text:
Maria Carlota Vaz Patto
Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Portugal
Daniela Marone
Cereal Research Centre, CRA-CER, Italy
Teresa Millan
University of Cordoba, Spain
Daniel Fonceka
Centre de Coopération Internationale en Recherche Agronomique pour le Développement, Institut National de la Recherche Agronomique (INRA), France
Front. Plant Sci., 04 April 2017
Sec. Plant Breeding
Volume 8 - 2017 | https://doi.org/10.3389/fpls.2017.00467
Fine-Mapping the Branching Habit Trait in Cultivated Peanut by Combining Bulked Segregant Analysis and High-Throughput Sequencing
Galya Kayam, Yael Brand, Adi Faigenboim-Doron, Abhinandan Patil, Ilan Hedvat and Ran Hovav*
Department of Field Crops, Plant Science Institute, Agricultural Research Organization, Bet-Dagan, Israel
The growth habit of lateral shoots (also termed "branching habit") is an important descriptive and agronomic character of peanut. Yet, both the inheritance of branching habit and the genetic mechanism that controls it in this crop remain unclear. In addition, the low degree of polymorphism among cultivated peanut varieties hinders fine-mapping of this and other traits in non-homozygous genetic structures. Here, we combined high-throughput sequencing with a well-defined genetic system to study these issues in peanut. Initially, segregating F2 populations derived from a reciprocal cross between very closely related Virginia-type peanut cultivars with spreading and bunch growth habits were examined. The spreading/bunch trait was shown to be controlled by a single gene with no cytoplasmic effect. That gene was named Bunch1 and was significantly correlated with pod yield per plant, time to maturation and the ratio of "dead-end" pods. Subsequently, bulked segregant analysis was performed on 52 completely bunch, and 47 completely spreading F3 families. In order to facilitate the process of SNP detection and candidate-gene analysis, the transcriptome was used instead of genomic DNA. Young leaves were sampled and bulked. Reads from Illumina sequencing were aligned against the peanut reference transcriptome and the diploid genomes. Inter-varietal SNPs were detected, scored and quality-filtered. Thirty-four candidate SNPs were found to have a bulk frequency ratio value >10 and 6 of those SNPs were found to be located in the genomic region of linkage group B5. Three best hits from that over-represented region were further analyzed in the segregating population. The trait locus was found to be located in a ~1.1 Mbp segment between markers M875 (B5:145,553,897; 1.9 cM) and M255 (B5:146,649,943; 2.25 cM). The method was validated using a population of recombinant inbreed lines of the same cross and a new DNA SNP-array. This study demonstrates the relatively straight-forward utilization of bulk segregant analysis for trait fine-mapping in the low polymeric and heterozygous germplasm of cultivated peanut and provides a baseline for candidate gene discovery and map-based cloning of Bunch1.
Peanut (Arachis hypogaea L.) is an economically significant crop grown throughout the world. It is the second-most important cultivated grain legume and the fourth largest edible oilseed crop (Faostat, 1998). It is an unusual legume plant in that its flowers are borne aboveground, but the fruits develop underground. The plant is an indeterminate, annual herbaceous bush that is 15–70 cm tall and is comprised of an erect main shoot and a number of lateral shoots (branches) that begin at the base of the plant.
The growth angle of the lateral shoots (commonly referred to as "growth habit" or "branching habit") is one of the most important descriptive characteristics of peanut (Pittman, 1995). The wild polyploid peanut species A. monticola usually has a spreading phenotype, in which lateral branches strike or partially strike the ground. In domesticated peanut, four different, and easily distinguishable categories of branching habit are known: prostrate, spreading, bunch, and erect (Table 1). The description of a new peanut variety, especially one that is of the Virginia marketing-type, will almost always begin with the definition of its growth habit (e.g., bunch or spreading). In addition to assisting breeders and other researchers in identifying accessions for specific traits, branching habit has a great impact on peanut physiology, productivity, and crop management. Since fruiting in peanut occurs underground, the distance between the flowering buds and the ground is an important factor. Pegs of bunch/erect plants that do not reach the ground will not produce pods on time. However, the pods of bunch/erect plants develop at the same time, promoting early maturation. The growth habit of peanut affects the implementation of agrotechnology such as mechanical cultivation and disease management (Butzler et al., 1998).
Table 1. The four main types of growth habit in peanut (from Pittman, 1995).
Despite the agronomic importance of the growth habit of peanut, both the inheritance of this trait and the genetic mechanism that controls branching habit in this crop are not clear. The trait was studied in detail during the 1960s and 1970s and, during that period, two distinct phenotypic groups were usually considered: the runner growth habit wherein the side branches are prostrate, always growing peripherally from the main axis, trailing on the ground except for the tips, which may be somewhat ascending and the bunch growth habit, in which the laterals are also erect or ascending. Initially, a two-gene model for the control of growth habit was suggested, with the runner habit dominant to the erect habit (Hull, 1933; Patel et al., 1936; Coffelt, 1974), but investigators had difficulty classifying the intermediate and/or abnormal growth habits of F2 progeny of crosses between plants exhibiting different growth habits. A fundamental set of experiments conducted by Ashri (1964, 1968) indicated the existence of a genic-cytoplasmic interaction that controls growth habit in peanut. In a project involving a series of reciprocal crosses, differences in growth habit were recorded. A few more nuclear and cytoplasmic genes were later identified by the same group and those researchers concluded that cytoplasmic inheritance has a major effect on the branching habit of peanut (Ashri, 1975; Ashri and Levi, 1975). In more recent studies, the branching habit trait was genetically characterized and mapped by using inter-specific crossing system with an amphidiploid species (Fonceka et al., 2012a,b). The branching habit trait was phenotyped quantitatively by using a continuous scale from 1 (procumbent) to 6 (erect). The trait showed a wide range of morphologies, ranging from completely prostrate to totally erect, and several QTLs were found to control the trait, with the most significant located on linkage groups a07, b05, a10, and b10.
Like many polyploid species, cultivated peanut has experienced a genetic bottleneck, which, together with the effects of domestication, has greatly narrowed its genetic diversity and limited DNA polymorphism among subsequently derived Arachis lines (Kochert et al., 1991; Moretzsohn et al., 2013). As a result, peanut has a low degree of polymorphism among cultivated varieties. This limited polymorphism has hindered the development of molecular and genomic tools for use in domesticated peanut. With the introduction of the genome sequences of peanut ancestors Arachis duranensis and Arachis ipaensis (Bertioli et al., 2016), peanut is now "the orphan legume genome whose time has come" (Ozias-Akins, 2013). These genome sequences provide the resources necessary to move peanut genomics to the next level, facilitating the development of SNP-based marker technologies. In the past, the most widely used molecular markers were simple-sequence repeats (SSRs). Despite their widespread use on the intra-species cultivated level (e.g., Selvaraj et al., 2009), the utility of SSR studies of peanut is limited by their apparent low frequency of across the genome and the relatively low-throughput method of analysis. The use of high-throughput markers like SNPs is necessary for efficient use of genomic data for marker-assisted selection, quantitative trait locus mapping and genomic selection. Recently, several platforms have been developed to facilitate the use of SNP markers for gene-mapping in peanut, including genotyping by sequencing (Zhou et al., 2014) and genome resequencing-based SNP arrays (Clevenger et al., 2016; Pandey et al., 2017). However, despite these advances, these platforms are usually highly efficient only for homozygous populations, like recombinant inbreed lines (RILs) and introgression lines (ILs), which are relatively tedious and expensive to construct in peanut. These methods are usually less effective for trait-mapping in heterozygous genetic populations (e.g., F2 and F3 generations) due to the allopolyploid nature of the peanut genome. This is particularly true in cases of genetic populations that are based upon a cross between closely related parental genotypes, which are occasionally needed for better genetic dissection of specific traits.
In this study, we used a well-defined genetic system to further investigate the genetic nature of the branching habit trait of peanut. Initially, segregating F2 populations derived from a reciprocal cross between very closely-related Virginia marketing-type cultivars were analyzed. Against this particular background, the spreading/bunch trait, a well-known characteristic of the Virginia varieties, was found to be controlled by a single gene with no cytoplasmic effect. Subsequently, a combination of bulked segregant analysis and deep sequencing was developed to facilitate the SNP detection process and the fine-mapping of this gene. The processes were validated using a RIL population derived from the same cross and a new SNP array (Pandey et al., 2017). The relatively straight-forward utilization of this technique in ultra-low polymorphic and highly heterozygous peanut germplasm is demonstrated.
Plant Material and Data Collection
Segregating F2 and F3 populations derived from a reciprocal cross between very closely-related Virginia-type peanut cultivars were studied. The parental lines were cv. "Hanoch," a late-maturing spreading-type cultivar, and cv. "Harari," a medium-maturing bunch-type cultivar. The two parental lines share substantial genetic background, since cv. Harari was developed from an initial cross between cv. Hanoch and cv. Shulamit and an additional back-cross of cv. Hanoch with cv. Hillah (the outcome of Hanoch × Shulamit). In 2013, 314, and 252 F2 individuals from reciprocal Hanoch × Harari and Harari × Hanoch crosses, respectively, were grown under field conditions. The plot consisted of two rows, 75 cm apart, with 40 cm spacing between plants within each row. The experimental plots were sown alongside commercial plots under full irrigation. All agricultural practices were carried out according to local growing protocols as described previously (Gupta et al., 2014).
Growth habit was recorded at 80 days after sowing. At the end of the season, pods were harvested on an individual-plant basis. For each sample, the total pod yield, net pod yield (where immature and unhealthy pods are excluded), number of pods, total seed weight, "dead-end" ratio (relative number of pods with the remote seed aborted) and seed ratio (net seed weight per plant divided by the net pod weight per plant) traits were recorded as well. From each population, approximately 150 F3 families were grown in the subsequent season, with 16 seeds from each F2 individual sown. Branching habit was recorded at the family level (spreading/bunch/segregating) at 80 days after sowing. Plant maturity index was determined based on three random plants in the homozygous F3 families, as the number of fully matured pods out of the total number of pods at 140 days after sowing. To validate the bulk segregant analysis, 94 RILs (F6:F8), which originated from the same Hanoch × Harari cross by single seed descent, were analyzed. In 2016, 16 randomly-selected plants from each RIL were grown under field conditions and the growth habit of each plant was recorded 70 days after sowing.
RNA Isolation, Preparation of Libraries, and High-Throughput Sequencing
Bulked segregant analysis was performed on the F3 families that were found to be homozygous for the spreading or bunch growth habit. In total, 52 completely bunch and 47 completely spreading families were sampled. Young leaves were collected from all 16 individuals in each family. In each phenotypic group (spreading/bunch), all tissues from the families were bulked for the RNA extraction. Working on the RNA level was preferable to working on the DNA genomic level due to the large and relatively complex peanut genome and also facilitated the detection of candidate genes. Samples were taken of each of the ground tissues (400 mg each) and were used for RNA extraction using the hot-borate method, as described by Brand and Hovav (2010). The total RNA was used to prepare two RNA-Seq libraries, using TruSeq RNA Sample Preparation Kit v2 (Illumina) following the manufacturer's protocol as described previously (Gupta et al., 2016). Libraries were validated using DNA Screen Tape D1000 and the Tapestation 2200 (Agilent). RNA-Seq libraries were sequenced using an Illumina HiSeqTM2000 (single lane) at the sequencing center at the Technion in Haifa, Israel.
Data analyses followed the general guidelines for bulk segregant analysis using next-generation sequencing (Magwene et al., 2011) and the specific guidelines for polyploids (Trick et al., 2012), with several modifications. Raw reads were subjected to a cleaning procedure using the FASTX Toolkit (http://hannonlab.cshl.edu/fastx_toolkit/ index.htm) including: (1) trimming read-end nucleotides with quality scores <30 using fastq_quality_trimmer and (2) removing reads with less than 70% base pairs with quality score ≤ 30 using fastq_quality_filter. The sequences were mapped against the 4X tetraploid peanut transcript assembly reference (http://www.peanutbase.org/) and against two Arachis diploid genomes (A. duranensis and A. ipaensis; Bertioli et al., 2016; peanutbase.org) using Bowtie2 aligner (Langmead and Salzberg, 2012). The genome Analysis Toolkit (GATK) Unified Genotyper software version 2.5.2 (McKenna et al., 2010; DePristo et al., 2011) was used for the detection of SNPs. A custom Perl script was used to derive the symmetric difference of the two SNP sets. Polymorphisms between homologous genomes generate the same doubled code and should be common to both SNP sets. Yet, differences in the SNPs between cv. Hanoch and cv. Harari (varietal-specific SNPs) should generate doubled code for only one bulk and, therefore, be unique to the corresponding SNP set. In this manner, ~13,000 varietal-specific SNPs were retrieved between the two bulks. These SNPs were further filtered according to the number of reads for each SNP > 50, GATK quality value >100 and BFR >3. Also, genes with SNP densities higher than 5 SNPs/kb were eliminated to avoid possible paralogue SNPs.
Validation of the SNP Markers and the Bulk Segregant Analysis
For further validation of the SNPs, DNA was collected from the parental lines and 20 F3 progeny of the cross cv. Hanoch × cv. Harari using a DNA Easy kit (SIGMA Aldrich). The same leaves that were used for the RNA study were used for DNA extraction, but the DNA analysis was conducted on a single-plant basis instead of with bulk samples. To validate the three best SNP markers, the following primers were used: M35: F-TCTCTCTCTCTCACAGTCAC; R-CTTGCCGGCAAATAGAGCAT. M255: F--CAGATATGCAAGGCCTAACT; R-TGCCAGAGCAAGGAACATGT. M875: F-CCATCTGCAGTGAGAGTCAA; R-GTGATTCCTGCGTTCAAGTC. These primers were also used for further mapping of the trait in 182 F4 individuals derived from one F3 family segregating for the branching habit trait.
The fine-mapping of the branching habit gene carried out using the bulk-segregant approach was further validated by a custom Affymetrix Axiom SNP array (Pandey et al., 2017). For that analysis, DNA was collected from the two parental lines and each of 94 Recombinant Inbreed Lines (RILs) derived from the same cv. Hanoch × cv. Harari cross. Young leaves were collected from 12 random plants from each RIL and DNA was extracted with a specific kit (GenElute™; Sigma). DNA was quantified by Qubit (Invitrogene LTD) and diluted to 30 ng/uL according to the Affymetrix guidelines (http://www.affymetrix.com). The chip array calls were subjected to cluster-quality filtering, carried out according to Affymetrix guidelines, and additional filtering to select only SNPs that were polymorphic between the parental lines and segregated in a 1:1 ratio in the RILs.
Branching Habit Is Controlled by a Single Gene
The segregation patterns of the branching habit trait in the F1, F2, and F3 generations of the "Hanoch" X "Harari" cross are presented in Table 2. As shown, in this genetic background, the spreading/bunch pattern appears to correspond to a single-gene model of inheritance with no cytoplasmic effect. The allele that confers the spreading phenotype is dominant over the one that confers the bunch habit, as demonstrated by the spreading phenotype of all the F1 hybrids, the 3:1 segregation ratio among the F2 progeny and the 1:2:1 segregation ratio among the F3 families. The gene was named bunch1 and the classification of its corresponding phenotype was very easy and clear-cut, even as early as 50 days after sowing (Figure 1).
Table 2. Segregation pattern of the spreading/bunch trait in several generations derived from closely related peanut varieties.
Figure 1. Bunch and spreading phenotypes among (A) F2 individuals and (B) F3 families grown under field conditions, t 50 days after sowing.
Bunch1 Is Associated with Several Important Agronomic Traits
In addition to growth habit, other traits with agronomic importance were examined in the segregating populations. The associations between the bunch1 phenotype and each of these traits are presented in Figure 2. The bunch phenotype of bunch1 was significantly associated with a lower dead-end ratio. The bunch1 phenotype was also found to have a small, but significant [Prob (t) = 0.0022] effect on early maturation. On the other hand, the BUNCH1 phenotype (spreading) was significantly associated with higher total pod weight and a greater number of pods per plant.
Figure 2. The difference between the bunch (bunch1) and the spreading (BUNCH1) phenotypes of the bunch1 gene in terms of several agronomic traits. Phenotypes that are not labeled with the same letter are significantly different from one another (p < 0.05). (A) Net pod weight, (B) total pod weight, (C) seed content, (D) number of pods per plant, (E) dead-end ratio, and (F) percentage of mature pods at 10 days after sowing (DAS).
Identifying SNP Markers that Are Linked to Bunch1
In order to map bunch1 on the peanut genome, a bulk segregant analysis was performed. For that analysis, 52 completely bunch and 47 completely spreading F3 families were bulked RNA was extracted from each bulk and converted into two libraries suitable for Illumina sequencing. After a cleaning procedure, 72 million reads per library (on average) were aligned to a 4X transcript assembly (peanutbase.org) that contains 120,364 peanut transcripts from both the A and B genomes [60,814 transcripts represent the A genome (Arachis duranensis) while 59,551 transcripts represent the B genome (Arachis ipaensis)]. With about 98% of reads mapped to the reference assembly, the expression levels of 117,957 peanut genes were measured.
Pipelines for the SNP discovery and the analysis of bulk frequency ratio were constructed according to the general scheme that was previously suggested for polyploid wheat (Trick et al., 2012). After initial filtering, ~13,000 SNPs were found to be polymorphic by the two bulks. Subsequently, the bulk frequency ratio was determined for each SNP by calculating the frequency of each nucleotide of the SNP in each bulk and then dividing one bulk by another. If the SNP is a result of false-positive call of a homoeologus SNP, or if it is not linked to the trait, then both of the SNP nucleotides will be equally represented in the two bulks. However, if the SNP is linked to the gene, the frequencies of the SNP nucleotides in one bulk will be with significantly higher frequencies than the other bulk. In that manner, ~1,200 SNPs were found to have bulk frequency ratios of > 3, while 34 had bulk frequency ratios >10 (Figure 3A).
Figure 3. Bulk frequency ratio (BFR) analysis to identify SNPs and genome locations that are linked to Bunch1. (A) The distribution of BFR in ~13,000 varietal-specific SNPs. (B) The genomic locations of the 34 SNPs with the highest BFR (>10), indicating one over-represented region at the end of linkage group 5B (red).
The genomic location of the 34 SNPs with bulk frequency ratios >10 within the peanut genome was recorded (Figure 3B, Supplemental Table 1). One region at the end of linkage group 5B was found to be over-represented in this SNP group; six of the 34 were located between 5B:135,963,343 and 5B147,304,662, including the SNP with the highest bulk frequency ratio [M875 (EZ721696.1); bulk frequency ratio = 23]. Two of these 5B linkage group and another few hypothetical SNPs with high BFR ratio from different linkage groups were further analyzed for SNP classification with Sanger sequencing. The purpose of this step was to roughly validate the location of bunch1. Therefore, samples from the parental lines and 6 F2 individual plants (from which 3 spreading and 3 bunch F3 families were derived) were selected (Supplemental Table 1). In this initial analysis, the SNPs from B5 linkage group were found almost perfectly segregating with trait, while the SNPs from the other genomic locations found to be either homoeologus SNPs (and not varietal) or didn't segregate with the trait (Supplemental Table 1), indicating for relatively high false positive ratio for the BSA by GBS technique in this system.
SNP marker M875 and other two SNPs from the same genomic location that had high bulk frequency ratios [M35 (EZ721381.1), M255 (EZ748922.1)] were further analyzed (Figure 4A). Samples from 20 homozygous F3 families were checked (10 spreading and 10 bunch; Figure 4B). In this initial analysis, M875 was found to be completely linked to bunch1, while the two others, M35 and M255, were also linked to bunch1, but not completely. In the next phase, 182 F4 individuals that originated from heterozygous segregating F3 families were genotyped and phenotyped using markers M255 and M875. Bunch1 was found to be located in a ~1.1 Mbp segment between markers M875 (B5:145,553,897; 1.9 cM) and M255 (B5:146,649,943; 2.25 cM).
Figure 4. SNP validation and linkage analysis with Bunch1. (A) An example of Sanger sequencing validation of two SNP markers. (B) Analysis of linkage between the top three SNP markers and the phenotype of Bunch1. Samples from 10 completely spreading (BUNCH1/BUNCH1) and 10 completely bunch (Bunch1/Bunch1) F3 families were analyzed.
Further Validation of the Bulk Segregant Analysis Using a Peanut SNP Array
Final confirmation of the fine-mapping of the Bunch1 gene was obtained using a new Affymetrix Axiom SNP array (Pandey et al., 2017). Since the chip technology is not efficient enough to distinguish between the heterozygous and homozygous states in the polyploid, a RIL population, which was advanced from the same cv. Hanoch × cv. Harari hybridization (F6:8), was used. Ninety-four RILs and the two parental lines were used for the analysis. Genomic DNA was extracted and applied to the 58,233 SNP clusters of the chip. Out of all of these SNPs, which were designed based on a wide spectrum of diploid and tetraploid peanut species, only 615 passed through the filtering pipeline, including significant differences between the parental lines and 1:1 segregation among the 94 RILs. The genetic analysis of these SNPs and the phenotype of Bunch1 gene are presented in Figure 5A. Ten SNP markers from the array significantly (p < 0.01) co-segregated with the phenotype of Bunch1 (Figure 5A). The best-linked SNP marker (AXX147251194) had only 1 recombinant RIL out of the 94 checked RILs (p = e−50; R2 = 0.92). All significant SNP markers were located in one region at linkage 5B, indicating once again that a single locus is controlling the branching habit trait in this background. These SNPs were located in very close proximity to the three SNPs that were derived from the bulk segregant analysis (Figure 5B). Interestingly, none of the SNPs from the bulk segregant analysis were detected in the chip array and vice versa, indicating that more SNPs could possibly be found by bulk segregant analysis and used in future SNP-array designs for cultivated peanut.
Figure 5. Validation and further fine-mapping of Bunch1 using an Affymetrix Axiom SNP array. (A) Manhattan plot for the analysis of linkage between 615 Affymetrix Axiom SNPs and the Bunch1 phenotype. 1–10 = genome A; 11–20 = genome B (e.g., 15 = linkage group 5B). (B) Integrative map for the bulk segregant analysis and SNP-array analyses of the peanut linkage group B5 (from PeanutBase.org). Markers derived from the bulk segregant analysis are indicated in red. Yellow–gene models. Green–ESTs of genes. Pink–syntheny of this region with A. duranensis (Genome A).
The genetic/molecular mechanism that controls growth angle in plants has been the subject of several studies, mainly involving monocotyledons, particularly rice. Several abnormal tiller-angle mutants and their corresponding genes have been reported in rice,) such as LA1 (Li et al., 2007) and PIN2 (Chen et al., 2012). Two additional genes with opposite effects on tiller angle, Tiller Angle Control 1 (TAC1) and Prostrate Growth 1 (PROG1), have also been identified in rice (Yu et al., 2007; Tan et al., 2008). These genes have played critical roles in the domestication of rice. There are several reports regarding the molecular biology of the growth angle of lateral shoots of dicot species. Roychoudhry et al. (2013) described a model in which the set point angle of lateral branches of higher plants is controlled by an auxin-dependent antigravitropic mechanism. The molecular basis for the spatial pattern of tree branches was also studied in peach, resulting in the discovery of a new ortholog of the TAC1 gene, which controls the "pillar" tree phenotype (Dardick et al., 2013).
We explored the branching habit in the leguminous crop Arachis hypogaea and fine-mapped a major gene that controls this trait. The bunch1 gene was mapped to a relatively small genomic region that includes ~70 ORFs for gene models. Interestingly, BlastX analysis showed that none of the above-mentioned genes that control the growth angle in either monocots or dicots were present in the peanut genome or mapped in approximation to bunch1. The genetic controller of bunch1 may therefore be novel. Several candidate genes involved in plant hormone metabolism and light reception are located within that region and have been identified as possibly controlling bunch1. One of these may be a FAR1-Related sequence (B05:146200756.146203528) that encodes a family of proteins that are essential for phytochrome A-controlled far-red responses in Arabidopsis (Arabidopsis thaliana; (Lin and Wang, 2004). Another putative candidate gene is the 1-aminocyclopropane-1-carboxylate oxidase-like protein (ACC-oxidase; B05:146236653.146238358), which catalyzes the last step in ethylene biosynthesis. Ethylene biosynthesis may play an important role in determining peanut growth angle. Applying a relatively small amount of EPCA (an ethylene-releasing compound) caused the horizontal branches of runner-type plants to become erect (Ziv et al., 1976). Yet, these and other candidate genes must, of course, be further examined in light- and plant hormone-targeted studies, as well as subjected to verification by positional cloning and transformation.
The Bunch1 gene had strong associations with several traits in the segregating F2 and F3 populations. Plants with the bunch phenotype had, on average, earlier maturity values and fewer dead-end pods. Plants with the spreading phenotype had on average more pods per plant, but many of those pods were actually undeveloped. In the bunch type, especially when a wide planting spacing is used (as in our experiments), many flowers are too far from the ground and cannot reach to the soil. For that manner, only the pods that are close to the root will develop. However, those pods that reach the soil develop in a more synchronized manner among the bunch types than among the spreading types. This promotes uniform maturation, better pod filling and, eventually, fewer dead-end pods. Indeed, many of the dead-end pods in the spreading types (like cv. Hanoch) are from distal parts of the branches, where pods develop late in the season and do not fully mature. In a subset of the RIL population, the bunch phenotype of the branching gene is significantly correlated with greater resistance to white mold (caused by S. sclerotiorum; data not shown; submitted for publication). Therefore, it is suggested that the phenotype of the Bunch1 gene has an important agricultural role in Virginia-type peanuts.
As we explored the segregation patterns of several other crosses between Virginia-type related cultivars with different branching habits, we noticed that this model of the bunch1 gene is relatively common within Israeli peanut breeding germplasm. Crosses between cv. Hanoch (spreading) and cv. Hillah/Shulamit (bunch) resulted in a 3:1 spreading:bunch ratio (data not shown), tracing the origin of this trait back to the early 1970s. Crosses between cv. Harari (bunch) and a runner-type peanut line GK-7-Ol (spreading) also resulted in a 3:1 ratio (data not shown), indicating that the single-gene model for this trait is not confined to the Virginia-type germplasm. Yet, in crosses with Valencia-type peanut germplasm (plants with an erect branching habit), this system of single-gene inheritance was not found andthe branching habit was therefore relatively hard to classify. We conclude that the bunch1 phenotype is confined to the A. hypogaea ssp. hypogaea gene pool. However, other allelic variations of this gene may exist in the A. hypogaea ssb. fastigiata germplasm, since some of the bunch-habit Virginia-type lines in Israel (e.g., cvs. Shulamit, Hillah, etc.) have A. fastigiata origins. Moreover, the branching habit trait of peanut was analyzed in a previous study by Fonceka et al. (2012a), which involved mapping traits in a cross between amphidiploid A. ipaensis /A. duranensis and a Spanish-type cultivar (A. hypogaea ssb. fastigiata var vulgaris) with an erect growth habit. There was one significant QTL for the branching habit at the same location as Bunch1 (at the end of linkage group B5), explaining 16.2 of the total variation of the trait in the population. It is very likely that the reported QTL for branching habit and the locus of Bunch1 are the same. This demonstrates once again that the origin of the Bunch1 phenotype may be beyond the A. hypogaea ssp. hypogaea genetic background.
We have demonstrated the relatively straight-forward and easy utilization of bulk segregant analysis for the fine-mapping of a monogenic trait in the low-polymorphic and heterozygous germplasm of cultivated peanut. Also, although the bunch/spreading trait is very easy to classify, there may be some new uses for these new markers in peanut breeding, particularly in the validation of successful F1 hybrids (when the female of the cross is the dominant spreading type) and the selection of homozygous spreading families in early breeding generations. The fine-mapping of this trait also provides a baseline for the cloning of Bunch1, presumably one of the first map-based positional cloning ventures in current genetic research of peanut.
GK and YB were responsible for the molecular work; AF was responsible for the bioinformatic analysis; AP was responsible for RIL population analysis; IH conducted the field trials; RH managed the study and wrote the manuscript.
This study was funded by the Israel Peanut Production and Marketing Board. The authors wish to thank Mr. Oren Buchshtab and other workers of Dod Moshe LTD for their assistance with the field trials.
The Supplementary Material for this article can be found online at: https://www.frontiersin.org/article/10.3389/fpls.2017.00467/full#supplementary-material
Supplementary Table 1. List of the 34 hypothetical SNPs with the highest (>10) BFR. The table indicates the location of the SNP, the change of the nucleotide and the statistic score for the change. Also, the table presents further Sanger sequencing analysis of six SNPs with high BFR ratio. For this, six F2 individual plants were sampled, from which three spreading (green) and three bunch (yellow) F3 families were derived.
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Gupta, K., Hedvat, I., Faigenboim-Doron, A., Clevenger, J., Ozias-Akins, P., and Hovav, H. (2016). Transcriptome profiling of peanut developing seed with a focus on duplicate oil related pathways. Front. Plant Sci. 7:1491. doi: 10.3389/fpls.2016.01491
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Trick, M., Adamski, N. M., Mugford, S. G., Jiang, C. C., Febrer, M., and Uauy, C. (2012). Combining SNP discovery from next-generation sequencing data with bulked segregant analysis (BSA) to fine-map genes in polyploid wheat. BMC Plant Biol. 12:14. doi: 10.1186/1471-2229-12-14
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Zhou, X., Xia, Y., Ren, X., Chen, Y., Huang, L., Huang, S., et al. (2014). Construction of a SNP-based genetic linkage map in cultivated peanut based on large scale marker development using next-generation double-digest restriction-site-associated DNA sequencing (ddRADseq). BMC Genom. 15:351. doi: 10.1186/1471-2164-15-351
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Keywords: peanut, branching habit, bulked segregant analysis, fine mapping
Citation: Kayam G, Brand Y, Faigenboim-Doron A, Patil A, Hedvat I and Hovav R (2017) Fine-Mapping the Branching Habit Trait in Cultivated Peanut by Combining Bulked Segregant Analysis and High-Throughput Sequencing. Front. Plant Sci. 8:467. doi: 10.3389/fpls.2017.00467
Received: 28 December 2016; Accepted: 16 March 2017;
Published: 04 April 2017.
Maria Carlota Vaz Patto, Instituto de Tecnologia Quimica e Biologica and Universidade Nova de Lisboa, Portugal
Daniela Marone, Centre of Cereal Research-CREA-CER-Foggia, Italy
Teresa Millan, Universidad de Cordoba, Spain
Daniel Fonceka, Agricultural Research Centre For International Development, France
Copyright © 2017 Kayam, Brand, Faigenboim-Doron, Patil, Hedvat and Hovav. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
*Correspondence: Ran Hovav, ranh@agri.gov.il
Advances in legume research


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text:
2:00 – 4:30 p.m. Thursday, May 20, 2021
Members Present: Donna Cole; Jenny Dahlberg; Tim Dalby, chair; Stephanie Elkins; Mallory Musolf; Leslie Petty; Deb Shapiro; Lindsey Stoddard Cameron; Bill Tishler
Guests: Lotus Buckner, Lesley Fisher, Jessica Karls-Ruplinger, Darren Martin, Karen Massetti-Moran, Karl Scholz, Mark Walters
The minutes of May 6 and May 18 were approved.
Tim Dalby, ASEC Chair, reported that protocols for summer were discussed at the latest meeting of the chiefs of staff, governance chairs, and secretaries. The Badger Badge is no longer a requirement for building admittance, with the exception of some units on campus. Tim attended the last PPPC meeting in his capacity as liaison. The discussion was on remote work and areas of interest for the committee in the coming year. The ASEC Listening Hours continue to have steady attendance; topics raised include job security and issues regarding the Title and Total Compensation Project. UW System held a town hall on May 13 regarding the single payroll conversion. The session was specifically held for 12-month employees. Tim, Mallory, Jenny, Lesley, and Jake met with Mark Walters for their monthly meeting. Topics included remote work policy updates and single payroll implications.
Jake Smith, Secretary of the Academic Staff, reported that the Chancellor and the Provost are charging a caregiving task force to look at impacts from COVID on caregiving. ASEC decided to put out a call to all academic staff for possible participants. Jake reported on the status of the finalist sessions for the Vice Chancellor for Legal Affairs, Employee Assistance Office Director, and Chief Diversity Officer positions. The Spring CASI Session was held last week; topics included the remote work policy, single payroll impacts, and the TTC Project. The Ad Hoc Committee on Academic and University Staff Diversity, Equity, and Inclusion requested an extension to the deadline for the work of their committee to late summer/early fall. ASEC agreed that the committee should take the time it needs to complete the report. Jake reported that the letter to UW System on single payroll was sent yesterday. The Joint Committee on Finance will be taking up the UW System budget next week.
Bill Tishler reported on the last University Committee meeting. Chancellor Blank discussed potential campus health protocols that may result from the new CDC guidance.
Stephanie Elkins reported that the Communications Committee reviewed the latest iteration of the academic staff website. They also discussed the frequency and content of an academic staff newsletter.
Mallory Musolf reported on the UW System Reps meeting on May 7. At the joint governance session, Jeff Buhrandt addressed the budget, an update on System initiatives, and discussed communication on the importance of governance System-wide. For the academic staff meeting, the distance education project was a main topic of discussion. There is a System committee looking at this, and Provost Scholz is representing UW-Madison.
Leslie Petty reported that the Ad Hoc Committee on Academic and University Staff Diversity, Equity, and Inclusion met this week. A substantial amount of work remains on the report, and the committee will be working through it over the summer, with the idea of presenting the report to ASEC in late summer/early fall and the Assembly thereafter.
Donna Cole reported on the SMPH CASI's meeting. There were continued discussions on the implementation of the research professor title within the School, as well as discussions on issues related to hostile and intimidating behavior.
Deb Shapiro reported that the L&S CASI met on Tuesday. There remain a number of questions on return to onsite work. Questions were also raised about the appeal process for TTC. The committee also passed its new bylaws.
Guest: Karl Scholz, Provost
Provost Scholz commented on commencement and how successful an event it was. He expressed his appreciation for the wonderful job that employees have done in every dimension of the university, from proposals for funding, feedback from parents, instruction, Wisconsin Idea activities, and more. Following up on conversations from the last meeting and a shift in perspective to living with COVID on an ongoing basis, Provost Scholz reiterated the need to ensure that employees have vaccination information uploaded to University Health Services. At present, it appears that approximately 75% of employees have been vaccinated, and it's possible that that number is underreported. We also need to allow time for people's mindset to change, and starting to see people in-person in a variety of venues will help with this. It was good to find out what individuals are feeling, and the OHR pulse survey helped with this. The transition back to onsite work will be phased, and we need to be patient as well as address anxieties as they arise. We don't yet definitively know what the health protocols for instruction will look like in the fall. We are likely to exceed the goal for enrollment, and Enrollment Management has done excellent work to bring students in at a difficult time.
Motion to Convene in Closed Session Pursuant to Wis. Stats. 19.85(1)(c), and (f) to Discuss Appointments to Academic Staff Assembly Districts and Election for ASEC Chair and ASEC Vice Chair (Cole). Seconded. Approved.
Motion to Reconvene in Open Session (Dahlberg). Seconded. Approved.
· Appointments – for vote
Motion to reappoint continuing representatives and alternates in odd-numbered districts to the Academic Staff Assembly (Stoddard Cameron). Seconded. Approved.
· ASEC Chair and Vice Chair – for vote
Tim Dalby was elected as ASEC Chair for 2021-22. Mallory Musolf was elected as ASEC Vice Chair for 2021-22.
Guests: Mark Walters, Chief Human Resources Officer, Office of Human Resources; Karen Massetti-Moran, Director of Total Rewards, Office of Human Resources; Lotus Buckner, Director of Talent Management, Office of Human Resources; Jessica Karls-Ruplinger, Chief of Staff, Office of Human Resources
OHR has had an active engagement with campus community, receiving hundreds of comments about the remote work policy. OHR has made adjustments to the policy based on that feedback. The primary concern was with the tone regarding the importance of onsite work. Adjustments have been made to emphasize the importance of remote and hybrid models in the future. This is the first iteration of this policy, and adjustments will continue to be made as things evolve. A response to the Assembly's resolution on remote work will also be coming. A key change in the policy also includes a definition of workplace flexibilities. Faculty and graduate assistants have been added to the policy for out-of-state and international remote work only; sabbaticals are excluded from the policy. The effective date for re-execution of existing remote work agreements is August 31. Two new factors for considering whether a position could include remote work include the effective function of the unit and adverse impacts on similar positions. There is also a provision for schools/colleges/divisions to look at additional factors. Reimbursable expenditures can also be identified within remote work agreements. The policy outlines more of what the process looks like regarding modification or revocation of agreements, including a "reasonable notice" period instead of a hard "30-day notice" period. An appeal process is also laid out within the policy, which goes up to the dean/director level. ASEC recommended that the explicit link to the policy on workplace flexibilities be put back in. The former telecommuting policy will continue to show in the policy library until August 1, when the remote work policy goes into effect. ASEC also asked OHR to think about ways to address feedback from survey respondents who said that they did not have flexibilities for remote work pre-pandemic, even though those flexibilities existed in policy.
ASEC thanked OHR for incorporating its feedback on the single payroll checklist and other resources. There will also be a pay calculator put in place for the July and August checks, so that employees can see what the gap looks like for their pay. OHR is planning to post more information as well. UW-Madison will also be doing a short-term interest-free loan program and a communication plan for that program for employees who may need assistance in covering the July gap. The application period is expected to launch in June and go through August 15. Repayment of the loan would start in October.
Provost: Center for Teaching, Learning, and Mentoring (with John Zumbrunnen), questions about instructor workload and enrollment; role on UWS committee on distance education
Interim VCFA: DoIT-AIMS Merger, issues with order status in ShopUW+ and workload on under $300 purchases, future of ATP (pushback of timeline to 2025 implementation), process improvement on virtual/CRM platforms (Salesforce as example)

redpajama_set_name: RedPajamaCommonCrawl


Every author secretly thinks their books are uniquely special, but the vast majority of books don't sell. I haven't been blogging regularly for eight months, and I still get urgent emails from authors, wondering why their sales are slumping. They ask if it's their covers, or if the market is crashing, or if they aren't doing the right kind of marketing.
But none of them ever ask if they self-pubbed too soon, before the book was Grade A. Writers are a pretty insecure bunch, but I've never met one who blames their sales on their bad writing.
I gotta say, it's seductive to think I could self-pub these three books instantly, make some money, not worry about the anticipated three star average (I encourage writers to not look at their reviews, and I usually don't), and immediately move on to something I know will sell better.
But I won't do that. I'll put in the time and make these books better. Money is nice. Having more time is nice. However, the nicest thing of all is having pride in my work.
Source: A Newbie's Guide to Publishing: How Good is Good Enough?
An interesting take on the creative process. Are you in it for the money or for the work?
Librarians are needed!Just how bad is multitasking?


text:
If a site plan approval is required, the applicant must submit plans in conformance with Article 11, Site Plan Review of the City's Zoning Ordinance. Complete Site Plan Packages are submitted to the CED department for review. Plans are reviewed and a determination is made regarding the plans compliance with applicable requirements. The plan may be approved administratively, (by the CED Director), or forwarded to the Planning Commission for approval.
Should I submit my site plan application and plans at City Hall? Site plan applications and plans are preferred to be submit through the online application (HERE). Special arrangements can be made for plans to be submit via a USB drive at City Hall. Paper or CD copies of plans will not be accepted, unless requested by staff.
How do I determine if my site plan needs Planning Commission or City Council approval? Site plans are reviewed according to the requirements of Article II in the City's Zoning Ordinance (HERE). Sec. 24-141 generally outlines which uses and structures require Planning Commission or City Council review.
How I determine what the uses and permitted and what a site's zoning district a site is designated as? Fill out a Zoning Determination Request (HERE). Staff will help confirm the site's zoning district and help guide you through the development process.
Where can I find the City's zoning map? Review the City of Ferndale Zoning Map HERE. If a site is zoned Central Business District (CBD), please also review the CBD Zoning District map HERE.
Where can I find the City's Zoning Ordinance? Review the City of Ferndale Zoning Ordinance HERE.
What is the typical timing for a site plan review that requires consideration by the Planning Commission? Site plans shall be submit a minimum of six (6) weeks in advance of a meeting. A staff review will be completed first to determine potential revisions or variances, and make a recommendation on a potential Planning Commission meeting date. Site plans that are submit with special land use and rezoning applications shall be submit for staff review 10 weeks in advance of a meeting. Site plans that are submit with Planned Unit Development (PUD) applications shall be submit for staff review 14 weeks in advance of a meeting.
If my project requires a zoning variance application, do I need to submit a variance application prior to a site plan? Staff recommends submitting a full site plan to complete a full review of an application to determine any possible variances. A project is typically required to have the Board of Zoning Appeals consider a variance application prior to Planning Commission review.
Christine Ross


text:
Q: Using jsonpath to get parent node Using node JSONPath, how can I get the parent node name from child node value

{
  "store": {
    "book": [
      {
        "id":"1",
        "author": "Nigel Rees",
        "title": "Sayings of the Century",
        "price": 8.95
      },
      {
        "id":"2",
        "author": "Evelyn Waugh",
        "title": "Sword of Honour",
        "price": 12.99
      }
    ]
  }
}

I use this expression to identify the child node based on value, I want to use this child node to find the parent node
$.[?(@.id =="1")]

A: You do not specify which implementation of JSON Path, but with both Gatling (Scala) and JayWay (Java) you can use nested filters to filter by children while returning the parent, grandparent or whatever.  Here is a sample:
With this JSON:
{
  "a": {
    "b": {
        "c": {
            "d": {
                "e": "foo"
            }
        },
        "something": "bar"
    }
  }
}

And this path:
$.a.b[?(@.c[?(@.d[?(@.e == "foo")])])].something

Returns:
[ "bar" ]

I am able to retrieve b by using expressions to get to lower nodes as the filter.
Some other implementations error out on these expressions.

A: Unfortunately, JSONpath does not support search by parameters in the contents of a child object:
http://goessner.net/articles/JsonPath/

A: With JayWay it works but the query provided by Jayson is wrong, because if you change the value to "foo2" it will still return "bar".
This can be tested on http://jsonpath.herokuapp.com/.
Query:
$.a.b[?(@.c.d.e=="foo")].something

Returns:
["bar"]

Query:
$.a.b[?(@.c.d.e=="foo2")].something

Returns:
[]

redpajama_set_name: RedPajamaStackExchange


The Eckhart House Our records show that this inn is closed.
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Wheeling bed and breakfast travel guide for romantic, historic and adventure b&b's. Browse through the iLoveInns.com database of Wheeling, West Virginia bed and breakfasts and country inns to find detailed listings that include room rates, special deals and area activities. You can click on the 'check rates and availability' button to contact the innkeeper.
Being nestled at the banks of the Ohio River makes the city of Wheeling one of West Virginia's best tourist destinations. In fact, tourism is one of the city's main sources of income as can be seen by the thriving Wheeling bed and breakfast inn businesses. Upon entering the state from the east, one would immediately see the Wheeling Suspension Bridge, which is called the gateway to the West. Visitors in the city never miss the Kruger Street Toy and Train Museum, with its gallery primarily dedicated to the history of toy trains. The Oglebay Institute Glass Museum, on the other hand, has a collection of a variety of artworks. Those who want to see dog races and try their luck can go to the Wheeling Island Race Track and Gaming Center. Courage House Museum provides visitors with glass shows while the Stifel Fine Arts Center is the venue for live performances. By The Lawrencefield Bed and Breakfast.


text:
Essentially, while you might be bogged down deciding whether
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Monday March 11th, 2013 | David Akana
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With his title "Israeli Boxing Champion" and a one- way ticket, Merhav Mohar takes off for the U.S. Leaving behind anxious parents, lots of friends and more than a few fans he sets out to realize his dream of becoming the boxing champion of the world.
The documentary follows Merhav to New York where he works out daily at Gleason's Gym in Brooklyn. His trainer, Hector Rocca, has turned out a number of world champions and has decided to bet on Merhav to be his next success.
After a major triumph in a match in Las Vegas Merhav gets to the most important fight of his career - his first fight for the world championship. He steps into the ring wrapped in an Israeli flag.


If you are in your 50s and considering buying a home, one of the available options is communities specially built for seniors. These offer many helpful features and services that specifically cater to the interests and needs of those in their post-retirement years.
The characteristics of properties in these projects are, however, by design, unlike those of in regular housing projects. So, it helps to understand the key factors to consider when buying. For example, the location of the property, access to infrastructure such as schools and shops and amenities may be important when buying a regular house. A senior home, due to the nature of the requirements, must be measured with a different yardstick.
One main difference is the details you need to get into in the agreement. A house in a senior living community often comes in different ownership models. Also, there are many services that come with the property. The period of engagement for the contract may be 20-30 years. So, it is important to understand the terms of the contract with the developer.
For one, the structure of the ownership and the various restrictions placed on you may be complex to comprehend. For example, even in an outright ownership model, there may be restrictions on renting out the place — your tenant must be over 50 years of age to stay in the property. Similar rules may exist when selling the property. And if your children inherit the house, but do not meet the minimum age required to live in the community, they should sell or rent out. It also helps to pay attention to the exit clause, in case you want to move out due to some circumstances.
There may also be more intricate rules if it is on a long-term lease model. As the contract may be for 20 years or so, any change in ownership or structure may adversely impact your position. So, it is good to understand the termination clauses — when you may be asked to terminate, or the conditions in which you can ask for termination.
The contract may also cover other terms on prices and escalations. It is good to get the list of one-time, fixed and variable costs (paid based on usage). These have big implications on your finances, as ongoing expenses form a sizeable outgo.
A 2017 study on retirement homes by Moneylife Foundation for HDFC showed that nearly two-thirds of the survey respondents had not signed a contractual agreement that clearly outlined the service terms and their rights. This is a big risk not worth taking.
Another factor that sets a senior housing project apart is access to services. Often, seniors opt for houses in these communities for their bouquet of services such as food, laundry, security, housekeeping and maintenance. There are also medical facilities, community activities in parks or a club house, gyms, ample open space, and various classes on health and fitness. Many communities also provide access to transportation services, within the premises and outside.
The Moneylife survey found many retirement homes without their own ambulances or even wheelchairs; some were in remote areas with poor access to medical facilities. The survey, not surprisingly, showed that over half of the residents were dissatisfied with the services provided and 61 per cent were unhappy with the medical services.
So, while selecting, you must check whether the community can tap qualified service-providers, the selection process, ongoing quality supervision and audits. As service costs tend to escalate a lot (typically 1.5-2 times the rate of inflation), you must understand what measures are in place to control costs. So, you must evaluate senior housing as you would a service contract — with attention to quality and cost of service. Another important aspect is the credibility of the developer who may typically also be the service provider. In a regular housing development, ensuring the quality of construction and the financial strength of the developer to complete the project is often enough; this consideration needs to go much further in a senior living project.
For one, the developments tend to be large projects with hundreds of houses, club houses, parks and other common facilities. Sales happen much later and at a slower pace; so, the financial strength of the developer to deliver must be assessed thoroughly.
Also, things do not end with construction; there needs to be ongoing support to offer services. If the developer is mired in larger financial issues, this may not be a priority for them. So, the residents may feel the pinch of late or non-payment to service providers or staff.
The Ministry of Housing and Urban Affairs (MoHUA) recently drafted a set of guidelines for retirement homes. It has information on building code requirements, common facilities, ways to ensure transparency in fund utilisation, disclosures required, and the right of seniors such as having a say in price escalations.
It also gives a sample agreement that can be used among the buyer, the developer and the service provider. You can look at this (on the MoHUA website) during your selection process.


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Cheryl 'considering other routes' to have more kids because she doesn't want to be in a relationship
Sarah DeenSaturday 13 Apr 2019 10:57 pm
Cheryl is 'considering her options' to have more children (Picture: Getty)
Cheryl wants to be a mum again and is 'considering other options' to make it happen without being in a relationship.
Cheryl, 35, says she hasn't found the right man yet but isn't going to let that stop her from having more kids.
The singer hinted she might use a sperm donor in the future.
She explained: 'I've got a friend who has had two children using donors. Not everything has to be conventional.
'You can spend ages looking for the right man, waiting for the perfect time to get pregnant, then the right man might turn out to be the wrong man. There are definitely other routes I would consider.'
Cheryl is mum to son Bear with ex-boyfriend Liam Payne.
She revealed that she is 'stricter' on their son than the One Direction star, who she split from last year.
The former Girls Aloud member praised the 25-year-old's parenting skills.
She added that their break-up involved no 'screaming and shouting' and that they 'manage to get on' for the sake of Bear.
Cheryl split from boyfriend Liam Payne last year. The couple share son Bear (Picture: Getty)
She told The Telegraph: 'Liam is a great dad. He's much softer than me. But it is what it is, and I'm OK about it.'
The former couple reunited at the Global Awards last month where they shared a 'big hug'.
An eyewitness told The Mirror: 'They were smiling and talking and they went in for a big hug. Liam put his arms around her and kissed her on the cheek.'
Liam has since been linked with supermodel Naomi Campbell, depsite the two of them not actually being pictured together.
Meanwhile Cheryl has been focusing on her upcoming album, with new track Let You due next month.
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Cheryl feeling herself in white corset as she attends Fashion Week after 36th birthday


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Latin Music Departed of 2018
By Hector Aviles	 On Dec 31, 2018
Like most years, I like to take a moment to honor some of the artists that contributed to Latin music and passed away in 2018.
I listed some of them below:
1. Roberto Anglero (May): although Anglero was a percussionist and singer, I think his major contributions were as a songwriter.
He wrote songs for many artists, but one of his main ones was El Gran Combo. One of his most memorable was " La Salsa de Hoy".
As an army soldier he saw the injustices of racism, which years later he wrote and sang on the satirical song "Si Dios Fuera Negro".
He made a funny cameo in one of his last compositions, when Jose Lugo recorded him in a short intro for his song "Parece Que Uno Se Va A Morir", which was part of Gyasabara's "Poetic Justice" album.
2. Anthony Cruz (May): one of the pioneers of the Salsa Romántica movement of the 1990's. Great singer.
3. Joseito Mateo (June): a Music legend in Dominican Republic and Latin America. Joseito was a versatile merengue singer who could sing just about anything.
Joseito Mateo basically helped launch El Gran Combo, and therefore, a piece of Salsa history.
He happens to be the singer in the very first recording of El Gran Combo, when producer Alvarez Guedes asked the newly formed band to backup his already famous singer in the album "Menéame los Mangos".
4. Jose "Chegui" Ramos (June): a singer with a tremendous voice, the young Puerto Rican came to fame with the group Batacumbele.
He later became a sought-after chorus singer for his magnificent voice. He departed way to early.
5. Antonio Barasorda (tenor PR) (July): the Puerto Rican tenor died this year at age 72. Barasorda studied in Spain and went on to perform in venues like the Metropolitan Opera House in New York and the Festival Casals in Puerto Rico.
He also performed in many other venues and operas around the world.
6. Eladio Pérez (July): the Puerto Rican percussionist was perhaps better know for being the conga player of Eddie Palmieri's Orchestra during the 70's.
After he relocated back to Puerto Rico, he briefly re-joined Palmieri when he also moved to Puerto Rivo in the 80's. Eladio also worked with Charlie Palmieri and Roberto Roena during his career.
Eventually ha formed his own band.
7. Emilio "Millito" Cruz (July): a great Puerto Rican cuatro and guitar player, Millito was one of the most recognized talents of the instrument starting in the 70's.
Millito performed with many artists, and replaced Pedro Guzmán in the famed Puerto Rican folk group Haciendo Punto en Otro Son.
8. Jack Constanzo (August): an American percussionist, he gained fame as "Mr. Bongo" starting in the 1940's. At that time, he visited Havana, Cuba and learned to play Afro-Cuban rhythms in the bongo and conga.
Jack performed early in his career with Desi Arnaz and Rene Tousett, after having been a dancer with his wife.
He later on went on to perform with Stan Kenton and Nat King Cole, among others before forming his own Afro-Cuban band.
9. Tony Sanchez (November): another great percussionist, Tony excelled in the trap drums, which he learned to play from his father.
Tony Sanchez was bandmate of my uncle Ruben Escabi with Pete Santoni y su Combo.
He got to play in the Hotel Normadie in the late 50's to early 60's. He also played with Noro Morales, and a plethora of reknown bands and artists.
Sanchez was a founding member of the Puerto Rico All Stars. He also played with Bobby Valentin, particularly in the "Va a la Carcel" albums.
In this last song, we get to appreciate both, Tony Sanchez in the trap drums, and Eladio Perez on congas.
May they all rest in peace!
2018departedlatin music
Palmieri, Santa Rosa & Olivera Deliver Great Salsa in "Mi Luz Mayor"
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Located in the heart of the residential area of Cimiez, in a gated domain, exceptional contemporary architect house of about 270 m2. This villa is distinguished by a refined style. Composed of a large reception area and 5 bedrooms, the whole enjoys very high quality finishes. The living room opening onto a large terrace facing south. Garage, Summer Kitchen, Wine Cellar.
The landscaped grounds of approximately 1000 m2, benefits from a large swimming pool.
Ideal family home, close to shops, schools, clinics and motorway access 5 minutes.


2 vietnamese spring rolls filled with pork and vegetables in rice paper with nuoc mam sauce.
Summer rolls with scampi, rice noodles, lettuce and asian herbs with peanut sauce dip.
Tofu over red and green cabbage, glass noodles, carrots, onions and bean sprouts in house vinaigrette. Topped with herbs, peanuts and shallots.
Tender poached shredded chicken over red and green cabbage, glass noodles, carrots, onions and bean sprouts in house vinaigrette. Topped with herbs, peanuts and shallots.
Rice noodles, fried spring rolls, lettuce, asian herbs, peanuts and shallots. Served with fish sauce.
Vietnamese noodle soup with rice sticks in beef broth and 5 different herbs. With thin sliced beef, onion, bean sprouts and asian herbs.
Vietnamese noodle soup with rice sticks in chicken broth and 5 different herbs. With thin poached chicken, onion, bean sprouts and asian herbs.
Rice sticks mixed with soya sauce dressing, chives, bean sprouts and BBQ-pork on top.
Rice noodles, stir fried beef in lemongrass and chili, lettuce, asian herbs, peanuts and shallots. Served with fish sauce.
Rice filled with chopped salmon, seaweed salad, avocado, cabbage and sesame. Served with our homemade soya dressing.
Rice filled with chopped salmon and tuna salad, avocado, cabbage, seaweed salad and roasted sesame. Served with our soya ponzu dressing.
Rice filled with fried scampi and light grilled salmon, tobikko, spring onion, sesame and cabbage. Served with spicy shrimp sauce and eel sauce.
Fried tofu spring rolls with sweet chili sauce. Vegan.
Summer rolls with tofu, ricenoodles, potato, lettuce and asian herbs with peanut sauce dip.
Vietnamese noodle soup with rice sticks in root vegetables broth and 5 different herbs. With tofu, onion, bean sprouts and asian herbs.
Spicy aromatic soup with shiitake mushroom, lemongrass, tofu, onion and asian herbs. Served with rice noodles.
Rice noodles, vegan spring rolls, lettuce, asian herbs, peanuts and shallots. Served with fish sauce.
Cabbage, tofu, avocado, apple, seaweed salad, cashew and cucumber. Served with our soya ponzu dressing.


Maharashtra is known for the rock-cut caves or shrines hidden in the mountain ranges spread across its landscape. Often we feel that to experience an archaeological site we would have to travel to an obscure looking place and hunt for a route to reach an architectural marvel. The Pataleshvara Caves are situated alongside one of the busiest roads of Pune city, on the Jangli Maharaj Road, but still manage to remain aloof and preserve the historicity of the place. Let's explore these caves, dated approximately to the seventh or eighth century. What do the caves tell us through their architecture, and how can we look at them in the urban chaos of today? Discussions surrounding the changing landscape in the area, a journey through time, history and some interesting stories are waiting to be heard at this marvellous site.
The walk involves interior and exterior site-seeing, hence it is recommended to wear suitable/comfortable shoes, and carry a cap/hat and a bottle of water.
Please maintain decorum throughout the walk as it is a living shrine and is substantially busy at most times.
The site is on the main road and has no authorised parking facilities. Public parking facilities are available nearby.


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29 Weird Phobias That Really Do Exist
This list of phobias proves that practically anything can be terrifying to someone.
By Markham Heid
Everyone's afraid of something—or a lot of somethings. But fear isn't the same thing as a true phobia.
"A phobia consists of a persistent fear or avoidance of a specific stimulus," says Kate Wolitzky-Taylor, PhD, an associate faculty member with the Anxiety and Depression Research Center at UCLA. "Usually the stimulus is a thing or a situation—like bees or heights." But there's a second component to phobias. "In addition, the fear causes significant distress or somehow impairs the person's life," Wolitzky-Taylor says.
So maybe you get creeped out by snakes or tight spaces—both of which are common, she says. Your fear may rise to the level of a phobia if you can't even see snakes on TV without losing sleep, or you have to quit your job because getting to your office requires that you ride in a cramped elevator.
Where do these phobias come from? Your DNA may play a role. Wolitzky-Taylor says many people have genes that raise their risk for anxiety-related disorders—an umbrella term that includes phobias. But even if you don't have those phobia genes, you can develop one through learning or "conditioning." For example, if your parents always told you to be fearful of spiders, that could snowball into a phobia—especially if you also had a negative experience with a spider.
Avoiding the thing that scares you can also increase its power over you. "Avoidance strengthens anxiety and keeps it going, and that's a recipe for phobia," Wolitzky-Taylor says. Quite literally, we can be afraid of just about anything. Keep reading to learn about the weirdest phobias out there.
Taphophobia
Taphophobia is the fear of being buried alive. Research suggests it arose (and probably peaked) during the 1700s at a time when plague was rampant, and people feared doctors or medical professionals would mistakenly pronounce them dead. Taphophobia was such a phenomenon that escapable "safety coffins" were a thing, and corpses were often stored for long periods before burial to ensure they wouldn't wake up.
Eisoptrophobia
Eisoptrophobia is the fear of mirrors or, more specifically, of seeing your own reflection in a mirror. Looking into a mirror can cause people with eisoptrophobia shame or distress, and can lead to depression, according to a 2014 case study detailing one 55-year-old woman's 30-year struggle with this phobia.
Ombrophobia
Ombrophobia is the fear of rain. It falls into a category researchers term "natural environment phobias," which also includes hurricanes (lilapsophobia), snow (chionophobia), cold (cryophobia), and wind (ancraophobia). People with these phobias may be more likely to have some kind of formal weather-related education, which saddles them with "a greater understanding of the potential dangers associated with severe weather," according to the authors of one recent study.
Phonophobia
Phonophobia is an "abnormal" and "unwarranted" fear of sound, according to research from Malaysia. Researchers say these are often normal, everyday sounds that cannot possibly harm a person's hearing or cause pain—stuff like doors closing or loud conversation. Phonophobia sometimes overlaps with a condition called "hyperacusis," which is an abnormally strong reaction to sound, stemming from the part of your brain that processes noise.
Numerophobia
Numerophobia is the fear of numbers, though not in the sense that a person believes giant 1's or 0's are hiding under the bed. (Although with all the recent hand-wringing about technology, maybe they should.) Instead, numerophobia usually pops up as a fear of doing math or dealing with numbers. There's some evidence that numerophobic doctors may ignore new evidence-based best practices because of their aversion to numbers and statistics.
Myrmecophobia
Myrmecophobia is a fear of ants. The term is usually applied to species of plants that recoil or don't respond well in the presence of these insects. But research touching on phobias and insect "swarms" suggests that some people are also freaked out by ants. Some experts are also looking at swarm-based fears in order to predict how people may react negatively in the future if they're surrounded by large numbers of robots. (Seriously.)
Thalassophobia
Thalassophobia is a fear of the ocean or deep, open water. While thoughts of sea creatures lurking below the waves plays into this phobia, it's often rooted in a fear of the unknown, writes Seán Harrington in the book Beasts of the Deep. "The deep sea offers us an oppressive and foreboding context—a space unexplored, unknowable and overwhelming," he writes.
Ephebiphobia
Ephebiphobia is a fear of teenagers or adolescents. Cultural and demographic researchers have long understood that nearly every generation of adults harbors mild forms of this phobia—meaning they look at teens as "out of control" or backwards in some way. For adults with more severe forms, spending time with teens can help dispel these fears.
Past Life Phobias
There's a good amount of research on people who believe in reincarnation, and who claim they remember details from their past lives. One University of Virginia study found that some children who claim to recall their previous lives suffer "unusual" phobias that they say are related to those past lives—and specifically the way they died. For example, a child might have an intense fear of water that she attributes to drowning in her past life.
Aerophobia
Aerophobia is the fear of fresh air or the movement of air—such as drafts or breezes. Along with a fear of water, fear of air is one common symptom of rabies. Blowing or fanning air on a person with rabies can trigger muscle spasms and an "intense startling reaction," shows one 2008 study.
Alektorophobia
Alektorophobia is the fear of chickens or hens. One 2016 case study tells the story of an 18-year-old woman who had a frightening encounter with a hen when she was a child. Later, whenever she saw a hen or chicken, she worried about being pecked or attacked. She even started experiencing intense anxiety when dining out and seeing dishes that contained chicken.
Amathophobia
Amathophobia is a fear of dust. Maybe not surprisingly, research has found it occasionally shows up among librarians—people who are constantly around dust-covered old books.
Hippopotomonstro - sesquippedaliophobia
Hippopotomonstro - sesquippedaliophobia (which we had to hyphenate to have it fully appear on the screen) is, no joke, a fear of long words—whoever coined the name for this phobia has a sick sense of humor. It's usually used in the context of patients with dyslexia, who may struggle with unusually long words. It's also sometimes called "sesquipedalophobia."
Globophobia
Globophobia is the fear of balloons. "After being sensitized in childhood by exploding balloons, patients tend to avoid proximity to balloons," writes the author of one recent BMJ report. That researcher says these patients often run into trouble when they have kids and encounter balloons at other children's birthday parties.
Phobophobia
Phobophobia is the fear of fear itself. It can lead to anxiety attacks and what some researchers describe as "free-floating anxiety."Basically, these people are so hyper-aware of the ways becoming afraid will affect them that they struggle to function out of a fear that they'll experience fear.
Zoophobia
Zoophobia is the fear of all animals. (The same term also applies to a category of more specific animal fears, such as a fear of dogs or snakes.) In some cases, this fear can cause people to stay housebound for fear of encountering an animal. Symptoms also include "excessive sweating, trouble controlling muscles, dizziness, fainting, elevated heart rate, rapid, and shallow breathing," when encountering an animal, according to a recent study.
Emetophobia
Emetophobia is the fear of vomiting. Researchers sometimes categorize this fear as a "social phobia," because patients may fear vomiting because it would be embarrassing if it happened in front of other people. Looking at certain foods or feeling stomach activity can trigger the phobia, according to a Columbia University study.
Nomophobia is the fear of being without a smartphone. Of course, this is a relatively new phobia. A recent study from Italy defines its characteristics as feeling "anxious or nervous" at the thought of losing your smartphone, or worrying about losing network coverage or battery power
Urophobia
Urophobia is the fear of urinating. For those with bladder-control issues, the fear can stem from concern over having an "accident" in public. Some people also experience this fear when peeing around others (commonly known as "stage fright"), which some experts consider a specific subcategory of social anxiety.
Somniphobia
Somniphobia is the fear of "initiating sleep," or falling asleep. Some people experience sleep paralysis as they drift off—basically, not being able to move even through they're still partly awake—and this can lead to somniphobia, according to a 2016 study from the University of Minnesota.
Ranidaphobia
Ranidaphobia is a fear of frogs (not to be confused with bufonophobia, which is a fear of toads). Unsurprisingly, a fear of frogs can cause people to avoid hiking in nature or visiting natural history museums.
Panophobia is the fear of everything, and is a term sometimes used to describe patients with generalized anxiety disorders. One recent study found that many panophobics are often frightened by darkness or harmless noises. But, more broadly, people with panaphobia have fear stemming from no identifiable cause.
Athazagoraphobia is the fear of forgetting or being forgotten. Having some experience with amnesia, or knowing someone who has endured amnesia, can trigger this fear, according to one 2012 study from Germany.
Vestiphobia is the fear of clothing. In a lot of cases, this fear focuses on a specific type of garment. One case study detailed a military veteran who experience extreme anxiety whenever he needed to wear part of his uniform. Some other sufferers have issues with tight-fitting clothing—or with any clothing at all.
Cibophobia
Cibophobia is the fear of food. And while that sounds a little ridiculous at first, one case study found this fear developed when a patient experienced painful muscle spasms after eating or swallowing.
Erythrophobia
Erythrophobia is the fear of blushing. "Blushing can become a stubborn and incapacitating symptom that often severely restricts the patient's social life," say the authors of a study in Psychosomatic Medicine. Unfortunately for people with this phobia—and for anyone who feels a little embarrassed about blushing—trying to suppress a blush usually makes it worse, the study authors say.
Frigophobia
Frigophobia is the fear of being cold. It may be especially common within Chinese culture because the Eastern philosophical principles of yin and yang hold that cold exposure is associated with heart ache, stomachache and illness, as well as a loss of vitality. One 45-year-old frigophobia sufferer described even mildly cold air "piercing" her skin and causing her headaches, body aches, and other symptoms, a case study found.
Deipnophobia
Deipnophobia is the fear of dining or, more specifically, dinner parties. It's not at all clear how this phobia develops—but it is listed in the Oxford Dictionary of Psychology, so some people have experienced it.
Ergophobia
Ergophobia is the fear of work. According to one recent study on disability claims, people with ergophobia feel intense stress and anxiety surrounding their the work environment, impairing their ability to function properly. This anxiety can become severe and cause full-blown panic attacks.
How should you deal with a phobia?
Exposure therapy is by far the most effective way to deal with a phobia. "It involves gradual confrontation of the thing you're afraid of," says Wolitzky-Taylor. "The basic idea is to disconfirm a person's belief about what will happen."
So let's say you're afraid of dogs. Exposure therapy might involve standing in a room with a dog, and then gradually moving closer to it and, eventually, petting it, she explains. Meanwhile, you'd be talking with your therapist about what exactly you're worried about—say, the dog biting you or jumping on you. The therapist would point out that these fears aren't coming to fruition, which would help recondition you to feel less fear.
"Sometimes negative things happen, and it can actually make therapy more effective," Wolitzky-Taylor says. She explains that usually these events—the dog jumping on you—aren't as bad as your mind thought they would be. So that negative experience actually helps shrink your fears down to size.
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