Reference Genes Selection for Quantitative Real-time PCR in Chinese Cabbage-Cabbage Translocation Lines

doi:10.7668/hbnxb.2018.05.008

Abstract

Abstract: In order to analyze the gene expression more accurately in Chinese cabbage by quantitative Real-time PCR,Chinese cabbage translocation lines added No.4 chromosome fragments from cabbage under Chinese cabbage background were used as materials in this experiment. The leaves of Chinese cabbage-cabbage translocation lines in different growth stages,including seedling stage,rosette stage,folding stage and heading stage,floral buds of different sizes and leaves of folding stage treated with auxin(Indole-3-acetic acid,IAA)and auxin inhibitor(2,3,5-triiodobenzoic acid,TIBA)were selected to evaluate the expressions stability of genes. Expression stability of twenty one candidate reference genes were detected by qRT-PCR,including Apr, BcTIP41, U34559, EF1α, TUB4, CYP, DNAJ, HIS, TUA5, UKN1, SKIP16, CAC, ACTIN, ACTIN-1, ACTIN-2, GAPDH, UBC30, UBQ, PPR, PP2A, MDH. geNorm and NormFinder were used to analyze above these results,which showed that the qRT-PCR most suitable reference genes of Chinese cabbage-cabbage translocation lines had some differences in different development periods,different tissue of the material,and under condition of different hormone treatment. The UKN1 and TUB4 genes were most stable expression in vegetative growth stage(from seedling to heading)of Chinese cabbage-cabbage translocation lines. In folding stage of Chinese cabbage-cabbage translocation lines,the most stable expression reference genes were BcTIP41 and ACTIN after treatment with auxin IAA,and the most stable expression reference genes were UKN1 and BcTIP41 after treatment with auxin inhibitor TIBA. DNAJ, ACTIN and PP2A genes showed the most stable expression in the floral buds with six size levels of Chinese cabbage-cabbage translocation lines. These results will lay the foundation for the accurate analysis of the gene expression of Chinese cabbage-cabbage translocation lines,and further provided a reference for the selection of the reference gene in other plants of Brassica at different developmental stages and hormone treatments.

Key words: Chinese cabbage, Translocation lines, qRT-PCR, Reference gene, Different developmental stages, Hormone treatment

CLC Number:

CUI Feifei, MENG Chuan, WANG Yanhua, ZHAO Jianjun, CHEN Xueping, SHEN Shuxing, GU Aixia. Reference Genes Selection for Quantitative Real-time PCR in Chinese Cabbage-Cabbage Translocation Lines[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2018, 33(5): 60-67. doi: 10.7668/hbnxb.2018.05.008.

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