Biotechnology Bulletin ›› 2021, Vol. 37 ›› Issue (8): 284-293.doi: 10.13560/j.cnki.biotech.bull.1985.2020-1467

Previous Articles     Next Articles

Construction of a Strain with Fluorescence Labeling of Cytoskeleton in Colletotrichum gloeosporioides

LIU Na(), LIU Shi-ke, WANG Qian-nan()   

  1. Hainan Key Laboratory for Sustainable Utilization of Tropical Bioresources,College of Tropical Crops,Hainan University,Haikou 570228
  • Received:2020-12-01 Online:2021-08-26 Published:2021-09-10
  • Contact: WANG Qian-nan E-mail:1032047781@qq.com;wangqiannan@hainanu.edu.cn

Abstract:

Colletotrichum gloeosporioides spp. is fungal pathogen that devastates crop plants worldwide. The structure,reorganization and polarization of cytoskeleton play essential roles in growth,morphogenesis and pathogenicity of filamentous fungi. However,the organization and dynamics of cytoskeleton during hyphae growth and infection of C. gloeosporioides is known little. To construct fluorescent labeled strains in the microfilament and microtubule skeleton of C. gloeosporioides,Lifeact-EGFP,MBD-EGFP and CgTUB1-EGFP labeled recombinant strains were successfully constructed by random recombination and homologous recombination. Laser confocal microscopy showed that Lifeact-EGFP clearly labeled the microfilament skeleton of C. gloeosporioides,and CgTUB1-EGFP clearly labeled the microtubule skeleton;whereas MBD-EGFP was not applicable for microtubule labeling. Further phenotypic analysis suggested that the fluorescence labeling of cytoskeleton by Lifeact-EGFP and CgTUB1-EGFP did not impact the normal physiological function. These 2 labeled strains can be used for further experiment,and lays a foundation for further investigating the dynamics of cytoskeleton and the functions of cytoskeleton-binding proteins.

Key words: Colletotrichum gloeosporioides, cytoskeleton, fluorescence labeling