Reference gene selection for qRT-PCR in Caragana korshinskii Kom. under different stress conditions
Reference gene selection for qRT-PCR in Caragana korshinskii Kom. under different stress conditions
复制标题
柠条 qRT-PCR 参考基因选择。
DOI：
10.1007/s11033-014-3086-9
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发表时间：
2014-01
影响因子：
2.8
通讯作者：
Li Guojing
中科院分区：
文献类型：
--
作者：
Yang Qi;Yin Jiajia;Li Gao;Qi Liwang;Yang Feiyun;Wang Ruigang;Li Guojing
锦鸡儿，广泛分布于我国西北地区和蒙古国，是豆科植物中重要的饲用灌木，具有很高的经济和生态价值。C.对各种逆境的耐受能力强，红松科是植物逆境研究的重要物种。本研究从ACT、GAPDH、EF 1 α、UBQ、TUA、CAP、TUB、TUB 3、SKIP 1、SKIP 5 -1和SKIP 5 -2等11个候选参考基因中筛选出适合进行实时荧光定量PCR（qRT-PCR）的参考基因。对干旱、高温、低温、盐、阿坝和高pH处理下的129份样品进行了分析，并利用geNORM、Normandy和BestKeeper等软件进行了参考基因的评价和选择。在不同的胁迫条件下，选择了不同的适宜参考基因。在所有129个样本中，发现GAPDH、EF 1 α和SKIP 5 -1是最稳定的参考基因，并且EF 1 α+ SKIP 5 -1是最稳定的参考基因组合。TUA、TUB和SKIP 1在某些胁迫条件下的表达变异较大，不适合作为参考基因。使用稳定和不稳定的参考基因检测CkWRKY 1的相对表达水平，并确认其适用性。这些结果为不同胁迫条件下的柑橘qRT-PCR提供了稳定的参考基因和参考基因组合。红松科CkWRKY 1基因在C. Korshinskii。
Caragana korshinskii Kom., which is widely distributed in the northwest China and Mongolia, is an important forage bush belonging to the legume family with high economic and ecological value. Strong tolerance ability to various stresses makes C. korshinskii Kom. a valuable species for plant stress research. In this study, suitable reference genes for quantitative real-time reverse transcription PCR (qRT-PCR) were screened from 11 candidate reference genes, including ACT, GAPDH, EF1α, UBQ, TUA, CAP, TUB, TUB3, SKIP1, SKIP5-1 and SKIP5-2. A total of 129 samples under drought, heat, cold, salt, ABA and high pH treatment were profiled, and software such as geNORM, NormFinder and BestKeeper were used for reference gene evaluation and selection. Different suitable reference genes were selected under different stresses. Across all 129 samples, GAPDH, EF1α and SKIP5-1 were found to be the most stable reference genes, and EF1α+SKIP5-1 is the most stable reference gene combination. Conversely, TUA, TUB and SKIP1 were not suitable for using as reference genes owing to their great expression variation under some stress conditions. The relative expression levels of CkWRKY1 were detected using the stable and unstable reference genes and their applicability was confirmed. These results provide some stable reference genes and reference gene combinations for qRT-PCR under different stresses in C. korshinskii Kom. for future research work, and indicate that CkWRKY1 plays essential roles in response to stresses in C. korshinskii.
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影响因子：
5.3
作者：
Artico S;Nardeli SM;Brilhante O;Grossi-de-Sa MF;Alves-Ferreira M
通讯作者：
Alves-Ferreira M
影响因子：
1.1
作者：
Xuemin Wang;Zan Wang;Jie Dong;Meizhen Wang;Hong-wen Gao
通讯作者：
Xuemin Wang;Zan Wang;Jie Dong;Meizhen Wang;Hong-wen Gao
影响因子：
2.7
作者：
Dheda, K;Huggett, JF;Zumla, A
通讯作者：
Zumla, A
影响因子：
--
作者：
Huijuan Guo;Xueli He;Yingpeng Li
通讯作者：
Huijuan Guo;Xueli He;Yingpeng Li
影响因子：
1.8
作者：
Y. Wu;X. Hu;Y. R. Wang
通讯作者：
Y. Wu;X. Hu;Y. R. Wang