四川农业大学学报 ›› 2016, Vol. 34 ›› Issue (04): 402-405.doi: 10.16036/j.issn.1000-2650.2016.04.002

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利用SLAF-seq技术开发新寡核苷酸探针鉴定小麦背景中的黑麦染色体

刘洪坤1, 唐宗祥2   

  1. 1. 南充职业技术学院, 四川 南充 637000;
    2. 四川农业大学植物遗传育种省级重点实验室, 成都 611130
  • 收稿日期:2016-09-05 出版日期:2016-12-31 发布日期:2017-02-15
  • 通讯作者: 唐宗祥,博士,教授,主要从事作物遗传育种研究工作,E-mail:zxtang@sicau.edu.cn。 E-mail:zxtang@sicau.edu.cn
  • 作者简介:刘洪坤,副教授,E-mail:893146765@qq.com。
  • 基金资助:
    国家自然科学基金项目(31471498)

Development of Novel Oligonucleotide Probes for Identifying Rye Chromosomes in Wheat Background

LIU Hong-kun1, TANG Zong-xiang2   

  1. 1. Nanchong Vocational and Technical College, Nanchong 637000, Sichuan, China;
    2. Province Key Laboratory of Plant Breeding and Genetics, Sichuan Agricultural University, Chengdu 611130, China
  • Received:2016-09-05 Online:2016-12-31 Published:2017-02-15

摘要: [目的] 通过荧光原位杂交(FISH)技术能有效地检测小麦背景中的黑麦染色体,进一步发现利用寡核苷酸探针和非变性FISH(ND-FISH)技术可以提高黑麦染色体的检测效率。[方法] 通过借助SLAF-seq(specific length amplified fragment sequencing)和生物信息学方法开发寡核苷酸探针。[结果] 开发了3种新的寡核苷酸探针Oligo-2874、Oligo-5296和Oligo-9965,它们可用于ND-FISH分析。通过与小麦背景中的黑麦染色体端部和亚端部特异杂交,从而达到识别黑麦染色体的目的。[结论] 这些寡核苷酸探针可以用于快速高效地鉴定小麦背景中的黑麦染色体,丰富了小麦-黑麦杂交后代FISH分析的探针源。研究结果也表明SLAF-seq技术可以用来开发小麦近缘种属特异的重复序列FISH分析探针。

关键词: 小麦, 黑麦, 寡核苷酸探针, 非变性FISH, SLAF-seq

Abstract: [Objective] Fluorescence in situ hybridization(FISH) technology can be used to identify rye chromosomes effectively in wheat backgrounds. Oligonucleotide probes and non-denaturing FISH(NDFISH) can increase the efficiency of detecting rye chromosomes. [Method] In this study, SLAF-seq(specific length amplified fragment sequencing)and bioinformatics methods were used to develop novel oligonucleotide probes. [Results] Three novel oligonucleotide probes Oligo-2874, Oligo-5296 and Oligo-9965 were developed and they can be used for ND-FISH assays. The three probes only hybridize to telomeres and sub-telomeres of rye chromosomes. [Conclusion] Therefore, these oligonucleotide probes can discriminate rye chromosomes in wheat backgrounds quickly and efficiently, and they enrich the probe sources for FISH analysis of wheat-rye hybrids. The results also indicate that SLAF-seq can be used to develop specific repetitive DNA probes for FISH analysis of relatives of wheat.

Key words: wheat, rye, oligonucleotideprobe, non-denaturingFISH, SLAF-seq

中图分类号: 

  • S512.5