'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
打开文本图片集
中图分类号：S541 文献标识码：A 文章编号：1007-0435（2026）04-1146-12
Abstract：The stability and applicability ofreference 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 Rerl，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 wellas under waterlogging stress. The analysis employed quantitative realtime PCR（qRT-PCR）and four softwares，including geNorm，NormFinder，BestKeeper，and RefFinder. The results showed that，at the seedling stage，Rerl， 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，Rerl，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 RerI . 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.
Keywords：Alfalfa;Reference genes;Abiotic stress;Expression stability
紫花苜蓿（MedicagosatiuaL.）是豆科苜蓿属多年生优质牧草，广泛分布在我国干旱和半干旱区，因其富含蛋白质和维生素而被称为“牧草之王”1-2]。（剩余15871字）