Construction of a T-DNA Insertional Mutant Library of Colletotrichum gloeosporioides of Mango and Its Molecular Analysis

Mangifera anthracnose,caused by Colletotrichum gloeosporioides,is one of the most damaging phytopathogens of mango(Mangifera indica L) in the world.A highly efficient Agrobacterium tumefaciens-mediated transformation of C. gloeosporioides was established. Strain SC2 of C. gloeosporioides was mutated through T-DNA insertion mediated by A. tumefaciens strain Agl-1. The transformation efficiency ranged from a low of 300 to a high of 980 transformants/106 conidia,and 4 680 transformants were obtained. The PCR amplification found that 38 tested transformants randomly selected were all resistant to hygromycin B and contained hph gene,indicating that the transformants were T-DNA inserted mutants. Southern blotting analysis showed the 20 T-DNA inserted mutants with single copy insertion accounted for 75% (15 mutants) of the total.