| Internal genes generally are used in real time quantitative PC R,as a standard reference genes,q RT-PCR because of its rapid,sensitive and quantitative characteristics,has become the common use of the analysis of the level of gene expression,Test process,however,because of the differences of the quality and quantity of RNA,c DNA synthesis efficiency and PCR amplification,the results commonly occurs serious deviation,An ideal internal gene systemwhich as the target gene expression quantity of reference objects have not yet fully established in vegetables,which leads to fundamental experimental deviation.At the same time,some recent studies have shown the traditional housekeeping gene expression is not constantin different physiological condition and d ifferent cell types,which leads to traditional housekeeping genes are questionable in real-time fluorescent quantitative study.This study through the near edge reference species,common internal gene transcription,vegetables leaf,stem tip and root is based on the set of data.This study elected 17 internal candidate genes.Useing three softwareconsist of Ge Norm,Best Keeper and Norm Finder analysis the stability of internal candidate gene expression,Seeking some stable genes and combination in all conditions.getting the results of the study In a total of 94 samplespool are:In the control group,according to the results of Ge Norm analysis:the most stable three genes are PP2AA3 、 ACT8 、 CYP2 in medium variety,the late variety is same;Best Keeper shown: the most stable three genes are PP2AA3、18S-RNA、PP2A-1 in medium variety,the late variety are PP2AA3、18SRNA、DNAJ;Norm Finder shown:the most stable three genes are PP2AA3、C YP2、ACT8 in medium variety and late variety;according to the three softwore get that PP2AA3、CYP2、ACT8 are the best stability.In the ratio of ammonia nitrate(1:3),through Ge Norm analysis the most stable three genes are PP2AA3、CYP2、ACT8 in the "69 days",and "80 days",Through variation of V deserves known the best combination of genes are PP2AA3 and C YP2;Norm Finder shown:the most stable three genes are PP2A-1、PP2AA3、CYP2 in medium variety and late variety;Best Keeper shown: the most stable three genes are PP2AA3、18SRNA、ACT3 in medium variety,the late variety are PP2A-1、PP2AA3、Tubulin.according to the three softwore get that PP2AA3、CYP2、ACT8 are the best stability.In the high temperature treatment,Ge Norm shown: the most stable three genes are ACT8、PP2A-1、GAPDH in “69 days” variety,the “80 days” variety are ACT8、ACT7、PP2AA3;Through variation of V deserves known the best combination of genes are ACT8、PP2A-1and ACT8、ACT7、PP2AA3;Norm Finder shown:the most stable three genes are ACT8、CYP2、ACT7 in “69 days” variety,the “80 days” variety are ACT8、ACT7、PP2AA3;Best Keeper shown:the most stable three genes are CYP2、ACT8、18SRNA in “69 days” variety,the “80 days” variety are ACT8、PP2A-1、UBQ10.according to the three softwore get that ACT8、ACT7、PP2A-1 are the best stability.In the low temperature treatment,Ge Norm shown:the most stable three genes are ACT2、ACT7、PP2AA3 in “69 days” variety,the “80 days” variety are PP2A-1、PP2AA3、GAPDH;Through variation of V deserves known the best combination of genes are ACT2、ACT7 and PP2A-1、PP2AA3;Norm Finder shown:the most stable three genes are CYP2、ACT3 、 PP2AA3 in “69days” variety,the “80 days” variety are PP2AA3 、GAPDH;Best Keeper shown: the most stable three genes are ACT3、PP2AA3、18SRNA in “69 days” variety,the “80 days” variety are PP2A-1、ACT3、PP2AA3.according to the three softwore get that PP2A-1、ACT3、PP2AA3 are the best stability.In the propiconazole treatment,Ge Norm shown:the most stable three genes are PP2A-1、PP2AA3、CYP2 in “69 days” variety,the “80 days” variety are GAPDH、18SRNA、ACT2;Through variation of V deserves known the best combination of genes are PP2A-1、PP2AA3、CYP2and GAPDH、18SRNA;Norm Finder shown:the most stable three genes are PP2A-1、PP2AA3、CYP2 in “69 days” variety,the “80 days” variety are GAPDH、18SRNA、ACT2;Best Keeper shown:the most stable three genes are PP2AA3、EF-TU、UBQ10 in “69 days” variety,the “80 days” variety are DNAJ、18SRNA、GAPDH.according to the three softwore get that PP2A-1、GAPDH、PP2AA3、CYP2 are the best stability.In the dark,Ge Norm shown: the most stable three genes are CYP2、GAPDH、PP2AA3 in “69 days” variety,the “80 days” variety are CYP2、GAPDH、PP2AA3;Through variation of V deserves known the best combination of genes are CYP2、GAPDH;Norm Finder shown:the most stable three genes are CYP2、PP2AA3、PP2A-1 in “69 days” variety,the “80 days” variety are CYP2、ACT8、GAPDH;Best Keeper shown:the most stable three genes are CYP2、ACT8、GAPDH in “69 days” variety,the “80 days” variety are Tubulin、EF-TU、PP2AA3.according to the three softwore get that CYP2、GAPDH、ACT8 are the best stability. |
