Moleculer Cloning And Primary Functional Study Of GmAGL15 Relative To Seed Development In Soybean

AGL15(AGAMOUS-like 15) is the only MADS domain regulatory factor identified to date that is expressed in a highly preferential manner in developing plant embryos.In this study,an AGL15-like gene was obtained from soybean by homology-based cloning strategy,here named as GmAGL15.Sequence analysis indicated that Arabidopsis AGL15 and GmAGL15 shared 61%identity at amino acid level,especially 84.3%in the MADS domains,63.2%in the K domains.RT-PCR analysis of GmAGL15 showed that it was expressed only in embryos.A real-time PCR was performed to see the expression profiling of GmAGL15 in developing seeds,and the results indicated that the expression of GmAGL15 appeared two times of peak values,at 15 DAF(days after flowering) and at 30 DAF.GmAGL15 was expressed at approximately 10-fold higher levels at the 30DAF than during any other time points of seed developing phases.Based on these data,we suggest that GmAGL15 is likely to be an important component of the regulatory circuitry directing seed specific processes in soybean developing seeds.The full length of GmAGL15 genomic is 5757bp,full length of GmAGL15 cDNA is 1253bp,There are 7 introns in GmAGL15,A lot of introns were deleted from AGL15 genes through PCR,A series of subsequent expression vectors have been used for tobacco transformation which were constructed using GATEWAY technique,and then a series of subsequent expression vectors,to have been used for tobacco transformation.The plantlets of soybean inoculated by wild type Agrobacterium rhizogenes K599,and hairy root was induced on inoculated site in vivo.The frequencies of hairy root induction from wound cotyledons of soybean were 75%,The established system should be ideal for studying soybean root function.