OsGA3ox通过合成不同活性GA调控水稻育性及株高

doi:10.16288/j.yczz.23-058

遗传 ›› 2023, Vol. 45 ›› Issue (9): 845-855.doi: 10.16288/j.yczz.23-058

• 研究报告 • 上一篇

OsGA3ox通过合成不同活性GA调控水稻育性及株高

郝小花^1,²(), 胡爽², 赵丹², 田连福², 谢子靖², 吴莎², 胡文俐², 雷晗², 李东屏²()

1.湖南文理学院生命与环境科学学院，植物产品精深加工科技创新团队，常德 415000
2.湖南师范大学生命科学学院，长沙 410081

收稿日期:2023-03-15 修回日期:2023-06-14 出版日期:2023-09-20 发布日期:2023-07-19
通讯作者: 李东屏 E-mail:14771417@qq.com;dli@hunnu.edu.cn
作者简介:郝小花，博士，副教授，研究方向：植物生理与分子生物学。E-mail: 14771417@qq.com
基金资助:
国家自然科学基金面上项目(32071930);湖南省自然科学基金面上项目(2022JJ30421);湖南省自然科学基金面上项目(2021JJ30445);长沙市自然科学基金项目(kq2014076)

OsGA3ox genes regulate rice fertility and plant height by synthesizing diverse active GA

Xiaohua Hao^1,²(), Shuang Hu², Dan Zhao², Lianfu Tian², Zijing Xie², Sha Wu², Wenli Hu², Han Lei², Dongping Li²()

1. Plant Product Deep Processing Technology Innovation Team, College of Life and Environmental Sciences, Hunan University of Arts and Science, Changde 415000, China
2. College of Life Science, Hunan Normal University, Changsha 410081, China

Received:2023-03-15 Revised:2023-06-14 Online:2023-09-20 Published:2023-07-19
Contact: Dongping Li E-mail:14771417@qq.com;dli@hunnu.edu.cn
Supported by:
National Natural Science Foundation of China(32071930);Hunan Province Natural Science Foundation Project(2022JJ30421);Hunan Province Natural Science Foundation Project(2021JJ30445);Changsha City Natural Science Foundation Project(kq2014076)

摘要/Abstract

摘要：

赤霉素(gibberellin，GA)是一种重要的激素，参与调控植物多种生长发育过程。GA生物合成通路已基本阐明，其中赤霉素3β羟化酶(gibberellin 3β-hydroxylase，GA3ox)是多种活性GA合成的关键酶。水稻中有2个GA3ox基因(OsGA3ox1和OsGA3ox2)，其生理功能虽有初步研究，但它们在合成活性GA调控水稻发育过程中是如何分工协作尚不清楚。本研究通过CRISPR/Cas9技术获得基因编辑突变体ga3ox1和ga3ox2，发现ga3ox1花粉育性显著下降，而ga3ox2株高显著变矮，表明OsGA3ox1是花粉正常发育必需的，而OsGA3ox2是茎叶伸长必需的。组织表达分析表明，OsGA3ox1主要在未开的花中表达，OsGA3ox2主要在未伸长的叶中表达。进一步对野生型(WT)和两个ga3ox突变体未开的花、未伸长的叶及根中的GA进行检测分析，发现OsGA3ox1在花中催化GA₉形成GA₇与花粉育性密切相关；OsGA3ox2在未伸长的叶中催化GA₂₀形成GA₁调控株高；OsGA3ox1在根中催化GA₁₉形成GA₂₀，调控GA₃的生成。总之，OsGA3ox1和OsGA3ox2响应发育信号，在不同组织分工协作合成内源GA，调控水稻生长和发育。本研究为阐明OsGA3ox1和OsGA3ox2在GA生物合成中的作用以及深入理解OsGA3ox的功能提供参考。

关键词: 水稻, 赤霉素, 赤霉素3β羟化酶基因, 功能分析

Abstract:

Gibberellin (GA) is an important hormone, which is involved in regulating various growth and development. GA biosynthesis pathway and synthetase have been basically clarified. Gibberellin 3β hydroxylase (GA3ox) is the key enzyme for the synthesis of various active GA. There are two GA3ox genes (OsGA3ox1 and OsGA3ox2) in rice, and their physiological functions have been preliminarily studied. However, it is not clear how they work together to synthesize active GA to regulate rice development. In this study, the knockout mutants ga3ox1 and ga3ox2 were obtained by CRISPR/Cas9 technology. The pollen fertility of ga3ox1 decreased significantly, while the plant height of ga3ox2 decreased significantly. It shows that OsGA3ox1 is necessary for normal pollen development, while OsGA3ox2 is necessary for stem and leaf elongation. Tissue expression analysis showed that OsGA3ox1 was mainly expressed in unopened flowers, while OsGA3ox2 was mainly expressed in unexpanded leaves. The GA in different tissues of wild type (WT), and two ga3ox mutants were detected. It was found that pollen fertility is most closely related to the content of GA₇, and plant height is most closely related to the content of GA₁. It was found that OsGA3ox1 catalyzes GA₉ to GA₇ in flowers, which is closely related to pollen fertility; OsGA3ox2 catalyzes the GA₂₀ to GA₁ in unexpanded leaves, thereby regulating plant height; OsGA3ox1 catalyzes the GA₁₉ to GA₂₀ in roots, regulating the generation of GA₃. OsGA3ox1 and OsGA3ox2 respond to developmental and environmental signals, and cooperate to synthesize endogenous GA in different tissues to regulate rice development. This study provides a reference for clarifying its role in GA biosynthesis pathway and further understanding the function of OsGA3ox.

Key words: Oryza sativa L., gibberellin (GA), OsGA3ox, functional analysis

郝小花, 胡爽, 赵丹, 田连福, 谢子靖, 吴莎, 胡文俐, 雷晗, 李东屏. OsGA3ox通过合成不同活性GA调控水稻育性及株高[J]. 遗传, 2023, 45(9): 845-855.

Xiaohua Hao, Shuang Hu, Dan Zhao, Lianfu Tian, Zijing Xie, Sha Wu, Wenli Hu, Han Lei, Dongping Li. OsGA3ox genes regulate rice fertility and plant height by synthesizing diverse active GA[J]. Hereditas(Beijing), 2023, 45(9): 845-855.

图/表 7

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引物名称	引物序列(5′→3′)	用途
OsGA3ox1-RT-PCR	F：CGAGACCGAGCGGAAGA	半定量RT-PCR和实时荧光定量PCR
OsGA3ox1-RT-PCR	R：GCCGACGACGACGATGA	半定量RT-PCR和实时荧光定量PCR
OsGA3ox2-RT-PCR	F：TTCTCCAAGCTCATGTGGTC	半定量RT-PCR和实时荧光定量PCR
OsGA3ox2-RT-PCR	R：GCATCTCCTTGTGAAACTC	半定量RT-PCR和实时荧光定量PCR
OsActin2-RT-PCR	F：ATGTGCCAGCTATGTATGTC	半定量RT-PCR和实时荧光定量PCR
OsActin2-RT-PCR	R：CGTTCAGCAGTGGTAGTGA	半定量RT-PCR和实时荧光定量PCR
OsGA3ox1-Cas9-1	F：GCCGGAGTCGCACGTGTGGA	构建CRISPR/Cas9基因编辑载体
OsGA3ox1-Cas9-1	R：TCCACACGTGCGACTCCGGC	构建CRISPR/Cas9基因编辑载体
OsGA3ox1-Cas9-2	F：CGATGAGAGCTCTGGGCGAG	构建CRISPR/Cas9基因编辑载体
OsGA3ox1-Cas9-2	R：CTCGCCCAGAGCTCTCATCG	构建CRISPR/Cas9基因编辑载体
OsGA3ox2-Cas9-1	F：GCTCTGCTTCGACTTCCGGG	构建CRISPR/Cas9基因编辑载体
	R：CCCGGAAGTCGAAGCAGAGC	构建CRISPR/Cas9基因编辑载体
OsGA3ox2-Cas9-2	F：GAGAAGATGCGCGCCGTCCG	构建CRISPR/Cas9基因编辑载体
OsGA3ox2-Cas9-2	R：CGGACGGCGCGCATCTTCTC	构建CRISPR/Cas9基因编辑载体
OsGA3ox1-GUS	F：GGTTTTCATGCCATGCCAAT	构建GUS表达载体
OsGA3ox1-GUS	R：GCATGAACTCGTTGGCTA	构建GUS表达载体
OsGA3ox2-GUS	F：ATAGTCCTCGGCAAGAAG	构建GUS表达载体
OsGA3ox2-GUS	R：CGACGACGACGACGAT	构建GUS表达载体

OsGA3ox通过合成不同活性GA调控水稻育性及株高

OsGA3ox genes regulate rice fertility and plant height by synthesizing diverse active GA

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