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Agrobacterium-Mediated Transformation of Diaporthe schini Endophytes Associated with Vitis labrusca L. and Its Antagonistic Activity Against Grapevine Phytopathogens
Ist Teil von
Indian journal of microbiology, 2019-06, Vol.59 (2), p.217-224
Ort / Verlag
New Delhi: Springer India
Erscheinungsjahr
2019
Quelle
Alma/SFX Local Collection
Beschreibungen/Notizen
Fungus-caused diseases are among the greatest losses in grapevine culture. Biological control of pathogens by endophytes may be used to decrease fungicide application rates and environmental impacts. Previously,
Diaporthe
sp. B46-64 and C27-07 were highlighted as antagonists of grapevine phytopathogens. Herein, molecular multigene (ITS-TUB-TEF1) identification and phylogenetic analysis allowed the identification of these endophytes as belonging to
Diaporthe schini
species.
Agrobacterium tumefaciens
-mediated transformation was employed for obtaining 14 stable and traceable
gfp
- or
DsRed
-expressing transformants, with high transformation efficiency: 96% for the pFAT-GFP plasmid and 98% for pCAM-DsRed plasmid. Transformants were resistant to hygromycin B with gene
hph
confirmed by polymerase chain reaction and proved to be mitotically stable, expressing the fluorescent phenotype, with morphological differences in the colonies when compared with wild strains. In vitro antagonism tests revealed an increased antagonistic activity of some transformant strains. The current genetic transformation of
D. schini
mediated by
A. tumefaciens
proved to be an efficient technique within the randomized insertion of reporter genes for the monitoring of the strain in the environment.