


text:
Imran Khan: Cricket hero who may well be Pakistan's subsequent PM
Symbol copyright AFP
Imran Khan is a former international cricket star who promises a "new Pakistan" and an finish to corruption if he wins the overall election.
The charismatic aristocrat who captained Pakistan to an international Cup victory in 1992 has lengthy shed his famous person playboy image and now styles himself as a pious, populist, anti-poverty reformer.
Mr Khan, 65, struggled for years to turn popular make stronger into electoral gains. He launched his Pakistan Tehreek-e-Insaf (PTI) in 1996 however he used to be the one candidate in the birthday celebration to win a seat in 2002. The PTI boycotted the 2008 vote.
Although he has long been one in every of Pakistan's very best-identified faces the world over, he spent years on the political sidelines. That led to teasing – for a few Imran Khan become "Imran Khan't".
It took till the ultimate general election in 2013 for his celebration to turn out to be a significant player, whilst it narrowly missed changing into the second largest nationally.
If his long look ahead to the top process is finally to finish, he wishes an an electoral swing of epic proportions.
The PTI currently holds just below one-10th of the 342 seats in the National Assembly and will wish to make massive profits within the an important province of Punjab, which has greater than half the 272 immediately-elected seats.
it is also the facility base of the ousted former prime minister Nawaz Sharif and his Pakistan Muslim League-Nawaz (PML-N) party.
'Playboy image'
Oxford-trained Mr Khan had an illustrious profession in global cricket spanning twenty years from the nineteen seventies.
Image copyright AFP Image caption Imran Khan was once at the helm for one in every of his u . s .'s nice triumphs
He additionally advanced a name as one thing of a playboy at the London nightclub circuit, even though he denies that he ever drank alcohol or engaged in any activities that may be regarded as irrelevant for a conservative Pakistani Muslim.
Many say his subsequent forays into the fields of philanthropy and politics have been fuelled by way of a want to positioned to make use of the leadership qualities and goodwill he received as a cricketer.
His pin-up seems to be and private life have ensured he is been a favourite of the world's media for decades.
In 1995, at the age of 43, he married the 21-year-vintage British socialite, Jemima Goldsmith – the daughter of certainly one of the world's richest men at the time, Sir James Goldsmith.
The marriage produced boys but was once dissolved in 2004. The split used to be amicable, and Mr Khan seems to have maintained a pleasant courting together with his ex-spouse.
Image copyright PA Image caption Imran Khan and Jemima Goldsmith's union used to be dissolved in 2004
A second marriage in 2015, to journalist Reham Khan, lasted not up to a year. the former BBC climate presenter alleges she was once bullied via his supporters and has written a tell-all memoir.
It has been making waves in Pakistan, where its newsletter was placed on hold by way of the courts in the run-as much as the election.
Mr Khan wed again in 2018 in a low-profile rite in Lahore.
His 3rd spouse Bushra Watto, a mom of 5, was once defined as his religious adviser, and observers say the match plays neatly together with his public shows of devotion to Islam.
Why Pakistan's election matters Fears of 'creeping coup' in Pakistan Who're the 'angels' and the 'electables'?
As a political candidate, Imran Khan's views have continuously shifted or been imprecise. Many accuse him of U-turns. What he would truly do in energy is basically uncertain.
Symbol copyright AFP Image caption Imran Khan supporters assume their time has come
He upholds liberalism but at the similar time appeals to Islamic values and anti-West sentiment, especially when it comes to perceived interference in Pakistan's inner affairs. He campaigned vocally in opposition to US drone moves in opposition to militants in Pakistan's tribal spaces as part of the "warfare on terror".
He has criticised a few of the Taliban's violence however last 12 months his celebration's executive in Khyber Pakhtunkhwa province gave $3m (£2.3m) to the notorious Haqqania madrassa. Its head, Maulana Sami-ul Haq, is known as the "father of the Taliban". Imran Khan has had to struggle off name callings he have to be referred to as "Taliban Khan".
However it's his campaign towards corruption and dynastic politics in Pakistan, and a promise to boost a complete new magnificence of "blank" politicians, that turns out to have chimed with his supporters.
He has sought to ride a wave of disillusionment at Pakistan's antique political order, particularly a number of the city middle magnificence and young voters, who are tired of dwelling in a rustic with an financial system and foreign money at the slide, and water and tool supply in constant trouble.
the primary aim of his anti-corruption campaign has been his arch-rival Nawaz Sharif.
The Panama Papers leak in 2015 revealed a couple of of Mr Sharif's kids had hyperlinks to offshore companies, which have been allegedly used to channel budget and buy foreign assets.
That moment provided Imran Khan along with his big likelihood – and he seized it.
His campaign towards the Sharifs saw him threaten to blockade the capital along with his supporters. Nawaz Sharif was ousted from the place of business of high minister in 2017 and is now behind bars, sentenced to 10 years in jail by way of an anti-corruption courtroom. The Sharifs maintain the costs are politically inspired.
Imran Khan scents victory in Pakistan vote Point Of View: Pakistan's dirtiest election in years
together with his main rival down however now not out, Imran Khan redoubled his efforts to woo politicians with dependable vote banks to his camp. Critics say lots of the so-called "electables" were coerced into defecting, charges the army and PTI deny.
Mr Khan rejects continual claims that the robust military has manipulated the poll in improve of his marketing campaign.
He told the BBC in Would Possibly: "The Current military leader, Gen Bajwa, might be probably the most professional-democratic man we have now ever observed."
How he may get on with the army within the longer term if he makes it to energy remains an open question.
If he actually wants to tackle a few of Pakistan's root problems, he would possibly to find himself on a collision path with the status quo, as earlier governments have discovered.
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Through May 19, the Figge Art Museum is hosting the special exhibition Louis Comfort Tiffany: Treasures from the Driehaus Collection, a lavish overview of Tiffany's mastery of color and form in a wide range of materials and decorative styles.
Drawn from the collection of the Richard H. Driehaus Museum in Chicago, the exhibition features 62 exemplary works, spanning over 30 years of Tiffany's prolific career. From small blown glass vases to breathtaking stained-glass windows and a spectacular selection of lamps, the exhibition demonstrates the craftsmanship and inventiveness of Tiffany Artisans.
One of America's most renowned artists, Louis Comfort Tiffany worked in nearly all the media available to artists and designers in the late nineteenth and early twentieth centuries—glass, ceramic, metalwork, jewelry and painting. Tiffany's technical brilliance in a wide variety of media enabled him to convey his awe of the natural world through a range of objects, from common household items to one-of-a-kind masterpieces. His work was enthusiastically collected by art museums and private collectors throughout his lifetime and continues to be highly sought after today.
Tiffany's rise coincided with the emergence of the Aesthetic Movement in America. The phenomenon of Aestheticism considered "beauty" as a social and artistic force. The movement's embrace of the decorative arts opened the doors to artists and craftsmen like Tiffany, who believed in enhancing the lives of their clients with beautifully designed rooms and furnishings. Although no one recognizable style typified the Aesthetic Movement, Tiffany's work showed a preference for forms drawn from nature and the influence of art and design from Asia and North Africa.
Louis Comfort Tiffany: Treasures from the Driehaus Collection is organized by the Richard H. Driehaus Museum and is toured by International Arts & Artists, Washington, DC. It is supported by The MacArthur Funds for Arts and Culture at The Richard H. Driehaus Foundation and sponsored locally by Arconic, US Bank, Xenotronics, MidAmerican Energy, Schafer Interiors, Cathy Weideman and John Gardner. Media sponsor: Quad-City Times.
The Figge is open Tuesday through Saturday from 10 am to 5 pm, and Sunday from noon to 5 pm. On Thursday evenings, the museum is open from 5 to 9 pm with free admission. The Figge Café is open for lunch from 11 am to 2 pm Tuesday through Friday, and on Thursday evenings with a special appetizer menu. The first Thursday of each month is Free Senior Day, with exhibition tours and a free film, and special discounts for seniors in the Museum Store and Café. The Figge is located at 225 West 2nd Street in downtown Davenport. For more information, go to www.figgeartmuseum.org.