Diversity, Novelty and Anti-tumor Activity of Actinomyces from Coastal Mangrove in Leizhou, Zhanjiang
LI Jun-Jie1, LIN Man-Jia4, CHEN Hai-Ming1,2,3, LIU Yue-Li1, LIANG Xiao-Yan1, HUANG Yong-Mei1,2,3,*, LUO Hui1,2,3,*
1 Marine Biomedical Research Institute, the Key Lab of Zhanjiang for R&D Marine Microbial Resources in the Beibu Gulf Rim, Guangdong Medical University, Zhanjiang 524023, China; 2 Southern Marine Science and Engineering Guangdong Laboratory (Zhanjiang), Zhanjiang 524023, China; 3 The Marine Biomedical Research Institute of Guangdong Zhanjiang, Zhanjiang 524023, China; 4 First Clinical Medical College, Guangdong Medical University, Zhanjiang, 524023, China
Abstract：Most of mangroves grow in intertidal zones with special and changeable environments, which are conducive for the growth of rare Actinomyces and production of active compounds. In order to explore the diversity, novelty and medicinal potential of Actinomyces resources from coastal mangrove in Leizhou, Zhanjiang, China. Pour plate method and 10 different media were used in this study to isolate Actinomyces from mangrove rhizosphere soil samples. After purification and culture, PCR amplification and sequencing and phylogenetic analysis of 16S rRNA of Actinomyces strains were performed. At the same time, according to the comparison results of 16S rRNA and the colony morphological characteristics of strains, portion of Actinomyces strains were selected for small-scale fermentation, all broths without mycelia were extracted by ethyl acetate and dried by rotary evaporator before the preparation of fermentation crude extract methanol solution. Anti-tumor activity of fermentation crude extracts against human lung cancer cell line A549 and human liver cancer cell line HepG2 were analyzed by Cell Counting Kit-8 (CCK-8). According to the result, a total of 154 Actinomycetes were isolated from 8 rhizosphere soil samples collected from 3 different sampling sites in coastal areas of Zhanjiang Mangrove National Nature Reserve, Guangdong Province, distributing in 11 orders, 15 families and 19 genera, with the dominant genera were Micromonspora (22.7%, 35 strains), followed by Microbacterium (20.8%, 32 strains) and Streptomyces (10.4%, 16 strains). And there were 7 potential new species distributing in 4 genera with 16S rRNA sequence similarity less than 98.65%. In the anti-tumor activity screening of fermentation crude extracts, 53 strains of Actinomyces showed anti-tumor activity against at least one tumor cell line, accounting for 79.1% of 67 strains. The above results suggested that, the diversity of Actinomyces and potential medicinal resources were rich in coastal mangrove areas of Leizhou. This study bases a sufficient strain foundation for the further improvement of the cognition of Actinomyces distribution in Leizhou coastal areas as well as the subsequent research on other biological activities such as antibacterial activities and related mechanisms.
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