Identification of a Novel S-RNase Gene and S-Genotypes of Four Pear (Pyrus pyrifolia) Cultivars

Lin Zhang, Jiao Hu, X. Tan, Hongxu Long, D. Yuan, Xiu-gen Li
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Abstract

Gametophytic self-incompatibility (GSI) is a genetically controlled mechanism to prevent inbreeding and promote out-crossing. Pyrus pyrifolia is a commercially important fruit tree that exhibits GSI. It is necessary to identify S-genotypes of cultivars for determination of cross-compatible combination prior to performing pear plantation and breeding programs. In this study, an important local cultivar 'Huobali' extensively used as parent in pear breeding programs, and its three progenies 'Mantianhong', 'Hongsucui' and 'Meirensu' were used for S-genotyping analysis by PCR-based molecular method. Agarose gel electrophoresis showed that two fragments were produced in each of the three progenies, while three in the 'Huobali'. After restriction digestion and sequencing analysis, the S-alleles in the four cultivars were determined. The allele of 345 bp from 'Huobali' was determined as a novel S-RNase allele that was tentatively denominated as S44-RNase. RT-PCR revealed that the S44-RNase was expressed specifically in the styles, which is consistent with the expression pattern of S-RNases. Comparison of genomic and cDNA sequences revealed an intron of 148 bp for the S44-RNase. At the amino acid level, it shared 65% to 93% similarity with other Maloideae S-RNases. This study will be helpful in pear production and breeding programs.
4个梨品种S-RNase新基因及其s基因型的鉴定
配子体自交不亲和(GSI)是一种防止近交和促进异交的遗传控制机制。梨叶是一种商业上重要的果树,具有GSI。在实施梨树种植和育种计划之前,有必要对品种进行s基因型鉴定,以确定杂交亲和性组合。本研究以在梨育种中广泛应用的重要地方栽培品种‘火巴梨’及其3个子代‘满天红’、‘红素萃’和‘美仁素’为材料,采用基于pcr的分子分型方法进行了s基因分型分析。琼脂糖凝胶电泳结果显示,3个子代各产生2个片段,而“火八”子代产生3个片段。经酶切和测序分析,确定了4个品种的s等位基因。从“火巴里”获得的345 bp等位基因被确定为一个新的S-RNase等位基因,暂定为S44-RNase。RT-PCR结果显示,S44-RNase在花型中特异表达,与s - rnase的表达模式一致。基因组序列和cDNA序列比较显示S44-RNase的内含子长度为148 bp。在氨基酸水平上,它与其他马洛亚科S-RNases的相似性为65% ~ 93%。本研究对梨的生产和育种具有一定的指导意义。
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