De Mori Gloria, Falchi Rachele, Testolin Raffaele, Bassi Daniele, Savazzini Federica, Dondini Luca, Tartarini Stefano, Palmisano Francesco, Minafra Angelantonio, Spadotto Alessandro, Scalabrin Simone, Geuna Filippo
Department of Agricultural, Food, Environmental and Animal Sciences, University of Udine, Udine, Italy.
Department of Agricultural and Environmental Sciences (DISAA), University of Milan, Milan, Italy.
Front Plant Sci. 2019 Dec 4;10:1576. doi: 10.3389/fpls.2019.01576. eCollection 2019.
Sharka, a common disease among most stone fruit crops, is caused by the (PPV). Resistant genotypes have been found in apricot ( L.), one of which-the cultivar 'Lito' heterozygous for the resistance-has been used to map a major quantitative trait locus (QTL) on linkage group 1, following a pseudo-test-cross mating design with 231 individuals. In addition, 19 SNP markers were selected from among the hundreds previously developed, which allowed the region to be limited to 236 kb on chromosome 1. A 'Lito' bacterial artificial chromosome (BAC) library was produced, screened with markers of the region, and positive BAC clones were sequenced. Resistant (R) and susceptible (S) haplotypes were assembled independently. To refine the assembly, the whole genome of 'Lito' was sequenced to high coverage (98×) using PacBio technology, enabling the development of a detailed assembly of the region that was able to predict and annotate the genes in the QTL region. The selected cultivar 'Lito' allowed not only to discriminate structural variants between the two haplotypic regions but also to distinguish specific allele expression, contributing towards mining the locus. In light of these findings, genes previously indicated (i.e., genes) to have a possible role in PPV resistance were further analyzed, and new candidates were discussed. Although the results are not conclusive, the accurate and independent assembly of R and S haplotypes of 'Lito' is a valuable resource to predict and test alternative transcription and regulation mechanisms underpinning PPV resistance.
李痘病是大多数核果类作物中常见的一种病害,由李痘病毒(PPV)引起。在杏(Prunus armeniaca L.)中发现了抗性基因型,其中一个抗性杂合的品种“Lito”,在与231个个体进行假测交交配设计后,被用于在第1连锁群上定位一个主要的数量性状位点(QTL)。此外,从之前开发的数百个标记中选择了19个SNP标记,这使得该区域在1号染色体上被限定在236 kb。构建了一个“Lito”细菌人工染色体(BAC)文库,用该区域的标记进行筛选,并对阳性BAC克隆进行测序。抗性(R)和感病(S)单倍型被独立组装。为了优化组装,使用PacBio技术对“Lito”的全基因组进行了高覆盖度(98×)测序,从而能够开发出该区域的详细组装,进而预测和注释QTL区域中的基因。所选品种“Lito”不仅能够区分两个单倍型区域之间的结构变异,还能区分特定等位基因的表达,有助于挖掘该位点。鉴于这些发现,对之前指出的(即NBS-LRR基因)可能在PPV抗性中起作用的基因进行了进一步分析,并讨论了新的候选基因。尽管结果尚无定论,但“Lito”的R和S单倍型的准确且独立的组装是预测和测试PPV抗性潜在的替代转录和调控机制的宝贵资源。