[1]于海伟,李铭杨,张军,等.大豆GmNCED1-2的非生物胁迫诱导表达及其转基因烟草鉴定[J].江苏农业学报,2023,(04):931-938.[doi:doi:10.3969/j.issn.1000-4440.2023.04.002]
　YU Hai-wei,LI Ming-yang,ZHANG Jun,et al.Expression of soybean GmNCED1-2 induced by abiotic stress and the identification of GmNCED1-2 transgenic tobacco[J].,2023,(04):931-938.[doi:doi:10.3969/j.issn.1000-4440.2023.04.002]
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大豆GmNCED1-2的非生物胁迫诱导表达及其转基因烟草鉴定()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2023年04期
页码:
931-938
栏目:
遗传育种·生理生化
出版日期:
2023-08-30

文章信息/Info

Title:
Expression of soybean GmNCED1-2 induced by abiotic stress and the identification of GmNCED1-2 transgenic tobacco
作者:
于海伟1; 李铭杨1; 张军2; 李珊珊1; 张梅娟1; 马天意1; 赵艳1; 兰红宇3; 翟莹1
(1.齐齐哈尔大学生命科学与农林学院，黑龙江齐齐哈尔161006；2.黑龙江省农业科学院畜牧兽医分院，黑龙江齐齐哈尔161005；3.黑龙江省农业科学院齐齐哈尔分院，黑龙江齐齐哈尔161006)
Author(s):
YU Hai-wei1; LI Ming-yang1; ZHANG Jun2; LI Shan-shan1; ZHANG Mei-juan1; MA Tian-yi1; ZHAO Yan1; LAN Hong-yu3; ZHAI Ying1
(1.College of Life Science and Agro-Forestry， Qiqihar University， Qiqihar 161006， China；2.Branch of Animal Husbandry and Veterinary of Heilongjiang Academy of Agricultural Sciences， Qiqihar 161005， China；3.Qiqihar Branch of Heilongjiang Academy of Agricultural Sciences， Qiqihar 161006， China)
关键词:
大豆; NCED基因; 非生物胁迫; 转基因烟草; 脱落酸
Keywords:
soybean; NCED gene; abiotic stress; transgenic tobacco; abscisic acid
分类号:
S565.1
DOI:
doi:10.3969/j.issn.1000-4440.2023.04.002
文献标志码:
A
摘要:
9-顺式-环氧类胡萝卜素双加氧酶（NCED）是脱落酸（ABA）合成途径中的关键酶。本研究从大豆中克隆GmNCED1-2，其开放阅读框全长1 818 bp，编码605个氨基酸，预测蛋白质相对分子质量6.693×104，等电点800。GmNCED1-2蛋白含有1个RPE65超家族结构域、2个保守序列MIAHPKxDP和HDFAITE以及４个保守的组氨酸残基。GmNCED1-2蛋白的亚细胞定位预测结果显示其位于叶绿体中。蛋白质系统进化分析结果表明GmNCED1-2蛋白与CrNCED1蛋白和MhNCED3蛋白的亲缘关系较近。实时荧光定量PCR结果表明GmNCED1-2在叶中的表达量最高。脱落酸、干旱、高盐、高温和低温胁迫下GmNCED1-2的表达量均升高。顺式作用元件预测分析结果表明GmNCED1-2启动子含有2个ABRE，3个ARE，1个TC-rich repeats和1个TGACG-motif顺式作用元件。将GmNCED1-2构建植物表达载体并转化烟草，获得3棵转基因烟草植株。烟草根长和生物量测定结果显示，GmNCED1-2的过表达抑制了转基因烟草根的伸长及植株的生长。
Abstract:
The 9-cis-epoxycarotenoid dioxygenase (NCED) is the key rate-limiting enzyme in abscisic acid (ABA) synthesis pathway. GmNCED1-2 was cloned from soybean in this research. The open reading frame (ORF) of GmNCED1-2 was 1 818 bp in length, encoding 605 amino acids, with a predicted protein molecular weight of 6.693×104 and a theoretical isoelectric point of 8.00. GmNCED1-2 protein contained one RPE65 superfamily domain, two conserved sequences MIAHPKxDP and HDFAITE, and four conserved histidine residues. The subcellular localization of GmNCED1-2 protein was predicted to be located in chloroplasts. Phylogenetic analysis showed that GmNCED1-2 protein was closely related to CrNCED1 protein and MhNCED3 protein. The results of real-time quantitative PCR showed that the expression level of GmNCED1-2 was the highest in leaves. The expression of GmNCED1-2 increased under abscisic acid, drought, salt, heat and cold stresses. Cis-element prediction showed that the promoter region of GmNCED1-2 contained two ABRE, three ARE, one TC-rich repeats and one TGACG-motif. GmNCED1-2 was constructed into plant expression vector and transformed into tobacco. Three transgenic tobacco plants were obtained. The root length and biomass of wild type tobacco and GmNCED1-2 transgenic tobacco were measured. The results showed that the overexpression of GmNCED1-2 inhibited the root elongation and plant growth of transgenic tobacco.

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备注/Memo

备注/Memo:
收稿日期：2022-08-12 基金项目：黑龙江省省属高等学校基本科研业务费科研项目（145109506）；国家自然科学基金项目（32101694）作者简介：于海伟（1979-），女，内蒙古赤峰人，学士，高级实验师，主要从事大豆分子遗传育种研究。(E-mail)Yuhaiwei2020@163.com 通讯作者：翟莹， (E-mail)fairy39809079@126.com
更新日期/Last Update: 2023-09-12