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棉花学报 ›› 2019, Vol. 31 ›› Issue (2): 156-162.doi: 10.11963/1002-7807.txqml.20190322

• 研究与进展 • 上一篇    下一篇

棉纤维DNA的提取及其在品种溯源中的尝试

田新权#(),付小琼#(),时萌,方丹,徐双娇,马磊*()   

  1. 棉花生物学国家重点实验室/农业农村部棉花产品质量安全风险评估实验室/中国农业科学院棉花研究所,河南 安阳 455000
  • 收稿日期:2018-04-25 出版日期:2019-03-15 发布日期:2019-03-15
  • 通讯作者: 马磊 E-mail:tianxq0918@163.com;m13663726262@163.com;malei2002@163.com
  • 作者简介:田新权(1990—),男,硕士, tianxq0918@163.com。|付小琼, m13663726262@163.com
  • 基金资助:
    国家现代农业产业技术体系(CARS-15-25);国家农产品质量安全风险评估专项(GJFP201801003);棉花生物学国家重点实验室自主课题(CB2017C15)

Extraction of Cotton Fiber DNA and Its Application in Traceability

Tian Xinquan#(),Fu Xiaoqiong#(),Shi Meng,Fang Dan,Xu Shuangjiao,Ma Lei*()   

  1. State Key Laboratory of Cotton Biology/Laboratory of Quality and Safety Risk Assessment for Cotton Products, Ministry of Agriculture and Rural Affairs/ Institute of Cotton Research of the Chinese Academy of Agricultural Sciences, Anyang, Henan 455000,China
  • Received:2018-04-25 Online:2019-03-15 Published:2019-03-15
  • Contact: Ma Lei E-mail:tianxq0918@163.com;m13663726262@163.com;malei2002@163.com

摘要:

【目的】针对我国进口棉花原料以及收储棉花的品种产地溯源监控技术体系缺乏的问题,探索了适用于棉纤维DNA的提取方法,旨在建立以棉纤维DNA为介质鉴定棉花品种真实性及产地溯源的体系。【方法】通过改进和优化提取方法,提取了开花后不同时间棉纤维及不同年份皮棉的DNA,以其作为模板,进行了常规聚合酶链式反应(Polymerase chain reaction,PCR)扩增;并选取13对简单重复序列(Simple sequence repeat,SSR)引物对鲁1127、石抗126和瑞杂816棉纤维DNA进行SSR-PCR扩增。【结果】此DNA提取法可提取出不同发育时期和不同年份的棉纤维DNA,随着纤维发育成熟及年代增加,所提取的DNA含量尽管有所降低,但仍能满足下游试验需求;所提取的棉纤维DNA可进行常规PCR扩增,且PCR扩增条带清晰;以13对棉花SSR引物对鲁1127、石抗126和瑞杂816棉纤维DNA进行SSR-PCR扩增,均可扩增出清晰的多态性谱带,且易于区分。【结论】该提取法适合提取棉花纤维的基因组DNA,且满足于常规PCR扩增和SSR分子标记。本研究证实基于棉纤维DNA分子标记用于棉花品种溯源切实可行,并有望为保障我国棉花产业安全提供技术支持。

关键词: 棉花纤维; DNA; SSR分子标记; 品种溯源

Abstract:

[Objective] To address the questions of lacking monitoring system for origins of varieties, this study explored a DNA extraction method suitable for cotton fiber, aiming to establish an identification system that uses cotton fiber DNA as a medium to trace the authenticity and cultivar of cotton varieties. [Method] By improving and optimizing the extraction method, the DNAs of cotton fibers at different days post anthesis and lint from different years were extracted and used as templates for routine polymerase chain reaction (PCR) amplification. 13 pairs of simple sequence repeat(SSR) primers were selected for SSR-PCR amplification using the DNA of Lu 1127, Shikang 126 and Ruiza 816. [Result] The DNA extraction method can extract cotton fiber DNAs of different developmental stages and different years. With the development of fiber, although the extracted DNA content is reduced, it can still meet the needs of downstream experiments. The extracted cotton fiber DNAs can be subjected to conventional PCR amplification, the PCR amplified bands were clear; 13 pairs of cotton SSR primers were used for SSR-PCR amplification of cotton fiber DNAs of Lu 1127, Shikang 126 and Ruiza 816, and a clear polymorphic band could be amplified and easily distinguished. [Conclusion] This extraction method is suitable for extracting genomic DNA of cotton fiber, and is satisfied with conventional PCR amplification with SSR markers. It is confirmed that the cotton fiber DNA molecular markers are feasible for tracing the source of cotton varieties, and it is expected to provide technical support for ensuring the safety of cotton industry.

Key words: cotton fiber; DNA; SSR molecular marker; varieties traceability