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journal1 ›› 2018, Vol. 34 ›› Issue (6): 937-944.DOI: 10.16409/j.cnki.2095-039x.2018.06.019

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Analysis of Fungal Diversity on Grape Leaves after Spraying Bacterial Agents by PCR-DGGE

GUO Jiping1, MA Guang1, ZHANG Zhiqiang1, SU Changqing1, QI Shanhou2   

  1. 1. Department of Life Science, Hengshui University, Hebei 053000, China;
    2. Hengshui University Hospital, Hengshui 053000, China
  • Received:2018-05-05 Online:2018-12-08 Published:2018-12-08

Abstract: PCR-DGGE technology was used to explore the variation of fungal diversity on the surface of grape leaves after spraying Microbacterium testaceum Y24 and Bacillus amyloliquefaciens N22 agents. The 18S rDNA high variation area of the fungus with the size of 400 bp was amplified with the sample genome DNA as the template. After DGGE, the map and the marker strips were analyzed. The bands were cut, reclaimed, cloned and sequenced to determine the characteristic value and composition of fungal diversity. The DGGE map analysis showed that the species richness, Shannon Wiener index of the leaf samples spraying Y24 was the highest, and the two indexes of samples spraying N22 was not significantly changed compared with those of the control. The sequence alignment of 25 bands marked on the DGGE map showed that the fungi on the grape leaves were mainly unculturable Alternaria alternata, Pleospora herbarum, Candida parapsilosis and unculturable eukaryotes. The specific fungal groups of leaf spraying Y24 were unculturable fungi, Cryptococcus sp. and unculturable eukaryotes. The fungal diversity on leaves treated with Y24 was more abundant than the control and N22 agent, and there was no difference between N22 and control.

Key words: grape leaves, bacteria agent, fungi, diversity, PCR-DGGE

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