Molecular Characterization of Down-regulated Gene At2g47500 Associated with myb26/mail sterile35 Mutant

doi:10.7668/hbnxb.2014.05.013

Abstract

Abstract: We have used mutants that are defective in the production of pollen as a means to study anther and pollen development.This has identified genes that are involved in pollen development and are likely to function in the same network.Previously,the Arabidopsis thaliana MYB26/MALE STERILE35(MS35)gene regulated endothecial cell development within the anther and acts upstream of the lignin biosynthesis pathway.Gene(At2g47500)related to microtubule identified from microarray analysis of ms35/myb26 mutant showed down-regulated gene expression in mutant were characterized by bioinformatics analysis.Insertional mutants was identified in knockout lines of Arabidopsis GenBank.Homozygous lines screened were then tested for altered expression of the genes of interest by reverse-transcriptase(RT)-PCR.Lines that show down-regulation were phenotypically characterized for changes in development.RT-PCR expression analysis suggested that theAt2g47500 was expressed in the anthers and confirmed using transformed promoter:GUS reporter constructs into wild type Arabidopsis plants by Agrobacterium transformation and analysis.Over-express At2g47500 in wild type plant and compensating in knock out lines,there was no much effect on general plant growth and specifically on pollen development and only recover expression in mutant.Quantity real time-PCR analysis some genes related to microtubule in mutant,expression as normal and similar to compare with some genes related to the secondary wall expression in myb26 buds(immature young and old stage).At2g47500 impact on pollen development is not obvious,it turns into mutant,the amount of gene expression,no performance difference.To sum up,the gene plays a secondary role in the anther development,and regulated by the main gene in anther.

Key words: At2g47500, ms35/myb26, Molecular identification, GUS reporter, Over-express of At2g47500

CLC Number:

FAN Xin-ping, QIAO Yong-sheng, NIU Xi-wu, Zoe A Wilson. Molecular Characterization of Down-regulated Gene At2g47500 Associated with myb26/mail sterile35 Mutant[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2014, 29(5): 71-79. doi: 10.7668/hbnxb.2014.05.013.

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