Screening Of Proteins Interacting With PhMYC2 And Identification Of JAZ Family In Petunia

Petunia hybrida Vilm.,an important bedding flower,has become a modle plant for flower development study because of its showy flower,short lifecycle,convenient gene transformation system and clear genetic background.A transcription factor,PhMYC2 that could regulate the expression level of PhGRP which had a significant impact on anther development was cloned.The JAZ family is known to regulate the expression level of MYC2.So we studied PhMYC2 and PhJAZ further,and the key results are as follows:1.The transcription level and transcription activation analysis of PhMYC2.We analyzed the expression level of PhMYC2 in the different tissues development stages of flower buds and anthers.It had a highest expression level in leaves,higher level in flowers,while relative week level was detected in the roots and stems.In the different stages of flower buds,the expression level of PhMYC2 were detected significantly in 0.2b and 2.5b;PhMYC2 expressed in the earlier stages of anthers and had a peak in the 0.5a.It couldn’t was detected in the anaphase of anthers.Except the TAD,which played an important role on the transcription activation,there was another domain controlling the transcription activation,corporately.The 100 aa at the N-terminal of this protein was responsible for this phenomenon.At last we construct the bait vector,C562BD.2.The construction and screening of the Y2H library.We collected the RNA from the anthers of the flower buds that length were 0.2cm,0.3cm,0.5cm,1.0cm and 1.5cm to construct the two-hybrid library.The titer of this library was 2.2x10~7cfu.The inserted fragment ranged 400bp from 2000bp.The rate of recombination was about 100%.Then the vector,C562BD,was used as the bait to screen the library.We gained 13sequences that were 9 genes,and blasted them on the web of Petunia hybrid genome.3.Identification the JAZ family of petunia.We identified 16 and 12 JAZ genes from two ancestor of petunia,P.inflata and P.axillaris,respectively.This genes could be clustered into 4 classed.The genes that had near genetic relation usually had similar motifs and gene structures.The JAZs of P.axillaris had balanced expression in the roots,stems,leaves,flowers expect a few specific expression gene.Most of this genes had the phenomenon that the relative expression was significant in the early stages in the flower buds,while that couldn’t be detective in the initial stage of anthers.