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【目的】明确小麦LEC1（TaLEC1）基因的时空表达模式及等位变异，探讨TaLEC1基因与抗穗发芽的关系。【方法】采用反转录PCR（reverse transcription-PCR， RT-PCR）技术克隆了小麦TaLEC1的3个部分同源基因的完整编码框；采用荧光定量PCR分析了TaLEC1的3个部分同源基因在不同组织和种子发育过程中的表达模式。利用在线数据库对3个TaLEC1的互作蛋白进行了预测，并对其在不同小麦品种中的等位变异进行分析。【结果】成功克隆了TaLEC1的3个部分同源基因的cDNA，分别命名为TaLEC1-6AL、TaLEC1-6BL和TaLEC1-6DL，开放阅读框均为741 bp，编码246个氨基酸。3个基因有相似的组织特异性表达模式，均在花后10 d叶中表达量最高。种子发育过程中，在抗穗发芽‘淮麦0360’中表达呈单峰曲线，花后10 d表达量最高。在易穗发芽‘中优9507’中，表达呈双峰曲线，峰值出现在花后10和20 d。3个基因表达水平依次为TaLEC1-6AL>TaLEC1-6DL>TaLEC1-6BL。TaLEC1可能与Hap5-like（h5l-1B）蛋白和多个CBFD＿NFYB＿HMF 域包含蛋白存在互作。TaLEC1-6AL单倍型多达5种，其中抗穗发芽‘扬麦158’属于单倍型I，易穗发芽‘中优9507’属于单倍型Ⅲ。【结论】TaLEC1基因，尤其是TaLEC1-6AL，可能在穗发芽及种子发育过程中起重要的调控作用。
Abstract：【Objective】To clarify the spatiotemporal expression pattern and allelic variation of the LEC1 genes and investigate the relationship between LEC1 genes and pre-harvest spouting (PHS) resistance in wheat.【Method】The complete coding boxes of the three homologous genes of wheat TaLEC1 was cloned by RT-PCR (reverse transcription-PCR). The expression patterns of TaLEC1 homologous genes in different tissues and seed development stage were analyzed by quantitative RT-PCR (qRT-PCR). The interacting proteins of TaLEC1 were predicted， and the allelic variations of TaLEC1 in different wheat varieties were analyzed by using online databases.【Result】The results showed that the three cDNAs of TaLEC1 homologous genes were successfully cloned， designated as TaLEC1-6AL， TaLEC1-6BL and TaLEC1-6DL. Sequence analysis showed that the open reading frame was 741bp， encoding 246 amino acids. The three homologous genes of TaLEC1 had similar tissue-specific expression patterns， and their expression levels were the highest in the 10d leaves. During seed development， the expression of the three genes in PHS-resistant Huaimai0360 presented a single peak curve， with the highest expression level at 10 days after flowering (DAF). In PHS-susceptible Zhongyou9507， the expression showed a bimodal curve， and the peaks appeared at 10 and 20 DAF， respectively. The expression levels among the three genes were TaLEC1-6AL > TaLEC1-6DL > TaLEC1-6BL. The three TaLEC1 may interact with Hap5-like protein (h5l-1B) and multiple CBFD＿NFYB＿HMF domain-containing proteins. There are as many as five haplotypes of TaLEC1-6AL， among which the PHS-resistant Yangmai158 belongs to haplotype I， and the PHS-susceptible Zhongyou9507 belongs to haplotype Ⅲ. 【Conclusion】It is speculated that TaLEC1 genes especially TaLEC1-6AL may play an important regulatory role in pre-harvest sprouting and grain development.
[1] Tai L， Wang H J， Xu X J， et al. Pre-harvest sprouting in cereals: genetic and biochemical mechanisms[J]. Journal of Experimental Botany， 2021， 72(8): 2857-2876.
[2] Kwong R W， Bui A Q， Lee H， et al. LEAFY COTYLEDON1-LIKE defines a class of regulators essential for embryo development[J]. The Plant Cell， 2003， 15(1): 5-18.
[3] 高剑波， 潘建梅， 陆钦武， 等. 小麦田间穗发芽的影响因素及防止对策[J]. 种子， 2006， 25(6): 75-77.
[4] 张海峰， 卢荣禾， 王建龙. 冬小麦穗发芽抗性及其遗传研究[J]. 西北植物学报， 1992， 12(2): 95-103.
[5] 陈路路. 小麦种子发育过程中活力变化及成熟脱水保护机制研究[D]. 泰安: 山东农业大学， 2017.
[6] 董慧雪， 陈倩， 郭晓江， 等. 小麦穗发芽抗性机制及抗性育种研究[J]. 中国农业科学， 2024， 57(7): 1237-1254.
[7] Ali F， Qanmber G， Li F G， et al. Updated role of ABA in seed maturation， dormancy， and germination[J]. Journal of Advanced Research， 2022， 35: 199-214.
[8] Yamamoto A， Kagaya Y， Toyoshima R， et al. Arabidopsis NF-YB subunits LEC1 and LEC1-LIKE activate transcription by interacting with seed-specific ABRE-binding factors[J]. The Plant Journal， 2009， 58(5): 843-856.
[9] Rikiishi K， Maekawa M. Seed maturation regulators are related to the control of seed dormancy in wheat (Triticum aestivum L.)[J]. PLoS One， 2014， 9(9): e107618.
[10] Pelletier J M， Kwong R W， Park S， et al. LEC1 sequentially regulates the transcription of genes involved in diverse developmental processes during seed development[J]. Proceedings of the National Academy of Sciences of the United States of America， 2017， 114(32): E6710-E6719.
[11] Lotan T， Ohto M A， Yee K M， et al. Arabidopsis LEAFY COTYLEDON1 is sufficient to induce embryo development in vegetative cells[J]. Cell， 1998， 93(7): 1195-1205.
[12] Zhang S B， Wong L， Meng L， et al. Similarity of expression patterns of knotted1 and ZmLEC1 during somatic and zygotic embryogenesis in maize (Zea mays L.)[J]. Planta， 2002， 215(2): 191-194.
[13] 陈金军， 王旭军， 陈建荣， 等. 拟南芥胚胎发生基因LEC1对水稻胚胎发育的遗传学效应[J]. 湖南师范大学自然科学学报， 2008， 31(3): 88-92.
[14] Tan H L， Yang X H， Zhang F X， et al. Enhanced seed oil production in canola by conditional expression of Brassica napus LEAFY COTYLEDON1 and LEC1-LIKE in developing seeds[J]. Plant Physiology， 2011， 156(3): 1577-1588.
[15] Huang M K， Hu Y L， Liu X， et al. Arabidopsis LEAFY COTYLEDON1 controls cell fate determination during post-embryonic development[J]. Frontiers in Plant Science， 2015， 6: 955.
[16] 肖静， 田纪春. 小麦(T.aestivum L.)D基因组的研究进展[J]. 分子植物育种， 2008， 6(3): 537-541.
Xiao J， Tian J C. Reviewed on D genome of T.aestivum L.[J]. Molecular Plant Breeding， 2008， 6(3): 537-541.
[17] Ramírez-González R H， Borrill P， Lang D， et al. The transcriptional landscape of polyploid wheat[J]. Science， 2018， 361(6403): eaar6089.
[18] Chen Z J. Genetic and epigenetic mechanisms for gene expression and phenotypic variation in plant polyploids[J]. Annual Review of Plant Biology， 2007， 58: 377-406.
[19] Yu H， Lin T， Meng X B， et al. A route to de novo domestication of wild allotetraploid rice[J]. Cell， 2021， 184(5): 1156-1170.e14.
[20] 刘豪， 王艳丽， 孟晓丹， 等. 小麦TaLEC1基因的克隆及其表达特性分析[J]. 西北植物学报， 2019， 39(5): 904-910.
Liu H， Wang Y L， Meng X D， et al. Cloning and expression analysis of TaLEC1 gene from wheat[J]. Acta Botanica Boreali-Occidentalia Sinica， 2019， 39(5): 904-910.
[21] Wang W X， Wang Z H， Li X T， et al. SnpHub: an easy-to-set-up web server framework for exploring large-scale genomic variation data in the post-genomic era with applications in wheat[J]. GigaScience， 2020， 9(6): giaa060.
[22] Hao C Y， Jiao C Z， Hou J， et al. Resequencing of 145 landmark cultivars reveals asymmetric sub-genome selection and strong founder genotype effects on wheat breeding in China[J]. Molecular Plant， 2020， 13(12): 1733-1751.
[23] Cheng S F， Feng C， Wingen L U， et al. Harnessing Landrace diversity empowers wheat breeding[J]. Nature， 2024， 632(8026): 823-831.
[24] 孟晓丹. 小麦穗发芽相关基因TaFUS3和TaLEC1的克隆与遗传转化[D]. 郑州: 河南农业大学， 2014.
[25] Finkelstein R， Reeves W， Ariizumi T， et al. Molecular aspects of seed dormancy[J]. Annual Review of Plant Biology， 2008， 59: 387-415.
[26] Tsukagoshi H， Morikami A， Nakamura K. Two B3 domain transcriptional repressors prevent sugar-inducible expression of seed maturation genes in Arabidopsis seedlings[J]. Proceedings of the National Academy of Sciences of the United States of America， 2007， 104(7): 2543-2547.
[27] Yadav D， Shavrukov Y， Bazanova N， et al. Constitutive overexpression of the TaNF-YB4 gene in transgenic wheat significantly improves grain yield[J]. Journal of Experimental Botany， 2015， 66(21): 6635-6650.
[28] Sinha S， Kim I S， Sohn K Y， et al. Three classes of mutations in the a subunit of the CCAAT-binding factor CBF delineate functional domains involved in the three-step assembly of the CBF-DNA complex[J]. Molecular and Cellular Biology， 1996， 16(1): 328-337.
[29] Irikova T， Grozeva S， Denev I. Identification of baby boom and leafy Cotyledon genes in sweet pepper (Capsicum annuum l.) genome by their partial gene sequences[J]. Plant Growth Regulation， 2012， 67(2): 191-198.
[30] 杨博慧， 粱月秀， 柴文婷， 等. 高粱NF-Y转录因子序列特征、表达及生物胁迫响应[J]. 植物病理学报， 2025， 55(2): 237-250.
Yang B H， Liang Y X， Chai W T， et al. Sequence characteristics， expression and response to biological stress of NF-Y transcription factors in sorghum(Sorghum bicolor)[J]. Acta Phytopathologica Sinica， 2025， 55(2): 237-250.
[31] Cui Z B， Wang X W， Dai Y D， et al. Transcription factor OsNF - YC1 regulates grain size by coordinating the transcriptional activation of OsMADS1 in Oryza sativa L[J]. The Plant Journal， 2024， 119(3): 1465-1480.
[32] Kumimoto R W， Siriwardana C L， Gayler K K， et al. NUCLEAR FACTOR Y transcription factors have both opposing and additive roles in ABA-mediated seed germination[J]. PLoS One， 2013， 8(3): e59481.
[33] Liu X， Hu P W， Huang M K， et al. The NF-YC–RGL2 module integrates GA and ABA signalling to regulate seed germination in Arabidopsis[J]. Nature Communications， 2016， 7: 12768.
[34] Du Y X， Ye C， Han P J， et al. The molecular mechanism of transcription factor regulation of grain size in rice[J]. Plant Science， 2025， 354: 112434.
[35] 陈嘉乐， 于清涛， 郑琛凡， 等. 水稻OsNF-YC10自然变异及其与谷粒宽度的相关性[J]. 中国水稻科学， 2025， 39(4): 552-562.
Chen J L， Yu Q T， Zheng C F， et al. Natural variation of Os NF-YC10 and its correlation with grain width in rice[J]. Chinese Journal of Rice Science， 2025， 39(4): 552-562.
[36] Bai A N， Lu X D， Li D Q， et al. NF-YB1-regulated expression of sucrose transporters in aleurone facilitates sugar loading to rice endosperm[J]. Cell Research， 2016， 26(3): 384-388.
[37] Bello B K， Hou Y X， Zhao J， et al. NF-YB1-YC12-bHLH144 complex directly activates W_(x) to regulate grain quality in rice (Oryza sativa L.)[J]. Plant Biotechnology Journal， 2019， 17(7): 1222-1235.
[38] Xu J J， Zhang X F， Xue H W. Rice aleurone layer specific OsNF-YB1 regulates grain filling and endosperm development by interacting with an ERF transcription factor[J]. Journal of Experimental Botany， 2016， 67(22): 6399-6411.
[39] Wang J D， Wang J， Huang L C， et al. ABA-mediated regulation of rice grain quality and seed dormancy via the NF-YB1-SLRL2-bHLH144 Module[J]. Nature Communications， 2024， 15: 4493.
[40] Liu Y C， Xi W， Wang X L， et al. TabHLH95-TaNF-YB1 module promotes grain starch synthesis in bread wheat[J]. Journal of Genetics and Genomics， 2023， 50(11): 883-894.
[41] Nelson D E， Repetti P P， Adams T R， et al. Plant nuclear factor Y (NF-Y) B subunits confer drought tolerance and lead to improved corn yields on water-limited acres[J]. Proceedings of the National Academy of Sciences of the United States of America， 2007， 104(42): 16450-16455.
[42] Stephenson T J， McIntyre C L， Collet C， et al. Genome-wide identification and expression analysis of the NF-Y family of transcription factors in Triticum aestivum[J]. Plant Molecular Biology， 2007， 65(1): 77-92.
[43] Zhao Y J， Ma C Y， Zheng M J， et al. Transcription factor TaNF-YB2 interacts with partners TaNF-YA7/YC7 and transcriptionally activates distinct stress-defensive genes to modulate drought tolerance in T. Aestivum[J]. BMC Plant Biology， 2024， 24(1): 705.
[44] Zhang C Y， Jian M Y， Li W J， et al. Gibberellin signaling modulates flowering via the DELLA-BRAHMA-NF-YC module in Arabidopsis[J]. The Plant Cell， 2023， 35(9): 3470-3484.
[45] Li S L， Hu Y， An C， et al. The amino acid residue E96 of Ghd8 is crucial for the formation of the flowering repression complex Ghd7-Ghd8-OsHAP5C in rice[J]. Journal of Integrative Plant Biology， 2023， 65(4): 1012-1025.
[46] 朱冬梅， 王慧， 刘大同， 等. 小麦籽粒灌浆与脱水特性[J]. 中国农业科学， 2019， 52(23): 4251-4261.
Zhu D M， Wang H， Liu D T， et al. Characteristics of grain filling and dehydration in wheat[J]. Scientia Agricultura Sinica， 2019， 52(23): 4251-4261.
[47] 何贤芳， 赵莉， 刘泽， 等. 安徽省主栽小麦品种(系)脱水及穗发芽特性研究[J]. 滁州学院学报， 2016， 18(2): 70-74.
基本信息:
DOI：10.16445/j.cnki.1000-2340.20260709.001
中图分类号:S512.1
引用信息:
[1]孟晓丹,李惠米,张盈盈,等.小麦TaLEC1的3个部分同源基因的克隆、表达特性及等位变异分析[J].河南农业大学学报().DOI:10.16445/j.cnki.1000-2340.20260709.001.
基金信息:
河南省重大科技专项（241100110300）; 中原学者工作站项目（224400510001）
2026-07-10
2026-07-10
2026-07-10