四种木本油料内参基因筛选及Actin基因时空表达分析

Screening of Reference Genes in Four Woody-oil Trees and Spatio-temporal Expression Analysis of Actin Gene

以油茶、沙棘、文冠果和油用牡丹的成熟种子为材料,利用实时荧光定量PCR分析了7个内参候选基因GAPDH、Actin、TUB、eEF1α、RPL4、UBQ和e IF4A在四种木本油料中的表达稳定性,并进一步验证了Actin基因在沙棘不同发育期种子和果肉中的表达。结果表明,6个内参候选基因GAPDH、TUB、eEF1α、RPL4、UBQ和e IF4A的表达水平在四种本油料种子间存在明显差异(p<0.05),内参候选Actin基因在四种木本油料种子中的表达较稳定(p>0.05)。Actin基因在沙棘种子和果实的不同发育期表达稳定(p=0.380和p=0.603),且在果肉和种子间在各个时期不存在明显差异(p>0.05)。这不仅可为沙棘果实不同器官基因表达差异研究提供内参基因,而且可为四种木本油料基因表达研究内参基因的选择提供科学参考。

The mature seeds of four woody-oil trees（C amellia oleifera,Hippophae rhamnoides,Xanthoceras sorbifolia and Paeonia suffruticosa） were selected as materials,the expression stability of seven candidate reference genes GAPDH,Actin,TUB,eEF1α,RPL4,UBQ and eIF4A were analyzed by qRT-PCR.We also further tested the expression of Actin gene in seeds and pulp of different developmental stages of sea buckthorn（Hippophae rhamnoides）.The results showed that there were significant differences（p〈0.05） in the expressions of GAPDH,TUB,eEF1α,RPL4,UBQ and e IF4A among seeds of four woody-oil trees,but the expressions of Actin gene were relatively stable（p〉0.05） in four seeds.The expressions of Actin gene were stable in different developmental stages of seeds and pulp of sea buckthorn（p=0.380 and p=0.603,respectively）,and there was no significant difference in expressions between pulp and seeds at different development stages（p〉0.05）.These data not only could provide the reference gene for the analysis of different gene expressions in different organs of sea buckthorn fruits,but also provide scientific reference for the selection of reference genes in studying the gene expression of four woody-oil trees.