Antidisease Antisenescence And Apetalous Brassica Napus Through Plant Genetic Engineering

Sclerotinia stem rot that caused by Sclerotinia sclerotiorum is the most devastating disease of rapeseed (Brassica napus L.) in China especially in Yangtze River Area and leads to great yield loss every year. Glucanase and AP24, which come from tobacco both, have broad spectrum of antifungal activity. AP24 and Glu were combined with CaMV35S promoter and were transferred into Brassica napus via Agrobacterium tumefaciens. We took two different inoculation methods during two different periods to estimate the resistance to Sclerotinia sclerotiorum of T2 and Ta progenies. That is the detached leaves inoculation and inoculation of stem by tooth sticks clinged with pathogene. The plants exhibited an increased resistance against the fungal attack compared with the no transgenic plants during two inoculations. The candidate plants were conducted with RT-PCR and Northern blot analysis. The result indicated the increase in the transcriptional level of foreign genes.A method for in situ transformation-in planta transformation was also preceded. We attempted this new transformation method on a variety of Brassica napus, Hua Shuang No.3, and collected 1,800 seed grains after treated with Agrobacterium tumefaciens containing the chimeric PsAGn-IPT gene. The harveated seeds were screened on MS medium with Kanamycine. The Km -resistant seedlings were transplanted into soil in a greenhouse then in the field and developed into 70 adult plants. So, 3.8% of in-planta seeds were grown to mature plants. PCR analysis showed that 16 among 70 plants had been integrated into the PsAGn-IPT. Several agronomy traits on the 16 TO plants were investigated including the plant height, green leaf number on main stems, flower number, biomass, seed yield and weight of seed grains etc. The highest green leaf number that the plant can reached of each plant varied much, from 15 to 56 among the 16 plants. The plant height ranged from 1.23 to 1.83m. The weight of seed grains varied from 2.27 to 3.95g/1000grains. The flower number ranged from 161 to 605. Biomass changed much from 21.4 to 104.22g. We chosed a nicer plant from TO generation, and the following field investigation also displayed favorable characters such as the green leaf number, biomass etc.With the same in planta transformation method we got the transgenic gagal plants. During the flowering time the apetalous flower can be found. But some bad characters were detected at the same time, which was depicted as following: the shorter plant shape, the fewer branch, little flower, less seed yield et al.