小麦TaTIFY11c-4A的克隆及遗传效应分析

doi:10.3864/j.issn.0578-1752.2025.17.001

中国农业科学 ›› 2025, Vol. 58 ›› Issue (17): 3357-3371.doi: 10.3864/j.issn.0578-1752.2025.17.001

• 作物遗传育种·种质资源·分子遗传学 • 上一篇下一篇

小麦TaTIFY11c-4A的克隆及遗传效应分析

贾玉静¹^,²(), 李超男², 潘志雄¹^,², 杨德龙¹^,^*(), 毛新国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/农作物基因资源与基因改良国家重大科学工程，北京 100081

收稿日期:2025-03-14 接受日期:2025-05-09 出版日期:2025-09-01 发布日期:2025-09-02
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: 贾玉静，E-mail：jiayujinglearn@163.com。
基金资助:
农业生物育种国家科技重大专项(2023ZD0407101); 中国农业科学院青年创新专项(Y2023QC01); 甘肃省种业攻关项目(GYGG-2024-2)

Cloning and Genetic Effect Analysis of TaTIFY11c-4A in Wheat

JIA YuJing¹^,²(), LI ChaoNan², PAN ZhiXiong¹^,², YANG DeLong¹^,^*(), MAO XinGuo²^,^*(), JING RuiLian²

¹ College of Life Science and Technology, Gansu Agricultural University/State Key Laboratory of Aridland Crop Science, Lanzhou 730070
² Institute of Crop Sciences, Chinese Academy of Agricultural Sciences/State Key Laboratory of Crop Gene Resources and Breeding/National Key Facility for Crop Gene Resources and Genetic Improvement, Beijing 100081

Received:2025-03-14 Accepted:2025-05-09 Published:2025-09-01 Online:2025-09-02

摘要/Abstract

摘要：

【目的】TIFY是植物特有的一类参与调控生长发育和逆境胁迫应答的转录因子家族，在小麦中开展TaTIFY11c-4A的克隆并验证其遗传效应，为小麦高产分子育种提供依据。【方法】以小麦品种旱选10号为材料，克隆TaTIFY11c-4A，并在不同种质中检测其等位基因的变异；利用qRT-PCR检测TaTIFY11c-4A的组织表达模式，研究其对不同激素信号和逆境胁迫的响应，并通过在烟草中瞬时表达明确TaTIFY11c-4A亚细胞定位；开发TaTIFY11c-4A多态性标记并检测其在自然群体中的分布，通过候选基因关联分析的方法验证其与表型的相关性。同时，研究不同等位基因在不同年代及地域的分布和频率。通过对TaTIFY11c-4A和TaSRL1-4A单倍型进行联合分析，挖掘优异基因型组合。【结果】克隆了小麦TaTIFY11c-4A，该基因包含3个外显子和2个内含子，编码198个氨基酸，具有TIFY和Jas结构域；该基因在小麦幼苗的根、根基和叶中均有表达，在小麦孕穗期根部和叶中表达量较高。TaTIFY11c-4A启动子区含有激素应答、胁迫响应和胚乳发育相关的多种顺式作用元件，响应ABA、IAA、MeJA等植物激素和干旱、高盐、低温、高温等胁迫信号。在TaTIFY11c-4A上游启动子区-405 bp位置检测到1个SNP（G/A），依据该SNP开发了目的基因的分子标记并进行关联分析。结果显示，TaTIFY11c-4A等位基因与干旱、高温等多环境下小麦的株高、千粒重和分蘖期根深均显著相关。与SNP-G相比，携带SNP-A等位基因小麦种质平均株高较矮、千粒重较高，但分蘖期扎根较浅，在小麦育种历程中受到正向选择。TaTIFY11c-4A-SNP-A和TaSRL1-4A-SNP-C基因型对株高的降低和千粒重的提高具有加性效应。【结论】TaTIFY11c-4A编码一个核定位JAZ蛋白，在小麦植株各组织中均有表达，响应ABA、IAA、MeJA信号和干旱、极端温度和高盐等非生物胁迫；TaTIFY11c-4A-SNP与干旱、高温等多种环境下的株高、千粒重和分蘖期根深相关，并且SNP-A等位基因在育种历程中受到了正向选择。TaTIFY11c-4A和TaSRL1-4A的优异等位变异及组合单倍型为培育高产抗逆小麦品种提供基因资源。

关键词: 小麦, TaTIFY11c-4A, 分子标记, 关联分析, 株高, 千粒重, 根深

Abstract:

【Objective】The TIFY family, a plant-specific group of transcription factors, plays critical roles in regulating growth, development, and stress responses. This study aimed to clone TaTIFY11c-4A in wheat, validate its genetic effects, and provide a theoretical basis for high-yield molecular breeding of wheat.【Method】The wheat cultivar Hanxuan 10 was used to clone TaTIFY11c-4A and allelic variations were detected in germplasms. The tissue-specific expression patterns of TaTIFY11c-4A and its responses to various hormones and stresses were analyzed via quantitative real-time PCR (qRT-PCR). The subcellular localization of TaTIFY11c-4A was determined through transient expression in tobacco. A molecular marker targeting the polymorphic site in TaTIFY11c-4A was developed to assess the genotypes in the natural population, and association analysis was performed to evaluate the correlations between the genotypes and phenotypes. Additionally, the spatial and temporal distribution of different genotypes were analyzed. Synergistic effects of TaTIFY11c-4A and TaSRL1-4A haplotypes were explored to identify superior genotype.【Result】TaTIFY11c-4A was successfully cloned, comprising three exons and two introns, encoding a 198-amino acid protein with conserved TIFY and Jas domains. TaTIFY11c-4A is expressed in roots, root bases and leaves at the seedling stage, and highly expressed in roots and leaves at the booting stage. There are multiple cis-acting elements related to hormone responses, stress adaptation, and endosperm development in the promoter of TaTIFY11c-4A. Its expression responds to plant hormones (ABA, IAA, MeJA) and abiotic stresses (drought, high salinity, low and high temperature). A SNP (G/A) was identified in its promoter at -405 bp. A molecular marker was developed based on the SNP and association analysis revealed significant correlations between TaTIFY11c-4A alleles and plant height, thousand grain weight under multiple environments such as drought and high temperature, and root depth at tillering stage. Compared with genotype SNP-G, wheat germplasms carrying the SNP-A allele exhibited shorter plants, higher thousand grain weight, and shallower roots at tillering stage, and have been positively selected in the wheat breeding process. TaTIFY11c-4A-SNP-A and TaSRL1-4A-SNP-C genotypes synergistically reduced plant height and enhanced thousand grain weight.【Conclusion】TaTIFY11c-4A encodes a nuclear-localized JAZ protein. It is expressed in various tissues of wheat and involved in responses to ABA, IAA, MeJA, as well as abiotic stresses such as drought, extreme temperature, and high salinity. The TaTIFY11c-4A-SNP is associated with plant height and thousand grain weight under multiple environments, and root depth. SNP-A allele has been positively selected in the wheat breeding process. The superior genotypes and combinations of TaTIFY11c-4A and TaSRL1-4A provide genetic resources for breeding high-yield and stress-resistant wheat cultivars.

Key words: wheat, TaTIFY11c-4A, molecular marker, association analysis, plant height, thousand grain weight, root depth

贾玉静, 李超男, 潘志雄, 杨德龙, 毛新国, 景蕊莲. 小麦TaTIFY11c-4A的克隆及遗传效应分析[J]. 中国农业科学, 2025, 58(17): 3357-3371.

JIA YuJing, LI ChaoNan, PAN ZhiXiong, YANG DeLong, MAO XinGuo, JING RuiLian. Cloning and Genetic Effect Analysis of TaTIFY11c-4A in Wheat[J]. Scientia Agricultura Sinica, 2025, 58(17): 3357-3371.

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引物名称Primer name	正向引物Forward primer (5′-3′)	反向引物Reverse primer (5′-3′)
TaTIFY11c-4A-qRT	CACGTACATACCTCGGCACC	CTGCTCCCTCATGTACTGGC
TaTUB-qRT	CGTGCTGTCTTTGTAGATCTCG	GACCAGTGCAGTTGTCTGAAAG
TaTIFY11c-4A-1300-GFP	CCAAATCGACTCTAGAATGGG AGCCGAGCAGCAGCA	TGCTCACCATGGTACCGTTGA GCTGGAGCGCGGG
TaTIFY11c-4A	ACCAAGGATAGCAACACTC	AACGGCAACAATCAAGGA
TaTIFY11c-4A-SacⅠ	ACGAAGTCTCAATGGACGGAGACCTAG	GGGGGTGGTGGAAGAAATGCCTGAG

环境 Environment	P（千粒重） P-value (Thousand grain weight)	P（株高） P-value (Plant height)	环境 Environment	P（千粒重） P-value (Thousand grain weight)	P（株高） P-value (Plant height)
E1	0.03915*	0.00599**	E9	0.02229*	0.01174*
E2	0.01354*	0.00141**	E10	0.03127*	0.00881**
E3	0.00081***	0.01997*	E11	0.32457	0.00192**
E4	0.03561*	0.00371**	E12	0.00305**	0.00783**
E5	0.04201*	0.02537*	E13	0.00097***	0.00555**
E6	0.10469	0.00875**	E14	0.00169**	0.00637**
E7	0.04312*	0.0136*	E15	0.05409	0.01093*
E8	0.02590*	0.01771*	E16	0.13964	0.00726**

小麦TaTIFY11c-4A的克隆及遗传效应分析

Cloning and Genetic Effect Analysis of TaTIFY11c-4A in Wheat

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编号 Number	材料名称 Accession name	基因型 Genotype	编号 Number	材料名称 Accession name	基因型 Genotype
1	PANDAS	SNP-A	17	临抗5108 Linkang 5108	SNP-G
2	安85中124-1 An 85 Zhong 124-1	SNP-G	18	白齐麦 Baiqimai	SNP-G
3	偃展1号 Yanzhan 1	SNP-A	19	昌乐5号 Changle 5	SNP-G
4	霸王鞭 Bawangbian	SNP-G	20	红和尚 Hongheshang	SNP-G
5	北京10号 Beijing 10	SNP-G	21	北京8686 Beijing 8686	SNP-G
6	北京14号 Beijing 14	SNP-G	22	04-044	SNP-G
7	沧州小麦 Cangzhouxiaomai	SNP-G	23	04-030	SNP-G
8	长武131 Changwu 131	SNP-A	24	春22 9th-25 Chun 22 9th-25	SNP-G
9	长6878 Chang 6878	SNP-G	25	紫秆白芒先 Ziganbaimangxian	SNP-G
10	大荔1号 Dali 1	SNP-G	26	京品10号 Jingpin 10	SNP-G
11	单R8093 Dan R8093	SNP-G	27	春04 9th-5-1 Chun 04 9th-5-1	SNP-G
12	丰抗13 Fengkang 13	SNP-G	28	春45 9th-50-1 Chun 45 9th-50-1	SNP-G
13	冀麦41 Jimai 41	SNP-G	29	内乡188 Neixiang 188	SNP-G
14	冀麦6号 Jimai 6	SNP-G	30	京411 Jing 411	SNP-G
15	晋2148-7 Jin 2148-7	SNP-G	31	中国春 Chinese Spring	SNP-G
16	京核8922 Jinghe 8922	SNP-A	32	白糙麦 Baicaomai	SNP-G

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