过表达钾转运蛋白基因trkH提高玉米的钾营养

doi:10.7525/j.issn.1673-5102.2020.01.019

摘要/Abstract

摘要： 钾是植物生长发育必须的营养元素，参与多种生理生化过程。土壤缺钾严重影响了玉米的产量和品质，通过遗传改良的方式提高玉米的钾营养是解决这个问题的有效途径。本实验室前期从海洋微生物宏基因组DNA中克隆得到细菌钾转运蛋白基因trkH，在酵母和烟草中验证了功能。为进一步分析trkH基因在玉米中的功能，以Hi-Ⅱ基因型玉米为转化受体，采用农杆菌介导法将trkH基因转入玉米，获得具有Baster抗性的独立转化苗21株。PCR检测结果表明trkH基因已成功转入到玉米基因组中。Bar试纸条检测结果表明Bar基因在蛋白水平上成功表达。将部分T₀代转基因植株与玉米骨干自交系PH6WC杂交，获得8个T₁代株系，半定量RT-PCR检测结果表明转基因植株在转录水平上均能表达。三叶一心期喷洒除草剂进行筛选，选择比例接近1：1的L3、L5和L7转基因株系进行PCR检测，统计检测结果并进行χ²测验，结果表明符合孟德尔分离定律。L3、L5和L7转基因株系玉米苗期的钾耗竭实验结果表明过表达trkH基因能够提高转基因玉米的K⁺吸收，为培育钾营养高效玉米新品种奠定基础。

关键词: trkH, 玉米, 遗传转化, 钾吸收

Abstract: Potassium is a necessary nutrient for plant growth and development, which is involved in various physiological and biochemical processes. Potassium deficiency seriously affects the quality and yield of maize. To improve K⁺ nutrition of maize by genetic modification is one of the effective approaches to solve this problem. The bacterial K⁺ transporter gene trkH was cloned from marine microbial metagenomic DNA, and its function was verified in yeast and tobacco. To further analyze the function of trkH gene in maize, the trkH gene was transferred into Hi-Ⅱ variety by Agrobacterium-mediated transformation method, and 21 independent baster resistant seedlings were obtained. By PCR, the trkH gene was successfully introduced into maize genome. By Bar strip test, the Bar gene was expressed at the protein level. Partial T₀ transgenic plants were crossed with the maize backbone inbred line PH6WC and eight hybrid progeny were obtained. By semi-quantitative RT-PCR, trkH gene was expressed at the transcriptional level. Herbicide was sprayed at the three-leaf and one-heart stage, and L3, L5 and L7 lines with a ratio close to 1:1 were selected for PCR detection. Chi-square was performed with PCR data and the results indicate that the ratio is in accordance with Mendelian segregation law. K⁺ depletion experiment showed that overexpression of trkH gene improved K⁺ absorption of L3, L5 and L7 transgenic seedlings, which would lay the foundation of breeding new maize variety with high efficient potassium nutrition.

Key words: trkH, maize, genetic transformation, K⁺ uptake

中图分类号:

S513

丁宝娟, 安利佳, 苏乔. 过表达钾转运蛋白基因trkH提高玉米的钾营养[J]. 植物研究, 2020, 40(1): 141-147.

DING Bao-Juan, AN Li-Jia, SU Qiao. Overexpression of K Transporter Gene trkH in Enhancing K Nutrition in Maize[J]. Bulletin of Botanical Research, 2020, 40(1): 141-147.

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