Article(id=1276616109801476481, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1276616049617408127, articleNumber=null, orderNo=null, doi=10.3969/j.issn.1000-2561.2025.08.009, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1741363200000, receivedDateStr=2025-03-08, revisedDate=null, revisedDateStr=null, acceptedDate=1744300800000, acceptedDateStr=2025-04-11, onlineDate=1782298598956, onlineDateStr=2026-06-24, pubDate=1756051200000, pubDateStr=2025-08-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1782298598956, onlineIssueDateStr=2026-06-24, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1782298598956, creator=13701087609, updateTime=1782298598956, updator=13701087609, issue=Issue{id=1276616049617408127, tenantId=1146029695717560320, journalId=1235980609244409860, year='2025', volume='46', issue='8', pageStart='1785', pageEnd='2029', issueExtLink='null', onlineDate='null', pubDate='1756051200000', pubDateStr='2025-08-25', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1782298584608, creator='13701087609', updateTime=1782298660748, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1276616369089147039, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1276616049617408127, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1276616369089147040, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1276616049617408127, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=1864, endPage=1873, ext={EN=ArticleExt(id=1276616110199935363, articleId=1276616109801476481, tenantId=1146029695717560320, journalId=1235980609244409860, language=EN, title=Trace Detection and Distribution Dynamics of Externally Applied Ethrel in Rubber Tree Tapping Cuts, columnId=1236256434120348225, journalTitle=Chinese Journal of Tropical Crops, columnName=Plant Cultivation, Physiology & Biochemistry, runingTitle=null, highlight=null, articleAbstract=

Ethrel is widely used as a stimulant in rubber tree plantations, which plays an important role in increaisng latex yield effectively and reducing labor requirements for tapping. However, the stimulatory effect of ethrel exhibits a pronounced dose-response relationship, excessive application may lead to adverse effects, such as tapping panel dryness (TPD). In this study, rubber tree clone Reyan 73397 was used as the experimental material, and an ultra-high performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for the trace determination of ethrel in the bark was developed. The distribution of ethrel at various time points and different parts of the bark following its application was investigated. The samples were extracted using a methanol∶1% formic acid aqueous solution (9∶1,V/V), and directly analyzed after filtration. The samples were separated by waters CORTECS HILIC C18 column (100 mm×2.1 mm, 1.6 μm) with mobile phase A (0.01% ammonia solution) and mobile phase B (acetonitrile∶water 80∶20, V/V) at a flow rate of 1.0 mL/min. The detection was performed under electrospray ion source, negative ion scanning and multiple reaction monitoring (MRM) mode. Quantitative analysis was conducted using external standard calibration curves, quantitative ion was 107. The results showed a good linear relationship for ethrel within the range of 0.02 μg/mL to 1.00 μg/mL, with coefficient of determination (R2) of 0.9997. Recoveries ranged from 85.3% to 101.2% at spiked levels 0.1 µg/kg to 1.0 µg/kg, with relative standard deviations of 1.14%-7.28%, and limits of quantification (LOQ) of 0.1 µg/kg. This method was applied to study the distribution characteristics of ethrel in stimulated bark rings, both above and below the ethrel-treated 5 cm and 10 cm areas, after applying different concentrations of ethrel (0.5%, 2.0%,4.0%) over 0-96 h. The results revealed that ethrel was unevenly distributed in the rubber tree, the concentration at the stimulated area was significantly higher than that in other areas, areas closer to the stimulated bark were higher than that in distant areas, and the areas below the stimulated bark were higher than that above it. The concentration of ethrel at the stimulated area showed a significant positive correlation with the stimulated ethrel concentration. Additionally, the concentration displayed an initial increase followed by a gradual decline over time. For the three concentrations, peak levels were observed between 12 h and 24 h post-stimulation, after which a gradual decrease occurred, with relatively high concentrations still detectable at 96 h. Compared to the indirect measurement of ethylene release rates by gas chromatography, this method has many significant advantages, including simplicity, rapidity, excellent reproducibility, and high sensitivity, thereby providing reliable technical support for the quantitative analysis of ethrel in rubber trees. The study preliminarily revealed the absorption, distribution and transport patterns of externally stimulated ethrel on rubber tree tapping cuts. It was found that the diffusion and transport of ethrel were limited, with the compound primarily concentrated at the stimulation site. The findings would provide a theoretical basis for further elucidating the dose-dependent effects of ethrel on yield stimulation and guiding its safe and efficient application.

, authors=null, authorsList=Wenfeng YANG, Ruishen FAN, Honghua GAO, Fang WEI, Jiong WAN, Jian QIU, authorCompany=null, correspAuthors=Jian QIU, 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=1276616113974808973, articleId=1276616109801476481, tenantId=1146029695717560320, journalId=1235980609244409860, language=CN, title=橡胶树割线外施乙烯利的痕量检测及其吸收和分布规律研究, columnId=1236256434313286224, journalTitle=热带作物学报, columnName=作物栽培与生理生化, runingTitle=null, highlight=null, articleAbstract=

乙烯利作为橡胶生产中广泛应用的增产剂,在促进橡胶树高效采胶中发挥了重要作用。然而,乙烯利刺激增产具有明显的剂量效应,过度施用会造成橡胶树死皮等副作用。本研究以热研73397橡胶树为试验材料,建立了基于超高效液相色谱串联质谱技术(UPLC-MS/MS)的树皮中乙烯利痕量检测方法,并系统研究了外施乙烯利后不同时间和树干不同部位树皮的乙烯利含量分布规律。样品前处理采用甲醇∶1%甲酸(9∶1,V/V)溶液匀浆浸提,过滤后直接进行定量分析。样品经Waters CORTECS HILIC C18色谱柱(100 mm×2.1 mm,1.6 μm)分离,以0.01%氨水溶液、乙腈∶水(80∶20,V/V)为流动相,流速为1.0 mL/min,在电喷雾离子源、负离子扫描和多反应监测模式(MRM)下进行检测,外标法定量,定量离子为107。结果表明:乙烯利在0.02~1.00 µg/mL浓度范围内线性关系良好(R2=0.9997),在0.1~1.0 µg/kg添加水平下,回收率为85.3%~101.2%,相对标准偏差为1.14%~7.28%,检出限为0.1 µg/kg。应用该方法研究了橡胶树涂施不同浓度乙烯利(0.5%、2.0%、4.0%)0~96 h后割线及其上下5、10 cm树皮中乙烯利的分布特征。结果显示:外施的乙烯利在橡胶树中分布不均匀,施药部位的含量显著高于其他部位,且近施药部位含量高于远施药部位,施药部位下方的含量高于上方。施药部位的乙烯利含量与外施乙烯利浓度呈显著正相关,且随着施药时间的延长呈先增后降的趋势,3种浓度均在施药后12~24 h达到峰值,随后逐渐下降,96 h仍能检测到较高浓度的乙烯利。与气相色谱法间接测定乙烯释放速率相比,本研究建立的检测方法具有简便快速、重现性好、灵敏度高等优势,为橡胶树中乙烯利的定量分析提供了可靠的技术支撑。研究初步揭示了橡胶树割线外施乙烯利后的吸收、分布及运输规律,发现其扩散运输不明显,主要集中在施药部位,可为深入解析乙烯利刺激增产的剂量效应及指导其安全高效施用提供理论依据。

, authors=

杨文凤(1983—),女,硕士,副研究员,研究方向:橡胶树采胶技术研究及推广应用。

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* 仇键(QIU Jian),E-mail:
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杨文凤(1983—),女,硕士,副研究员,研究方向:橡胶树采胶技术研究及推广应用。

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杨文凤(1983—),女,硕士,副研究员,研究方向:橡胶树采胶技术研究及推广应用。

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(in Chinese), articleTitle=Review on the analysis and testing method of typical plant growth regulators in the environment, refAbstract=null), Reference(id=1276616156391805466, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616109801476481, doi=null, pmid=null, pmcid=null, year=1991, volume=null, issue=4, pageStart=69, pageEnd=74, url=null, language=null, rfNumber=[42], rfOrder=72, authorNames=范思伟, 杨少琼, journalName=热带作物研究, refType=null, unstructuredReference=范思伟, 杨少琼. 巴西橡胶的乙烯生理学[J]. 热带作物研究, 1991(4): 69-74., articleTitle=巴西橡胶的乙烯生理学, refAbstract=null), Reference(id=1276616156446331419, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616109801476481, doi=null, pmid=null, pmcid=null, year=1991, volume=null, issue=4, pageStart=69, pageEnd=74, url=null, language=null, rfNumber=[42], rfOrder=73, authorNames=FAN S W, YANG S Q, journalName=Chinese Journal of Tropical Agriculture, refType=null, unstructuredReference=FAN S W, YANG S Q. 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(in Chinese), articleTitle=Analysis of physiological characteristics effected by ethrel stimulation on clone Reyan 7-33-97 of young tapped rubber tree, refAbstract=null), Reference(id=1276616156865761822, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616109801476481, doi=null, pmid=null, pmcid=null, year=2020, volume=51, issue=1, pageStart=133, pageEnd=139, url=null, language=null, rfNumber=[44], rfOrder=76, authorNames=仇键, 校现周, 高宏华, 杨文凤, 魏芳, 吴明, 罗世巧, journalName=南方农业学报, refType=null, unstructuredReference=仇键, 校现周, 高宏华, 杨文凤, 魏芳, 吴明, 罗世巧. 六天一刀割制对热研7-33-97幼龄开割橡胶树产量胶乳生理及死皮病发生的影响[J]. 南方农业学报, 2020, 51(1): 133-139., articleTitle=六天一刀割制对热研7-33-97幼龄开割橡胶树产量胶乳生理及死皮病发生的影响, refAbstract=null), Reference(id=1276616157167751711, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616109801476481, doi=null, pmid=null, pmcid=null, year=2020, volume=51, issue=1, pageStart=133, pageEnd=139, url=null, language=null, rfNumber=[44], rfOrder=77, authorNames=QIU J, XIAO X Z, GAO H H, YANG W F, WEI F, WU M, LUO S Q, journalName=Journal of Southern Agriculture, refType=null, unstructuredReference=QIU J, XIAO X Z, GAO H H, YANG W F, WEI F, WU M, LUO S Q. Effects of tapping per six days system on the yield, latex physiology and tapping panel dryness of young tapped Hevea brasiliensis var. Reyan 7-33-97[J]. Journal of Southern Agriculture, 2020, 51(1): 133-139. (in Chinese), articleTitle=Effects of tapping per six days system on the yield, latex physiology and tapping panel dryness of young tapped Hevea brasiliensis var. Reyan 7-33-97, refAbstract=null)], funds=[Fund(id=1276616137928479175, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616109801476481, awardId=320MS111, language=CN, fundingSource=海南省自然科学基金项目(320MS111), fundOrder=null, country=null), Fund(id=1276616139123855816, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616109801476481, awardId=1630022022016, language=CN, fundingSource=中央级公益性科研院所基本科研业务费专项(1630022022016), fundOrder=null, country=null), Fund(id=1276616139207741897, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616109801476481, awardId=CARS-33-ZP4, language=CN, fundingSource=现代农业产业技术体系建设专项(CARS-33-ZP4), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1276616114813669775, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616109801476481, xref=null, ext=[AuthorCompanyExt(id=1276616114822058384, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616109801476481, companyId=1276616114813669775, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=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 / Hainan Key Laboratory for Cultivation & Physiology of Tropical Crops / State Key Laboratory Incubation Base for Cultivation and Physiology of Tropical Crops, Haikou, Hainan 571101, China), AuthorCompanyExt(id=1276616114843029905, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616109801476481, companyId=1276616114813669775, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=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/海南省热带作物栽培生理学重点实验室/省部共建国家重点实验室培育基地,海南海口 571101)])], figs=[ArticleFig(id=1276616130877854137, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616109801476481, language=EN, label=Fig. 1, caption=Full scan product ion spectrums of ethrel, figureFileSmall=F/IGgASRLpPlxi1PsDeZqA==, figureFileBig=kZSm/n3EiHTE9eWmRR8RVw==, tableContent=null), ArticleFig(id=1276616131616051642, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616109801476481, language=CN, label=图1, caption=乙烯利的子离子全扫描质谱图, figureFileSmall=F/IGgASRLpPlxi1PsDeZqA==, figureFileBig=kZSm/n3EiHTE9eWmRR8RVw==, tableContent=null), ArticleFig(id=1276616132526215611, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616109801476481, language=EN, label=Fig. 2, caption=Total ion chromatogram of ethrel, figureFileSmall=Xhgvjn+D/WpNowKr4CAJwQ==, figureFileBig=OB6UyVfO5EBebkFOJaXFiQ==, tableContent=null), ArticleFig(id=1276616132933063100, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616109801476481, language=CN, label=图2, caption=乙烯利的总离子流图, figureFileSmall=Xhgvjn+D/WpNowKr4CAJwQ==, figureFileBig=OB6UyVfO5EBebkFOJaXFiQ==, tableContent=null), ArticleFig(id=1276616133339910589, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616109801476481, language=EN, label=Fig. 3, caption=Changes of ethrel content in rubber tree bark under different concentrations and stimulation time of ethrel, figureFileSmall=5790E8n6vejH0qmFVizokg==, figureFileBig=XUcPnYn8viJs/EgzZFq+Bg==, tableContent=null), ArticleFig(id=1276616134996660670, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616109801476481, language=CN, label=图3, caption=不同浓度乙烯利不同刺激时间下橡胶树树皮中乙烯利含量的变化, figureFileSmall=5790E8n6vejH0qmFVizokg==, figureFileBig=XUcPnYn8viJs/EgzZFq+Bg==, tableContent=null), ArticleFig(id=1276616135344787903, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616109801476481, language=EN, label=Fig. 4, caption=Changes of ethrel content at stimulation area in rubber tree bark, figureFileSmall=afp+o/VXWPtUimMwm8Rwqw==, figureFileBig=aJyoxqBmZWNNZAJcYNLgEg==, tableContent=null), ArticleFig(id=1276616135449645504, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616109801476481, language=CN, label=图4, caption=施药区域树皮乙烯利含量的变化

不同小写字母表示不同时间差异显著(P<0.05)。

, figureFileSmall=afp+o/VXWPtUimMwm8Rwqw==, figureFileBig=aJyoxqBmZWNNZAJcYNLgEg==, tableContent=null), ArticleFig(id=1276616135927796161, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616109801476481, language=EN, label=Tab. 1, caption=

UPLC-MS/MS parameters of ethrel

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分析物Analyte母离子Precursor ion (m/z)子离子Product ion (m/z)碰撞能量Collision energy/eV去簇电压Declustering potential/V
乙烯利143107*-10.0-50
79-22.0-52
), ArticleFig(id=1276616136603079106, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616109801476481, language=CN, label=表1, caption=

乙烯利测定的主要质谱参数

, figureFileSmall=null, figureFileBig=null, tableContent=
分析物Analyte母离子Precursor ion (m/z)子离子Product ion (m/z)碰撞能量Collision energy/eV去簇电压Declustering potential/V
乙烯利143107*-10.0-50
79-22.0-52
), ArticleFig(id=1276616136724713923, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616109801476481, language=EN, label=Tab. 2, caption=

Results of tests for recovery

, figureFileSmall=null, figureFileBig=null, tableContent=
加标水平Spiked level/(µg·kg-1)回收率Recovery/%相对标准偏差Relative standard deviation/%
0.1094.21.87
0.50101.21.14
1.0085.37.28
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回收率试验结果

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加标水平Spiked level/(µg·kg-1)回收率Recovery/%相对标准偏差Relative standard deviation/%
0.1094.21.87
0.50101.21.14
1.0085.37.28
), ArticleFig(id=1276616137131561413, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616109801476481, language=EN, label=Tab. 3, caption=

Linear equation parameters of ethrel content and stimulant concentration at stimulation area

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时间Time/h线性方程Regression equation相关系数R2
0y=17.414x+2.1300.984
6y=18.593x+3.8580.989
12y=19.198x+14.0170.950
24y=16.770x+13.9060.969
48y=16.441x+12.3250.954
96y=17.931x+5.4970.997
), ArticleFig(id=1276616137475494342, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616109801476481, language=CN, label=表3, caption=

橡胶树施药区域乙烯利含量与施药浓度线性方程参数

, figureFileSmall=null, figureFileBig=null, tableContent=
时间Time/h线性方程Regression equation相关系数R2
0y=17.414x+2.1300.984
6y=18.593x+3.8580.989
12y=19.198x+14.0170.950
24y=16.770x+13.9060.969
48y=16.441x+12.3250.954
96y=17.931x+5.4970.997
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橡胶树割线外施乙烯利的痕量检测及其吸收和分布规律研究
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杨文凤 , 范睿深 , 高宏华 , 魏芳 , 万炯 , 仇键 *
热带作物学报 | 作物栽培与生理生化 2025,46(8): 1864-1873
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热带作物学报 |作物栽培与生理生化 2025 , 46 (8) : 1864 -1873
橡胶树割线外施乙烯利的痕量检测及其吸收和分布规律研究
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杨文凤, 范睿深, 高宏华, 魏芳, 万炯, 仇键*
作者信息
  • 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/海南省热带作物栽培生理学重点实验室/省部共建国家重点实验室培育基地,海南海口 571101
通讯作者:
* 仇键(QIU Jian),E-mail:
Trace Detection and Distribution Dynamics of Externally Applied Ethrel in Rubber Tree Tapping Cuts
Wenfeng YANG, Ruishen FAN, Honghua GAO, Fang WEI, Jiong WAN, Jian QIU*
Affiliations
  • 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 / Hainan Key Laboratory for Cultivation & Physiology of Tropical Crops / State Key Laboratory Incubation Base for Cultivation and Physiology of Tropical Crops, Haikou, Hainan 571101, China
出版时间: 2025-08-25 doi: 10.3969/j.issn.1000-2561.2025.08.009
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乙烯利作为橡胶生产中广泛应用的增产剂,在促进橡胶树高效采胶中发挥了重要作用。然而,乙烯利刺激增产具有明显的剂量效应,过度施用会造成橡胶树死皮等副作用。本研究以热研73397橡胶树为试验材料,建立了基于超高效液相色谱串联质谱技术(UPLC-MS/MS)的树皮中乙烯利痕量检测方法,并系统研究了外施乙烯利后不同时间和树干不同部位树皮的乙烯利含量分布规律。样品前处理采用甲醇∶1%甲酸(9∶1,V/V)溶液匀浆浸提,过滤后直接进行定量分析。样品经Waters CORTECS HILIC C18色谱柱(100 mm×2.1 mm,1.6 μm)分离,以0.01%氨水溶液、乙腈∶水(80∶20,V/V)为流动相,流速为1.0 mL/min,在电喷雾离子源、负离子扫描和多反应监测模式(MRM)下进行检测,外标法定量,定量离子为107。结果表明:乙烯利在0.02~1.00 µg/mL浓度范围内线性关系良好(R2=0.9997),在0.1~1.0 µg/kg添加水平下,回收率为85.3%~101.2%,相对标准偏差为1.14%~7.28%,检出限为0.1 µg/kg。应用该方法研究了橡胶树涂施不同浓度乙烯利(0.5%、2.0%、4.0%)0~96 h后割线及其上下5、10 cm树皮中乙烯利的分布特征。结果显示:外施的乙烯利在橡胶树中分布不均匀,施药部位的含量显著高于其他部位,且近施药部位含量高于远施药部位,施药部位下方的含量高于上方。施药部位的乙烯利含量与外施乙烯利浓度呈显著正相关,且随着施药时间的延长呈先增后降的趋势,3种浓度均在施药后12~24 h达到峰值,随后逐渐下降,96 h仍能检测到较高浓度的乙烯利。与气相色谱法间接测定乙烯释放速率相比,本研究建立的检测方法具有简便快速、重现性好、灵敏度高等优势,为橡胶树中乙烯利的定量分析提供了可靠的技术支撑。研究初步揭示了橡胶树割线外施乙烯利后的吸收、分布及运输规律,发现其扩散运输不明显,主要集中在施药部位,可为深入解析乙烯利刺激增产的剂量效应及指导其安全高效施用提供理论依据。

橡胶树  /  乙烯利  /  超高效液相色谱串联质谱  /  分布规律

Ethrel is widely used as a stimulant in rubber tree plantations, which plays an important role in increaisng latex yield effectively and reducing labor requirements for tapping. However, the stimulatory effect of ethrel exhibits a pronounced dose-response relationship, excessive application may lead to adverse effects, such as tapping panel dryness (TPD). In this study, rubber tree clone Reyan 73397 was used as the experimental material, and an ultra-high performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for the trace determination of ethrel in the bark was developed. The distribution of ethrel at various time points and different parts of the bark following its application was investigated. The samples were extracted using a methanol∶1% formic acid aqueous solution (9∶1,V/V), and directly analyzed after filtration. The samples were separated by waters CORTECS HILIC C18 column (100 mm×2.1 mm, 1.6 μm) with mobile phase A (0.01% ammonia solution) and mobile phase B (acetonitrile∶water 80∶20, V/V) at a flow rate of 1.0 mL/min. The detection was performed under electrospray ion source, negative ion scanning and multiple reaction monitoring (MRM) mode. Quantitative analysis was conducted using external standard calibration curves, quantitative ion was 107. The results showed a good linear relationship for ethrel within the range of 0.02 μg/mL to 1.00 μg/mL, with coefficient of determination (R2) of 0.9997. Recoveries ranged from 85.3% to 101.2% at spiked levels 0.1 µg/kg to 1.0 µg/kg, with relative standard deviations of 1.14%-7.28%, and limits of quantification (LOQ) of 0.1 µg/kg. This method was applied to study the distribution characteristics of ethrel in stimulated bark rings, both above and below the ethrel-treated 5 cm and 10 cm areas, after applying different concentrations of ethrel (0.5%, 2.0%,4.0%) over 0-96 h. The results revealed that ethrel was unevenly distributed in the rubber tree, the concentration at the stimulated area was significantly higher than that in other areas, areas closer to the stimulated bark were higher than that in distant areas, and the areas below the stimulated bark were higher than that above it. The concentration of ethrel at the stimulated area showed a significant positive correlation with the stimulated ethrel concentration. Additionally, the concentration displayed an initial increase followed by a gradual decline over time. For the three concentrations, peak levels were observed between 12 h and 24 h post-stimulation, after which a gradual decrease occurred, with relatively high concentrations still detectable at 96 h. Compared to the indirect measurement of ethylene release rates by gas chromatography, this method has many significant advantages, including simplicity, rapidity, excellent reproducibility, and high sensitivity, thereby providing reliable technical support for the quantitative analysis of ethrel in rubber trees. The study preliminarily revealed the absorption, distribution and transport patterns of externally stimulated ethrel on rubber tree tapping cuts. It was found that the diffusion and transport of ethrel were limited, with the compound primarily concentrated at the stimulation site. The findings would provide a theoretical basis for further elucidating the dose-dependent effects of ethrel on yield stimulation and guiding its safe and efficient application.

rubber tree  /  ethrel  /  UPLC-MS/MS  /  distribution patterns
杨文凤, 范睿深, 高宏华, 魏芳, 万炯, 仇键. 橡胶树割线外施乙烯利的痕量检测及其吸收和分布规律研究. 热带作物学报, 2025 , 46 (8) : 1864 -1873 . DOI: 10.3969/j.issn.1000-2561.2025.08.009
Wenfeng YANG, Ruishen FAN, Honghua GAO, Fang WEI, Jiong WAN, Jian QIU. Trace Detection and Distribution Dynamics of Externally Applied Ethrel in Rubber Tree Tapping Cuts[J]. Chinese Journal of Tropical Crops, 2025 , 46 (8) : 1864 -1873 . DOI: 10.3969/j.issn.1000-2561.2025.08.009
巴西橡胶树(Hevea brasiliensis)为大戟科橡胶树属的落叶乔木,是全球商用天然橡胶最主要的来源[1]。割胶是天然橡胶生产中的关键技术环节,也是目前从橡胶树乳管组织中获取胶乳产品的唯一直接手段[2]。乙烯利(ethrel或ethephon,ETH,乙烯释放剂)作为各植胶国生产上广泛应用的橡胶树产量刺激剂,在提高胶乳产量、提升割胶劳动效率以及减少割胶用工需求方面发挥了重要作用[3-5]。然而,乙烯利刺激橡胶树的增产效果具有显著的剂量效应[6-8],在适当的乙烯利刺激浓度范围内与干胶产量呈正相关[9-10],但过度施用会导致胶乳长流、干含急剧下降甚至引发橡胶树死皮等副作用[11-13]。随着胶工短缺现象日益加剧,割胶技术低频化的推进以及乙烯利施用浓度的增加,这些负面效应更加凸显[2]。本课题组通过初步研究发现,外施乙烯利后剂量效应的产生可能与乙烯利在橡胶树体内的时空分布有关(未发表资料)。因此,研究乙烯利在橡胶树体内的吸收、分布及运输规律,对解析其剂量效应产生机制,指导乙烯利的合理施用以及实现橡胶树的安全高效生产具有重要意义。
建立乙烯利痕量检测方法是研究乙烯利在植株体内吸收、分布及运输规律的基础。然而,乙烯利作为一种强极性小分子化合物,具有易溶于水、挥发性低、无生色和荧光基团、对紫外光敏感且易分解的特性,直接进入常规仪器时响应值较低,因此需要超高灵敏度的检测方法[14-16]。目前,常用的乙烯利检测方法主要有离子色谱法、气相色谱法、气相色谱-质谱法以及液相色谱-质谱法[17-18]。其中,离子色谱法操作简单,但抗干扰能力差且灵敏度较低[19];气相色谱法易产生假阳性结果[20];气相色谱-质谱法需要通过衍生化处理,样品前处理步骤繁琐,定量准确性受衍生化产物稳定性及乙烯释放量的限制[21]。相比之下,超高效液相色谱-质谱联用法(UPLC-MS/MS)结合了高效液相色谱和质谱技术的优势,具有样品前处理简便快捷、特异性强、灵敏度高以及定性能力好等优势,已成为乙烯利定量分析的首选方法[22-23],并在棉花、坚果、中药材、水果以及蔬菜等领域得到广泛应用[24-28]。在橡胶树中,尚无采用UPLC-MS/MS法直接测定橡胶树中乙烯利含量的研究报道,目前仅见杨少琼[29]、许闻献等[30]、曹建华等[31]、邓军等[32-33]采用气相色谱法间接测定橡胶树离体树皮及割胶后胶乳中乙烯的释放速率来推测乙烯利含量。该方法基于乙烯利在pH>4条件下分解释放乙烯的特性[34],样品前处理复杂,且假设橡胶树产生的乙烯完全来源于外施乙烯利,忽略了割胶伤害和施用外源乙烯利诱导产生的内源乙烯,可能导致假阳性结果。此外,关于外施乙烯利在植株体内的运转分配,早期研究曾采用14C同位素示踪技术在小麦[35]、茶树[36]、棉花[37]及瓠瓜[38]等植物中进行探索,发现不同植物对乙烯利的吸收分布存在显著差异,且同一植株中各器官的积累量也不均匀。关于乙烯利在橡胶树中的吸收分布研究,已有AUDLEY等[39]采用14C同位素示踪技术,PARANJOTHY等[40]、杨少琼[29]、许闻献等[30]通过间接测定乙烯释放速率推测其吸收分布的报道。但上述方法由于放射性风险、高成本和操作复杂性限制了其在乙烯利吸收分布研究中的广泛应用。
本研究建立了基于直接提取法结合UPLC-MS/MS技术的橡胶树乙烯利痕量检测方法,并利用该方法测定了割线外施不同浓度乙烯利后的残留量及其吸收分布状况,进一步明确了乙烯利刺激的高效作用部位和高效刺激时间。研究结果可为生产中指导乙烯利的高效安全施用提供理论依据。
选取中国热带农业科学院试验场十二队(19°31′10″N, 109°29′41″E)开割3 a未涂乙烯利刺激的橡胶树热研73397为试验材料,试验以单株为处理对象,每个处理重复3次。
超高效液相色谱系统(美国Waters公司),AB SCIEX API4000-质谱系统(美国AB SCIEX公司),研磨机、高速匀浆机(德国IKA公司)。
乙烯利标准品(纯度96%,AccuStandard公司);C18色谱柱(Waters CORTECS,Waters公司);甲醇、乙腈(色谱纯,Fisher试剂公司);甲酸、氨水(色谱纯,上海凌峰化学试剂有限公司);有机相针式过滤器(13 mm×0.22 μm,上海安谱实验科技有限公司);40%乙烯利水剂(绍兴东湖高科股份有限公司);实验用水均为超纯水。
于2023年5月开割前用割胶刀割除供试橡胶树割线上回枯部分树皮,待新割线干后,分别于采样前0(割胶前刺激,刺激后马上割胶)、6、12、24、48、96 h在割线处每株次涂施2 g不同浓度乙烯利水剂(0.5%、2.0%、4.0%),选取树干不同部位(施药区上方10 cm、施药区上方5 cm、施药区下方5 cm、施药区下方10 cm)为纵直分布的采样点。由同一胶工在每株胶树的5个纵直分布采样点分别用胶刀切割下树皮,同时选取同林段相邻的无乙烯利处理的3株胶树割线处的树皮为空白样,切下的树皮装袋密封后放入冰盒中立即带回实验室置于-80 ℃冰箱保存,备用。
配制1 L甲醇∶1%甲酸(9∶1, V/V)溶液作为提取液,即:900 mL甲醇,99 mL水及1 mL甲酸。
精确称取乙烯利标准品10 mg,用甲醇溶解并定容至100 mL容量瓶中,配制成100 μg/mL乙烯利标准储备液,置于冰箱中冷藏保存。用甲醇逐级稀释标准储备液至浓度分别为0.20、1.00、2.50、5.00、10.00 μg/mL乙烯利标准溶液。准确吸取各浓度的乙烯利标准溶液1 mL于10 mL的容量瓶中,用提取液定容至刻度,摇匀,配制成质量浓度分别为0.02、0.10、0.25、0.50、1.00 μg/mL的乙烯利系列标准曲线溶液。
精确称取树皮样品5.0 g置于研磨机在液氮中研磨30 s至粉末状,加入提取液10 mL,高速匀浆1 min后转移至15 mL离心管,于4 ℃浸提过夜,用注射器吸取2 mL上清液经0.22 μm过滤器后置于样品瓶中待分析测定。
液相色谱条件:Waters CORTECS HILIC C18色谱柱(2.1 mm×100 mm, 1.6 μm),柱温为40 ℃,进样体积为10.0 μL,流速为0.2 mL/min,流动相A为0.01%氨水溶液,流动相B为乙腈∶水(80∶20, V/V),采用梯度洗脱。洗脱程序:0~2.0 min,90% A;2.0~4.5 min,90% A~20% A;4.5~5.0 min,20% A~90% A。
质谱条件:电喷雾负离子(ESI-)扫描,多反应监测(MRM)模式,喷雾电压(IS)为-4500 V,离子源温度(TEM)为600 ℃。采用外标法,以乙烯利保留时间定性,以峰面积定量。采用负离子全扫描模式进行扫描,确定其母离子,并对其已确定的母离子进行二级质谱Product Ion扫描,得到子离子。根据响应强度高、基线噪声低、抗干扰能力强的原则,最终选择1个母离子和2个子离子。并在MRM模式下对所分析物离子对的碰撞能量、去簇电压等质谱参数进行优化,相关质谱参数见表1
采用Excel 2019软件处理试验数据,采用SPSS 18.0软件进行数据分析,采用Origin 2021软件作图。
乙烯利是一种强极性的二元酸,在极性溶剂中具有较高的溶解度,因此常采用水、甲醇或酸化甲醇等作为乙烯利的提取溶剂[16]。本研究选用甲醇∶1%甲酸(9∶1, V/V)混合溶剂作为提取液,样品加入提取液研磨后高速匀浆、浸提过夜,过滤后直接采用超高效液相色谱-串联三重四级杆质谱仪进行乙烯利检测,定量离子m/z为106.7(图1)。其提取离子流色谱图如图2所示,乙烯利的分离效果良好,峰形尖锐且对称,乙烯利的保留时间为0.95 min。
将配制的乙烯利标准工作液按照上述色谱-质谱条件,浓度从低到高分别进样测定。以乙烯利浓度(µg/mL)为横坐标,定量离子的峰面积为纵坐标,绘制标准曲线,得到其标准工作曲线方程:y=5.033×106x-1.132×104,乙烯利在0.02~1.00 µg/mL浓度范围内呈良好的线性关系,相关系数(R2)为0.9997。在上述条件下,按照信噪比(S/N)≥3计算出方法的检出限(LOD)为0.1 µg/kg,S/N≥10计算出定量限为(LOQ)为0.3 µg/kg。
在无乙烯利刺激的空白树皮样品中,进行乙烯利标准溶液的加标回收试验。加标水平分别为0.1、0.5、1.0 µg/kg,采用相应处理方法对样品进行处理,每个添加水平重复测定6次,乙烯利在橡胶树树皮中的加标回收率及相对标准偏差见表2。由表2可知,乙烯利在橡胶树树皮样品中的平均回收率为85.3%~101.2%,相对标准偏差为1.14%~7.28%。上述数据表明,所建立的检测方法精密度和准确度较好,可以满足后续定量检测分析的要求。
利用上述乙烯利痕量检测方法,分析不同乙烯利浓度(0.5%、1.0%、4.0%)刺激后,橡胶树不同部位树皮中乙烯利含量的变化(图3)。乙烯利不同浓度或不同处理时间,施药区域树皮的乙烯利含量均明显高于其他部位;距离施药区域5 cm处的树皮乙烯利含量均高于10 cm处的树皮;此外,无论距离施药区域5 cm还是10 cm,施药区域下方树皮的乙烯利含量均高于上方。结果表明,外施乙烯利在橡胶树皮中的分布呈现不均匀性,主要残留集中在施药区域的割线处,其余部分通过扩散或蒸腾作用在施药区域上、下10 cm范围内分布,且距离施药区域越近乙烯利含量越高,施药区域下方的含量高于上方。
不同刺激时间下树皮中乙烯利含量的变化因乙烯利浓度和采样部位的不同而有所差异(图3)。施药区域上方树皮中乙烯利含量均较低,除0.5%乙烯利处理,施药区域上方5 cm树皮中的乙烯利含量随刺激时间的延长呈先升后降趋势外,其他浓度处理的变化趋势不明显。在乙烯利富集的施药区域,乙烯利含量随刺激时间的延长整体呈先升后降的倒“V”型趋势。而在施药区域下部的树皮中,0.5%乙烯利处理的施药区域下部5 cm树皮和4.0%乙烯利处理的下部10 cm树皮中的乙烯利含量呈先降后升的“V”型趋势;而在其他条件下乙烯利含量则呈先升后降再升的“N”型趋势;且施药后96 h施药区域下方5 cm和10 cm树皮中乙烯利含量均高于初始值(0 h)。表明乙烯利在橡胶树皮中的动态分布具有明显的浓度依赖性和空间特异性。
橡胶树不同部位树皮中的乙烯利含量随外施乙烯利浓度增加而增加的趋势,其中施药区域乙烯利含量的动态变化对刺激浓度和刺激时间的响应尤为显著(图4)。刺激后0 h,由于乙烯利在树皮表面停留时间较短,树体吸收和扩散尚不充分,检测到的乙烯利含量较低,从6 h开始随刺激时间的延长乙烯利含量逐渐升高。在0.5%乙烯利刺激下,树皮中的乙烯利含量在施药后24 h达到峰值;而2.0%和4.0%乙烯利处理的峰值则提前至施药后12 h;此外,3种浓度处理后96 h的乙烯利含量均高于初始值(0 h),而4.0%乙烯利处理后96 h的乙烯利含量高于处理后48 h。
相关性分析表明,乙烯利处理后0~96 h树皮中的乙烯利含量与外施乙烯利浓度均呈极显著正相关(P<0.01),线性回归方程见表3。然而,刺激时间与乙烯利含量的相关性未达显著水平。结果表明,施药部位树皮中的乙烯利含量随刺激时间的延长呈现明显的峰值,乙烯利浓度越高,峰值出现的越早,而且乙烯利在树皮中具有一定的持久性。
乙烯利的痕量分析检测中,检测方法的选择和样品前处理是2个关键环节,其中样品前处理的效率直接影响分析方法的可靠性、准确性和分析通量[41]。本研究根据乙烯利的强极性特性,选用极性溶剂甲醇∶1%甲酸(9∶1, V/V)作为提取溶剂,通过匀浆浸提过夜后直接提取,乙烯利的回收率达85.3%~101.2%。该方法样品前处理操作简便,对待测组分提取较为完全,单个样品的检测时间缩短至5 min,显著提高了检测效率。与邓军等[33]报道的1.12 µL/L的检出限相比,本研究中的检出限为0.1 µg/kg,不仅提高了检测的准确性,还避免了气相色谱法易出现的假阳性问题。此外,本研究中的检测方法具有较高的精密度和准确度,能够更好地满足橡胶树中乙烯利含量的定量分析需求。
早期14C乙烯利示踪研究表明,乙烯利在植株体内的运转可分为三部分:一部分长时间保留在处理部位;一部分快速进入植株体内,逐步分解释放乙烯发挥其生理功能或通过扩散转移至其他部位;另一部分则直接从处理部位通过输导组织运输至其他器官再分解释放或代谢为其他物质[34]。杨少琼[29]发现橡胶树涂施乙烯利后,其扩散范围可达施药区上下2 m。许闻献等[30]则认为乙烯利在橡胶树体内主要通过扩散转移,扩散分布范围为90 cm,扩散分布的不均匀性造成了刺激产量效应的动态范围。本研究发现,在橡胶树割线处涂施不同浓度的乙烯利后其大部分残留在施药部位,其余部分进入橡胶树体后在施药部位上、下10 cm范围内均有分布,且距离施药部位越近,乙烯利含量越高。
关于乙烯利在橡胶树体内的运转方向一直以来存在争议。PARANJOTHY等[40]认为乙烯利的向下运动不明显;而AUDLEY等[39]、杨少琼[29]发现乙烯利向上扩散的速度快于向下扩散,认为乙烯利主要是通过蒸腾液流向上运输;许闻献等[30]发现外源乙烯利在上、下方向的扩散范围与扩散速度基本相同,而且施药后树干不同部位的乙烯利分布与产量分布一致;SETHURAL等发现乙烯利的药效范围主要集中在施药部位的下方[30],与本研究结果相对一致,即施药部位下方的乙烯利含量高于上方。这种差异可能与所选取的试验材料、检测方法的精确度及灵敏度有关。本研究和PARANJOTHY等[40]采用的是离体树皮作为样品,而杨少琼[29]采用的是割胶后的胶乳样品,AUDLEY等[39]则采用橡胶树幼苗树皮。此外,乙烯利在橡胶树体内的运转分布及原位分解释放受温度、橡胶树物候、pH等多因素影响,样品采集时段、采样方法等的差异也可能导致结果不一致。
本研究还发现橡胶树对外源乙烯利的吸收呈明显的时间差异,乙烯利含量随刺激时间的延长呈先增后降的趋势,乙烯利吸收高峰期出现在刺激后12~24 h。这表明橡胶树对外源乙烯利的生理响应具有一定滞后性[42],与魏芳等[43]、仇键等[44]研究认为橡胶树响应外源乙烯利产生排胶相关生理反应的高峰期在刺激后24~96 h的结果相吻合。此外,乙烯利含量与乙烯利施用浓度之间呈显著线性正相关。
本研究发现外施乙烯利在橡胶树树皮中的分布具有广泛性、局域性和不均匀性等特点,通过系统分析施药区域乙烯利的时空动态变化规律,为指导橡胶生产中乙烯利的高效安全施用提供理论依据。然而,由于乙烯利在植物体内的不稳定性,未来研究还需进一步开发高效、无损的原位实时检测技术,以建立乙烯利释放动态的同步监测体系,这将有助于深入解析乙烯利刺激橡胶树增产的分子机制和开发橡胶树精准高效的乙烯利施用技术。
  • 海南省自然科学基金项目(320MS111)
  • 中央级公益性科研院所基本科研业务费专项(1630022022016)
  • 现代农业产业技术体系建设专项(CARS-33-ZP4)
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2025年第46卷第8期
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doi: 10.3969/j.issn.1000-2561.2025.08.009
  • 接收时间:2025-03-08
  • 首发时间:2026-06-24
  • 出版时间:2025-08-25
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  • 收稿日期:2025-03-08
  • 录用日期:2025-04-11
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海南省自然科学基金项目(320MS111)
中央级公益性科研院所基本科研业务费专项(1630022022016)
现代农业产业技术体系建设专项(CARS-33-ZP4)
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    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/海南省热带作物栽培生理学重点实验室/省部共建国家重点实验室培育基地,海南海口 571101

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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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