Genome-wide identification and characterization of Puccinia striiformis-responsive lncRNAs in Triticum aestivum

Wheat stripe rust (yellow rust) caused by Puccinia striiformis f. sp. tritici (Pst) is a serious biotic stress factor limiting wheat production worldwide. Emerging evidence demonstrates that long non-coding RNAs (lncRNAs) participate in various developmental processes in plants via post-transcription regulation. In this study, RNA sequencing (RNA-seq) was performed on a pair of near-isogenic lines—rust resistance line FLW29 and rust susceptible line PBW343—which differed only in the rust susceptibility trait. A total of 6,807 lncRNA transcripts were identified using bioinformatics analyses, among which 10 lncRNAs were found to be differentially expressed between resistance and susceptible lines. In order to find the target genes of the identified lncRNAs, their interactions with wheat microRNA (miRNAs) were predicted. A total of 199 lncRNAs showed interactions with 65 miRNAs, which further target 757 distinct mRNA transcripts. Moreover, detailed functional annotations of the target genes were used to identify the candidate genes, pathways, domains, families, and transcription factors that may be related to stripe rust resistance response in wheat plants. The NAC domain protein, disease resistance proteins RPP13 and RPM1, At1g58400, monodehydroascorbate reductase, NBS-LRR-like protein, rust resistance kinase Lr10-like, LRR receptor, serine/threonine-protein kinase, and cysteine proteinase are among the identified targets that are crucial for wheat stripe rust resistance. Semiquantitative PCR analysis of some of the differentially expressed lncRNAs revealed variations in expression profiles of two lncRNAs between the Pst-resistant and Pst-susceptible genotypes at least under one condition. Additionally, simple sequence repeats (SSRs) were also identified from wheat lncRNA sequences, which may be very useful for conducting targeted gene mapping studies of stripe rust resistance in wheat. These findings improved our understanding of the molecular mechanism responsible for the stripe rust disease that can be further utilized to develop wheat varieties with durable resistance to this disease

Agro-morphological characterization of lentil germplasm of Indian National Genebank and Development of a core set for efficient utilization in lentil improvement programs

Lentil (Lens culinaris Medik.) is one of the major cool-season pulse crops worldwide. Its increasing demand as a staple pulse has led to the unlocking of diverse germplasm collections conserved in the genebanks to develop its superior varieties. The Indian National Genebank, housed at the Indian Council of Agricultural Research (ICAR)-National Bureau of Plant Genetic Resources, New Delhi, India, currently has 2,324 accessions comprising 1,796 indigenous and 528 exotic collections. This study was conducted to unveil the potential of lentil germplasm by assessing its agro-morphological characteristics and diversity, identifying trait-specific germplasm, and developing a core set. The complete germplasm set was characterized for two years, i.e., 2017-2018 and 2018-2019, and data were recorded on 26 agro-morphological traits. High phenotypic variability was observed for nine quantitative and 17 qualitative traits. A core set comprising 170 accessions (137 Indian and 33 exotic) was derived based on the characterization data as well as geographical origin using a heuristic method and PowerCore software. This core set was found to be sufficiently diverse and representative of the entire collection based on the comparison made using Shannon-Weaver diversity indices and χ2 test. These results were further validated by summary statistics. The core set displayed high genetic diversity as evident from a higher coefficient of variance in comparison to the entire set for individual traits and overall Shannon-Weaver diversity indices (entire: 1.054; core: 1.361). In addition, the total variation explained by the first three principal components was higher in the core set (70.69%) than in the entire collection (68.03%). Further, the conservation of pairwise correlation values among descriptors in the entire and core set reflected the maintenance of the structure of the whole set. Based on the results, this core set is believed to represent the entire collection, completely. Therefore, it constitutes a potential set of germplasm that can be used in the genetic enhancement of lentils

Morpho-biochemical characterization of a RIL population for seed parameters and identification of candidate genes regulating seed size trait in lentil (Lens culinaris Medik.)

The seed size and shape in lentil (Lens culinaris Medik.) are important quality traits as these influences the milled grain yield, cooking time, and market class of the grains. Linkage analysis was done for seed size in a RIL (F5:6) population derived by crossing L830 (20.9 g/1000 seeds) with L4602 (42.13 g/1000 seeds) which consisted of 188 lines (15.0 to 40.5 g/1000 seeds). Parental polymorphism survey using 394 SSRs identified 31 polymorphic primers, which were used for the bulked segregant analysis (BSA). Marker PBALC449 differentiated the parents and small seed size bulk only, whereas large seeded bulk or the individual plants constituting the large-seeded bulk could not be differentiated. Single plant analysis identified only six recombinant and 13 heterozygotes, of 93 small-seeded RILs (<24.0 g/1000 seed). This clearly showed that the small seed size trait is very strongly regulated by the locus near PBLAC449; whereas, large seed size trait seems governed by more than one locus. The PCR amplified products from the PBLAC449 marker (149bp from L4602 and 131bp from L830) were cloned, sequenced and BLAST searched using the lentil reference genome and was found amplified from chromosome 03. Afterward, the nearby region on chromosome 3 was searched, and a few candidate genes like ubiquitin carboxyl-terminal hydrolase, E3 ubiquitin ligase, TIFY-like protein, and hexosyltransferase having a role in seed size determination were identified. Validation study in another RIL mapping population which is differing for seed size, showed a number of SNPs and InDels among these genes when studied using whole genome resequencing (WGRS) approach. Biochemical parameters like cellulose, lignin, and xylose content showed no significant differences between parents and the extreme RILs, at maturity. Various seed morphological traits like area, length, width, compactness, volume, perimeter, etc., when measured using VideometerLab 4.0 showed significant differences for the parents and RILs. The results have ultimately helped in better understanding the region regulating the seed size trait in genomically less explored crops like lentils

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Not AvailableIt is expected the predictive performance of genomic prediction methods may be adversely affected in the presence of outliers. In agriculture science an outlier may arise due to wrong data imputation, outlying response, and in a series of trials over the time or location. Although several statistical procedures are already there in literature for identification of outlier but identification of true outlier is still a challenge especially in case of high dimensional genomic data. Here we have proposed an efficient approach for detecting outlier in high dimensional genomic data, our approach is p-value based combination methods to produce single p-value for detecting the outliers. Robustness of our approach has been tested using simulated data through the evaluation measures like precision, recall etc. It has been observed that significant improvement in the performance of genomic prediction has been obtained by detecting the outliers and handling them accordingly through our proposed approach using real data.Not Availabl

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Not AvailableBioinformatics plays a significant role in the development of many fields of biological science and plant genetic resources (PGR) is one of them. With the advent of high throughput sequencing technology, bioinformatics continues to make considerable progress in biology by providing scientists with access to the genomic information. There are many areas of plant genetic resources such as development of core set, trait associated gene discovery, genetic diversity analysis, Genome Wide Association Studies (GWAS), phylogenetic and evolutionary analysis, database development and its management etc. where bioinformatics plays important roles. Main role of bioinformatics is to provide computational algorithm and software tools to accelerate the research of PGR. Bioinformatics is a new paradigm in the genomic research, which provides PGR research a great thrust. However, there are many areas such as pan genomics, multi locus GWAS, Genomic Selection with epistasis effects where bioinformatics can play better role.Not Availabl

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Not AvailableIt is expected the predictive performance of genomic prediction methods may be adversely affected in the presence of outliers. In agriculture science an outlier may arise due to wrong data imputation, outlying response, and in a series of trials over the time or location. Although several statistical procedures are already there in literature for identification of outlier but identification of true outlier is still a challenge especially in case of high dimensional genomic data. Here we have proposed an efficient approach for detecting outlier in high dimensional genomic data, our approach is p-value based combination methods to produce single p-value for detecting the outliers. Robustness of our approach has been tested using simulated data through the evaluation measures like precision, recall etc. It has been observed that significant improvement in the performance of genomic prediction has been obtained by detecting the outliers and handling them accordingly through our proposed approach using real data.Not Availabl

Estimation of Error Variance in Genomic Selection for Ultrahigh Dimensional Data

Estimation of error variance in the case of genomic selection is a necessary step to measure the accuracy of the genomic selection model. For genomic selection, whole-genome high-density marker data is used where the number of markers is always larger than the sample size. This makes it difficult to estimate the error variance because the ordinary least square estimation technique cannot be used in the case of datasets where the number of parameters is greater than the number of individuals (i.e., p > n). In this article, two existing methods, viz. Refitted Cross Validation (RCV) and kfold-RCV, were suggested for such cases. Moreover, by considering the limitations of the above methods, two new methods, viz. Bootstrap-RCV and Ensemble method, have been proposed. Furthermore, an R package “varEst” has been developed, which contains four different functions to implement these error variance estimation methods in the case of Least Absolute Shrinkage and Selection Operator (LASSO), Least Squares Regression (LSR) and Sparse Additive Models (SpAM). The performances of the algorithms have been evaluated using simulated and real datasets

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Not AvailableGenomic selection is a very recent area of study in case of molecular breeding of livestock or crop species. There are various statistical models available for genomic selection. The performances of these models depend on several factors like sampling population, genetic architecture of target species, statistical models as well as missing genotypes. Missing genotype is very common problem in high throughput sequencing data. These missing genotypes are necessary to be imputed in order to implement the genomic selection models. Different statistical models of genomic selection behave differently in imputed data. So, it is highly imperative to evaluate the performances of statistical models under different levels of imputations to know the behavior of the models. In this article, performance of three statistical models viz. Sparse Additive Models (SpAM), Hilbert-Schmidt Independence Criterion Least Absolute Shrinkage and Selection Operator (HSIC LASSO) and Integrated Model for genomic selection are compared after incorporating the various degree of imputation (0, 2, 5 and 10%) in the real data. Results indicate that integrated model is found to be more robust against the level of imputation of the genotypic data.Not Availabl