农杆菌直接转化粳稻种子的高效方案:克服愈伤组织再生的复杂性,避免体细胞无性系突变的发生。

IF 2 3区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS
Md Maksudul Haque, Yutaro Okumura, Shin-Ichiro Kidou
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引用次数: 0

摘要

植物的分子育种和基因功能研究需要较高的转化效率。农杆菌介导的转化对许多植物的分子研究做出了重大贡献,但在没有农杆菌的水稻中效率低下且不一致。水稻的转化效率仍然很低。因此,本研究旨在建立一种简单高效的农杆菌转化水稻的方法。实验采用两个外源基因(CISP1-GFP和CISP2-GFP)和农杆菌菌株(EHA105)。然后,对在减压浸渍条件下吸水发芽的水稻种子进行农杆菌侵染,生长后形成的种子中平均有14% (CISP1-GFP为12%,CISP2-GFP为16%)携带外源基因,并通过PCR、Western blot、GFP荧光和tailpcr证实了这一点。由于这种方法不涉及水稻植株的愈伤组织形成和再分化,因此不需要特殊的设备和复杂的操作,仅需3个月即可获得转化体。因此,该方法有望简化水稻的遗传操作,促进水稻的分子育种。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Highly efficient direct seed transformation protocol for japonica rice (Oryza sativa L.) by Agrobacterium tumefaciens: overcoming the complexity of callus regeneration and avoiding the occurrence of somaclonal-mutations.

Molecular breeding and gene function studies in plants require high transformation efficiency. Agrobacterium-mediated transformation has contributed significantly to molecular research in many plants, but is inefficient and inconsistent in rice that do not host Agrobacterium. Transformation efficiency in rice remains low. Therefore, this study aimed to establish a simple and efficient transformation method for rice using Agrobacterium. Two foreign genes (CISP1-GFP and CISP2-GFP) and Agrobacterium strain (EHA105) was used in the experiments. Then, Agrobacterium infection of rice seeds that had absorbed water and germinated under reduced pressure infiltration conditions showed that an average of 14% of the seeds formed after growth (12% with CISP1-GFP and 16% with CISP2-GFP) carried the foreign gene, and it was also confirmed by PCR, Western blot, GFP fluorescence and TAIL-PCR. Since this method does not involve callus formation or re-differentiation of rice plants, no special equipment or complicated operations are required, and transformants can be obtained in only three months. Therefore, this method is expected to simplify rice genetic manipulation and promote molecular breeding of rice.

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来源期刊
Transgenic Research
Transgenic Research 生物-生化研究方法
CiteScore
5.40
自引率
0.00%
发文量
38
审稿时长
4-8 weeks
期刊介绍: Transgenic Research focusses on transgenic and genome edited higher organisms. Manuscripts emphasizing biotechnological applications are strongly encouraged. Intellectual property, ethical issues, societal impact and regulatory aspects also fall within the scope of the journal. Transgenic Research aims to bridge the gap between fundamental and applied science in molecular biology and biotechnology for the plant and animal academic and associated industry communities. Transgenic Research publishes -Original Papers -Reviews: Should critically summarize the current state-of-the-art of the subject in a dispassionate way. Authors are requested to contact a Board Member before submission. Reviews should not be descriptive; rather they should present the most up-to-date information on the subject in a dispassionate and critical way. Perspective Reviews which can address new or controversial aspects are encouraged. -Brief Communications: Should report significant developments in methodology and experimental transgenic higher organisms
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