AtCPS V326M突变显著影响赤霉素合成

doi:10.16288/j.yczz.21-405

遗传 ›› 2022, Vol. 44 ›› Issue (3): 245-252.doi: 10.16288/j.yczz.21-405

AtCPS V326M突变显著影响赤霉素合成

赵三增¹(), 孔丹宇², 辛培勇³, 褚金芳^3,⁵, 万迎朗¹, 凌宏清^4,⁵(), 刘毅^2,⁴()

1. 海南大学热带作物学院,海口 570228
2. 中国科学院庐山植物园,江西庐山 332900
3. 中国科学院遗传与发育生物学研究所,国家植物基因研究中心(北京),中国科学院种子创新研究院,北京 100101
4. 中国科学院遗传与发育生物学研究所,植物细胞与染色体工程国家重点实验室,中国科学院种子创新研究院,北京 100101
5. 中国科学院大学现代农学院,北京 100039;

收稿日期:2021-11-26 修回日期:2022-01-19 出版日期:2022-03-20 发布日期:2022-02-22
通讯作者: 凌宏清,刘毅 E-mail:834395875@qq.com;hqling@genetics.ac.cn;yiliu609@outlook.com
作者简介:赵三增,在读硕士研究生,专业方向：分子遗传学。E-mail: 834395875@qq.com
基金资助:
国家自然科学基金青年项目资助(31900171)

AtCPS V326M significantly affect the biosynthesis of gibberellins

Sanzeng Zhao¹(), Danyu Kong², Peiyong Xin³, Jinfang Chu^3,⁵, Yinglang Wan¹, Hong-Qing Ling^4,⁵(), Yi Liu^2,⁴()

1. College of Tropical Crops, Hainan University, Haikou 570228, China
2. Lushan Botanical Garden, Chinese Academy of Sciences, Lushan 332900, China
3. National Centre for Plant Gene Research (Beijing), Innovation Academy for Seed Design, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China
4. State Key Laboratory of Plant Cell and Chromosome Engineering, Innovation Academy for Seed Design, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China
5. College of Advanced Agricultural Sciences, University of Chinese Academy of Sciences, Beijing 100039, China;

Received:2021-11-26 Revised:2022-01-19 Online:2022-03-20 Published:2022-02-22
Contact: Ling Hong-Qing,Liu Yi E-mail:834395875@qq.com;hqling@genetics.ac.cn;yiliu609@outlook.com
Supported by:
Supported by the National Natural Science Foundation of China(31900171)

摘要/Abstract

摘要：

赤霉素(gibberellins, GA)是植物激素之一,调控植物生长和发育。植物体中赤霉素合成量直接影响植物的形态和生物量。在赤霉素合成途径中,柯巴基焦磷酸合酶基因(copalyl diphosphate synthase, CPS)是第一个合酶基因,该基因突变会严重影响赤霉素合成量。本研究通过对根和下胚轴缩短、晚花、丛生、矮化的拟南芥(Arabidopsis thaliana)突变体ga1-168进行图位克隆,鉴定出AtCPS的一个等位基因AtCPS-168。该等位基因的突变位点是AtCPS基因的第2768位核苷酸,鸟嘌呤(G)突变为腺嘌呤(A),导致AtCPS蛋白萜类合酶(Terpene_synth)结构域中的第326位缬氨酸(V)突变成蛋氨酸(M)。通过等位分析确定AtCPS-168是AtCPS的等位基因。遗传互补实验显示AtCPS V326M突变导致植物丛生、矮化等发育缺陷表型。赤霉素含量测定结果证明AtCPS V326M突变导致赤霉素的合成量减少。外施赤霉素实验结果表明,外施赤霉素能恢复拟南芥突变体ga1-168赤霉素合成量降低导致的植株丛生、矮化等发育缺陷表型。因此,本研究为通过定点突变赤霉素合成酶基因的特定位点改变赤霉素含量来塑造植物理想株高和株型提供理论指导。

关键词: 赤霉素, AtCPS基因, 根长, 株型, 株高

Abstract:

Gibberellins are a class of typical phytohormones, which regulate plant growth and development. The contents of gibberellins dramatically affect the morphology and biomass of plant. The encoding protein of copalyl diphosphate synthase gene (CPS) catalyzes the first-step in the biosynthetic pathway of gibberellins. The mutation in this gene may significantly affect the contents of gibberellins in plants. In this study, we found an EMS-triggered mutant, ga1-168, showing short roots, short hypocotyls, late flowering and dwarf. Map-based cloning revealed that the causal gene of ga1-168 was AtCPS-168, an allele of AtCPS gene. The encoding protein of AtCPS-168 was AtCPS V326M which was resulted from a single-point mutation (guanine to adenine at nucleotide 2768) of AtCPS gene. Protein domain analysis showed that V326 was located in the Terpene_synth domain. The allelism test demonstrated that AtCPS-168 was an allele of AtCPS gene. The transgenic complementation of ga1-168 indicated that AtCPS V326M led to the dwarf and bushy phenotype of ga1-168. The endogenous gibberellins contents analysis suggested that the gibberellins contents of ga1-168 were much lower than that of wild-type. The exogenous GA₃ application assay uncovered that application of GA₃ can complement the dwarf and bushy phenotype of ga1-168 caused by low endogenous gibberellins contents. Therefore, this study suggested that it is an elegant way to create the ideal plant architecture and height by site-directed mutating the gibberellin biosynthetic genes.

Key words: gibberellins, AtCPS gene, root length, plant architecture, plant height

赵三增, 孔丹宇, 辛培勇, 褚金芳, 万迎朗, 凌宏清, 刘毅. AtCPS V326M突变显著影响赤霉素合成[J]. 遗传, 2022, 44(3): 245-252.

Sanzeng Zhao, Danyu Kong, Peiyong Xin, Jinfang Chu, Yinglang Wan, Hong-Qing Ling, Yi Liu. AtCPS V326M significantly affect the biosynthesis of gibberellins[J]. Hereditas(Beijing), 2022, 44(3): 245-252.

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用途	引物名称	序列(5′→3′)
SSLP 分子标记	CIW5 LP	GGTTAAAAATTAGGGTTACGA
	CIW5 RP	AGATTTACGTGGAAGCAAT
	F4C21 LP	GAGATTGCTCGGTTTGATTG
	F4C21 RP	ACAAGGATACGGTGTTAGAG
	T4I9 LP	ACAGAATCTGATGGAATATG
	T4I9 RP	AACTATTGTGCGTAAACATC
CAPS 分子标记	T5J8 Dde I LP	AGAGTGGTTTGAATCTGAAG
	T5J8 Dde I RP	CTGTCCTAGCTTATAACTTG
	T5J8 Hpy178 III LP	ATAGCTATGTGCAGAACAAG
	T5J8 Hpy178 III RP	TATCATACTGAGATACTCCG

	GA₁₂	GA₁₅	GA₃₄	GA₄	GA₉	GA₂₀	GA₂₄	GA₄₄	GA₅₁
Col-0	0.603±0.048	0.505±0.007	1.000±0.071	0.466±0.026	0.207±0.014	0.399±0.013	1.550± 0.081	0.220±0.026	0.732±0.059
ga1-168	0.019±0.004	0.246±0.015	0.149±0.018	ND	ND	ND	ND	ND	ND

AtCPS V326M突变显著影响赤霉素合成

AtCPS V326M significantly affect the biosynthesis of gibberellins

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