摘要
为了筛选苦豆子不同组织中适应非生物胁迫的内参基因,本研究采用实时荧光定量PCR技术,分析了苦豆子中GAPDH、ACT、TUA、ELF1α、ELF1β、G6PDH、UBC、F-box、UBQ、CYP等10个内参基因在根、茎、叶中表达量的变化,并应用ge Norm、Norm Finder、Best Keeper软件以及在线工具Ref Finder对其在不同非生物胁迫下根、茎、叶中的表达稳定性进行了分析。结果表明,苦豆子内参基因F-box、TUA和ELF1β分别是冷胁迫处理下根、茎、叶中最稳定的内参基因;干旱胁迫处理下根、茎、叶中最稳定的内参基因分别是ELF1α、ELF1β和ELF1β;而UBC、UBC和ELF1α则是盐胁迫处理下根、茎、叶中最稳定的内参基因。此外,本研究还评价了苦豆子抗旱基因MYB的表达量,发现不同胁迫处理下获得的内参基因稳定性的排序是正确可靠的。这一研究为今后苦豆子耐逆基因的表达分析提供了参考基因和理论依据。
In this study, the expression levels of ten reference genes are analyzed in the root, stem and leaf of Sophora alopecuroides with real-time quantitative PCR, including GAPDH, ACT, TUA, ELF1α, ELF1β, G6PDH, UBC, F-box, UBQ and CYP. Their expression stabilities were evaluated using geNorm, NormFinder, BestKeeper and online tool RefFinder in order to screen the internal reference genes adapting to abiotic stress in different tissues of S. alopecuroides. The results showed that F-box, TUA and ELF1β were the most stable reference genes in root, stem and leaf of S. alopecuroides under cold stress, respectively. ELF1α, ELF1βand ELF1β were the most stable reference genes in roots, stems and leaves under drought stress, respectively. UBC, UBC and ELF1α were the most stable reference genes in roots, stems and leaves under salt stress. In addition, the expression of drought resistance gene MYB in S. alopecuroides is evaluated in this study. It has found that the stability ranking of internal reference genes obtained under different stress treatments is correct and reliable, which provided a reference gene and theoretical basis for the expression analysis of stress tolerance genes in S. alopecuroides for the future study.
作者
苏丹丹
刘玉萍
张雨
郑长远
王亚男
苏旭
孙成林
SU Dandan;LIU Yuping;ZHANG Yu;ZHENG Changyuan;WANG Yanan;SU Xu;SUN Chenglin(School of Life Sciences,Qinghai Normal University,Xining 810008,China;Key Laboratory of Medicinal Animal and Plant Resources of the Qinghai-Tibet Plateau in Qinghai Province,Qinghai Normal University,Xining 810008,China;Academy of Plateau Science and Sustainability,Xining 810016,China)
出处
《植物生理学报》
CAS
CSCD
北大核心
2022年第7期1295-1306,共12页
Plant Physiology Journal
基金
青海省自然科学基金面上项目(2022-ZJ-913)
祁连山国家公园青海研究中心开放课题。
关键词
苦豆子
内参基因
实时荧光定量PCR
组织器官
非生物胁迫
Sophora alopecuroides
reference gene
real-time quantitative PCR
tissue and organ
abiotic stress
