Article(id=1276601511035663199, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1276601397818814642, articleNumber=null, orderNo=null, doi=10.3969/j.issn.1000-2561.2025.01.001, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1720713600000, receivedDateStr=2024-07-12, revisedDate=1724688000000, revisedDateStr=2024-08-27, acceptedDate=null, acceptedDateStr=null, onlineDate=1782295118340, onlineDateStr=2026-06-24, pubDate=1737734400000, pubDateStr=2025-01-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1782295118340, onlineIssueDateStr=2026-06-24, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1782295118340, creator=13701087609, updateTime=1782295118340, updator=13701087609, issue=Issue{id=1276601397818814642, tenantId=1146029695717560320, journalId=1235980609244409860, year='2025', volume='46', issue='1', pageStart='1', pageEnd='245', issueExtLink='null', onlineDate='null', pubDate='1737734400000', pubDateStr='2025-01-25', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1782295091347, creator='13701087609', updateTime=1782295207335, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1276601884408418422, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1276601397818814642, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1276601884408418423, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1276601397818814642, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=1, endPage=9, ext={EN=ArticleExt(id=1276601511509619553, articleId=1276601511035663199, tenantId=1146029695717560320, journalId=1235980609244409860, language=EN, title=Antagonistic Effects of Jasmonic Acid and Ethylene on Rubber Biosynthesis in Hevea brasiliensis, columnId=1236256430337085821, journalTitle=Chinese Journal of Tropical Crops, columnName=Omics & Biotechnology, runingTitle=null, highlight=null, articleAbstract=

Based on the fact that ethephon (an ethylene releaser) stimulation can significantly increase latex yield per tapping, it is traditionally believed that rubber biosynthesis is positively regulated by ethylene. Based on the critical role of jasmonic acid signaling in promoting the biosynthesis of secondary metabolites in plants, jasmonic acid signaling is also thought to play a role in the positive feedback regulation of natural rubber biosynthesis. Recent studies suggest that tapping-enhanced rubber biosynthesis is closely related to the activation of jasmonate signaling in laticifer cells of rubber tree. However, it remains unclear whether there is a crosstalk between the jasmonic acid signaling pathway and the ethylene signaling pathway in the regulation of rubber biosynthesis. For this purpose, in the present study, in vitro rubber biosynthesis efficiency of whole latex and small rubber particles (SRPs) from short rested trees treated by either methy jasmonates or ethylene were detected by using substrate 13C-MVA. qPCR was used to analyze the expression of four jasmonic acid signaling genes, HbCOI1, HbJAZ1, HbMYC1, HbMYC2, four ethylene-responsive element binding factors (ERFs) genes HbERFⅢa, HbERFⅦa, HbERFⅨc, HbERFⅩa, four rubber biosynthesis genes, HbHMGR1,HbSRPP1, HbREF1, HbHRT2 in laticifer cells after epicormic shoots were treated with either methy jasmonates or ethylene. The results showed that exogenous methyl jasmonate treatment significantly up-regulated the expression levels of jasmonic acid signaling genes HbCOI1, HbJAZ1, HbMYC1, HbMYC2, and rubber biosynthetic protein genes HbHMGR1,HbSRPP1, HbREF1, HbHRT2 in laticifer cells of rubber tree, and promoted rubber biosynthesis, but had little effect on the expression of ethylene-responsive element binding factor genes HbERFⅢa, HbERFⅦa, HbERFⅨc, HbERFⅩa, or even inhibited the expression. On the contrary, exogenous ethylene treatment significantly up-regulated the expression levels of ethylene-responsive element binding factor genes HbERFⅢa, HbERFⅦa, HbERFⅨc, HbERFⅩa in laticifer cells of rubber tree, but had little effect on the expression of jasmonic acid signaling genes HbCOI1, HbJAZ1, HbMYC1,HbMYC2 and rubber biosynthetic protein genes HbHMGR1, HbSRPP1, HbREF1, HbHRT2 or even inhibited the expression and rubber biosynthesis. The results suggest that there exists an antagonistic effects of jasmonic acid and ethylene on rubber biosynthesis in H. brasiliensis, which would provide a theoretical basis for elucidating the crosstalk of jasmonic acid signaling and ethylene signaling on regulation of natural rubber production.

, authors=null, authorsList=Shuguang YANG, Jinquan CHAO, Yan LI, Shaohua WU, Shixin ZHANG, Xiaomin DENG, Minjing SHI, Weimin TIAN, authorCompany=null, correspAuthors=Weimin TIAN, 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=1276601512721773419, articleId=1276601511035663199, tenantId=1146029695717560320, journalId=1235980609244409860, language=CN, title=茉莉酸和乙烯对巴西橡胶树橡胶生物合成的拮抗作用, columnId=1236256430517440904, journalTitle=热带作物学报, columnName=组学与生物技术, runingTitle=null, highlight=null, articleAbstract=

基于乙烯利(一种乙烯释放剂)刺激采胶可以显著增加每刀次的胶乳产量,以往认为橡胶生物合成主要受乙烯正调控。基于茉莉酸信号在促进植物次生代谢物质的生物合成中所起的关键作用,茉莉酸信号也被认为在天然橡胶生物合成的正反馈调节中起作用。最近的研究表明,割胶促进橡胶树合成天然橡胶与激活乳管细胞的茉莉酸信号途径密切相关。然而,茉莉酸信号途径和乙烯信号途径在橡胶生物合成调控过程中是否存在交互作用仍然不清楚。为此,本研究用茉莉酸甲酯和乙烯处理短期停割树,以13C-MVA为底物测定全胶乳和小橡胶粒子的体外橡胶生物合成效率,用qPCR技术分析茉莉酸甲酯和乙烯处理条件下4个茉莉酸信号途径关键环节基因HbCOI1HbJAZ1HbMYC1HbMYC2,4个乙烯-响应元件结合因子(ERFs)基因HbERFⅢaHbERFⅦaHbERFⅨcHbERFⅩa,4个橡胶生物合成关键蛋白基因HbHMGR1HbSRPP1HbREF1HbHRT2在萌条乳管细胞中的表达。结果表明:外源茉莉酸甲酯处理显著上调橡胶树乳管细胞中茉莉酸信号途径关键环节基因HbCOI1HbJAZ1HbMYC1HbMYC2和橡胶生物合成关键蛋白基因HbHMGR1HbSRPP1HbREF1HbHRT2的表达水平,促进橡胶生物合成,但对乙烯-响应元件结合因子基因HbERFⅢaHbERFⅦaHbERFⅨcHbERFⅩa的表达影响甚微,甚至抑制它们的表达。相反,外源乙烯处理显著上调橡胶树乳管细胞中乙烯-响应元件结合因子基因HbERFⅢaHbERFⅦaHbERFⅨcHbERFⅩa的表达水平,但对茉莉酸信号途径关键环节基因HbCOI1HbJAZ1HbMYC1HbMYC2和橡胶生物合成关键蛋白基因HbHMGR1HbSRPP1HbREF1HbHRT2的表达影响甚微,甚至抑制它们的表达,抑制橡胶生物合成。结果证明了茉莉酸和乙烯对巴西橡胶树橡胶生物合成的拮抗作用,将为阐明茉莉酸信号和乙烯信号的增产机理供理论依据。

, authors=

杨署光(1982—),男,硕士,副研究员,研究方向:橡胶树乳管发育及橡胶生物合成调控。

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* 田维敏(TIAN Weimin),E-mail:
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杨署光(1982—),男,硕士,副研究员,研究方向:橡胶树乳管发育及橡胶生物合成调控。

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杨署光(1982—),男,硕士,副研究员,研究方向:橡胶树乳管发育及橡胶生物合成调控。

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2.中国热带农业科学院三亚研究院,海南三亚 572000
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2.Institute of Sanya Research Institute, Chinese Academy of Tropical Agricultural Sciences, Sanya, Hainan 572000, China
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Different capital letters indicate extremely significant difference (P<0.01); Different lowercase letters indicate significant difference (P<0.05).

, figureFileSmall=JfH4HD12pyfsz9h70U0wFw==, figureFileBig=Z4zSeZzkUvpx+0ByFVhfzA==, tableContent=null), ArticleFig(id=1276601838728245682, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276601511035663199, language=CN, label=图1, caption=MeJA和ET处理对latex和SRPs体外橡胶生物合成效率的影响

不同大写字母表示组间差异极显著(P<0.01);不同小写字母表示组间差异显著(P<0.05)。

, figureFileSmall=JfH4HD12pyfsz9h70U0wFw==, figureFileBig=Z4zSeZzkUvpx+0ByFVhfzA==, tableContent=null), ArticleFig(id=1276601839101538739, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276601511035663199, language=EN, label=Fig. 2, caption=Effects of MeJA and ET treatment on expression of jasmonate signaling pathway genes in latex

Different capital letters indicate extremely significant difference (P<0.01), different lowercase letters indicate significant difference (P<0.05).

, figureFileSmall=M6FLLVr19K96IIDLXL2jVQ==, figureFileBig=RWTDcArcMXsvGKR4VfTx1w==, tableContent=null), ArticleFig(id=1276601839168647604, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276601511035663199, language=CN, label=图2, caption=MeJA和ET处理对胶乳中茉莉酸信号途径基因表达的影响

不同大写字母表示组间差异极显著(P<0.01),不同小写字母表示组间差异显著(P<0.05)。

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Different capital letters indicate extremely significant difference (P<0.01), different lowercase letters indicate significant difference (P<0.05).

, figureFileSmall=njOVR0+DpXiKbyr2eMkE6Q==, figureFileBig=Kj7Gg2yU2yjHZuOWxgJJww==, tableContent=null), ArticleFig(id=1276601839319642550, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276601511035663199, language=CN, label=图3, caption=MeJA和ET处理对胶乳中ERFs基因表达的影响

不同大写字母表示组间差异极显著(P<0.01),不同小写字母表示组间差异显著(P<0.05)。

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Different capital letters indicate extremely significant difference (P<0.01), different lowercase letters indicate significant difference (P<0.05).

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不同大写字母表示组间差异极显著(P<0.01),不同小写字母表示组间差异显著(P<0.05)。

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茉莉酸和乙烯对巴西橡胶树橡胶生物合成的拮抗作用
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杨署光 1 , 晁金泉 1, 2, 3 , 李言 4 , 吴绍华 1, 2, 3 , 张世鑫 1, 2, 3 , 邓小敏 1 , 史敏晶 1 , 田维敏 4, *
热带作物学报 | 组学与生物技术 2025,46(1): 1-9
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热带作物学报 |组学与生物技术 2025 , 46 (1) : 1 -9
茉莉酸和乙烯对巴西橡胶树橡胶生物合成的拮抗作用
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杨署光1, 晁金泉1, 2, 3, 李言4, 吴绍华1, 2, 3, 张世鑫1, 2, 3, 邓小敏1, 史敏晶1, 田维敏4, *
作者信息
  • 1.中国热带农业科学院橡胶研究所/农业农村部橡胶树生物学与遗传资源利用重点实验室/省部共建国家重点实验室培育基地-海南省热带作物栽培生理学重点实验室,海南海口 571101
  • 2.中国热带农业科学院三亚研究院,海南三亚 572000
  • 3.热带作物生物育种全国重点实验室,海南三亚 572000
  • 4.中国科学院西双版纳热带植物园,云南西双版纳 666303
通讯作者:
* 田维敏(TIAN Weimin),E-mail:
Antagonistic Effects of Jasmonic Acid and Ethylene on Rubber Biosynthesis in Hevea brasiliensis
Shuguang YANG1, Jinquan CHAO1, 2, 3, Yan LI4, Shaohua WU1, 2, 3, Shixin ZHANG1, 2, 3, Xiaomin DENG1, Minjing SHI1, Weimin TIAN4, *
Affiliations
  • 1.Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences / Key Laboratory of Biology and Genetic Resources of Rubber Tree, Ministry of Agriculture and Rural Affairs / State Key Laboratory Incubation Base for Cultivation & Physiology of Tropical Crops, Haikou, Hainan 571101, China
  • 2.Institute of Sanya Research Institute, Chinese Academy of Tropical Agricultural Sciences, Sanya, Hainan 572000, China
  • 3.National Key Laboratory for Tropical Crop Breeding, Sanya, Hainan 572000, China
  • 4.Xishuangbanna Tropical Botanical Garden, Chinese Academy of Sciences, Xishuangbanna, Yunnan 666303, China
出版时间: 2025-01-25 doi: 10.3969/j.issn.1000-2561.2025.01.001
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基于乙烯利(一种乙烯释放剂)刺激采胶可以显著增加每刀次的胶乳产量,以往认为橡胶生物合成主要受乙烯正调控。基于茉莉酸信号在促进植物次生代谢物质的生物合成中所起的关键作用,茉莉酸信号也被认为在天然橡胶生物合成的正反馈调节中起作用。最近的研究表明,割胶促进橡胶树合成天然橡胶与激活乳管细胞的茉莉酸信号途径密切相关。然而,茉莉酸信号途径和乙烯信号途径在橡胶生物合成调控过程中是否存在交互作用仍然不清楚。为此,本研究用茉莉酸甲酯和乙烯处理短期停割树,以13C-MVA为底物测定全胶乳和小橡胶粒子的体外橡胶生物合成效率,用qPCR技术分析茉莉酸甲酯和乙烯处理条件下4个茉莉酸信号途径关键环节基因HbCOI1HbJAZ1HbMYC1HbMYC2,4个乙烯-响应元件结合因子(ERFs)基因HbERFⅢaHbERFⅦaHbERFⅨcHbERFⅩa,4个橡胶生物合成关键蛋白基因HbHMGR1HbSRPP1HbREF1HbHRT2在萌条乳管细胞中的表达。结果表明:外源茉莉酸甲酯处理显著上调橡胶树乳管细胞中茉莉酸信号途径关键环节基因HbCOI1HbJAZ1HbMYC1HbMYC2和橡胶生物合成关键蛋白基因HbHMGR1HbSRPP1HbREF1HbHRT2的表达水平,促进橡胶生物合成,但对乙烯-响应元件结合因子基因HbERFⅢaHbERFⅦaHbERFⅨcHbERFⅩa的表达影响甚微,甚至抑制它们的表达。相反,外源乙烯处理显著上调橡胶树乳管细胞中乙烯-响应元件结合因子基因HbERFⅢaHbERFⅦaHbERFⅨcHbERFⅩa的表达水平,但对茉莉酸信号途径关键环节基因HbCOI1HbJAZ1HbMYC1HbMYC2和橡胶生物合成关键蛋白基因HbHMGR1HbSRPP1HbREF1HbHRT2的表达影响甚微,甚至抑制它们的表达,抑制橡胶生物合成。结果证明了茉莉酸和乙烯对巴西橡胶树橡胶生物合成的拮抗作用,将为阐明茉莉酸信号和乙烯信号的增产机理供理论依据。

巴西橡胶树  /  茉莉酸甲酯  /  乙烯  /  生物合成  /  拮抗作用

Based on the fact that ethephon (an ethylene releaser) stimulation can significantly increase latex yield per tapping, it is traditionally believed that rubber biosynthesis is positively regulated by ethylene. Based on the critical role of jasmonic acid signaling in promoting the biosynthesis of secondary metabolites in plants, jasmonic acid signaling is also thought to play a role in the positive feedback regulation of natural rubber biosynthesis. Recent studies suggest that tapping-enhanced rubber biosynthesis is closely related to the activation of jasmonate signaling in laticifer cells of rubber tree. However, it remains unclear whether there is a crosstalk between the jasmonic acid signaling pathway and the ethylene signaling pathway in the regulation of rubber biosynthesis. For this purpose, in the present study, in vitro rubber biosynthesis efficiency of whole latex and small rubber particles (SRPs) from short rested trees treated by either methy jasmonates or ethylene were detected by using substrate 13C-MVA. qPCR was used to analyze the expression of four jasmonic acid signaling genes, HbCOI1, HbJAZ1, HbMYC1, HbMYC2, four ethylene-responsive element binding factors (ERFs) genes HbERFⅢa, HbERFⅦa, HbERFⅨc, HbERFⅩa, four rubber biosynthesis genes, HbHMGR1,HbSRPP1, HbREF1, HbHRT2 in laticifer cells after epicormic shoots were treated with either methy jasmonates or ethylene. The results showed that exogenous methyl jasmonate treatment significantly up-regulated the expression levels of jasmonic acid signaling genes HbCOI1, HbJAZ1, HbMYC1, HbMYC2, and rubber biosynthetic protein genes HbHMGR1,HbSRPP1, HbREF1, HbHRT2 in laticifer cells of rubber tree, and promoted rubber biosynthesis, but had little effect on the expression of ethylene-responsive element binding factor genes HbERFⅢa, HbERFⅦa, HbERFⅨc, HbERFⅩa, or even inhibited the expression. On the contrary, exogenous ethylene treatment significantly up-regulated the expression levels of ethylene-responsive element binding factor genes HbERFⅢa, HbERFⅦa, HbERFⅨc, HbERFⅩa in laticifer cells of rubber tree, but had little effect on the expression of jasmonic acid signaling genes HbCOI1, HbJAZ1, HbMYC1,HbMYC2 and rubber biosynthetic protein genes HbHMGR1, HbSRPP1, HbREF1, HbHRT2 or even inhibited the expression and rubber biosynthesis. The results suggest that there exists an antagonistic effects of jasmonic acid and ethylene on rubber biosynthesis in H. brasiliensis, which would provide a theoretical basis for elucidating the crosstalk of jasmonic acid signaling and ethylene signaling on regulation of natural rubber production.

Hevea brasiliensis Muell. Arg.  /  methy jasmonates  /  ethylene  /  biosynthesis  /  antagonism
杨署光, 晁金泉, 李言, 吴绍华, 张世鑫, 邓小敏, 史敏晶, 田维敏. 茉莉酸和乙烯对巴西橡胶树橡胶生物合成的拮抗作用. 热带作物学报, 2025 , 46 (1) : 1 -9 . DOI: 10.3969/j.issn.1000-2561.2025.01.001
Shuguang YANG, Jinquan CHAO, Yan LI, Shaohua WU, Shixin ZHANG, Xiaomin DENG, Minjing SHI, Weimin TIAN. Antagonistic Effects of Jasmonic Acid and Ethylene on Rubber Biosynthesis in Hevea brasiliensis[J]. Chinese Journal of Tropical Crops, 2025 , 46 (1) : 1 -9 . DOI: 10.3969/j.issn.1000-2561.2025.01.001
巴西橡胶树(Hevea brasiliensis Muell. Arg)的胶乳主要是由树干中的次生乳管合成和贮藏[1]。次生乳管数量的多少与胶乳的产量密切相关[2-3]。生产中通过割胶,即机械伤害切割树皮中的乳管使胶乳流出(即排胶),从而收集胶乳制备天然橡胶(nature rubber,NR)。因此,乳管数量、排胶时间和2次割胶之间的胶乳再生(包括天然橡胶生物合成)是天然橡胶产量的3个主要限制因素[3]。外源茉莉酸甲酯(methy jasmonates,MeJA)能促进橡胶树的次生乳管分化[4]。乙烯(ethylene,ET)对橡胶树的次生乳管分化无影响[5],但乙烯利(一种乙烯释放剂)刺激能显著延长每刀次割胶的棑胶时间[6]。茉莉酸和乙烯对橡胶生物合成的效应仍存在争议,基于乙烯利刺激采胶可以显著增加每刀次的胶乳产量,以往认为橡胶生物合成主要受乙烯正调控[7],基于茉莉酸信号在促进植物次生代谢物质的生物合成中所起的关键作用,茉莉酸信号也被认为在天然橡胶生物合成的正反馈调节中起作用[8-10]。最近的研究表明,割胶促进橡胶树合成天然橡胶与激活乳管细胞的茉莉酸信号途径密切相关[11]。然而,茉莉酸信号途径和乙烯信号途径在橡胶生物合成调控过程中是否存在交互作用仍然不清楚。
割胶促进胶乳再生是生产上的一个熟知的事实[12],在有伤口存在的情况下外施茉莉酸[4,11]和乙烯[13]能成功激活橡胶树对茉莉酸和乙烯的响应。因此,对割胶树作适当停割并结合外源茉莉酸甲酯和乙烯处理是研究茉莉酸和乙烯对巴西橡胶树橡胶生物合成效率的影响的可行方案。利用该系统,本研究以13C-MVA(mevalonolactone-2-13C)为底物,评价茉莉酸和乙烯对全胶乳(latex)和小橡胶粒子(small rubber particles,SRPs)的体外橡胶生物合成效率的影响。橡胶生物合成效率的变化必然涉及到橡胶生物合成关键蛋白基因表达的改变,考虑到基因表达的变化更灵敏以及为避免机械伤害和割胶诱导的内源茉莉酸[4,11]和乙烯[14]对试验结果的干扰,在无伤害的条件下研究外源茉莉酸和乙烯对橡胶生物合成关键蛋白基因表达的影响是比较稳妥的策略。基于此,本研究用外源茉莉酸甲酯和乙烯处理一年生橡胶树稳定期萌条的节间,用qPCR技术分析茉莉酸甲酯和乙烯处理条件下相关基因在萌条乳管细胞中的表达。有效的MeJA和ET处理能激活橡胶树乳管细胞中的JA信号途径[11]和ET信号途径[13],因此,分别通过检测4个茉莉酸信号途径关键环节蛋白冠菌素不敏感1 HbCOI1(coronatine insensitive 1,COI1)、茉莉酸ZIM结构域蛋白HbJAZ1(jasmonate ZIM-domain,JAZ)、髓细胞组织增生蛋白(myelocytomatosis proteins,MYC)HbMYC1、HbMYC2,4个乙烯-响应元件结合因子HbERFⅢaHbERFⅦaHbERFⅨcHbERFⅩa基因(ethylene-responsive element binding factors,ERFs)的表达变化检验外源茉莉酸甲酯和乙烯处理的有效性,进一步分析外源茉莉酸甲酯和乙烯处理对4个橡胶生物合成关键蛋白3-羟基-3-甲基戊二酸单酰辅酶A还原酶(3-Hydroxy-3-Methylglutaryl coenzyme A reductase,HMGR)HbHMGR1、小橡胶粒子膜蛋白(small rubber particle protein,SRPP)HbSRPP1、橡胶延伸因子(rubber elongation factor,REF)HbREF1、橡胶转移酶(Hevea brasiliensis rubber transferase,HRT)HbHRT2基因表达的影响。旨在为阐明茉莉酸和乙烯的增产机理提供理论依据。
以生产中按照S/2·d3的割胶制度割胶的八年生橡胶树无性系CATAS7-33-97为材料,在第二割年的5—10月割胶后,于11月初选取12株大小、长势基本一致的橡胶树开展试验。
每株树为1个生物学重复。将其中的9株停割15 d,在第15天上午,用0.5%的乙烯处理3株停割树(Rested-ET),用0.07%的茉莉酸甲酯处理3株停割树(Rested-MeJA),3株停割树作为停割对照(Rested-CK),3株正常割胶(Tapped)。处理3 d后割胶采集胶乳。冰浴收集前2 min流出的胶乳。取一部分胶乳在4 ℃、10 000 r/min条件下离心15 min,取中层的C-乳清(含小橡胶粒子),用0.45 µm的针头过滤器过滤,收集滤液获得小橡胶粒子。以13C-MVA(mevalonolactone-2-13C)(Sigma Aldrich,Lot#MBBB5201V)为底物,以PDB(Pee Dee Belemnite)为标准品,参考已报道的方法[11]测定全胶乳和小橡胶粒子的体外橡胶生物合成效率,测定结果以NR样品的C稳定性同位素组成δ13C(‰)表示。
以大小、长势、物候期基本一致的CATAS7-33-97无性系一年生橡胶树稳定期萌条为材料,分别以0.05%乙烯水溶液(ET)、含0.07%茉莉酸甲酯的7.14%乙醇溶液(MeJA)、7.14%乙醇溶液(EtOH)处理顶蓬节间,对照(CK)不作任何处理。分别在处理后2 h、6 h、1 d、3 d、5 d,用刀片切割韧皮部采胶,每个时间点收集5株的混合样,用于提取胶乳总RNA。根据报道的cDNA序列[13]设计4个乙烯-响应元件结合因子(ERFs)基因的qPCR引物,HbERF aⅢ(scaffold-0026_3462933,F: 5′-CGCATTTGGCTTGGGAGTTATG-3′,R: 5′-CAAGTTCAGGGAAGTTGAGGTG-3′)、HbERFⅦa(scaffold0566_721951,F: 5′-CGTACCAGCACCGCAAACAT-3′,R: 5′-CTCGGGCTCCAATCCCAGA-3′)、HbERFⅨc(scaffold-1015_293137,F: 5′-ACAGAGGTGTTAGGAGGCGT-3′,R: 5′-GCAGCTTGGTCATAAGCCAGA-3′)、HbERFⅩa(scaffold0246_1294475,F: 5′-CCGCATAAAGCAGCAAGAGTC-3′,R: 5′-CGGTTGAAGACCGATAGGGAT-3′)。胶乳总RNA的提取、cDNA合成、4个茉莉酸信号途径关键环节基因HbCOI1(EU136026)、HbJAZ1(GQ369508)、HbMYC1(GU434304)、HbMYC2(HM061097),4个橡胶生物合成关键基因HbHMGR1(X54659)、HbSRPP1(AJ223388.1)、HbREF1(AY430052)、HbHRT2(AB064661)的qPCR引物合成、qPCR基因表达分析方法与先前的报道[15-16]一致。根据Q=2△Cq=2min Cq-sample Cq计算基因的表达值(Q),以Hb18S(AY435212.1)作为内参基因,根据E=Q目的基因/Q内参基因分析目的基因的相对表达值(E)。
采用Excel 2021软件进行数据整理,NR的碳同位素值δ13C和基因相对表达量为平均数±标准偏差,使用SPSS 17.0软件的Duncan(D)检验进行单因素方差分析。
用外源MeJA和ET处理停割15 d的割胶树,处理3 d后割胶采集胶乳,以13C-MVA为底物,评价MeJA和ET对全胶乳和小橡胶粒子体外橡胶生物合成效率的影响。结果表明,在第二割年,按S/2·d3的割胶制度割胶60刀左右以后,割胶树(Tapped)的橡胶生物合成效率处于较高水平,并且胶乳(latex)和小橡胶粒子(SRPs)的橡胶生物合成效率水平相当,说明由于胶乳的大量流失,胶乳的合成和积累跟不上排胶,橡胶粒子的发育不充分,此时胶乳中的橡胶粒子以SRPs为主,橡胶生物合成活跃;短期停割后(Rested)胶乳和SRPs的橡胶生物合成效率显著低于割胶树,与停割树相比,ET处理抑制而MeJA处理促进胶乳和SRPs的橡胶合生物成效率(P>0.05),无论是胶乳还是SRPs,MeJA处理的橡胶生物合成效率均显著高于ET处理,MeJA处理的SRPs的橡胶生物合成效率甚至提高到割胶树的水平。上述结果表明,JA促进橡胶树的橡胶生物合成,而ET抑制橡胶树的橡胶生物合成(图1)。
用外源MeJA和ET处理一年生橡胶树稳定期萌条的节间。以18S基因为内参,qPCR分析MeJA、ET对胶乳中4个茉莉酸信号途径关键基因HbCOI1HbJAZ1HbMYC1HbMYC2表达的影响(图2)。结果表明,EtOH(MeJA的溶剂)处理对HbJAZ1的基因表达几乎无影响,对HbMYC2的基因表达无明显的规律,对HbCOI1HbMYC1的基因表达有一定的上调效应。ET处理对HbCOI1HbJAZ1基因的表达有一定的上调作用,对HbMYC1基因的表达无明显的规律,对HbMYC2基因的表达有明显的下调作用。MeJA显著上调HbCOI1HbJAZ1HbMYC1HbMYC2基因的表达,呈现出敏感持续高表达(HbCOI1HbJAZ1)或持续高表达(HbMYC1HbMYC2)的趋势,几乎在所有时间点,MeJA处理的基因表达水平均显著高于EtOH和ET处理。这些结果表明,在无伤害的条件下,外源MeJA处理能有效激活橡胶树乳管细胞中的JA信号途径,而外源ET处理对橡胶树乳管细胞中JA信号途径的活化影响甚微。
用外源ET和MeJA处理一年生橡胶树稳定期萌条的节间。以18S基因为内参,qPCR分析ET、MeJA对胶乳中4个乙烯-响应元件结合因子(ERFs)基因HbERFⅢaHbERFⅧaHbERFⅨcHbERFⅩa表达的影响(图3)。结果表明,EtOH处理对HbERFⅢaHbERFⅨc基因的表达有一定的上调效应,对HbERFⅧa基因的表达无明显的规律,对HbERFⅩa基因的表达有一定的下调效应。MeJA处理对HbERFⅢaHbERFⅨcHbERFⅩa基因的表达水平有一定的促进作用,对HbERFⅦa基因的表达水平有一定的抑制作用。ET显著上调HbERFⅢaHbERFⅧaHbERFⅨcHbERFⅩa基因的表达,呈现出瞬时超高表达的特点,ET处理的最高表达水平均显著高于MeJA和EtOH处理。这些结果表明,在无伤害的条件下,外源ET处理能有效激活橡胶树乳管细胞中的ET信号途径,而外源MeJA处理对橡胶树乳管细胞中ET信号途径的活化影响甚微。
进一步用qPCR分析MeJA、ET对胶乳中4个橡胶生物合成关键基因HbHMGR1HbSRPP1HbREF1HbHRT2表达的影响(图4)。结果表明,EtOH处理6h对HbHMGR1HbSRPP1HbREF1HbHRT2基因的表达均有一定的上调作用,这种作用在HbHMGR1HbSRPP1中持续时间更长。ET处理2h对HbHMGR1HbSRPP1HbREF1HbHRT2基因的表达均有一定的上调作用,但ET处理5 d后4个基因的表达均明显下调。MeJA显著上调HbHMGR1HbSRPP1HbREF1HbHRT2基因的表达,呈现出敏感持续高表达的趋势,在所有时间点,MeJA处理的基因表达水平均显著高于EtOH和ET处理。这些结果表明,JA信号参与橡胶树橡胶生物合成的转录调节,而ET信号在此过程中并不起主要作用。
COI1、JAZ和MYC是茉莉酸信号传导途径的3个核心环节[17],割胶上调乳管细胞中茉莉酸生物合成关键酶HbLOX的基因表达[18],促进内源茉莉酸的合成和积累[11,19],上调茉莉酸信号途径基因HbCOI1HbJAZsHbMYCs和橡胶生物合成关键蛋白基因HbFPS1HbHRT1HbHRT2HbREFHbSRPP1的表达[11,20],增加HbFPS1和HbSRPP1蛋白的含量[11],促进橡胶生物合成[11,20]。茉莉酸处理上调胶乳中茉莉酸信号途径基因(COI1、JAZ、MYC)的表达[11,21-22],表明乳管细胞可以响应茉莉酸信号;而且茉莉酸上调橡胶生物合成相关基因的表达[11,20],显著上调HbHMGR1正调控因子HbCZF1的基因表达[23],但对HbSRPP1负调控因子HbWRKY1的基因表达几乎无影响[24],促进橡胶生物合成[11]。HbCOI1与HbJAZs、HbJAZs与HbJAZs、HbJAZs与HbMYCs、HbMYCs与HbMYCs蛋白之间能广泛地发生互作[20,25],而且HbMYCs是HbREF、HbSRPP、HbHRT2等橡胶生物合成关键蛋白的转录因子[20,26],HbJAZ1和HbJAZ3与HbSRPP1[27-28]蛋白也能相互作用。在缺乏COR(JA类似物)时,HbJAZ3蛋白与HbMYC2a互作,抑制HbMYC2a对橡胶生物合成关键蛋白HbSRPP1HbFPS1基因的转录激活[11]。在COR存在的条件下,HbJAZ3与HbCOI1结合,释放出的HbMYC2a转录激活橡胶生物合成关键蛋白HbSRPP1和HbFPS1[11]。乳管细胞中的茉莉酸信号转导途径HbCOI1-HbJAZ3-HbMYC2-HbFPS1/HbSRPP1直接证明了茉莉酸对天然橡胶生物合成的调控[11]
基于乙烯利刺激割胶可以显著增加每刀次的胶乳产量,以往认为橡胶生物合成主要受乙烯促进[7]。虽然割胶会诱导橡胶树乳管细胞内源乙烯的合成[14],但乳管细胞中乙烯生物合成基因的表达远低于其他组织[13,29],这可能是由于乳胶的低氧条件不能满足乙烯生物合成对氧气的需求[2,30],导致乙烯合成能力较弱。在乙烯处理后,胶乳中乙烯信号通路(ETR、CTR1、EIN2EIN3/EIL1[13,31-32]及乙烯响应因子(ERFs[13]基因表达受影响,表明乳管细胞可以响应乙烯信号。然而,乙烯处理后乳管中的差异表达基因(DEGs)不包括橡胶生物合成相关基因[13],施用乙烯对橡胶生物合成相关基因的表达几乎无影响[33-36];乙烯上调HbSRPP1负调控转录因子HbWRKY1[24]、HbMADS4[37]的基因表达,下调HbSRPP1的基因表达[24],而对HbHMGR1正调控因子HbCZF1的基因表达影响很小[23],甚至降低REF和SRPP蛋白的含量[38],抑制橡胶生物合成[11]。虽然HbFPS1HbSRPP1基因的启动子含有乙烯响应元件,但胶乳中高丰度表达的HbEIN3基因(KR013139)[39]HbFPS1HbSRPP1基因的转录几乎无影响[11]。因此,乙烯信号本身在激活橡胶生物合成方面可能并不起直接作用。生产上使用乙烯刺激能显著地提高单次割胶的橡胶产量,主要是延长了排胶持续时间[6],同时增强了蔗糖分配[40]、乳管细胞内的水份运输[41]、糖酵解和C3循环固碳能力[35]
在本研究中,MeJA处理上调茉莉酸信号途径相关基因HbCOI1HbJAZ1HbMYC1HbMYC2和橡胶生物合成关键蛋白基因HbHMGR1HbSRPP1HbREF1HbHRT2的表达,可能是MeJA促进橡胶粒子将13C-MVA中的13C高效掺入NR中的原因之一。ET处理激活ET-响应元件结合因子(ERFs)基因的表达,但对橡胶生物合成关键蛋白基因HbHMGR1HbSRPP1HbREF1HbHRT2及其转录调节相关基因HbCOI1HbJAZ1HbMYC1HbMYC2的表达无影响,可能是ET抑制latex和SRPs的体外橡胶生物合成效率的主要原因。
综上所述,本研究进一步区分出JA和ET对橡胶树橡胶生物合成的直接影响,JA促进橡胶树的橡胶生物合成,而ET抑制橡胶树的橡胶生物合成。研究结果为深入研究橡胶树橡胶生物合成调控机制提供参考资料,为阐明茉莉酸信号和乙烯信号的增产机理提供理论依据。
  • 海南省自然科学基金项目(321MS0806)
  • 云南省重大科技专项计划项目(202402AE09001901)
  • 海南省重点研发计划项目(ZDYF2022XDNY252)
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doi: 10.3969/j.issn.1000-2561.2025.01.001
  • 接收时间:2024-07-12
  • 首发时间:2026-06-24
  • 出版时间:2025-01-25
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  • 收稿日期:2024-07-12
  • 修回日期:2024-08-27
基金
海南省自然科学基金项目(321MS0806)
云南省重大科技专项计划项目(202402AE09001901)
海南省重点研发计划项目(ZDYF2022XDNY252)
作者信息
    1.中国热带农业科学院橡胶研究所/农业农村部橡胶树生物学与遗传资源利用重点实验室/省部共建国家重点实验室培育基地-海南省热带作物栽培生理学重点实验室,海南海口 571101
    2.中国热带农业科学院三亚研究院,海南三亚 572000
    3.热带作物生物育种全国重点实验室,海南三亚 572000
    4.中国科学院西双版纳热带植物园,云南西双版纳 666303

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* 田维敏(TIAN Weimin),E-mail:
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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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