Article(id=1302192646089367970, tenantId=1146029695717560320, journalId=1301850032934322245, issueId=1302192562882761358, articleNumber=null, orderNo=null, doi=10.3864/j.issn.0578-1752.2026.16.004, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1777996800000, receivedDateStr=2026-05-06, revisedDate=null, revisedDateStr=null, acceptedDate=1782748800000, acceptedDateStr=2026-06-30, onlineDate=1788396520397, onlineDateStr=2026-09-03, pubDate=1786809600000, pubDateStr=2026-08-16, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1788396520397, onlineIssueDateStr=2026-09-03, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1788396520397, creator=13701087609, updateTime=1788396520397, updator=13701087609, issue=Issue{id=1302192562882761358, tenantId=1146029695717560320, journalId=1301850032934322245, year='2026', volume='59', issue='16', pageStart='3465', pageEnd='3698', issueExtLink='null', onlineDate='null', pubDate='1786809600000', pubDateStr='2026-08-16', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=1, specialIssue=null, createTime=1788396500558, creator='13701087609', updateTime=1788405251849, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1302229268860264480, tenantId=1146029695717560320, journalId=1301850032934322245, issueId=1302192562882761358, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1302229268860264481, tenantId=1146029695717560320, journalId=1301850032934322245, issueId=1302192562882761358, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=3509, endPage=3518, ext={EN=ArticleExt(id=1302192646315860387, articleId=1302192646089367970, tenantId=1146029695717560320, journalId=1301850032934322245, language=EN, title=Ethylene and Gibberellin Synergistically Regulate Coleoptile Elongation in Rice, columnId=1302192590426763393, journalTitle=Scientia Agricultura Sinica, columnName=TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY·AGRICULTURE INFORMATION TECHNOLOGY, runingTitle=null, highlight=null, articleAbstract=

【Objective】 Ethylene and gibberellin are key phytohormones regulating rice coleoptile elongation, however, the interaction of ethylene and gibberellin in coleoptile elongation remains unclear. This study aimed to investigate the molecular mechanism of ethylene and gibberellin in regulating coleoptile elongation.【Method】 This study utilized transgenic rice lines of the core components of ethylene and gibberellin signaling pathways as experimental materials. The coleoptile lengths were examined with or without different phytohormone treatments. The expression levels of downstream genes of OsEIN2/OsEIL2 were detected by RT-qPCR. Yeast two-hybrid and pull-down assays were further employed to verify the interaction between OsSLR1 and OsEIL2. 【Result】 Exogenous phytohormones treatment revealed that both ethylene and gibberellin promoted coleoptile elongation, and they exhibited a synergistic effect. Paclobutrazol (PAC, a GA biosynthesis inhibitor) treatment or overexpression of the gibberellin inactivation gene OsGA2ox3 weakened the promoting effect of ethylene on coleoptile elongation, whereas exogenous application of gibberellin partially relieved the 1-MCP (1-Methylcyclopropene) inhibited coleoptile elongation, indicating that ethylene and gibberellin shared a common regulatory pathway in controlling coleoptile elongation. Further studies revealed that OsSLR1, a negative regulator of gibberellin signaling, interacted with OsEIL2. Moreover, OsEIN2/OsEIL2-regulated genes, such as OsERF63, OsERF73, OsHKT2; 1 and OsGY1 were also regulated by OsSLR1. The OsSLR1 loss-of-function mutant slr1 exhibited a long coleoptile phenotype, and the promoting effect of ethylene on coleoptile elongation was further enhanced in slr1, suggesting that OsSLR1 was involved in ethylene-mediated coleoptile elongation and signal transduction. 【Conclusion】 In summary, this study revealed that OsSLR1 served as a crosstalk node for the synergistic regulation of coleoptile elongation by ethylene and gibberellin, and it interacted with OsEIL2 to co-regulate downstream signaling pathways and coleoptile elongation. This study not only enriched the understanding of the phytohormone interaction network in coleoptile growth, but also provided a theoretical basis and useful genes for the breeding of rice varieties suitable for direct seeding cultivation.

, authors=Bin WU1, BingKun GE2, TianYu QIN2, GuiQing XIAO1, Hua QIN2, authorsList=Bin WU, BingKun GE, TianYu QIN, GuiQing XIAO, Hua QIN, authorCompany=null, correspAuthors=null, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, fund=null), CN=ArticleExt(id=1302192647590928814, articleId=1302192646089367970, tenantId=1146029695717560320, journalId=1301850032934322245, language=CN, title=乙烯与赤霉素协同调控水稻胚芽鞘伸长生长, columnId=1302192591009771651, journalTitle=中国农业科学, columnName=耕作栽培·生理生化·农业信息技术, runingTitle=null, highlight=null, articleAbstract=

【目的】 乙烯和赤霉素是调控水稻胚芽鞘伸长的关键激素,但二者在该过程中的交互作用机制尚不明确,本文旨在探究二者在调控胚芽鞘伸长中的分子机制。【方法】 利用乙烯和赤霉素信号通路核心组分的相关水稻材料,统计不同激素处理后胚芽鞘的长度,采用qPCR技术检测OsEIN2/OsEIL2下游基因表达,利用酵母双杂和Pull-down技术验证OsSLR1与OsEIL2互作。【结果】 乙烯和赤霉素均能促进胚芽鞘的伸长,且二者存在协同效应。赤霉素合成抑制剂多效唑(Paclobutrazol,PAC)处理或过表达赤霉素失活基因OsGA2ox3削弱了乙烯对胚芽鞘伸长的促进作用,外源补充赤霉素也能够部分解除1-MCP(1-Methylcyclopropene)对胚芽鞘的抑制,表明乙烯与赤霉素调控胚芽鞘的伸长存在共同的调控途径。进一步研究发现,赤霉素信号负调控因子OsSLR1与OsEIL2存在相互作用,且OsEIN2/OsEIL2调控的下游基因OsERF63OsERF73OsHKT2;1OsGY1也受到了OsSLR1的调控,OsSLR1功能缺失突变体slr1表现出长胚芽鞘的表型,且乙烯对胚芽鞘伸长的促进作用在slr1中进一步增强,表明OsSLR1参与调控乙烯促进的胚芽鞘生长和信号转导。【结论】 OsSLR1是乙烯和赤霉素协同调控胚芽鞘伸长生长的节点,通过与OsEIL2互作来共同调控下游信号通路和胚芽鞘的伸长生长,这不仅丰富了胚芽鞘生长中的植物激素互作网络,也为直播稻新品种的培育提供了理论依据和有用的基因。

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A: Coleoptile phenotypes of NIP treated with different hormones; B: Statistical analysis of coleoptile length. All significance symbols in the figure represent comparisons between each treatment group and the Mock group. *, ** and **** indicate significant differences at P<0.05, P<0.005 and P<0.001, respectively. The same as below

, figureFileSmall=/5ba/D/wkjEYu43bcAV/1A==, figureFileBig=35SvMMPFqMX8R11rybYCiw==, tableContent=null), ArticleFig(id=1302192650166231515, tenantId=1146029695717560320, journalId=1301850032934322245, articleId=1302192646089367970, language=CN, label=图1, caption=不同激素处理NIP的胚芽鞘长度

A:不同激素处理下的NIP胚芽鞘表型;B:胚芽鞘长度统计。图中显著性标记均为各处理组与对照组比较所得,*、**和****分别表示在P<0.05、P<0.005和P<0.001水平差异显著。下同

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A, B: Coleoptile phenotypes of EIN2 and EIL2 overexpression lines and mutants with or without ethylene treatment

, figureFileSmall=GIKn+4mtaafMvmKGZ3eFyg==, figureFileBig=CwvsJTDpfEHsPFsPB9wkNw==, tableContent=null), ArticleFig(id=1302192650434666973, tenantId=1146029695717560320, journalId=1301850032934322245, articleId=1302192646089367970, language=CN, label=图2, caption=乙烯促进胚芽鞘的伸长生长依赖于乙烯信号通路

A、B:乙烯处理EIN2EIL2过表达和突变体材料的胚芽鞘表型

, figureFileSmall=GIKn+4mtaafMvmKGZ3eFyg==, figureFileBig=CwvsJTDpfEHsPFsPB9wkNw==, tableContent=null), ArticleFig(id=1302192650501775838, tenantId=1146029695717560320, journalId=1301850032934322245, articleId=1302192646089367970, language=EN, label=Fig. 3, caption=Coleoptile length of EIN2 and EIL2 overexpression lines and mutants under GA3 or PAC treatment, figureFileSmall=OCVymtzSehIfXLsRGQcJsA==, figureFileBig=RniaYyIqDhRBVg6qy5CIGg==, tableContent=null), ArticleFig(id=1302192650577273311, tenantId=1146029695717560320, journalId=1301850032934322245, articleId=1302192646089367970, language=CN, label=图3, caption=GA3或PAC处理下EIN2/EIL2过表达和突变体材料的胚芽鞘表型, figureFileSmall=OCVymtzSehIfXLsRGQcJsA==, figureFileBig=RniaYyIqDhRBVg6qy5CIGg==, tableContent=null), ArticleFig(id=1302192650648576480, tenantId=1146029695717560320, journalId=1301850032934322245, articleId=1302192646089367970, language=EN, label=Fig. 4, caption=Coleoptile length under ethylene and 1-MCP treatment

A: Coleoptile phenotypes of ZH11 and OsGA2ox3 overexpression plants treated with ethylene; B: Statistical analysis of coleoptile length in ZH11 and OsGA2ox3 overexpression plants; C: Coleoptile phenotypes of WT and slr1 mutants treated with ethylene and 1-MCP; D: Statistical analysis of coleoptile length in WT and slr1 mutants

, figureFileSmall=fO1qC8IwB9dDp4M6lLpyAQ==, figureFileBig=NssvO5KWLcdG6Jj+NusU8w==, tableContent=null), ArticleFig(id=1302192650719879649, tenantId=1146029695717560320, journalId=1301850032934322245, articleId=1302192646089367970, language=CN, label=图4, caption=乙烯和1-MCP处理胚芽鞘长度

A:乙烯处理ZH11和OsGA2ox3过表达植株胚芽鞘表型;B:ZH11和OsGA2ox3过表达植株胚芽鞘长度统计;C:乙烯和1-MCP处理WT和slr1突变体胚芽鞘表型;D:WT和slr1突变体胚芽鞘长度统计

, figureFileSmall=fO1qC8IwB9dDp4M6lLpyAQ==, figureFileBig=NssvO5KWLcdG6Jj+NusU8w==, tableContent=null), ArticleFig(id=1302192650799571426, tenantId=1146029695717560320, journalId=1301850032934322245, articleId=1302192646089367970, language=EN, label=Fig. 5, caption=The interaction of EIL2 with SLR1 and the expression profiles of ethylene-responsive genes in different lines of EIN2, EIL2, and SLR1

A: Yeast two-hybrid assay confirms EIL2-SLR1 interaction; B: GST pull-down assay verifies their direct binding in vitro

, figureFileSmall=r/Ob5Ddt7suVMlOc7uR2Wg==, figureFileBig=JwdCVD4hx4cnRm2+ffvRvw==, tableContent=null), ArticleFig(id=1302192650866680291, tenantId=1146029695717560320, journalId=1301850032934322245, articleId=1302192646089367970, language=CN, label=图5, caption=EIL2与SLR1互作及乙烯响应基因在EIN2、EIL2SLR1不同株系中的表达情况

A:酵母双杂交试验验证EIL2与SLR1的互作;B:GST pull-down试验验证EIL2与SLR1的体外直接结合

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乙烯与赤霉素协同调控水稻胚芽鞘伸长生长
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吴斌 1 , 葛秉坤 2 , 秦田雨 2 , 肖桂青 1 , 秦华 2
中国农业科学 | 耕作栽培·生理生化·农业信息技术 2026,59(16): 3509-3518
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中国农业科学 |耕作栽培·生理生化·农业信息技术 2026 , 59 (16) : 3509 -3518
乙烯与赤霉素协同调控水稻胚芽鞘伸长生长
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吴斌1 , 葛秉坤2, 秦田雨2, 肖桂青1 , 秦华2
作者信息
  • 1 湖南农业大学生物科学技术学院, 长沙 410128
  • 2 中国农业科学院生物技术研究所, 北京 100081
通讯作者:
肖桂青,E-mail:
秦华,E-mail:
作者简介:

吴斌,E-mail:

Ethylene and Gibberellin Synergistically Regulate Coleoptile Elongation in Rice
Bin WU1 , BingKun GE2, TianYu QIN2, GuiQing XIAO1 , Hua QIN2
Affiliations
  • 1 College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha 410128
  • 2 Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081
出版时间: 2026-08-16 doi: 10.3864/j.issn.0578-1752.2026.16.004
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【目的】 乙烯和赤霉素是调控水稻胚芽鞘伸长的关键激素,但二者在该过程中的交互作用机制尚不明确,本文旨在探究二者在调控胚芽鞘伸长中的分子机制。【方法】 利用乙烯和赤霉素信号通路核心组分的相关水稻材料,统计不同激素处理后胚芽鞘的长度,采用qPCR技术检测OsEIN2/OsEIL2下游基因表达,利用酵母双杂和Pull-down技术验证OsSLR1与OsEIL2互作。【结果】 乙烯和赤霉素均能促进胚芽鞘的伸长,且二者存在协同效应。赤霉素合成抑制剂多效唑(Paclobutrazol,PAC)处理或过表达赤霉素失活基因OsGA2ox3削弱了乙烯对胚芽鞘伸长的促进作用,外源补充赤霉素也能够部分解除1-MCP(1-Methylcyclopropene)对胚芽鞘的抑制,表明乙烯与赤霉素调控胚芽鞘的伸长存在共同的调控途径。进一步研究发现,赤霉素信号负调控因子OsSLR1与OsEIL2存在相互作用,且OsEIN2/OsEIL2调控的下游基因OsERF63OsERF73OsHKT2;1OsGY1也受到了OsSLR1的调控,OsSLR1功能缺失突变体slr1表现出长胚芽鞘的表型,且乙烯对胚芽鞘伸长的促进作用在slr1中进一步增强,表明OsSLR1参与调控乙烯促进的胚芽鞘生长和信号转导。【结论】 OsSLR1是乙烯和赤霉素协同调控胚芽鞘伸长生长的节点,通过与OsEIL2互作来共同调控下游信号通路和胚芽鞘的伸长生长,这不仅丰富了胚芽鞘生长中的植物激素互作网络,也为直播稻新品种的培育提供了理论依据和有用的基因。

水稻  /  胚芽鞘  /  乙烯  /  赤霉素  /  OsEIL2  /  OsSLR1

【Objective】 Ethylene and gibberellin are key phytohormones regulating rice coleoptile elongation, however, the interaction of ethylene and gibberellin in coleoptile elongation remains unclear. This study aimed to investigate the molecular mechanism of ethylene and gibberellin in regulating coleoptile elongation.【Method】 This study utilized transgenic rice lines of the core components of ethylene and gibberellin signaling pathways as experimental materials. The coleoptile lengths were examined with or without different phytohormone treatments. The expression levels of downstream genes of OsEIN2/OsEIL2 were detected by RT-qPCR. Yeast two-hybrid and pull-down assays were further employed to verify the interaction between OsSLR1 and OsEIL2. 【Result】 Exogenous phytohormones treatment revealed that both ethylene and gibberellin promoted coleoptile elongation, and they exhibited a synergistic effect. Paclobutrazol (PAC, a GA biosynthesis inhibitor) treatment or overexpression of the gibberellin inactivation gene OsGA2ox3 weakened the promoting effect of ethylene on coleoptile elongation, whereas exogenous application of gibberellin partially relieved the 1-MCP (1-Methylcyclopropene) inhibited coleoptile elongation, indicating that ethylene and gibberellin shared a common regulatory pathway in controlling coleoptile elongation. Further studies revealed that OsSLR1, a negative regulator of gibberellin signaling, interacted with OsEIL2. Moreover, OsEIN2/OsEIL2-regulated genes, such as OsERF63, OsERF73, OsHKT2; 1 and OsGY1 were also regulated by OsSLR1. The OsSLR1 loss-of-function mutant slr1 exhibited a long coleoptile phenotype, and the promoting effect of ethylene on coleoptile elongation was further enhanced in slr1, suggesting that OsSLR1 was involved in ethylene-mediated coleoptile elongation and signal transduction. 【Conclusion】 In summary, this study revealed that OsSLR1 served as a crosstalk node for the synergistic regulation of coleoptile elongation by ethylene and gibberellin, and it interacted with OsEIL2 to co-regulate downstream signaling pathways and coleoptile elongation. This study not only enriched the understanding of the phytohormone interaction network in coleoptile growth, but also provided a theoretical basis and useful genes for the breeding of rice varieties suitable for direct seeding cultivation.

rice  /  coleoptile  /  ethylene  /  gibberellin  /  OsEIL2  /  OsSLR1
吴斌, 葛秉坤, 秦田雨, 肖桂青, 秦华. 乙烯与赤霉素协同调控水稻胚芽鞘伸长生长. 中国农业科学, 2026 , 59 (16) : 3509 -3518 . DOI: 10.3864/j.issn.0578-1752.2026.16.004
Bin WU, BingKun GE, TianYu QIN, GuiQing XIAO, Hua QIN. Ethylene and Gibberellin Synergistically Regulate Coleoptile Elongation in Rice[J]. Scientia Agricultura Sinica, 2026 , 59 (16) : 3509 -3518 . DOI: 10.3864/j.issn.0578-1752.2026.16.004
【研究意义】胚芽鞘是单子叶植物特有的锥形组织,能够抵御土壤的摩擦和挤压,避免胚芽受损[1]。胚芽鞘通过快速伸长引导初生叶向上生长,为种子顺利破土开辟通道[2]。在单子叶植物中,胚芽鞘的完整性直接决定了幼苗能否顺利出土,进而启动后续的光合器官发育与形态建成过程[3-4]。在农业生产中,种子播种后能否成功出苗并形成健壮幼苗,直接关系到作物的群体结构和最终产量。胚芽鞘长度、机械强度、伸长速度等性状决定了作物从播种到出苗的能力。胚芽鞘长度与出苗率呈显著正相关关系,若种子播种深度超过胚芽鞘的长度,胚芽鞘将无法到达土壤表面,导致幼苗黄化甚至死亡[5-6]。随着全球气候变化和水资源短缺问题的加剧,以及农业生产机械化程度的提高,深播是获取深层土壤水分的必要措施,而在机械化播种过程中,播种深度往往难以精确控制,容易出现播深不均的问题。在此背景下,培育长胚芽鞘品种对保障作物出苗具有重要意义。【前人研究进展】乙烯(Ethylene,C2H4)作为一种关键的气态植物激素,广泛参与调控植物生长发育的多个生物学过程[7-9]。当乙烯被内质网定位的受体家族感知后,信号通过CTR1和EIN2进行转导,并由核心转录因子EIN3和EIL1放大信号,这些因子进一步激活乙烯响应基因的表达,从而触发乙烯反应[10]。乙烯能够调控胚芽鞘的生长[11-12]。在单子叶植物水稻中,土壤覆盖促进了胚芽鞘中乙烯的合成,减少胚芽鞘顶端活性氧的积累,最终促进胚芽鞘伸长[2]。除此之外,土壤覆盖带来的机械压力诱导幼苗乙烯合成后一个以COP1-EBF1/ 2-EIN3为核心的信号模块,根据土壤的“深度”(即光强的微弱变化)来精确调控乙烯信号的强度,最终通过下游不同通路协调下胚轴伸长与子叶的光合准备[13-14];乙烯也通过抑制OsGY1介导的茉莉酸生物合成,促进水稻黄化苗胚芽鞘和中胚轴的伸长[15]。这些研究共同揭示了乙烯在调控胚芽鞘伸长生长中的核心地位,进一步凸显了其在植物适应土壤环境中的重要作用。赤霉素(Gibberellic acid,GA)作为调控植物生长发育的关键激素之一,同样在胚芽鞘伸长过程中发挥着重要作用。通过对水稻胚芽鞘离体培养发现外源赤霉素处理能够促进胚芽鞘切段的伸长[16]。研究发现赤霉素生物合成抑制剂多效唑(PAC)处理能够显著抑制水稻胚芽鞘伸长,表明赤霉素在水稻胚芽鞘的伸长生长中发挥重要作用[17]。在小麦胚芽鞘相关研究中发现对赤霉素不敏感的小麦其胚芽鞘的伸长受到限制[18]。在大麦胚芽鞘相关研究中则发现,赤霉素诱导的淀粉酶活性不依赖于内源生长素,提示赤霉素在某些组织中可能具有独立于生长素的作用路径,这反映了赤霉素作用机制的复杂性[19]。赤霉素信号转导途径的关键负调控因子SLR1是水稻中的DELLA蛋白,在抑制下游基因转录、调控植物生长发育中发挥核心作用。SLR1通过与其他转录因子相互作用来抑制下游基因的表达,是连接多种激素信号网络的“分子枢纽”[20]。在赤霉素缺乏条件下,DELLA蛋白(如水稻中的SLR1)作为转录抑制因子在细胞核内积累,通过直接结合PIF等转录因子启动子抑制细胞伸长相关基因的表达[21]。SLR1与OsMADS23互作增强OsMADS23稳定性,导致OsMADS23介导的D14转录抑制增强[22]。当赤霉素存在时,DELLA蛋白被SCF E3泛素连接酶复合体泛素化降解,从而解除其对下游转录因子的抑制促进胚芽鞘细胞伸长[23-25]。乙烯和赤霉素之间的串扰在一些研究中已经得到证实。在拟南芥中,DELLA蛋白与EIN3/EIL1互作来抑制EIN3/EIL1对顶端弯钩形成关键基因HLS1的激活,赤霉素促进DELLA蛋白的降解,从而解除对EIN3转录活性的抑制,使得乙烯对HLS1的诱导作用显著增强,共同促进顶端弯钩的形成以保护子叶免受土壤损伤[26]。在水稻中,中胚轴(位于幼苗种子根基部与胚芽鞘节之间的结构)在推动茎尖穿透土壤表层时提供生长活力,对于直播稻的均匀出苗至关重要。研究发现覆土诱导的乙烯通过转录因子OsEIL1直接激活赤霉素合成基因OsSD1的表达,解除对OsPIF转录因子的抑制,进而驱动中胚轴细胞伸长帮助幼苗破土[27]。而在水稻初生根伸长的调控中,赤霉素与乙烯则呈现出拮抗关系:乙烯信号转录因子OsEIL1直接结合到多个赤霉素代谢基因OsGA2oxs的启动子上并激活其表达,从而降低根中活性赤霉素的含量,导致根尖分生区细胞增殖受阻,从而抑制根的伸长[28]。在小麦株高调控的研究中,乙烯信号转录因子TaEIL1抑制赤霉素合成基因TaGA3ox2的转录抑制因子TaERF1的表达,导致赤霉素水平升高,促进节间伸长和株高增加[29]。【本研究切入点】以上研究揭示了乙烯与赤霉素互作调控植物的生长发育机制的复杂性,但两者在水稻胚芽鞘生长中的调控关系还不清楚。【拟解决的关键问题】以外源激素处理乙烯和赤霉素信号通路核心组分的相关水稻材料,解析乙烯和赤霉素在调控水稻胚芽鞘伸长中的关系,明确SLR1-OsEIL2模块通过调控OsEIL2下游靶基因表达介导胚芽鞘伸长的分子路径,揭示乙烯与赤霉素协同调控水稻胚芽鞘伸长的内在机制,为选育适宜直播栽培的水稻新品种提供理论支撑。
水稻野生型NIP,ZH11,ein2、eil2-1、eil2-2、eil2-3、EIL2-RNAi、EIN2-OXEIL2-OXEIL2-OX-1EIL2-OX- 2EIL2-OX-3由实验室保存。Eva Green Mix购于加拿大ABM公司;RNA提取试剂盒购于北京康为世纪生物科技有限公司;反转录试剂盒购于南京诺唯赞生物科技有限公司;植物激素C2H4、GA3、PAC和1-MCP购于西格玛奥德里奇(上海)贸易有限公司。
PCR仪(北京东胜创新生物科技有限公司);PowerPac系列通用电泳仪(美国Bio-Rad司);7500实时荧光定量PCR仪(美国应用生物系统公司)。
将水稻种子用清水恒温37 ℃浸种24 h后,倒掉多余的水分。然后置于37 ℃继续培养,每天早晚各换1次水,直至种子露白。挑取萌发一致的种子种到铁架上,并将铁架放到密封的塑料盒中,放置于黑暗的培养架上,恒温28 ℃培养,后续均在此基础上添加植物生长调节剂处理。
基本处理分为4组:C2H4处理(1 μL·L-1)、1-MCP(1 μL·L-1)处理、GA3处理(2 μmol·L-1)、PAC处理(1 μmol·L-1);PAC、GA3处理对照组加入等体积无水乙醇,C2H4、1-MCP处理对照组仅加入无菌水。水稻培养2 d后拍照,观察并记录表型,取水稻胚芽鞘用于检测赤霉素合成基因和EIN2/EIL1下游基因的表达。3次生物学重复,每次重复处理30株苗。本研究所有试验均为2025年3月至10月在中国农业科学院作物科学研究所重大工程楼完成。
为研究水稻种子胚芽鞘的伸长状况,统计30粒萌发种子生长2 d的胚芽鞘长度,胚芽鞘取种子破壳处至胚芽鞘尖端之间长度。本研究统计胚芽鞘长度做3次生物学重复试验,每次重复30株幼苗。
取黑暗条件下恒温28 ℃培养2 d长的水稻胚芽鞘0.1 g,样品保存于-80 ℃冰箱中,用于RNA提取。每个株系设置3个生物学重复,使用超纯RNA提取试剂盒(北京康为世纪生物科技有限公司)提取水稻胚芽鞘的RNA,具体的操作步骤参照试剂盒说明书。
以提取的水稻胚芽鞘总RNA为模板,使用反转录试剂盒(诺唯赞生物科技有限公司)制备水稻cDNA,具体的试验步骤参照试剂盒说明书,制备的水稻cDNA于-20 ℃保存备用。
本研究使用的荧光染料为SYBR Green,使用的仪器为ABI 7500实时荧光定量PCR仪。qPCR反应体系(20 μL)为:模板1 μL,上、下游引物各1 μL,qPCR Master Mix 10 μL,双蒸水7 μL。qPCR反应程序为:95 ℃ 10 min:95 ℃ 15 s,60 ℃ 60 s,共40个循环。以水稻中的actin肌动蛋白基因作为qPCR反应的内参,每个样品做3次平行重复试验。
利用Image J软件测量水稻幼苗地上部的长度;采用GraphPad Prism 9软件进行数据统计分析,试验结果以“平均值±标准差”表示。组间差异采用单因素方差分析(ANOVA),显著性差异通过Tukey多重比较法进行检验。
乙烯(C2H4)和赤霉素(GA)均能促进胚芽鞘的伸长。为了探究二者在调控胚芽鞘伸长生长中的关系,本研究统计了不同植物生长调节剂处理下NIP的胚芽鞘长度,试验结果表明,C2H4处理显著促进了胚芽鞘的伸长,GA3处理同样表现出明显的促进效应,1-MCP、PAC处理则强烈抑制了胚芽鞘的伸长。C2H4、GA3共同处理时,表现出显著的协同效应,其胚芽鞘长度超过了任一激素单独处理的效果,而赤霉素合成抑制剂PAC和乙烯竞争性抑制剂1-MCP共同处理时,其对胚芽鞘的抑制超过了任一激素单独处理的效果(图1),表明乙烯与GA在调控水稻胚芽鞘伸长生长上存在协同效应。C2H4与PAC复合处理显示PAC显著抑制了乙烯的促进作用,而GA3与1-MCP复合处理显示GA3在一定程度上缓解了1-MCP的抑制效果,表明乙烯促进胚芽鞘的伸长生长需要GA3的参与,扰乱内源赤霉素合成会削弱乙烯对胚芽鞘的伸长生长的促进作用。以上结果表明乙烯与GA3均能促进水稻胚芽鞘的伸长生长,且两者在促进水稻胚芽鞘的伸长生长中可能存在相同的调控途径。
为了验证乙烯信号通路核心组分在乙烯调控胚芽鞘伸长生长中的作用,对实验室已有的乙烯信号通路核心调控因子EIN2EIL2的相关水稻材料进行乙烯处理,观察胚芽鞘的生长情况。发现在没有乙烯时,ein2突变体植株的胚芽鞘长度显著短于日本晴,而eil2-1、eil2-2、eil2-3突变体植株和EIL2-RNAi植株胚芽鞘长度与日本晴相比无明显差异,过表达EIN2EIN2-OX)与EIL2EIL2-OXOX-1OX-2OX-3)的植株的胚芽鞘长度显著长于日本晴。外源施加乙烯显著促进NIP胚芽鞘的伸长,这种促进作用在ein2、eil2-1、eil2-2、eil2-3突变体植株和EIL2-RNAi中则明显被抑制,而在EIN2-OXEIL2-OXOX-1OX-2OX-3植株中则显著增强(图2)。
为了探究赤霉素与乙烯在调控水稻胚芽鞘伸长生长中调控关系,本研究用GA3与赤霉素合成抑制剂PAC处理野生型、ein2、eil2突变体及EIN2、EIL2过表达材料,并观察胚芽鞘长度变化(图3)。结果显示,外源GA3处理显著促进各基因型材料胚芽鞘的伸长,而PAC处理则显著抑制胚芽鞘伸长,且GA3的促进作用和PAC的抑制作用在野生型、ein2、eil2突变体及EIN2、EIL2过表达株系中无显著差异(图3)。以上结果表明,赤霉素对胚芽鞘伸长的促进作用不依赖于乙烯信号核心组分EIN2和EIL2,结合图1中乙烯和赤霉素协同调控胚芽鞘的伸长生长的结果,证明赤霉素可能作用于EIL2下游组分来调控胚芽鞘的伸长生长。
为了进一步探究赤霉素与乙烯在调控水稻胚芽鞘伸长生长中调控关系,对野生型ZH11及3个OsGA2ox3过表达株系(OsGA2ox3-ox-1OsGA2ox3- ox-2OsGA2ox3-ox-3)进行乙烯处理。OsGA2ox3属于GA2ox家族,其功能是将具有生物活性的赤霉素转化为无活性形式,从而降低植物体内活性赤霉素水平[28]。结果发现,过表达OsGA2ox3显著抑制了胚芽鞘的伸长(图4-A、B),表明降低内源活性赤霉素含量导致胚芽鞘的生长受到了抑制,从而进一步证明了赤霉素对胚芽鞘的伸长生长的促进作用。外源C2H4处理促进了野生型和OsGA2ox3过表达株系胚芽鞘的伸长,但这种促进作用在OsGA2ox3过表达植株中要弱于野生型(图4-A、B),表明降低内源赤霉素水平削弱了乙烯促进的胚芽鞘生长,赤霉素可能作用于乙烯信号下游途径来调控胚芽鞘的生长。进一步用乙烯和1-MCP处理野生型(WT)及赤霉素信号通路的核心负调控因子OsSLR1的功能缺失突变体slr1,并观察胚芽鞘长度变化(图4-C、D)。结果发现,slr1突变体的胚芽鞘长度显著长于野生型,1-MCP处理显著抑制了WT和slr1突变体胚芽鞘的伸长,而乙烯处理则促进了两者的胚芽鞘伸长,且对slr1突变体的促进效果更为显著(图4-C、D),表明SLR1参与调控乙烯促进的胚芽鞘伸长生长,且SLR1可能是乙烯和赤霉素协同调控胚芽鞘伸长生长的互作节点。
已有研究表明SLR1通过与转录因子互作来调节其结合或转录活性,从而调控植物的生长发育进程[30-31]。本研究发现SLR1参与调控乙烯促进的胚芽鞘伸长生长,且乙烯信号核心组分EIN2和EIL2突变体能正常响应GA3和PAC处理,因此推测SLR1可能与EIL2互作来共同调控下游信号途径,从而影响胚芽鞘的伸长。为了验证这一猜想,首先通过酵母双杂交(Y2H)试验在酵母体内验证二者的互作。结果如图5-A所示:在SD/-Leu/-Trp培养基上,所有转化组合均能正常生长,证明质粒成功转入酵母细胞;而在SD/-Leu/-Trp/-His/-Ade培养基上,仅共转化AD-EIL2与BD-SLR的酵母菌株能够正常生长,且在X-Gal显色试验中呈现明显的β-半乳糖苷酶活性(蓝色菌落),而共转化AD+BD、AD-EIL2+BD、AD+BD-SLR1的酵母菌株不能在SD/-Leu/-Trp/-His/-Ade培养基上生长,表明EIL2与SLR1在酵母细胞内存在相互作用。在此基础上,进一步通过Pull-down试验验证二者的互作(图5-B)。以GST标签蛋白为阴性对照,GST-EIL2融合蛋白作为诱饵蛋白,His-SLR1为猎物蛋白进行体外孵育与亲和捕获。Western blot结果显示:anti-GST抗体检测证实在Input及IP样品中均能检测到GST与GST-EIL2蛋白,表明GST与GST-EIL2蛋白被有效捕获,W3洗涤样品无明显信号,证明洗涤充分、非特异性结合已被去除;anti-His抗体检测结果显示,仅在Input和GST-EIL2组的IP样品中可检测到His-SLR1的特异性条带,GST对照组IP样品中无对应信号,表明EIL2与SLR1存在直接的物理相互作用。综合酵母双杂交与Pull-down试验结果,本研究证实SLR1与EIL2存在特异性的相互作用,赤霉素与乙烯信号可通过OsSLR1-OsEIL2模块调控胚芽鞘伸长生长。
为了研究SLR1与EIL2互作对下游信号通路的影响,本研究利用qRT-PCR检测了乙烯marker基因OsERF63OsERF73及EIL2下游靶基因OsHKT2;1、OsGY1的表达(图5-C—E)。结果表明,eil2、ein2突变体株系中OsERF6、OsERF73OsHKT2;1 3个基因的表达水平均显著低于NIP,而OsGY1的表达水平高于NIP;在OsEIL2、OsEIN2过表达株系中OsERF63、OsERF73OsHKT2;1 3个基因的表达水平均显著高于NIP,而OsGY1的表达水平低于NIP(图5-C、D)。在slr1突变体中,OsOsERF63OsOsERF73OsHKT2;1的表达水平显著高于WT,而OsGY1的表达则显著低于WT(图5-E),与OsEIL2、OsEIN2过表达的效果类似。以上结果表明,OsSLR1与OsEIL2拮抗调控乙烯信号途径下游通路,OsSLR1可能通过与OsEIL2互作来抑制OsEIL2的转录活性,进而调控乙烯信号途径下游信号通路和胚芽鞘的生长。
胚芽鞘的快速伸长是单子叶植物幼苗成功破土并成功建立光合能力的关键,直接决定作物出苗效率与后期群体产量,尤其在水稻直播、机械化深播等轻简化栽培模式推广的当下,解析胚芽鞘伸长的调控机制,对培育耐深播、出苗整齐的水稻品种具有重要的理论与生产实践意义。乙烯与赤霉素作为调控植物生长发育的核心激素,二者在胚芽鞘伸长中的单独作用已得到证实,但二者的作用模式及分子调控网络仍不完善。本研究以水稻为研究材料,探究了乙烯与赤霉素协同调控水稻胚芽鞘伸长的机制,为水稻直播栽培与分子育种提供了新的靶点与思路。
本研究通过外源激素及抑制剂处理试验证实乙烯与GA3单独处理均能显著促进水稻胚芽鞘的伸长,二者共同处理时呈现显著的协同效应,其促进效果显著优于单一激素处理;而乙烯竞争性抑制剂1-MCP与GA合成抑制剂PAC单独处理均强烈抑制胚芽鞘伸长,且二者联合处理的抑制效应更显著。Lü等[27]报道乙烯-GA信号级联调控水稻中胚轴伸长,说明乙烯-GA信号通过多方面促进幼苗出土。本研究发现二者之间并非简单的级联模式,而是存在更加复杂的协同调控,研究发现PAC可部分解除乙烯的促进作用,GA3也可部分缓解1-MCP的抑制效应,暗示二者可能通过相同的信号通路共同推动胚芽鞘伸长。进一步qRT-PCR检测发现,乙烯通过EIN2/EIL模块正调控OsOsERF63OsOsERF73OsHKT2;1的表达,同时负调控OsGY1的表达;而赤霉素信号通路负调控因子SLR1负调控OsOsERF63OsOsERF73OsHKT2;1的表达,同时正调控OsGY1的表达,进一步证实两者在胚芽鞘生长中存在相同的调控途径。Xiong等[15]发现OsGY1介导的茉莉酸生物合成位于乙烯信号途径下游调控胚芽鞘的伸长,结合本研究结果可推测,乙烯和赤霉素可能协同调控OsGY1介导的茉莉酸生物合成来调控胚芽鞘的伸长。
赤霉素信号通路的核心负调控因子SLR1(DELLA蛋白)在乙烯-GA协同调控中的枢纽作用,是本研究的重要创新点。SLR1作为植物激素信号整合的关键节点[32],其经典功能是通过自身降解解除对GA响应基因的抑制;然而本研究发现,OsSLR1还可通过调控乙烯信号通路下游基因的表达,负向调控乙烯对胚芽鞘伸长的促进作用——osslr1功能缺失突变体中,乙烯诱导的胚芽鞘伸长效应更显著,且OsERF63OsERF73OsHKT2;1的表达量显著升高,OsGY1的表达量显著降低,这与EIN2EIL2过表达的效果类似,意味着SLR1可能是整合乙烯和赤霉素协同调控胚芽鞘伸长生长的节点,这一发现拓展了DELLA蛋白的功能,证实其不仅参与GA信号转导,还可通过与乙烯信号通路的交互,整合两种激素的调控作用。Hirano等[33]研究发现GA积累可通过GA-GID1-DELLA三聚体形成,促进SLR1泛素化降解。本研究进一步通过酵母双杂交与pull-down试验证实,SLR1与EIL2存在直接的互作,推测SLR1可能通过与EIL2结合,抑制其下游转录因子的转录活性,从而抑制乙烯信号的传递。而GA介导的SLR1降解可解除这种抑制,实现乙烯信号的增强,osslr1突变增强乙烯敏感性的研究,进一步印证了二者的协同关系。OsGA2ox3作为GA失活基因,其过表达可显著降低体内活性GA水平,本研究发现,OsGA2ox3过表达株系中,乙烯促进胚芽鞘伸长的效果显著弱于野生型,推测植物内源GA水平越高,植物对乙烯的敏感性越强。
结合当前水稻生产实际,直播栽培因省工节本、生育期短等优势,已成为水稻生产的主流模式之一,但土壤不平整、覆土厚度不均、干旱等问题,常导致水稻出苗不齐、成苗率低,而胚芽鞘长度是影响直播稻出苗能力的核心性状——胚芽鞘越长,幼苗破土能力越强,越能适应深播及逆境环境。本研究揭示的乙烯-GA协同调控机制,为培育长胚芽鞘水稻品种提供了明确的分子靶点:可通过调控EIN2EIL2基因的过表达,同时结合SLR1基因的适度沉默,一方面增强乙烯信号传递,另一方面削弱SLR1的抑制作用,协同促进胚芽鞘伸长;也可通过抑制OsGA2ox3的表达,减少活性GA的降解,增强植株对乙烯的敏感性,从而提升幼苗破土能力。此外,本研究发现的OsOsERF63OsOsERF73等下游响应基因,也可作为分子标记,用于水稻胚芽鞘长度性状的分子标记辅助育种,加快长胚芽鞘品种的培育进程,为解决直播稻出苗难题提供技术支撑。
综上,本研究系统揭示了乙烯与赤霉素协同调控水稻胚芽鞘伸长:乙烯经EIN2/EIL通路激活OsERF63、OsERF73等下游基因,正向调控胚芽鞘伸长;赤霉素诱导SLR1蛋白降解,解除SLR1对细胞伸长基因的抑制,同时也解除SLR1对OsEIL2的抑制以放大乙烯信号,二者依托EIL2-SLR1模块协同调控下游功能基因表达,促进胚芽鞘伸长,提升水稻幼苗破土能力与直播出苗效率。
乙烯和赤霉素是2种重要的植物激素,共同促进胚芽鞘伸长生长,乙烯和赤霉素通过EIL2-SLR1模块调控乙烯下游关键基因的表达促进水稻胚芽鞘的伸长生长,进而改善幼苗破土能力、提升直播水稻出苗率。
  • 国家自然科学基金(32472037)
  • 中国农业科学院青年创新专项(Y2024QC14)
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2026年第59卷第16期
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doi: 10.3864/j.issn.0578-1752.2026.16.004
  • 接收时间:2026-05-06
  • 首发时间:2026-09-03
  • 出版时间:2026-08-16
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  • 收稿日期:2026-05-06
  • 录用日期:2026-06-30
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国家自然科学基金(32472037)
中国农业科学院青年创新专项(Y2024QC14)
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    1 湖南农业大学生物科学技术学院, 长沙 410128
    2 中国农业科学院生物技术研究所, 北京 100081

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2种不同金属材料的力学参数

Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)

Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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