Establishment Of Banana And Longan Transgenic Receptor System And The Preliminary Study On The Transformation Of The PEAS Gene

In this experiment, a high efficient receptor system was established using the slices induced from the thin cell layer of Musa spp. cv. Tianbaojiao(AAA group) and the long-term maintained embryogenic call! (EC) induced from immature embryos of Dimocarpus longan Lour. cv. Honghezi, and then the preliminary study on the introduction of PEAS gene mediated by Agrobacterium tumefaciens was performed. The main results were described as follows:1) An appropriate receptor system for banana genetic transformation was established. The high regeneration system was established by thin-cell-layer culture.It was found that the high differentiation of banana shoots was obtained from the slices cultured at 24C in light for 5d after they had been cultured at 30*C in darkness for 15d and, which grew well and was suitable for the receptor system; the sensitivity of banana thin cell layer to carbenicillin was limited, and it did not restrain the growth of banana, which could be used as the bacteriostasis for banana transformation; and banana was sensitive to hygromycin B, and it remarkably restrained the growth of banana., which could be used as the screening agent, and the concentration of 50mg/L for hygromycin B was the effective concentration for banana transgenic selection.2) An appropriate receptor system for longan genetic transformation was established. Calli pre-cultured for 10-15 d were proved at their rapid-growth stage and their strong vitality stage through the measurement of EC fresh weight increase and suistable for transgenics. The sensitivity of longan EC to hygromycin B was evaluated through the method of the measurement of fresh weight increase combined with qualitative obserbation, which showed that high concentration of hygromycin B obviously depressed the growth of longan EC. The concentration of 50mg/L for hygromycin B was the effective concentration for longan EC transgenic selection.3) The preliminary study was performed on the transformation of banana and longan. The target gene PEAS was introduced to the thin cell layer of banana and EC of longan mediated by Agrobacterium tumefaciens with hpt gene. After infection the materials were selected on MS medium supplemented with hygromycin B, finally 10lines of banana and 5 cell lines of longan were obtained. PCR assays indicated that hpt gene had been integrated into both the genomes of the 2 lines from 15 tested banana plantlets and one line from 5 tested longan lines. Something about the assays of PEAS gene> Southern blotting, Northern blotting and the expression of the transplants in field were to be further studied.The establishment and the preliminary study on transformation of thin cell layer of banana and EC of longan would be the technique plat of genetic transformation of banana and longan, which could be used for genetic improvement of banana and longan.