The Establishment Of Transgenic System Of Amorphophallus Rivieri Species

Amorphophallus is a monocotyledonous, perennial herb, which has been exploited largely for food, medicinal, chemical uses because of rich polysaccharides in the corms. In recent years, the rapid development of processing industry has driven the Amorphophallus production and accelerated the spread of planting areas. Along with the extention of the crop, diseases are getting more important and becoming the major obstacle in the production. Because the rapid development of Amorphophallus production is based on a short listing of domestic cultivation in household level there have been neither theoretical varieties nor the technology of breeding. In addition, sexual reproduction of Amorphophallusi usually happens once every four years; and most of the seeds produced are pseudoseeds with non-sexually combined embryo. Therefore it is very difficult to breed new Amorphophallus varieties via the conventional technology.Gene engineering, as a new strategy for plant breeding, has been widely used for many species, but there is only one report about transgenic research on Amorphophallus. This study, based on the previous research on tissue culture, is aimed to establish the transgenic system of Amorphophallus, construct the vector which contains NtPRp27-like gene (pathogenesis-related protein gene), get transgenic plants via transformation by Agrobacterium tumefaciens and lay a foundation for further disease resistance improvement. The main results are as follows:1. Construction of the vector for transformation A new binary vector (pBI121-like-27) containing the target gene (NtPRp27-like) was constructed and transferred into different kinds of Agrobacterium tumefaciens strains: LBA4404, EHA105, and GV3101. The vector was proved by both enzyme digestion and PCR detection.2. The establishment of receptor system by using the callus of A. albus as explants By testing different materials, petiole from the field grown crop was the best one to induce callus, which was easy to proliferate. Medium Dl(MS+0.1 mg/L NAA+1.0 mg/L 6-BA) was the best for the differentiation of the callus and the percentage of plant regeneration was up to 39.1%. The test of antibiotic sensitivity revealed that the callus was sensitive to G418 but not to kanamycin. The suitable concentration to G418 for selecting was 30 mg/L, and to carbenicillin for bacterial inhibiting was 250 mg/L.3. The establishment of receptor system by using the petiole cuttings of in vitro plantlet of A. albus as explants The petiole cuttings(6 mm) of in vitro plantlets were inserted vertically with an upright position into A5 medium and obtained higher callus induction frequency, up to 85.0%. The percentage of plant regeneration reached 33.5%...