Identification of reference genes for expression analysis by real-time quantitative PCR in drought-stressed soybean

Pesquisa Agropecuária Brasileira

Renata Stolf-moreira²  Eliana Gertrudes De Macedo Lemos²  Ricardo Vilela Abdelnoor¹  Magda Aparecida Beneventi¹  Amanda Alves Paiva Rolla¹  Selma Dos Santos Pereira¹  Maria Cristina Neves De Oliveira¹  Alexandre Lima Nepomuceno¹  Francismar Corrêa Marcelino-guimarães¹ 

[1] ,Universidade Estadual PaulistaJaboticabal SP

关键词: Glycine max;    expression stability;    RT-qPCR;    water deficit;    Glycine max;    estabilidade de expressão;    RT-qPCR;    déficit hídrico;   

DOI  :  10.1590/S0100-204X2011000100008

来源: SciELO

【 摘 要 】

The objective of this work was to validate, by quantitative PCR in real time (RT-qPCR), genes to be used as reference in studies of gene expression in soybean in drought-stressed trials. Four genes commonly used in soybean were evaluated: Gmβ-actin, GmGAPDH, GmLectin and GmRNAr18S. Total RNA was extracted from six samples: three from roots in a hydroponic system with different drought intensities (0, 25, 50, 75 and 100 minutes of water stress), and three from leaves of plants grown in sand with different soil moistures (15, 5 and 2.5% gravimetric humidity). The raw cycle threshold (Ct) data were analyzed, and the efficiency of each primer was calculated for an overall analysis of the Ct range among the different samples. The GeNorm application was used to evaluate the best reference gene, according to its stability. The GmGAPDH was the least stable gene, with the highest mean values of expression stability (M), and the most stable genes, with the lowest M values, were the Gmβ-actin and GmRNAr18S, when both root and leaves samples were tested. These genes can be used in RT-qPCR as reference gene for expression analysis.

【 授权许可】

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