Biotechnology Bulletin ›› 2017, Vol. 33 ›› Issue (3): 100-105.doi: 10.13560/j.cnki.biotech.bull.1985.2017.03.015

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Labeling and Tracing of Green Fluorescent Protein in Fungal Endophyte with Growth-promoting Activity to Rice Seedlings

CHEN Nan, YU Fei, HE Yan-liu, BU Ning   

  1. College of Life Sciences,Shenyang Normal University,Shenyang 110034
  • Received:2016-07-06 Online:2017-03-26 Published:2017-03-07

Abstract: This work is to transform the gene of green fluorescent protein(gfp)into the fungal endophyte JP4-1 of Suaeda salsa,and to detect the colonization of the fungi in rice seedlings. The plasmid pCT74 with gfp was transformed into the protoplasts with PEG-CaCl2 mediated method and then integrated with the strain for transformants. Further,the transformants was used to infect rice seedlings,and finally the tracer to JP4-1 and its infection characteristics were detected with fluorescence microscope. Results showed that the transformants were quite stable with promising green fluorescence intensity after six successive subcultures. PCR amplification showed that the gfp was successfully transformed into strain JP4-1 and was expressed stably in rice seedlings. In conclusion,the JP4-1 transformants with stably-expressed gfp was acquired via transformation,the JP4-1 was colonized in the roots,shoots and leaves of rice seedlings,the location of colonization was intercellular,and the growth-promoting activity showed no difference with the wild type.

Key words: fungal endophyte, green fluorescent protein, protoplasts, transformation, colonization