小麦烯醇化酶基因TaENO1-5B在多种环境下对株高与穗粒数的调控

doi:10.3864/j.issn.0578-1752.2024.14.002

中国农业科学 ›› 2024, Vol. 57 ›› Issue (14): 2717-2731.doi: 10.3864/j.issn.0578-1752.2024.14.002

• 作物遗传育种·种质资源·分子遗传学 • 上一篇下一篇

小麦烯醇化酶基因TaENO1-5B在多种环境下对株高与穗粒数的调控

张紫慧¹^,²(), 张艳菲²^,³, 李龙², 李超男², 王景一², 杨德龙¹(), 毛新国¹^,²(), 景蕊莲²

¹ 甘肃农业大学生命科学技术学院/省部共建干旱生境作物学国家重点实验室，兰州 730070
² 中国农业科学院作物科学研究所/作物基因资源与育种全国重点实验室，北京 100081
³ 河南农业大学/国家小麦技术创新中心（筹），郑州 450046

收稿日期:2024-01-11 接受日期:2024-02-06 出版日期:2024-07-24 发布日期:2024-07-24
通信作者:
杨德龙，Tel：0931-7631875；E-mail：yangdl@gsau.edu.cn。
毛新国，Tel：010-82105829；E-mail：maoxinguo@caas.cn
联系方式: 张紫慧，E-mail：zzh980416@163.com。
基金资助:
国家重点研发计划(2021YFD1200603); 中央引导地方科技发展资金项目(23ZYQA0322); 甘肃省重大科技专项(22ZD6NA009)

Wheat Enolase Gene TaENO1-5B Involved in Regulating Plant Height and Grain Number Per Spike in Multiple Environments

ZHANG ZiHui¹^,²(), ZHANG YanFei²^,³, LI Long², LI ChaoNan², WANG JingYi², YANG DeLong¹(), MAO XinGuo¹^,²(), JING RuiLian²

¹ College of Life Science and Technology, Gansu Agricultural University/State Key Laboratory of Arid Land Crop Science, Lanzhou 730070
² Institute of Crop Sciences, Chinese Academy of Agricultural Sciences/State Key Laboratory of Crop Gene Resources and Breeding, Beijing 100081
³ Henan Agricultural University/National Wheat Technology Innovation Center (Preparatory), Zhengzhou 450046

Received:2024-01-11 Accepted:2024-02-06 Published:2024-07-24 Online:2024-07-24

摘要/Abstract

摘要：

【目的】烯醇化酶是糖酵解过程中的关键限速酶，在植物生长发育和逆境胁迫应答中发挥重要作用。揭示小麦烯醇化酶基因TaENO1-5B的功能，开发分子标记，为通过分子育种改良小麦提供基因资源。【方法】以小麦品种旱选10号为材料克隆TaENO1-5B，在SMART网站分析其编码蛋白的结构域，采用Phyre2软件预测蛋白的二级和三级结构；采用实时荧光定量PCR技术分析基因在小麦不同发育时期的组织表达水平，以及在植物激素处理和逆境胁迫下的表达模式；以30份遗传多样性丰富的小麦种质为材料分析基因序列多态性，开发分子标记；以323份小麦构成的自然群体为材料进行TaENO1-5B单倍型与表型性状的关联分析，利用小麦地方品种和现代育成品种群体分析优异单倍型在我国不同麦区受育种选择的趋势。【结果】 TaENO1-5B由17个外显子和16个内含子组成，编码446个氨基酸，含有保守的N端结构域和C端TIM（triose-phosphate isomerase）桶状结构域；TaENO1-5B在小麦幼苗期、拔节期、抽穗期和开花期的各个组织中均有表达，在根、根基和穗中表达量较高；TaENO1-5B启动子区含有多种顺式作用元件，包括脱落酸（ABA）、生长素（IAA）和茉莉酸甲酯（MeJA）等激素应答元件和干旱、低温胁迫相关的多种应答元件，TaENO1-5B的表达受植物激素和干旱、低温等胁迫的显著诱导；在TaENO1-5B的启动子区检测到4个SNP，基因区检测到3个SNP，组成3种单倍型Hap-5B-1、Hap-5B-2和Hap-5B-3。在干旱、高温等多种环境下，单倍型Hap-5B-2为优异单倍型，与矮秆、多小穗数和穗粒数显著相关，在我国小麦主产区的育种历史中受到了正向选择。基于TaENO1-5B启动子区SNP（2 399 bp，G/A）开发的KASP标记，与多种环境下的每穗小穗数显著相关。【结论】 TaENO1-5B响应植物激素信号及非生物胁迫，在干旱、高温等多种环境下，与株高、每穗小穗数和穗粒数显著相关，Hap-5B-2是矮秆及每穗小穗数和穗粒数较多的优异单倍型。根据TaENO1-5B变异位点开发的分子标记可用于小麦株高及相关产量性状的遗传改良。

关键词: 小麦, TaENO1-5B, 农艺性状, KASP标记, 单倍型

Abstract:

【Objective】 Enolase is a key rate-limiting enzyme in the process of glycolysis, and plays a crucial role in plant growth and development, and response to abiotic stress. The function of common wheat enolase gene TaENO1-5B was revealed and molecular markers were developed to provide genetic resources for improving wheat through molecular breeding. 【Method】 TaENO1-5B was cloned from wheat variety Hanxuan 10. The domains of its encoded protein were analyzed on the SMART website. The secondary and tertiary structures of the protein were predicted by Phyre2 software. Real-time quantitative fluorescent PCR was used to analyze the expression levels of the target gene in wheat tissues at different developmental stages and its expression patterns under phytohormone treatment and abiotic stress. Thirty wheat germplasm with rich genetic diversity were used as plant materials to analyze the gene sequence polymorphisms, and develop molecular markers. The association analysis between TaENO1-5B haplotypes and phenotypic traits was carried out in a natural population consisting of 323 wheat accessions. The trend of breeding selection of superior haplotype in different wheat production zones in China was analyzed by using a landrace population and a modern variety population. 【Result】 TaENO1-5B gene consists of 17 exons and 16 introns encoding 446 amino acids and contains a conserved N-terminal domain and a C-terminal TIM (triose-phosphate isomerase) barrel domain. TaENO1-5B was expressed in all tissues of wheat at seedling, jointing, heading and flowering stages, and the expression level was higher in roots, root bases and spikes. The TaENO1-5B promoter region contains a variety of cis-acting elements, including elements responding to plant hormones, such as abscisic acid (ABA), auxin (IAA), and methyl jasmonate (MeJA), as well as elements responding to drought and low temperature. The expression of TaENO1-5B was significantly induced by phytohormones and abiotic stress in wheat. Four SNPs were detected in the promoter region and three SNPs in the gene region of the TaENO1-5B gene, which constituted three haplotypes, i.e., Hap-5B-1, Hap-5B-2, and Hap-5B-3. Among them, Hap-5B-2 was a favorable haplotype highly associated with shorter plant height, more spikelets per spike, and more grains per spike under various environments such as drought and high temperature, and had been positively selected in the breeding history of major wheat production zones in China. The KASP (kompetitive allele-specific PCR) marker developed based on the SNP (2 399 bp, G/A) of the TaENO1-5B promoter region was significantly correlated with the spikelet number per spike in multiple environments. 【Conclusion】 TaENO1-5B gene responds to phytohormones and abiotic stress, and is significantly correlated with plant height, spikelet number per spike and grain number per spike under various environments such as drought and high temperature. Hap-5B-2 is a favorable haplotype with shorter plant height and more number of spikelets and grains per spike. Molecular markers developed based on the variation sites of TaENO1-5B gene sequence can be used for genetic improvement of plant height and related yield traits in wheat.

Key words: wheat, TaENO1-5B, agronomic trait, KASP marker, haplotype

张紫慧, 张艳菲, 李龙, 李超男, 王景一, 杨德龙, 毛新国, 景蕊莲. 小麦烯醇化酶基因TaENO1-5B在多种环境下对株高与穗粒数的调控[J]. 中国农业科学, 2024, 57(14): 2717-2731.

ZHANG ZiHui, ZHANG YanFei, LI Long, LI ChaoNan, WANG JingYi, YANG DeLong, MAO XinGuo, JING RuiLian. Wheat Enolase Gene TaENO1-5B Involved in Regulating Plant Height and Grain Number Per Spike in Multiple Environments[J]. Scientia Agricultura Sinica, 2024, 57(14): 2717-2731.

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参考文献 37

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序号 Number	种质 Germplasm	序号 Number	种质 Germplasm
1	沧麦6001 Cangmai 6001	16	泰山23 Taishan 23
2	冬协2号Dongxie 2	17	泰山24 Taishan 24
3	丰产1号Fengchan 1	18	太原566 Taiyuan 566
4	衡5229 Heng 5229	19	温麦6号（豫麦49）Wenmai 6 (Yumai 49)
5	葫芦头Hulutou	20	小白麦（京856）Xiaobaimai (Jing 856)
6	冀麦6号Jimai 6	21	西峰9号Xifeng 9
7	冀麦9号Jimai 9	22	西农688（西农213）Xinong 688 (Xinong 213)
8	冀麦30 Jimai 30	23	小山8号Xiaoshan 8
9	京品3号Jingpin 3	24	原冬847 Yuandong 847
10	京品11 Jingpin 11	25	运旱20410 Yunhan 20410
11	洛旱11号Luohan 11	26	运旱23-35 Yunhan 23-35
12	临旱935 Linhan 935	27	烟农21 Yannong 21
13	兰天15号Lantian 15	28	偃展一号Yanzhan 1
14	秦麦3号Qinmai 3	29	中7902 Zhong 7902
15	山农优麦2号Shannongyoumai 2	30	中作60064 Zhongzuo 60064

引物名称 Primer name	序列 Sequence (5′-3′)
qRT-TaENO1-5B-F	AGATGACTGAAGAATGTGGAGAGC
qRT-TaENO1-5B-R	GGTACATATCCGTCCCGTTCA
TaACTIN-F	CTCCCTCACAACAACAACCGC
TaACTIN-R	TACCAGGAACTTCCATACCAAC
TaENO1-5B-F	GGAGATAGAGGCTGGGGAGG
TaENO1-5B-R	AGATTTCACTACAAACTACATACGGAT
TaENO1-pro-5B-F	GGGCTCCCGATTGGCTT
TaENO1-pro-5B-R	TTGTGGGGACGAGGAAACTG
TaENO1-5B-SNP1-A1	GAAGGTGACCAAGTTCATGCTTCCGCCTCCTCCACCTTCATT
TaENO1-5B-SNP1-A2	GAAGGTCGGAGTCAACGGATTCCGCCTCCTCCACCTTCATC
TaENO1-5B-SNP1-C	ACTCGTCGGAGCCGAGGGAG
TaENO1-5B-SNP2-A1	GAAGGTGACCAAGTTCATGCTTTTTGCCTGCTAGGAATCCGTTC
TaENO1-5B-SNP2-A2	GAAGGTCGGAGTCAACGGATTTTATTTTGCCTGCTAGGAATCCGTTT
TaENO1-5B-SNP2-C	CCATCTGTGGATACACACTGTGAAAAC

顺式作用元件 Cis-acting regulatory element	数量 Number	生物学功能 Biological function
G-box	9	光响应元件cis-acting regulatory element involved in light responsiveness
ABRE	8	脱落酸应答元件cis-acting element involved in the abscisic acid responsiveness
TGACG-motif	3	茉莉酸甲酯应答元件cis-acting regulatory element involved in the MeJA-responsiveness
CGTCA-motif	3	茉莉酸甲酯应答元件cis-acting regulatory element involved in the MeJA-responsiveness
AuxRR-core	1	生长素应答元件cis-acting regulatory element involved in auxin responsiveness
TCA-element	1	水杨酸应答元件cis-acting element involved in salicylic acid responsiveness
MYB	2	MYB蛋白结合元件MYB protein binding element
TATA-box	29	低温胁迫应答元件cis-acting element involved in low-temperature responsiveness
DRE core	1	DREB结合元件DREB binding element

小麦烯醇化酶基因TaENO1-5B在多种环境下对株高与穗粒数的调控

Wheat Enolase Gene TaENO1-5B Involved in Regulating Plant Height and Grain Number Per Spike in Multiple Environments

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