Transformation And Primarily Functional Analysis Of Blast-resistance Gene Pi-y43(t) From Japonica Variety Yunyin

The blast resistance gene Pi-y43(t)was cloned from japonica variety with broad spectrum blast resistance,Yunyin.The blast-resistance gene Pi-y43(t)is alleles with the Pib.In addition,there is a pseudogene Pib2 near the loci in the japonica variety Yunyin genome.In order to verify the function of the blast resistance gene Pi-y43(t)of YY,the following studies were mainly carried out:1.The overexpression vector with Pi-y43(t)was constructed,the Nipponbare and LTH were transformed with the construct overexpressing the gene Pi-y43(t)mediated with Agrobacterium.The corresponding overexpressing transgenic plants were obtained.Meanwhile,the transgenic plants with blast-resistance Pi-y43(t)gene knockout were obtained from the variety Yunyin with CRISPR/Cas9.Moreover,the protein of Pi-y43(t)was positioned at the nucleus and cell membrane by protoplast co-transformation under the laser confocal scanning microscopy.2.The corresponding transgenic plants were analyzed.The transgenic plants were preliminarily identified by hygromycin primers and the blast-resistance Pi-y43(t)gene primers,respectively,the results indicated that the transgenic plants with the blast-resistance Pi-y43(t)gene were obtained mediated by Agrobacterium tumefaciens.Meanwhile,the transgenic plants with the blast-resistance Pi-y43(t)gene knockout were also obtained.3.The phenotype of transgenic plants were identified inoculated with spraying isolate Sichuan-43.The results indicated that the plants with overexpressing the bla st-resistance gene Pi-y43(t)were resistance to isolate Sichuan 43.The transgenic pl ants with the blast-resistance Pi-y43(t)gene knockout were susceptible to isolate Si chuan 43.