Cloning And Interactions Of HbJAZs Of Hevea Brasiliensis

Hevea brasiliensis is the main source of natural rubber. Laticifer is the main organ of latex synthesis and storage. Mechanical damage or treatment of exogenous jasmonate acid (JA) can induce laticifer differentiation. And JAZ protein is a key repressor that regulates the downstream transcription factors of JA singnal pathway. Identification of JAZ proteins is important to the mechanism of rubber production and laticifer diffferenciation. This work is trying to clone and characterize the JAZs of Hevea brasiliensis (HbJAZs) as follows:1 Combined with the cDNA sequences deposited in GenBank and RNA-seq data of our lab, five full-length cDNA sequences of new JAZ gene were predicted by in silico procedure for H. brasiliensis, of which four genes were successfully amplified by PCR and termed as HbJAZ3, HbJAZ5, HbJAZ6, and HbJAZ12, and the size of the coding regions is852bp,786bp,426bp,699bp.2 Using real time quantitative PCR, we found that that these genes were differentially expressed in different tissues such as latex, leaf, stem-tip, bark, stamen and pistil, we found that repression of these four genes are influenced by wounding treatment and jasmonic acid treatment. HbJAZ3 expresses most in stamen and pistil, HbJAZ5 expresses most in bark, HbJAZ6 expresses most in bark and stamen, HbJAZ12 expresses most in pistil.All the four genes are up regulated by wounding treatment. The expression of HbJAZ3 and HbJAZ12 is induced by JA treatment, while the expression of HbJAZ5 and HbJAZ6 is repressed by JA treatment.3 The bait and prey vectors for HbJAZ3,5,6,12 were constructed, respectively. Together with previously constructed bait and prey vectors of HbJAZs gene, the interactions within HbJAZs were analysed by yeast two hybrid. Results show that HbJAZ2, 5,6 interacted as homodimers, and heteromeric interactions were observed among HbJAZ1, 2,3,5,6,8,9,10,11,12.4 Interactions between HbCoi1 and all the HbJAZs were observed by yeast two hybrid using BD bait vector of HbCoi1 and AD prey vectors of HbJAZs. Results show that, HbCoi1 interacts with HbJAZ8 and HbJAZ9 when coronatine is added, suggesting that HbCoi1 regulated the downstream signal through interation with HbJAZs proteins.