Ali Jauhar, Aslam Umair M, Tariq Rida, Murugaiyan Varunseelan, Schnable Patrick S, Li Delin, Marfori-Nazarea Corinne M, Hernandez Jose E, Arif Muhammad, Xu Jianlong, Li Zhikang
International Rice Research Institute, Los Baños, Philippines.
Institute of Crop Science, University of the Philippines Los Baños, Los Baños, Philippines.
Front Plant Sci. 2018 Jun 22;9:849. doi: 10.3389/fpls.2018.00849. eCollection 2018.
This study demonstrates genotyping-by-sequencing-based single-nucleotide polymorphism (SNP)-typing in 11 early-backcross introgression populations of rice (at BCF), comprising a set of 564 diverse introgression lines and 12 parents. Sequencing using 10 Ion Proton runs generated a total of ∼943.4 million raw reads, out of which ∼881.6 million reads remained after trimming for low-quality bases. After alignment, 794,297 polymorphic SNPs were identified, and filtering resulted in LMD50 SNPs (low missing data, with each SNP, genotyped in at least 50% of the samples) for each sub-population. Every data point was supported by actual sequencing data without any imputation, eliminating imputation-induced errors in SNP calling. Genotyping substantiated the impacts of novel breeding strategy revealing: (a) the donor introgression patterns in ILs were characteristic with variable introgression frequency in different genomic regions, attributed mainly to stringent selection under abiotic stress and (b) considerably lower heterozygosity was observed in ILs. Functional annotation revealed 426 non-synonymous deleterious SNPs present in 102 loci with a range of 1-4 SNPs per locus and 120 novel SNPs. SNP-typing this diversity panel will further assist in the development of markers supporting genomic applications in molecular breeding programs.
本研究展示了在11个水稻早期回交渗入群体(处于BCF阶段)中基于测序的单核苷酸多态性(SNP)基因分型,该群体包括一组564个不同的渗入系和12个亲本。使用10次Ion Proton测序运行共产生了约9.434亿条原始读数,其中经过低质量碱基修剪后还剩下约8.816亿条读数。比对后,鉴定出794,297个多态性SNP,经过筛选得到了每个亚群体的LMD50 SNP(低缺失数据,每个SNP在至少50%的样本中进行了基因分型)。每个数据点都得到了实际测序数据的支持,无需任何填充,消除了SNP检测中因填充导致的错误。基因分型证实了新育种策略的影响,揭示出:(a)渗入系中的供体渗入模式具有特征性,不同基因组区域的渗入频率各不相同,这主要归因于非生物胁迫下的严格选择;(b)在渗入系中观察到杂合度显著降低。功能注释揭示了102个位点中存在426个非同义有害SNP,每个位点有1 - 4个SNP,以及120个新SNP。对这个多样性群体进行SNP分型将进一步有助于开发支持分子育种计划中基因组应用的标记。