Selection Of Reference Genes And Analysis Some Related Gene Expression In Soybean Under Aluminum Toxicity

Soybean (Glycine max L.) belong to legumes, plays an important role in the production and human life. Studies have shown that acid soil was the the main factor that affected the crop yield in Guangxi region, aluminum toxicity is the most. Using reference genes to normalized the expression of ALMT and MATE gene, related genes of glutathione metabolism and callose metabolism. In recent research showed that there is none of them can expressed stability in all experiment condition. It is necessary to vertify the reference’s stability in our experiment.In this study, the soybean variety Brazil No.10 (BX10) and soybean variety BD2 were cultivated in 1/5 Hoagland contained with 0 mmol/L,0.05 mmol/L AICl3. Root tips were collected at three sampling times(6 h,2 d,12 d after Al treatment). Using four methods(△Ct approach, GeNorm, Normfinder, BestKeeper) to analysis the stable gene in soybean from 22 candidate reference genes. Results are as follows:(1) In ACt approach’s result MTP was the most stable one, ABC gene was the worst. (2) In GeNorm’s result:Peptidase_S10 and Peptidase_S10 were the most stable genes in all samples, the UNK1 was the worst. Analysis the gene expression stable in BX10 and BD2 separately. In BX10 sample, Peptidase_S10 and CYP were the most, UNK1 was the worst. In BD2 sample, the most were RBO and UBC2, the worst was ABC gene. (3) In NormFinder’s result, UBC2 was the most stable one, UNK1 was the most unstable one in all samples. In BX10 samples, MTP was the best and UNKlwas the worst. ELF IA was the most stable one and Letin was the worst in BD2 samples. (4) The result of BestKeeper’s as follows:Pept_S16 was the best and ABC was the worst in all samples, TIF was the most stable one and UNK1 the worst in BX10, Pept_S16 the best and ABC the worst in BD2.Selection MTP and UBC2 as reference genes to normalized the expression of ALMT, MATE, glutathione metabolism related genes (hGSHS, hPCS, y-ECS, GPX, GR, GST)and callose metabolism related genes(Ca/S2, CalS3, CalS5, CalS7, CalS8, CalS9, CalS10, CalS11, CalS12, BG1, BG2). The result showed that the expression of ALMT, GST, CalS7, CalS8 and some genes changed significantly under aluminum toxicity compared with control. Illustrated that they participate mechanism of plant aluminum resistance. The same gene expression result remain distinction when using different reference gene. It is more reliable when using more reference genes to normalize gene expression.