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作物学报 ›› 2023, Vol. 49 ›› Issue (7): 1994-2001.doi: 10.3724/SP.J.1006.2023.24196

• 研究简报 • 上一篇    下一篇

马铃薯锌转运蛋白基因StZIP12调控锌吸收功能

索海翠(), 刘计涛, 王丽, 李成晨, 单建伟, 李小波*()   

  1. 广东省农业科学院作物研究所 / 广东省农作物遗传改良重点实验室, 广东广州 510640
  • 收稿日期:2022-08-26 接受日期:2022-11-28 出版日期:2023-07-12 网络出版日期:2022-12-12
  • 通讯作者: *李小波, E-mail: Lixiaobo1981@163.com
  • 作者简介:E-mail: suohaicui@163.com
  • 基金资助:
    本研究由国家自然科学基金青年基金项目(31701486)

Functional analysis of StZIP12 in regulating potato Zn uptake

SUO Hai-Cui(), LIU Ji-Tao, WANG Li, LI Cheng-Chen, SHAN Jian-Wei, LI Xiao-Bo*()   

  1. Crops Research Institute, Guangdong Academy of Agricultural Sciences / Guangdong Province Key Laboratory of Crop Genetic Improvement, Guangzhou 510640, Guangdong, China
  • Received:2022-08-26 Accepted:2022-11-28 Published:2023-07-12 Published online:2022-12-12
  • Contact: *E-mail: Lixiaobo1981@163.com
  • Supported by:
    The National Natural Science Foundation of China’s Youth Fund(31701486)

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摘要：

马铃薯作为世界第四大粮食作物, 是全球2/3人口的主食, 是锌生物强化策略的重要目标作物, 因此研究马铃薯锌吸收具有重要作用。锌转运蛋白调节植物锌吸收过程。本研究在前期研究基础上克隆了马铃薯锌转运蛋白基因StZIP12, 定量PCR结果表明该基因在马铃薯各组织中均有表达, 在新叶中表达量显著高于其他组织, 且受到低锌诱导表达; 利用异源酵母互补试验证实了StZIP12能够恢复锌吸收障碍酵母突变体ZHY3 (zrt1zrt2)的锌吸收功能; 利用转基因手段过表达StZIP12于马铃薯品种鄂薯3号, 在缺锌处理下, 与非转基因对照植株相比较, 过表达StZIP12提高了转基因马铃薯株高, 转基因组培苗的根长和总锌含量以及盆栽转基因植株块茎的锌含量增加, 其中2个株系块茎锌含量分别比对照提高22.00%和32.95%。以上结果证明了StZIP12在马铃薯锌的吸收过程中起着重要调节作用, 为马铃薯锌生物强化提供了理论依据。

关键词: 马铃薯, 锌转运蛋白, StZIP12, 锌吸收

Abstract:

Potato is the fourth largest food crop in the world. Two-thirds of the world’s population feed on potato, which is an important target crop for zinc biofortification strategy. It is of great importance to study zinc uptake in potato. Zinc transporters regulate zinc uptake in plants. In this study, we cloned potato zinc transporter StZIP12 on the basis of previous studies. Quantitative PCR results showed that StZIP12 was expressed in all potato tissues, and the relative expression level of StZIP12 in young leaves was significantly higher than that in other tissues. In addition, low zinc treatment could also induce StZIP12 expression. The heterologous yeast complementation test confirmed that StZIP12 could restore the zinc absorption function of ZHY3 (zrt1zrt2) yeast mutant. We obtained StZIP12 overexpressing transgenic plants in cultivar potato Eshu 3 by genetic transformation method. Under zinc deficiency treatment, the overexpression of StZIP12 increased plant height of transgenic plants, root length, and the total zinc content of transgenic seedlings, and the zinc content of tubers of pot-cultured transgenic plants. The zinc content of tubers of two lines was 22.00% and 32.95% higher than that of the control, respectively. These results prove that StZIP12 playes an important role in the absorption of zinc in potato, and provides a theoretical basis for the biofortification of potato zinc.

Key words: potato, zinc transporter, StZIP12, zinc absorption

图1

StZIP12基因的克隆及跨膜结构域分析 A: StZIP12基因的分离; 1、2、3泳道均为马铃薯StZIP12基因, M为DL2000 DNA marker。B: StZIP12序列分析, 灰色字母代表氨基酸序列。C: StZIP12跨膜结构域分析, 黄色区域代表跨膜结构部分, 灰色区域代表细胞膜。"

图2

StZIP12蛋白亚细胞定位在细胞膜上 标尺为50 μm。"

图3

StZIP12的表达分析 误差线代表标准误, 多重比较采用Duncan分析法, 不同小写字母代表显著水平(P < 0.05)。"

图4

StZIP12对酵母锌吸收障碍突变体功能互补 VC: 空载体对照; 10-1~10-4代表菌液稀释浓度。"

图5

转基因株系定量PCR检测 ES3: 非转基因对照; OE-26、OE-27、OE-28、OE-31、OE-32、OE-47、OE-67为过表达株系。"

图6

缺锌处理下马铃薯转基因组培苗的表型及锌含量 A: 植株表型及锌含量分析, 误差线代表标准误, 多重比较采用Duncan分析法, 不同小写字母代表显著水平(P < 0.05)。B: 植株表型图片; C: 根体积分级。CK: 正常锌;-Zn: 缺锌处理。"

图7

正常和缺锌处理下下转基因马铃薯植株表型、株高和锌含量 CK: 正常锌;-Zn: 缺锌处理。误差线代表标准误, 多重比较采用Duncan分析法, 不同小写字母代表显著水平(P < 0.05)。"

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