title: [Development of genetic transformation system of Valsa mali of apple mediated by PEG].
abstract: OBJECTIVE
  The genetic transformation of Valsa mali var. mali was developed by PEG-mediated protoplasts transformation.
  
  
  METHODS
  It was transformed by PEG-induced fusion of protoplasts. The plasmid pBIG2RHPH2-GFP-GUS carrying hph gene was used and Valsa mali var. mali 03-8 isolate was used as the host strain.
  
  
  RESULTS
  At 50 mg/mL driselase + 10 mg/mL lysing enzymes concentration, the mycelium of Valsa mali var. mali cultured in YEPD medium for 48 h was hydrolyzed in 10 mL enzymes liquid /0.5 g wet mycelium for 2 h. The protoplast yield was 4 x 10(7) CFU/mg. The transformation efficiency was 44 per g DNA. Analysis of the transformants by PCR and Southern blotting showed that the selectable marker gene hph was integrated effectively into the genome of Valsa mali var. mali. After 5 subculturing on PDA, 87.5% transformants could grow. This stability test of transformants suggested that the foreign gene hph was stable in heredity.
  
  
  CONCLUSIONS
  This transformation system is a valuable and important tool for the further study of the pathogenic gene of Valsa mali.
authors: National Key Laboratory of Crops Stress Biology for Dryland and College of Plant Protection, Northwest A&F University, Yangling 712100, China. gaojingshai@yahoo.cn
authors: Jing
authors: J
authors: Gao
authors: Yanbo
authors: Y
authors: Li
authors: Xiwang
authors: X
authors: Ke
authors: Zhensheng
authors: Z
authors: Kang
authors: Lili
authors: L
authors: Huang
mesh_terms: D001203:Ascomycota / Q000235:genetics*; D027845:Malus / Q000382:microbiology; D011092:Polyethylene Glycols / Q000737:chemistry*; D011523:Protoplasts / Q000378:metabolism; D014170:Transformation, Genetic
chemical_list: D011092:Polyethylene Glycols