Sydenham Scott L, Barnard Annelie
Agricultural Research Council-Small Grain, Bethlehem, South Africa.
Front Plant Sci. 2018 Feb 1;9:63. doi: 10.3389/fpls.2018.00063. eCollection 2018.
Pre-harvest sprouting (PHS) has been a serious production constraint for over two decades, especially in the summer rainfall wheat production regions of South Africa. It is a complex genetic trait controlled by multiple genes, which are significantly influenced by environmental conditions. This complicates the accurate prediction of a cultivar's stability in terms of PHS tolerance. A number of reports have documented the presence of major QTL on chromosomes 3A and 4A of modern bread wheat cultivars, which confer PHS tolerance. In this study, the SSR marker haplotype combination of chromosomes 3A and 4A of former and current South African cultivars were compared with the aim to select for improved PHS tolerance levels in future cultivars. A total of 101 wheat cultivars, including a susceptible cultivar and five international tolerant sources, were used in this study. These cultivars and donors were evaluated for their PHS tolerance by making use of a rain simulator. In addition, five seeds of each entry were planted out into seedling trays and leaf material harvested for DNA isolation. A modified CTAB extraction method was used before progressing to downstream PCR applications. Eight SSR markers targeted from the well-characterized 3A and 4A QTL regions associated with PHS tolerance, were used to conduct targeted haplotype analysis. Additionally, recently published KASP SNP markers, which identify the casual SNP mutations within the gene, were used to genotype the germplasm. The haplotype marker data and phenotypic PHS data were compared across all cultivars and different production regions. A relative change in observed phenotypic variation percentage was obtained per marker allele and across marker haplotype combinations when compared to the PHS susceptible cultivar, Tugela-DN. Clear favorable haplotypes, contributing 40-60% of the variation for PHS tolerance, were identified for QTL 3A and 4A. Initial analyses show haplotype data appear to be predictive of PHS tolerance status and germplasm can now be selected to improve PHS tolerance. These haplotype data are the first of its kind for PHS genotyping in South Africa. In future, this can be used as a tool to predict the possible PHS tolerance range of a new cultivar.
收获前发芽(PHS)在二十多年来一直是严重制约生产的因素,尤其是在南非夏季降雨的小麦产区。它是一个由多个基因控制的复杂遗传性状,受环境条件影响显著。这使得准确预测品种在PHS耐受性方面的稳定性变得复杂。许多报告记录了现代面包小麦品种3A和4A染色体上存在赋予PHS耐受性的主要QTL。在本研究中,比较了南非过去和现在品种3A和4A染色体的SSR标记单倍型组合,目的是在未来品种中选择提高PHS耐受性水平。本研究共使用了101个小麦品种,包括一个感病品种和五个国际耐受性种质。利用降雨模拟器对这些品种和种质的PHS耐受性进行了评估。此外,每个材料选取五粒种子种在育苗盘中,收获叶片材料用于DNA提取。在进行下游PCR应用之前,使用了改良的CTAB提取方法。从与PHS耐受性相关的特征明确的3A和4A QTL区域靶向了八个SSR标记,用于进行靶向单倍型分析。此外,最近发表的用于鉴定该基因内偶然SNP突变的KASP SNP标记,用于对种质进行基因分型。比较了所有品种和不同生产区域的单倍型标记数据和表型PHS数据。与PHS感病品种图盖拉-DN相比,每个标记等位基因和整个标记单倍型组合的观察表型变异百分比都有相对变化。确定了对QTL 3A和4A明确有利的单倍型,其对PHS耐受性变异的贡献率为40%-60%。初步分析表明,单倍型数据似乎可以预测PHS耐受性状态,现在可以选择种质来提高PHS耐受性。这些单倍型数据是南非首次用于PHS基因分型的此类数据。未来,这可以作为一种工具来预测新品种可能的PHS耐受性范围。