Article(id=1309573248384327788, tenantId=1146029695717560320, journalId=1308397196074385509, issueId=1309573247344144835, articleNumber=null, orderNo=null, doi=10.11733/j.issn.1007-0435.2026.04.002, pmid=null, cstr=null, oa=null, hot=0, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1744646400000, receivedDateStr=2025-04-15, revisedDate=1749744000000, revisedDateStr=2025-06-13, acceptedDate=null, acceptedDateStr=null, onlineDate=1790156193108, onlineDateStr=2026-09-23, pubDate=null, pubDateStr=null, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1790156193108, onlineIssueDateStr=2026-09-23, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1790156193108, creator=13701087609, updateTime=1790156193108, updator=13701087609, issue=Issue{id=1309573247344144835, tenantId=1146029695717560320, journalId=1308397196074385509, year='2026', volume='34', issue='4', pageStart='1137', pageEnd='1556', issueExtLink='null', onlineDate='null', pubDate='1776182400000', pubDateStr='2026-04-15', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1790156192860, creator='13701087609', updateTime=1790156800047, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1309575794117144683, tenantId=1146029695717560320, journalId=1308397196074385509, issueId=1309573247344144835, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1309575794117144684, tenantId=1146029695717560320, journalId=1308397196074385509, issueId=1309573247344144835, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=1146, endPage=1157, ext={EN=ArticleExt(id=1309573250347262063, articleId=1309573248384327788, tenantId=1146029695717560320, journalId=1308397196074385509, language=EN, title=Evaluation and Screening of Reference Genes in Alfalfa (Medicago sativa L. ‘Gongnong No.1’) Different Tissues under Waterlogging Stress, columnId=null, journalTitle=Acta Agrestia Sinica, columnName=null, runingTitle=null, highlight=null, articleAbstract=The stability and applicability of reference genes in alfalfa vary significantly depending on the cultivar, growth and development stages, and environmental conditions. In this study, alfalfa (Medicago sativa L.) cultivar ‘Gongnong No.1’ was used as material, and eight candidate reference genes, glyceraldehyde-3-phosphate dehydrogenase GAPDH, ubiquitin protein ligase 2a UBL-2a, Retention in Endoplasmic Reticulum Rer1, eukaryotic elongation factor-1 alpha eEF-1α, actin-depolymerizing factor ADF, ubiquitin carrier protein UBC, tubulin beta-1 chain TUB, and Actin were selected to evaluate their expression stability of different tissues in seedling and adult plant, as well as under waterlogging stress. The analysis employed quantitative real-time PCR (qRT-PCR) and four softwares, including geNorm, NormFinder, BestKeeper, and RefFinder. The results showed that, at the seedling stage, Rer1, Actin, and UBC were the most stable reference genes. At the tissue level, the most stable reference genes for leaves, stems, and roots were Actin, Rer1, and GAPDH, respectively. At the adult stage, ADF, Actin, and TUB exhibited the highest stability, while the most stable reference genes for leaves, stems, roots, and flowers were eEF-1α, Rer1, Actin, and UBL-2a, respectively. Under waterlogging stress, the three most stable reference genes in seedling leaves were TUB, ADF, and eEF-1α, whereas in seedling roots they were ADF, UBL-2a, and Rer1. In the adult stage, the most stable genes were ADF, Actin, and TUB in leaves, and UBL-2a, UBC, and ADF in roots. Overall, Rer1 and UBL-2a are recommended as reference genes for studies of tissue-specific expression, as their stability was less affected by developmental stage or tissue type. ADF and TUB are suitable reference genes for studies involving waterlogging stress., authors=XU Yi1,2, WANG Ting1, GUO Yu-peng3, MIAO Xing-fen2, ZHENG Xia1,4, SHI Peng-jun1,4, authorsList=XU Yi, WANG Ting, GUO Yu-peng, MIAO Xing-fen, ZHENG Xia, SHI Peng-jun, authorCompany=1 Institute of Bast Fiber Crops, Chinese Academy of Agricultural Sciences, Changsha, Hunan Province 410221, China;
2 College of Agriculture, Heilongjiang Bayi Agricultural University, Daqing, Heilongjiang Province 163319, China;
3 The College of Ecological Environment and Resources, Qinghai Minzu University, Xining, Qinghai Province 810007, China;
4 National Nanfan Research Institute, Chinese Academy of Agricultural Sciences, Sanya, Hainan Province 572024, China, 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=1309573248635986030, articleId=1309573248384327788, tenantId=1146029695717560320, journalId=1308397196074385509, language=CN, title=‘公农1号’苜蓿不同组织和涝害胁迫下内参基因的筛选评价, columnId=1309565024058835950, journalTitle=草地学报, columnName=研究论文, runingTitle=null, highlight=null, articleAbstract=紫花苜蓿(Medicago sativa L.)内参基因的稳定性和适用性因品种、生长发育及环境条件的不同差异较大。本研究以紫花苜蓿‘公农1号’为材料,使用qRT-PCR技术与geNorm,NormFinder,BestKeeper和RefFinder等软件,评估8个常见内参基因GAPDH,UBL-2a,Rer1,eEF-1α,ADF,UBC,TUB和Actin等不同时期的组织表达特异性以及涝害胁迫下的表达稳定性。结果表明,‘公农1号’幼苗期稳定的内参基因是Rer1,Actin,UBC,其中叶、茎和根中表达最稳定的内参基因分别是Actin,Rer1,GAPDH;成株期最稳定的内参基因是ADF,Actin,TUB,而成株期叶、茎、根和花中表达最稳定的分别是eEF-1α,Rer1,Actin和UBL-2a。涝害胁迫下,幼苗期叶片稳定性前三的内参基因为TUB,ADF和eEF-1α,根部为ADF,UBL-2a和Rer1;成株期叶中稳定性前三的内参基因为ADF,Actin和TUB,根部为UBL-2a,UBC和ADF。Rer1和UBL-2a的稳定性受发育时期和组织特异性影响较小,适合用作组织表达特异性研究的内参基因;ADF适合作为涝害胁迫研究的内参基因。, authors=徐艺1,2, 王婷1, 郭玉朋3, 苗兴芬2, 郑霞1,4, 石鹏君1,4, authorsList=徐艺, 王婷, 郭玉朋, 苗兴芬, 郑霞, 石鹏君, authorCompany=1 中国农业科学院麻类研究所, 湖南 长沙 410221;
2 黑龙江八一农垦大学农学院, 黑龙江 大庆 163319;
3 青海民族大学生态环境与资源学院, 青海 西宁 810007;
4 中国农业科学院国家南繁研究院, 海南 三亚 572024, correspAuthors=null, authorNote=徐艺(1996-),女,汉族,山东济宁人,硕士研究生,主要从事牧草种质资源创新与利用研究,E-mail:xuyi01010219@163.com;王婷(1998-),女,汉族,甘肃会宁人,硕士研究生,主要从事苜蓿逆境分子生物学研究,E-mail:82101235179@caas.cn, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=+Pa/qLrHvjc//nCrnuDAqA==, pdfFileSize=2382626, 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=湖南省自然科学基金-省市联合基金 (2022JJ50021); 中国农业科学院科技创新工程 (CAAS-ASTIP-IBFC); 三亚中国农业科学院国家南繁研究院“南繁专项” (YYLH2505))}, authors=null, keywords=[Keyword(id=1309573250519228529, tenantId=1146029695717560320, journalId=1308397196074385509, articleId=1309573248384327788, language=CN, orderNo=1, keyword=紫花苜蓿), Keyword(id=1309573250577948786, tenantId=1146029695717560320, 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DENG J S,FENG C,ZHANG L H,et al. Screening of eggplant qRT-PCR internal reference genes under different tissues and adversity stresses[J]. China Vegetables,2024(8):37-46 邓嘉硕,冯程,张泷昊,等. 不同组织与逆境胁迫下茄子qRT-PCR内参基因筛选[J]. 中国蔬菜,2024(8):37-46
SHI K,DONG H B,DU H L,et al. The chromosome-level assembly of the wild diploid alfalfa genome provides insights into the full landscape of genomic variations between cultivated and wild alfalfa[J]. Plant Biotechnology Journal,2024,22(6):1757-1772
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ZHANG W,CUI W T,DUAN X L,et al. Reference gene selection for qRT-PCR normalization in alfalfa under UV-B irradiation[J]. Journal of Nanjing Agricultural University,2015,38(2):248-254 张薇,崔为体,段星亮,等. UV-B胁迫下紫花苜蓿qRT-PCR内参基因的筛选[J]. 南京农业大学学报,2015,38(2):248-254
LIU Q N,WANG B J,XU W,et al. Genome-wide investigation of the PIF gene family in alfalfa (Medicago sativa L.) expression profiles during development and stress[J]. BMC Genomic Data,2024,25(1):79
CHEN X F,XU H L,WANG R X,et al. Effects of flooding stress on biological characteristics of different alfalfa varieties and evaluation of their waterlogging tolerance[J]. Grassland and Turf,2024,44(2):155-163 陈小芳,徐化凌,王瑞香,等. 9个紫花苜蓿品种对淹水胁迫的响应及耐涝性评价[J]. 草原与草坪,2024,44(2):155-163
ZHANG Q,LIU X F,ZHANG Z F,et al. Melatonin improved waterlogging tolerance in alfalfa (Medicago sativa) by reprogramming polyamine and ethylene metabolism[J]. Frontiers in Plant Science,2019,10:44
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XIE F L,XIAO P,CHEN D L,et al. miRDeepFinder: a miRNA analysis tool for deep sequencing of plant small RNAs[J]. Plant Molecular Biology,2012,80(1):75-84)
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草地学报 |研究论文 2026 , 34 (4) : 1146 -1157
‘公农1号’苜蓿不同组织和涝害胁迫下内参基因的筛选评价
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徐艺1,2, 王婷1, 郭玉朋3, 苗兴芬2, 郑霞1,4, 石鹏君1,4
作者信息
    1 中国农业科学院麻类研究所, 湖南 长沙 410221;
    2 黑龙江八一农垦大学农学院, 黑龙江 大庆 163319;
    3 青海民族大学生态环境与资源学院, 青海 西宁 810007;
    4 中国农业科学院国家南繁研究院, 海南 三亚 572024
作者简介:
徐艺(1996-),女,汉族,山东济宁人,硕士研究生,主要从事牧草种质资源创新与利用研究,E-mail:xuyi01010219@163.com;王婷(1998-),女,汉族,甘肃会宁人,硕士研究生,主要从事苜蓿逆境分子生物学研究,E-mail:82101235179@caas.cn
Evaluation and Screening of Reference Genes in Alfalfa (Medicago sativa L. ‘Gongnong No.1’) Different Tissues under Waterlogging Stress
  • XU Yi1,2, WANG Ting1, GUO Yu-peng3, MIAO Xing-fen2, ZHENG Xia1,4, SHI Peng-jun1,4
  • Affiliations
      1 Institute of Bast Fiber Crops, Chinese Academy of Agricultural Sciences, Changsha, Hunan Province 410221, China;
      2 College of Agriculture, Heilongjiang Bayi Agricultural University, Daqing, Heilongjiang Province 163319, China;
      3 The College of Ecological Environment and Resources, Qinghai Minzu University, Xining, Qinghai Province 810007, China;
      4 National Nanfan Research Institute, Chinese Academy of Agricultural Sciences, Sanya, Hainan Province 572024, China
    doi: 10.11733/j.issn.1007-0435.2026.04.002
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    紫花苜蓿(Medicago sativa L.)内参基因的稳定性和适用性因品种、生长发育及环境条件的不同差异较大。本研究以紫花苜蓿‘公农1号’为材料,使用qRT-PCR技术与geNorm,NormFinder,BestKeeper和RefFinder等软件,评估8个常见内参基因GAPDH,UBL-2a,Rer1,eEF-1α,ADF,UBC,TUB和Actin等不同时期的组织表达特异性以及涝害胁迫下的表达稳定性。结果表明,‘公农1号’幼苗期稳定的内参基因是Rer1,Actin,UBC,其中叶、茎和根中表达最稳定的内参基因分别是Actin,Rer1,GAPDH;成株期最稳定的内参基因是ADF,Actin,TUB,而成株期叶、茎、根和花中表达最稳定的分别是eEF-1α,Rer1,Actin和UBL-2a。涝害胁迫下,幼苗期叶片稳定性前三的内参基因为TUB,ADF和eEF-1α,根部为ADF,UBL-2a和Rer1;成株期叶中稳定性前三的内参基因为ADF,Actin和TUB,根部为UBL-2a,UBC和ADF。Rer1和UBL-2a的稳定性受发育时期和组织特异性影响较小,适合用作组织表达特异性研究的内参基因;ADF适合作为涝害胁迫研究的内参基因。
    紫花苜蓿  /  内参基因  /  非生物胁迫  /  表达稳定性
    The stability and applicability of reference genes in alfalfa vary significantly depending on the cultivar, growth and development stages, and environmental conditions. In this study, alfalfa (Medicago sativa L.) cultivar ‘Gongnong No.1’ was used as material, and eight candidate reference genes, glyceraldehyde-3-phosphate dehydrogenase GAPDH, ubiquitin protein ligase 2a UBL-2a, Retention in Endoplasmic Reticulum Rer1, eukaryotic elongation factor-1 alpha eEF-1α, actin-depolymerizing factor ADF, ubiquitin carrier protein UBC, tubulin beta-1 chain TUB, and Actin were selected to evaluate their expression stability of different tissues in seedling and adult plant, as well as under waterlogging stress. The analysis employed quantitative real-time PCR (qRT-PCR) and four softwares, including geNorm, NormFinder, BestKeeper, and RefFinder. The results showed that, at the seedling stage, Rer1, Actin, and UBC were the most stable reference genes. At the tissue level, the most stable reference genes for leaves, stems, and roots were Actin, Rer1, and GAPDH, respectively. At the adult stage, ADF, Actin, and TUB exhibited the highest stability, while the most stable reference genes for leaves, stems, roots, and flowers were eEF-1α, Rer1, Actin, and UBL-2a, respectively. Under waterlogging stress, the three most stable reference genes in seedling leaves were TUB, ADF, and eEF-1α, whereas in seedling roots they were ADF, UBL-2a, and Rer1. In the adult stage, the most stable genes were ADF, Actin, and TUB in leaves, and UBL-2a, UBC, and ADF in roots. Overall, Rer1 and UBL-2a are recommended as reference genes for studies of tissue-specific expression, as their stability was less affected by developmental stage or tissue type. ADF and TUB are suitable reference genes for studies involving waterlogging stress.
    Alfalfa  /  Reference genes  /  Abiotic stress  /  Expression stability
    徐艺, 王婷, 郭玉朋, 苗兴芬, 郑霞, 石鹏君. ‘公农1号’苜蓿不同组织和涝害胁迫下内参基因的筛选评价. 草地学报, 2026 , 34 (4) : 1146 -1157 . DOI: 10.11733/j.issn.1007-0435.2026.04.002
    XU Yi, WANG Ting, GUO Yu-peng, MIAO Xing-fen, ZHENG Xia, SHI Peng-jun. Evaluation and Screening of Reference Genes in Alfalfa (Medicago sativa L. ‘Gongnong No.1’) Different Tissues under Waterlogging Stress[J]. Acta Agrestia Sinica, 2026 , 34 (4) : 1146 -1157 . DOI: 10.11733/j.issn.1007-0435.2026.04.002

      湖南省自然科学基金-省市联合基金 (2022JJ50021); 中国农业科学院科技创新工程 (CAAS-ASTIP-IBFC); 三亚中国农业科学院国家南繁研究院“南繁专项” (YYLH2505)

    参考文献 引证文献
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    XIE C C,MA J,LI J,et al. Selection of quantitative real-time reverse transcription-PCR reference genes in lead-stressed bermudagrass roots treated with different concentrations of melatonin[J]. Pratacultural Science,2025,42(1):119-128 谢程程,马均,李璟,等. 不同浓度褪黑素缓解狗牙根铅胁迫实时荧光定量PCR内参基因的筛选[J]. 草业科学,2025,42(1):119-128
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    小菇属 Mycena 11 5.26
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