Plant Genome

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Development and Application of High‐Density Axiom Cajanus SNP Array with 56K SNPs to Understand the Genome Architecture of Released Cultivars and Founder Genotypes
Plant Genome - Tập 11 Số 3 - 2018
Rachit K. Saxena, Abhishek Rathore, Abhishek Bohra, Pooja Yadav, Roma Rani Das, Aamir W. Khan, Vikas Kumar Singh, Annapurna Chitikineni, I. P. Singh, C.V. Sameer Kumar, K. B. Saxena, Rajeev K. Varshney

As one of the major outputs of next‐generation sequencing (NGS), a large number of genome‐wide single‐nucleotide polymorphisms (SNPs) have been developed in pigeonpea [Cajanus cajan (L.) Huth.]. However, SNPs require a genotyping platform or assay to be used in different evolutionary studies or in crop improvement programs. Therefore, we developed an Axiom Cajanus SNP array with 56K SNPs uniformly distributed across the genome and assessed its utility in a genetic diversity study. From the whole‐genome resequencing (WGRS) data on 104 pigeonpea lines, ∼2 million sequence variations (SNPs and insertion–deletions [InDels]) were identified, from which a subset of 56,512 unique and informative sequence variations were selected to develop the array. The Axiom Cajanus SNP array developed was used for genotyping 103 pigeonpea lines encompassing 63 cultivars released between 1960 and 2014 and 40 breeding, germplasm, and founder lines. Genotyping data thus generated on 103 pigeonpea lines provided 51,201 polymorphic SNPs and InDels. Genetic diversity analysis provided in‐depth insights into the genetic architecture and trends in temporal diversity in pigeonpea cultivars. Therefore, the continuous use of the high‐density Axiom Cajanus SNP array developed will accelerate high‐resolution trait mapping, marker‐assisted breeding, and genomic selection efforts in pigeonpea.

A Modified TILLING Method for Wheat Breeding
Plant Genome - Tập 2 Số 1 - 2009
Chongmei Dong, Jessica Dalton‐Morgan, Kate Vincent, P. J. Sharp

The large genome and polyploidy of wheat (Triticum aestivum L.) makes it difficult to identify desirable genetic changes based on phenotypic screening due to gene redundancy. Forward genetics is, therefore, more difficult in wheat than in diploid plants. A modified TILLING (Targeting Induced Local Lesions IN Genomes) method including the harvest of five heads per M1 plant, storage of M2 seeds, using unlabeled primers and agarose gels for mutation detection, and crossing of useful mutants for desired grain quality was explored in this report. A soft wheat cultivar, QAL2000, and a hard wheat cultivar, Ventura, were mutagenized with ethyl methanesulfonate (EMS). Screening of the waxy genes Wx‐A1 and Wx‐D1 in 2348 EMS‐treated M2 plants allowed identification of 121 mutants, including silent, missense, and knockout (truncation) mutations. A complete waxy wheat was successfully bred in 18 mo by crossing two truncation mutants (Wx‐A1‐truncation and Wx‐D1‐truncation; Wx‐B1 is naturally null in both mutants). Screening of two puroindoline genes (Pina and Pinb) in QAL2000 identified 19 mutants. A hard grain variant of a soft cultivar was identified due to a mutation in Pinb caused by a premature stop codon. Background mutations were observed and further self‐fertilization or crossing with a wild type was performed to eliminate deleterious mutations. With the rapid accumulation of wheat genomics information, many potential target genes of interest can be screened for mutations in these TILLING populations.

Single Nucleotide Polymorphism Genotyping for Breeding and Genetics Applications in Chickpea and Pigeonpea using the BeadXpress Platform
Plant Genome - Tập 6 Số 2 - 2013
Manish Roorkiwal, S. L. Sawargaonkar, Annapurna Chitikineni, Mahendar Thudi, Rachit K. Saxena, Hari D. Upadhyaya, M. Isabel Vales, Oscar Riera‐Lizarazu, Rajeev K. Varshney

Single nucleotide polymorphisms (SNPs) are ideal molecular markers due to their higher abundance. Although several types of genotyping platforms for assaying large number of SNPs are available, in cases such as marker‐assisted selection, where few markers are required for genotyping a set of potential lines, high‐throughput SNP genotyping platforms (e.g., iScan or Infinium) may not be cost effective. In this scenario, GoldenGate assays based on VeraCode technology using Illumina BeadXpress seems to be the most cost‐effective platform. The objective of this study was to develop cost‐effective SNP genotyping platforms in chickpea (Cicer arietinum L.) and pigeonpea (Cajanus cajan L.). Two sets of SNPs, one each for chickpea (96 SNPs) and pigeonpea (48 SNPs), were developed and tested by genotyping 288 diverse genotypes from respective reference sets. The SNPs selected for the oligo pool assays had high transferability to crop wild relative species. The mean polymorphism information content value of assayed SNP markers was 0.31 and 0.32 in chickpea and pigeonpea, respectively. No unique pattern was observed in the chickpea reference set whereas two major groups were observed in the case of the pigeonpea reference set. The Illumina BeadXpress platform assays developed for chickpea and pigeonpea are highly informative and cost effective for undertaking genetic studies in these legume species.

Genomic Prediction of Barley Hybrid Performance
Plant Genome - Tập 9 Số 2 - 2016
Norman Philipp, Guozheng Liu, Yusheng Zhao, Sang He, Monika Spiller, Gunther Stiewe, Klaus Pillen, Jochen C. Reif, Li Zuo

Hybrid breeding in barley (Hordeum vulgare L.) offers great opportunities to accelerate the rate of genetic improvement and to boost yield stability. A crucial requirement consists of the efficient selection of superior hybrid combinations. We used comprehensive phenotypic and genomic data from a commercial breeding program with the goal of examining the potential to predict the hybrid performances. The phenotypic data were comprised of replicated grain yield trials for 385 two‐way and 408 three‐way hybrids evaluated in up to 47 environments. The parental lines were genotyped using a 3k single nucleotide polymorphism (SNP) array based on an Illumina Infinium assay. We implemented ridge regression best linear unbiased prediction modeling for additive and dominance effects and evaluated the prediction ability using five‐fold cross validations. The prediction ability of hybrid performances based on general combining ability (GCA) effects was moderate, amounting to 0.56 and 0.48 for two‐ and three‐way hybrids, respectively. The potential of GCA‐based hybrid prediction requires that both parental components have been evaluated in a hybrid background. This is not necessary for genomic prediction for which we also observed moderate cross‐validated prediction abilities of 0.51 and 0.58 for two‐ and three‐way hybrids, respectively. This exemplifies the potential of genomic prediction in hybrid barley. Interestingly, prediction ability using the two‐way hybrids as training population and the three‐way hybrids as test population or vice versa was low, presumably, because of the different genetic makeup of the parental source populations. Consequently, further research is needed to optimize genomic prediction approaches combining different source populations in barley.

Genome‐Wide Association Studies of Grain Yield Components in Diverse Sorghum Germplasm
Plant Genome - Tập 9 Số 2 - 2016
Richard Boyles, Elizabeth Cooper, Matthew T. Myers, Zachary Brenton, B. Rauh, Geoffrey P. Morris, Stephen Kresovich

Grain yield and its primary determinants, grain number and weight, are important traits in cereal crops that have been well studied; however, the genetic basis of and interactions between these traits remain poorly understood. Characterization of grain yield per primary panicle (YPP), grain number per primary panicle (GNP), and 1000‐grain weight (TGW) in sorghum [Sorghum bicolor (L.) Moench], a hardy C4 cereal with a genome size of ∼730 Mb, was implemented in a diversity panel containing 390 accessions. These accessions were genotyped to obtain 268,830 single‐nucleotide polymorphisms (SNPs). Genome‐wide association studies (GWAS) were performed to identify loci associated with each grain yield component and understand the genetic interactions between these traits. Genome‐wide association studies identified associations across the genome with YPP, GNP, and TGW that were located within previously mapped sorghum QTL for panicle weight, grain yield, and seed size, respectively. There were no significant associations between GNP and TGW that were within 100 kb, much greater than the average linkage disequilibrium (LD) in sorghum. The identification of nonoverlapping loci for grain number and weight suggests these traits may be manipulated independently to increase the grain yield of sorghum. Following GWAS, genomic regions surrounding each associated SNP were mined for candidate genes. Previously published expression data indicated several TGW candidate genes, including an ethylene receptor homolog, were primarily expressed within developing seed tissues to support GWAS. Furthermore, maize (Zea mays L.) homologs of identified TGW candidates were differentially expressed within the seed between small‐ and large‐kernel lines from a segregating maize population.

Genome‐Wide Association Study for Nine Plant Architecture Traits in Sorghum
Plant Genome - Tập 9 Số 2 - 2016
Jing Hua Zhao, Maria B. Mantilla Perez, Jieyun Hu, Maria G. Salas Fernandez

Sorghum [Sorghum bicolor (L) Moench], an important grain and forage crop, is receiving significant attention as a lignocellulosic feedstock because of its water‐use efficiency and high biomass yield potential. Because of the advancement of genotyping and sequencing technologies, genome‐wide association study (GWAS) has become a routinely used method to investigate the genetic mechanisms underlying natural phenotypic variation. In this study, we performed a GWAS for nine grain and biomass‐related plant architecture traits to determine their overall genetic architecture and the specific association of allelic variants in gibberellin (GA) biosynthesis and signaling genes with these phenotypes. A total of 101 single‐nucleotide polymorphism (SNP) representative regions were associated with at least one of the nine traits, and two of the significant markers correspond to GA candidate genes, GA2ox5 (Sb09 g028360) and KS3 (Sb06 g028210), affecting plant height and seed number, respectively. The resolution of a previously reported quantitative trait loci (QTL) for leaf angle on chromosome 7 was increased to a 1.67 Mb region containing seven candidate genes with good prospects for further investigation. This study provides new knowledge of the association of GA genes with plant architecture traits and the genomic regions controlling variation in leaf angle, stem circumference, internode number, tiller number, seed number, panicle exsertion, and panicle length. The GA gene affecting seed number variation (KS3, Sb06 g028210) and the genomic region on chromosome 7 associated with variation in leaf angle are also important outcomes of this study and represent the foundation of future validation studies needed to apply this knowledge in breeding programs.

Ridge Regression and Other Kernels for Genomic Selection with R Package rrBLUP
Plant Genome - Tập 4 Số 3 - Trang 250-255 - 2011
Jeffrey B. Endelman
Status and Prospects of Association Mapping in Plants
Plant Genome - Tập 1 Số 1 - 2008
Chengsong Zhu, Michael A. Gore, Edward S. Buckler, Jianming Yu

There is tremendous interest in using association mapping to identify genes responsible for quantitative variation of complex traits with agricultural and evolutionary importance. Recent advances in genomic technology, impetus to exploit natural diversity, and development of robust statistical analysis methods make association mapping appealing and affordable to plant research programs. Association mapping identifies quantitative trait loci (QTLs) by examining the marker‐trait associations that can be attributed to the strength of linkage disequilibrium between markers and functional polymorphisms across a set of diverse germplasm. General understanding of association mapping has increased significantly since its debut in plants. We have seen a more concerted effort in assembling various association‐mapping populations and initiating experiments through either candidate‐gene or genome‐wide approaches in different plant species. In this review, we describe the current status of association mapping in plants and outline opportunities and challenges in complex trait dissection and genomics‐assisted crop improvement.

AGHmatrix: R Package to Construct Relationship Matrices for Autotetraploid and Diploid Species: A Blueberry Example
Plant Genome - Tập 9 Số 3 - 2016
Rodrigo R. Amadeu, Catherine Cellon, James W. Olmstead, Antônio Augusto Franco Garcia, Márcio F. R. Resende, Patricio Muñoz

Progress in the rate of improvement in autopolyploid species has been limited compared with diploids, mainly because software and methods to apply advanced prediction and selection methodologies in autopolyploids are lacking. The objectives of this research were to (i) develop an R package for autopolyploids to construct the relationship matrix derived from pedigree information that accounts for autopolyploidy and double reduction and (ii) use the package to estimate the level and effect of double reduction in an autotetraploid blueberry breeding population with extensive pedigree information. The package is unique, as it can create A‐matrices for different levels of ploidy and double reduction, which can then be used by breeders to fit mixed models in the context of predicting breeding values (BVs). Using the data from this blueberry population, we found for all the traits that tetrasomic inheritance creates a better fit than disomic inheritance. In one of the five traits studied, the level of double reduction was different from zero, decreasing the estimated heritability, but it did not affect the prediction of BVs. We also discovered that different depths of pedigree would have significant implications on the estimation of double reduction using this approach. This freely available R package is available for autopolyploid breeders to estimate the level of double reduction present in their populations and the impact in the estimation of genetic parameters as well as to use advanced methods of prediction and selection.

CaDMR1 Cosegregates with QTL Pc5.1 for Resistance to Phytophthora capsici in Pepper (Capsicum annuum)
Plant Genome - Tập 7 Số 2 - 2014
William Z. Rehrig, Hamid Ashrafi, Theresa Hill, James P. Prince, Allen Van Deynze

A major problem for the pepper (Capsicum annuum) industry is the root rot disease caused by Phytophthora capsici (Pc), to which all commercial varieties suffer yield losses despite good management practices and available landraces with high levels of resistance. A high‐density map with 3887 markers was generated in a set of recombinant inbred lines (RIL) derived from the highly resistant Capsicum annuum accession Criollo de Morelos‐334 and Early Jalapeño. These lines have been systematically screened for Pc resistance against a set of isolates collected from Mexico, New Mexico, New Jersey, California, Michigan and Tennessee. Quantitative trait loci (QTL) associated with effective resistance across isolates have been identified and validated with SNP markers across additional segregating populations. By leveraging transcriptomic and genomic information, we describe CaDMR1, a homoserine kinase (HSK), as a candidate gene responsible for the major QTL on chromosome P5 for resistance to Pc. SNP markers for the resistant allele were validated to facilitate gene pyramiding schemes for recurrent selection in pepper.

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