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作物学报 ›› 2023, Vol. 49 ›› Issue (4): 906-916.doi: 10.3724/SP.J.1006.2023.21029

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

小麦转录因子TaMYB5-3B与株高和千粒重相关

朱治1,2(), 李龙2, 李超男2, 毛新国2, 郝晨阳2, 朱婷2, 王景一2,*(), 常建忠1,*(), 景蕊莲2   

  1. 1山西农业大学山西有机旱作农业研究院/有机旱作山西省重点实验室/省部共建有机旱作国家重点实验室(筹), 山西太原 030031
    2中国农业科学院作物科学研究所, 北京 100081
  • 收稿日期:2022-04-21 接受日期:2022-07-21 出版日期:2023-04-12 网络出版日期:2022-08-29
  • 通讯作者: *常建忠, E-mail: cjzyfx@163.com;王景一, E-mail: wangjingyi@caas.cn
  • 作者简介:E-mail: zz1752782610@163.com
  • 基金资助:
    山西农业大学省部共建有机旱作农业国家重点实验室自主研发项目(202105D121008-2-7);财政部和农业农村部国家现代农业产业技术体系建设专项(小麦, CARS-03)

Transcription factor TaMYB5-3B is associated with plant height and 1000- grain weight in wheat

ZHU Zhi1,2(), LI Long2, LI Chao-Nan2, MAO Xin-Guo2, HAO Chen-Yang2, ZHU Ting2, WANG Jing-Yi2,*(), CHANG Jian-Zhong1,*(), JING Rui-Lian2   

  1. 1Shanxi Institute of Organic Dryland Farming, Shanxi Key Laboratory of Organic Dry Farming, State Key Laboratory of Integrative Sustainable Dryland Agriculture (in Preparation), Shanxi Agricultural University, Taiyuan 030031, Shanxi, China
    2Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2022-04-21 Accepted:2022-07-21 Published:2023-04-12 Published online:2022-08-29
  • Contact: *E-mail: cjzyfx@163.com;E-mail: wangjingyi@caas.cn
  • Supported by:
    State Key Laboratory of Integrative Sustainable Dryland Agriculture, the Shanxi Agricultural University(202105D121008-2-7);China Agriculture Research System of MOF and MARA(Wheat, CARS-03)

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摘要：

MYB转录因子在植物生长发育过程中发挥着重要作用。本研究克隆了小麦3B染色体上的TaMYB5-3B基因, 基因组序列全长3005 bp, 其中编码区上游为2112 bp, 编码区为893 bp, 包含2个外显子和1个内含子, 编码一个R2R3-MYB蛋白。序列多态性分析表明, 在TaMYB5-3B的-2048、-1632、-1178、-1156、-504、-461、-433和61 bp处各有1个SNP位点, 分别是G/A转换、G/A转换、G/A转换、T/C转换、C/T转换、A缺失、T缺失和T/A颠换, 这8个SNP位点连锁。基于启动子区SNP-1632的变异开发分子标记, 检测小麦自然群体的基因型, 与表型性状进行关联分析, 结果显示TaMYB5-3B与株高、穗下节长和千粒重显著相关。TaMYB5-3B在群体中有2种单倍型Hap-3B-1和Hap-3B-2, 其中Hap-3B-2是植株较矮、千粒重较高的优异单倍型。在我国的小麦育种历程中Hap-3B-2受到了正向选择, 在育成品种中的频率逐步增加, 但仍然有进一步的应用潜力。研究结果为深入探讨小麦株高和产量的形成机制提供参考, 也为小麦株型和产量分子育种提供了基因资源与选择标记。

关键词: 分子标记, 株高, 千粒重, 单倍型, 关联分析, 小麦

Abstract:

MYB transcription factor plays an important role in plant growth and development. In this study, we cloned TaMYB5-3B gene on chromosome 3B in wheat. The full-length genome sequence is 3005 bp, including 2112 bp promoter region and 893 bp coding region. TaMYB5-3B coding region consists of two exons and one intron, which encodes a R2R3-MYB protein. The polymorphism of TaMYB5-3B was analyzed by sequencing 32 wheat accessions with wide variations. A total of eight SNPs were detected at -2048, -1632, -1178, -1156, -504, -461, -433, and 61 bp, respectively. They were eight SNPs linked by G/A conversion, G/A conversion, G/A conversion, T/C conversion, C/T conversion, A deletion, T deletion and T/A inversion, respectively. A pair of molecular markers were developed based on the promoter region SNP-1632 to detect the genotypes of wheat natural population. The association analysis of genotype and phenotypic traits showed that TaMYB5-3B was significantly associated with plant height (PH), peduncle length (PLE), and 1000-grain weight (TGW). Two haplotypes (Hap-3B-1 and Hap-3B-2) were detected in the population, in which Hap-3B-2 was an excellent haplotype with short PH and high TGW. Hap-3B-2 had been positively selected in the breeding, and its frequency in modern cultivars gradually increased with the advance of breeding years in China. Therefore, TaMYB5-3B could be used to further understand the mechanism of wheat plant height and grain yield formation, and its molecular markers may contribute to ideal plant architecture and grain yield breeding of wheat.

Key words: molecular markers, plant height, 1000-grain weight, haplotype, association analysis, wheat

表1

32份高多态性小麦材料的信息"

序号
Number		 材料
Accession name		 来源
Origin
1 安85中124-1 An 85 Zhong 124-1 中国北京Beijing, China
2 北京10号 Beijing 10 中国北京Beijing, China
3 北京14号 Beijing 14 中国北京Beijing, China
4 北京8686 Beijing 8686 中国北京Beijing, China
5 单R8093 Dan R8093 中国北京Beijing, China
6 丰抗13 Fengkang 13 中国北京Beijing, China
7 京411 Jing 411 中国北京Beijing, China
8 京核8922 Jinghe 8922 中国北京Beijing, China
9 京品10号 Jingpin 10 中国北京Beijing, China
10 04-030 中国北京Beijing, China
11 04-044 中国北京Beijing, China
12 晋2148-7 Jin 2148-7 中国福建Fujian, China
13 白齐麦 Baiqimai 中国甘肃Gansu, China
14 霸王鞭 Bawangbian 中国河北Hebei, China
15 沧州小麦 Cangzhouxiaomai 中国河北Hebei, China
16 冀麦6号 Jimai 6 中国河北Hebei, China
17 冀麦41 Jimai 41 中国河北Hebei, China
18 白糙麦 Baicaomai 中国河南Henan, China
19 内乡188 Neixiang 188 中国河南Henan, China
20 偃展1号 Yanzhan 1 中国河南Henan, China
21 紫杆白芒先 Ziganbaimangxian 中国河南Henan, China
22 昌乐5号 Changle 5 中国山东Shandong, China
23 长6878 Chang 6878 中国山西Shanxi, China
24 红和尚 Hongheshang 中国山西Shanxi, China
25 临抗5108 Linkang 5108 中国山西Shanxi, China
26 长武131 Changwu 131 中国陕西Shaanxi, China
27 大荔1号 Dali 1 中国陕西Shaanxi, China
28 中国春 Chinese Spring 中国四川Sichuan, China
29 9th-5-1 国际玉米小麦改良中心CIMMYT
30 9th-25 国际玉米小麦改良中心CIMMYT
31 9th-50-1 国际玉米小麦改良中心CIMMYT
32 PANDAS 意大利Italy

表2

本研究所用引物"

引物名称
Primer name		 引物序列
Primer sequence (5'-3')		 试验目的
Experimental purpose
TaMYB5-3B-F1 GCCAGATCCGTCAAGCAATTCATGT 克隆基因编码区 Cloning gene coding region
TaMYB5-3B-R1 GCAACGTTTCCCAACATGTGTGC 克隆基因编码区 Cloning gene coding region
TaMYB5-3B-F2 GCAAACGAGGGCTGAATACCAATCA 克隆基因启动子区 Cloning gene coding region
TaMYB5-3B-R2 ATGTTGCCGGAGCTCATCCACTA 克隆基因启动子区/测序 Cloning gene coding region
TaMYB5-3B-dCAPS-F ACCAACTACTTTGGGGGTGCAGA dCAPS分子标记 dCAPS molecular markers
TaMYB5-3B-dCAPS-R GCAACGTTTCCCAACATGTGTGC dCAPS分子标记 dCAPS molecular markers
TaMYB5-3B-cDNA-F GCCAGATCCGTCAAGCAATTCATGT 克隆cDNA/测序 Cloning cDNA/sequencing
TaMYB5-3B-cDNA-R TCTCAGTCTCAGAAAAAGCGTCCGA 克隆cDNA/测序 Cloning cDNA/sequencing
TaMYB-3B-seq-R CAACATGTGGCGTGAGTTCCTCTC 测序 Sequencing
TaMYB-3B-seq-F GAGAGGAACTCACGCCACATGTTG 测序 Sequencing

图1

TaMYB5蛋白序列比对及亲缘关系 A: TaMYB5氨基酸序列比对图。黑线指示R2和R3结构域; B: TaMYB5蛋白进化树。TaMYB5-3B用红点标注, Ta: 小麦; Td: 野生二粒小麦; Hv: 大麦; Bd: 二穗短柄草; Sb: 高粱; Zm: 玉米; Si: 谷子; Ph: 哈氏黍; Pv: 柳枝稷; Ob: 短花药野生稻; Os: 水稻。"

图2

TaMYB5-3B的核苷酸多态性和分子标记开发 A: TaMYB5-3B的结构示意图与多态性位点, 红色字母表示设计分子标记的SNP位点; B: 分子标记dCAPS-1632的开发, 红框和红点代表通过碱基T错配为C引入Bgl II酶切位点, 红色字母代表2种单倍型碱基差异; C: PCR产物用Bgl II酶切的结果。M: 100 bp DNA ladder。"

表3

小麦dCAPS-1632标记与农艺性状关联分析"

年份
Year		 地点
Site		 处理
Treatment		 性状Trait
株高PH 穗下节长PLE 千粒重TGW
2015 顺义Shunyi WW 0.01404* 0.01882* ns
WW+HS 0.00191** 0.01511* 0.01324*
DS ns ns 0.00139**
DS+HS 0.04595* 0.02753* 0.01404*
2016 顺义Shunyi WW 0.01204* 0.01945* ns
WW+HS 0.00600** 0.02717* ns
DS 0.00661** 0.00912** 0.01480*
DS+HS 0.01621* 0.00883** ns
昌平Changping WW 0.04117* ns 0.04570*
DS 0.03954* ns 0.00178**
2017 顺义Shunyi WW 0.00621** 0.01578* 9.93E-05***
WW+HS 0.00860** 0.03151* 0.00514**
DS 0.02229* ns 0.00405**
DS+HS 0.00712** 0.00773** ns
昌平Changping WW 0.02190* 0.01346* 0.00625**
DS 0.02983* 0.02417* 0.01491*

图3

TaMYB5-3B两种单倍型的农艺性状对比 A~C: TaMYB5-3B的2种单倍型在16种环境中株高(A)、穗下节长(B)和千粒重(C)的比较; D: 2种单倍型在3种环境中株高和千粒重的比较。E: 环境, E1: 15-SY-WW; E2: 15-SY-WW-HS; E3: 15-SY-DS; E4: 15-SY-DS-HS; E5: 16-SY-WW; E6: 16-SY-WW-DS; E7: 16-SY-DS; E8: 16-SY-DS-HS; E9: 16-CP-WW; E10: 16-CP-DS; E11: 17-SY-WW; E12: 17-SY-WW-HS; E13: 17-SY-DS; E14: 17-SY-DS-HS; E15: 17-CP-WW; E16: 17-CP-DS。15: 2015; 16: 2016; 17: 2017; 02: 2002; 05: 2005; 10: 2010。SY: 顺义; CP: 昌平; LY: 洛阳。WW: 水分充足; DS: 干旱胁迫; HS: 高温胁迫。数据显著性采用t检验。**P < 0.01; ***P < 0.001。误差值: ±SE。"

图4

TaMYB5-3B两种单倍型的频率与分布 A~B: TaMYB5-3B两种单倍型在中国10个小麦产区的157个地方品种(A)和348个现代育成品种(B)中的分布。I: 北方冬麦区; II: 黄淮冬麦区; III: 长江中下游麦区; IV: 西南冬麦区; V: 华南冬麦区; VI: 东北春麦区; VII: 北部春麦区; VIII: 西北春麦区; IX: 青藏春冬麦区; X: 新疆冬春麦区。"

图5

我国348个育成品种中TaMYB5-3B单倍型频率以及PH和TGW变化 A: 随年代推进两个单倍型Hap-3B-1和Hap-3B-2在群体3中的频率变化; B: 随年代推进PH和TGW在群体3中的变化。误差值: ±SE。PH和TGW的数据来自Hao等[23]。"

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