TY - JOUR
T1 - Efficient gene knockout in the maize pathogen Setosphaeria turcica using Agrobacterium tumefaciens-mediated transformation
AU - Xue, Chunsheng
AU - Wu, Dongliang
AU - Condon, Bradford J.
AU - Bi, Qing
AU - Wang, Weiwei
AU - Turgeon, B. Gillian
PY - 2013/6
Y1 - 2013/6
N2 - Setosphaeria turcica, a hemibiotrophic pathogenic dothideomycete, is the causal agent of Northern Leaf Blight of maize, which periodically causes significant yield losses worldwide. To explore molecular mechanisms of fungal pathogenicity and virulence to the host, an efficient targeted gene knockout transformation system using Agrobacterium tumefaciens was established with field collected strains. The starting materials, incubation time, induction medium type, Agrobacterium cell density, and method of co-incubation were optimized for deletion of 1,3,8-trihydroxynaphthalene reductase, a gene in the melanin biosynthesis pathway, as a test case. Four additional genes were deleted in two different S. turcica field isolates to confirm robustness of the method. One of these mutant strains was reduced in virulence compared with the wildtype strain when inoculated on susceptible maize. Transformation efficiency was ≈20 ± 3 transformants per 1 × 106 germlings and homologous recombination efficiency was 33.3 to 100%.
AB - Setosphaeria turcica, a hemibiotrophic pathogenic dothideomycete, is the causal agent of Northern Leaf Blight of maize, which periodically causes significant yield losses worldwide. To explore molecular mechanisms of fungal pathogenicity and virulence to the host, an efficient targeted gene knockout transformation system using Agrobacterium tumefaciens was established with field collected strains. The starting materials, incubation time, induction medium type, Agrobacterium cell density, and method of co-incubation were optimized for deletion of 1,3,8-trihydroxynaphthalene reductase, a gene in the melanin biosynthesis pathway, as a test case. Four additional genes were deleted in two different S. turcica field isolates to confirm robustness of the method. One of these mutant strains was reduced in virulence compared with the wildtype strain when inoculated on susceptible maize. Transformation efficiency was ≈20 ± 3 transformants per 1 × 106 germlings and homologous recombination efficiency was 33.3 to 100%.
KW - ECP6
UR - http://www.scopus.com/inward/record.url?scp=84877951770&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84877951770&partnerID=8YFLogxK
U2 - 10.1094/PHYTO-08-12-0199-R
DO - 10.1094/PHYTO-08-12-0199-R
M3 - Article
C2 - 23384859
AN - SCOPUS:84877951770
SN - 0031-949X
VL - 103
SP - 641
EP - 647
JO - Phytopathology
JF - Phytopathology
IS - 6
ER -