Functional Dissection Analysis Of The 370bp Sequence Promoter Of Myrosinase Associated Protein Gene

In the Cruciferae plant there is a special substrate-enzyme system, namelyglucosinolate - myrosinase system. In vivo, Myrosinases usually occur in complexeswith myrosinase-binging protein and myrosinase-associated protein, iMyAP is one ofmyrosinase-associated proteins present in vegetative organ, which can be promotedby iMyAP promoter induced by wounding, MeJA, JA and the ABA treatment. Theresult investigated by Ying Ruan showed that 370bp fragment is enough for finishingthe'induced function of whole promoter. The seedlings with 370bp fragment can startup the GUS gene expression after being treated by wounding and methyl jasmonicacid. It was suggested that the 370bp sequence was the core promoter of iMyAP genepromoter and had the GUS gene express specifically in the guard cell. In order to laygood foundation for finding out the new guard-cell specific motif, we dissected the370bp fragment of iMyAP gene promoter.We cloned three fragments from the iMyAP promoter. The first one is from-370bp to -1bp. The second is from -308bp to -1bp.And the third is from -150bp to-1bp. We inserted the three fragments just before GUS gene and transferred thefusions into Arabidopsis genome. Then the functional structure of this 370bpfragment was analyzed on the arabidopsis heredity background. We have transferredthese fragments into the Arabidopsis. The result reveals that these three promoterfragments can all promote the GUS gene expression without any inducement.MA are the seedlings with 370bp fragment, MB are the seedlings with 308bpfragment, and MC are the seedlings with 150bp fragment. MA are the dyed deepest inthese three transferred seedlings. And the GUS gene is specifically expressed in theguard cell of MA. The root and stem of MA are also dyed. The join of root and stemis dyed deepest in MA. But the 308bp fragment cannot promote the GUS geneexpress in the guard cell. 150bp fragment also cannot promote the GUS gene expressspecifically in the guard cell. From this result, we presumed that there is a motif in the-308bp to -370bp, and it is related with guard cell specifically expression. This resultlays foundation for finding new mitif, and the motif is related with the guard cell specifically expression.