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园艺学报 ›› 2015, Vol. 42 ›› Issue (7): 1251-1259.doi: 10.16420/j.issn.0513-353x.2015-0104

• 果树 • 上一篇    下一篇

甜樱桃‘拉宾斯’自交亲和性与SFB4′ 基因的关系研究

李洋,李长龙,王晶,闫国华,张晓明,李威,张开春,李天忠   

  1. 1中国农业大学农学与生物技术学院,北京 100193;2北京市农林科学院林业果树研究所,北京 100093;3北京市落叶果树工程技术研究中心,北京 100093;4农业部华北地区园艺作物生物学与种质创制重点实验室,北京 100093
  • 出版日期:2015-07-25 发布日期:2015-07-25
  • 基金资助:
    ‘十二五’国家科技支撑计划项目(2013BAD02B03);国家自然科学基金项目(31272123);国家自然科学基金青年基金项目(31401827);科技部科技基础性工作专项(2012FY110100);北京市科委重大项目(D131100000113001)

Research of Relationship Between Sweet Cherry Lapins Self-compatibility and SFB4′ Gene

LI Yang, LI Chang-Long, WANG Jing, YAN Guo-Hua, ZHANG Xiao-Ming, LI Wei, ZHANG Kai-Chun, LI Tian-Zhong   

  1. 1College of Agriculture and Biotechnology,China Agricultural University,Beijing 100193,China;2Institute of Forestry and Pomology,Beijing Academy of Agriculture and Forestry Sciences,Beijing 100093,China;3Beijing Engineering Research Center for Deciduous Fruit Trees,Beijing 100093,China;4Key Laboratory of Biology and Genetic Improvement of Horticultural Crops(North China),Ministry of Agriculture,Beijing 100093,China
  • Online:2015-07-25 Published:2015-07-25

摘要: 甜樱桃品种‘拉宾斯’与‘雷尼’均为S1S4基因型。调查其自花结实率,‘拉宾斯’为31.3%,表现自交亲和;‘雷尼’为0,表现为自交不亲和。将‘拉宾斯’与‘雷尼’相互授粉,‘拉宾斯’为父本时表现亲和,反之不亲和,推断‘拉宾斯’的自交亲和性由花粉侧突变导致;调查花粉萌发率,‘拉宾斯’为58.08%,‘雷尼’为57.82%。鉴定‘拉宾斯’自交后代及其与‘雷尼’杂交后代的S-RNase的基因型,发现所有后代中均出现S4基因型,部分后代中出现S1基因型,证明‘拉宾斯’花粉育性是正常的,其亲和性可能是由S4基因座上花粉侧的基因突变导致。测序发现‘拉宾斯’花粉SFB4′ 在911 bp处有4个碱基缺失。分析‘拉宾斯’花粉SFB4′ 与花柱S4-RNase的连锁性,发现两者连锁率为100%。以‘拉宾斯’为父本的带有SFB4′ 的杂交后代的自花结实率为23.3% ~ 61.4%,表现自交亲和,说明来源于‘拉宾斯’的SFB4′ 基因与自交亲和性有关。RT-PCR及测序显示‘雷尼’SFB4与‘拉宾斯’SFB4′ 均能正常转录,但4个碱基缺失导致其C端翻译提前终止,原核表达SFB4和SFB4′ 蛋白发现SFB4略大于SFB4′,推测是由于SFB4′ 的翻译提前终止导致的。以上结果表明:‘拉宾斯’的自交亲和性可能是由花粉侧SFB4′ C端的氨基酸缺失导致,该缺失可能使其丧失了与S-RNase的识别能力。

关键词: 甜樱桃, 自交亲和性, 花粉, SFB4′, 氨基酸缺失

Abstract: The sweet cherry Lapins and Rainier were S1S4 genotype,self-pollination rate of Lapinswas 31.3%,performed self-compatibility and that of Rainier was 0,showed self-incompatibility. In the reciprocal cross experiment,it showed compatible when Lapins was the male parent,however,it showed incompatible if using Rainier as male parent. This result suggested that the reason of the Lapins self-compatibility might be caused by pollen side mutation;The pollen germination rate of Lapins was 58.08% and that of Rainier was 57.82%. The S genotypes offspring of self-pollination and cross-pollination were identified by Rainier and the results showed that S4 genotype can be detected in all generations,but S1 genotype was in part of the offspring,this further proved that Lapins pollen can germinate normal,and the self-compatibility may be caused by a mutated gene on S4 genotype pollen side. Previous studies had found that Lapins pollen SFB4 has 4 nucleotide deletion in 911 bp,and this SFB4′ gene linked style S4-RNase closely in our experiment,the linkage rate was 100%;The self-fruit rate ranged from 23.3% to 61.4% in the offsprings contained SFB4′,suggest that Lapins self-compatible may be determined by SFB4′;RT-PCR and sequencing displayed that Lapins SFB4′ and Rainier SFB4 can transcription normally,but 4 bp DNA deletion should cause a truncated protein in C terminal,and SDS-PAGE showed that the SFB4-GST fusion protein was a bit bigger than the SFB4′-GST protein. The results above showed that:The self-compatible of Lapins may be caused by the pollen side SFB4′ mutation,amino acid deletion may lead SFB4′ loss the ability to recognize S-RNase.

Key words: sweet cherry, self-incompatibility, pollen, SFB4′, amino acid deletion

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