Cloning and Expression Analysis of 3′ End Region of HbbHLH Gene in Bark from Hevea Brasiliensis

[Objective] The aim was to lay the foundation for further understanding the molecular mechanism of the jasmonate acid inducing laticifer differentiation in Hevea brasiliensis.[Method] The EST sequence of HbbHLH gene in bark from H.brasiliensis was obtained from the first strand cDNA by reverse transcript mRNA from the bark of H.brasiliensis by using the primers designed according to the conservative region of bHLH transcription factor gene and the 3′ end region of this gene was cloned by rapid amplification cDNA ends(RACE) technique using nested primers designed according to the obtained sequence.Further more,the expressions of the HbbHLH gene treated with jasmonate acid after 4 h,8 h,1 d,2 d and 3 d were analyzed by semi-quantitative RT-PCR.[Result] A 1 084 bp cDNA fragment of the 3′ end region from HbbHLH gene was obtained by clone and the sequence analysis showed that the gene belonged to the HLH domain from MYC type.The expression analysis result showed that after the treatment with jasmonic acid,the expression level of HbbHLH gene was increased gradually within 8 hours,and kept stable at a high level after treatment with 1 d and 2 d,but decreased at 3 d.[Conclusion] The jasmonic acid had the regulatory effect on the expression of HbbHLH gene.