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基于激酶结构域序列分析的萝卜S单倍型分类与鉴定

Classification and Identification of S Haplotypes in Radish Based on Kinase Domain Sequence Analysis.

作者信息

Ni Meng, Yi Xiaofang, Wang Qin, Wang Juan, Wang Shuang, Liu Liwang, Xu Liang, Wang Yan

机构信息

State Key Laboratory of Crop Genetics and Germplasm Enhancement/Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crop in East China, Ministry of Agriculture and Rural Affairs, College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China.

出版信息

Plants (Basel). 2022 Sep 2;11(17):2304. doi: 10.3390/plants11172304.

Abstract

Radish is a typical self-incompatible crop. The rapid and accurate identification of S haplotypes can circumvent the blindness of the hybrid combination process, which is critical in radish heterosis utilization and the breeding of new varieties. In this study, based on the gene sequence which encodes the -locus receptor kinase (SRK) of radish, and the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis, the S haplotypes were identified among 79 cultivated radish genotypes. The PCR results indicated that 79 radish genotypes could be divided into 48 Class I, 13 Class II, and 17 Class I/II S haplotypes. Sequence alignment confirmed that the Class I materials contained 19 S haplotypes, of which three haplotypes ('NAU-S53', 'NAU-S54' and 'NAU-S55') were identified for the first time in radish. After digestion using the f I restriction endonuclease, the domain of DNA fragments of different genotypes showed high polymorphism. Homozygous materials S haplotypes could be quickly distinguished by the differences in the digested bands. Molecular identification of the S haplotype was highly consistent with the field pollination and pollen tube germination results. These results would provide an important approach for the rapid identification of radish S haplotypes and the efficient utilization of self-incompatibility in heterosis breeding.

摘要

萝卜是典型的自交不亲和作物。快速准确地鉴定S单倍型可以避免杂交组合过程中的盲目性,这在萝卜杂种优势利用和新品种选育中至关重要。本研究基于编码萝卜S位点受体激酶(SRK)的基因序列,通过聚合酶链反应-限制性片段长度多态性(PCR-RFLP)分析,在79份栽培萝卜基因型中鉴定了S单倍型。PCR结果表明,79份萝卜基因型可分为48种I类、13种II类和17种I/II类S单倍型。序列比对证实,I类材料包含19种S单倍型,其中三种单倍型(‘NAU-S53’、‘NAU-S54’和‘NAU-S55’)为萝卜首次鉴定。用f I限制性内切酶消化后,不同基因型DNA片段的结构域显示出高度多态性。通过消化条带的差异可以快速区分纯合材料的S单倍型。S单倍型的分子鉴定与田间授粉和花粉管萌发结果高度一致。这些结果将为快速鉴定萝卜S单倍型和高效利用杂种优势育种中的自交不亲和性提供重要方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d00/9459979/86dfae2ecd3b/plants-11-02304-g001.jpg

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