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Screening of taxol biosynthesis-related genes in taxol produced from Nodulisporium sylviforme HDF-68 by mRNA differential display

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Abstract

Differential expressed genes in taxol producing Nodulisporium sylviforme HDF-68 at different periods of fermentation were analyzed by mRNA differential display. A total of 38 fragments were found expressed differentially during taxol synthesis phase. Using reverse Northern blot analysis, the hybridization signals of 16 fragments were consistent with the results of differential-display reverse transcription-PCR (DDRT-PCR). Sequencing results showed that nucleic acid sequences of TBR3, TBR4 and TBR7 were extremely similar to those of human, and the genes of cytochrome oxidase subunit I in TBR11 and Neurospora crassa mitochondrial were similar. No other functions could be identified by homolog analysis of the rest of expressed sequence tag (EST) sequences. In total, 16 open reading frames (ORFs) were found in these 16 ESTs, and seven of them have homologous proteins in the NCBI protein database. However, only the functions of TBR10 and TBR11 homolog were known; others were hypothetical proteins. Furthermore, the results also showed that the limitation of information about amplification product in 3′-UTR region could be overcome through a single random primer, instead of the anchor primer to combine with random primer in DDTR-PCR. This study provides valuable information for identifying functionally related genes in taxol-producing endophytic fungi and clues on further defining the taxol biosynthetic pathway.

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Acknowledgments

We gratefully acknowledge the Engineering Research Center of Agricultural Microbiology Technology, Ministry of Education for providing the facilities to carry out this work. This work was supported in part by the National Natural Science Foundation of China (31370137 and 31270130), Program for New Century Excellent Talents in University (NCET-12-0707), Program for New Century Excellent Talents in Heilongjiang Provincial University (1251-NCET-005), Innovative Research Team for Agricultural Microbiology Fermentation Technology in Heilongjiang Provincial University (2012td009) and Research Program Heilongjiang Education Bureau (11551377).

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Authors and Affiliations

  1. Laboratory of Microbiology, College of Life Science, Heilongjiang University, Harbin, 150080, China

    Kai Zhao, Ye Xiao, Dan Liu, Yue Zhang, Xin Wang, Xiuliang Li & Tao Jin

  2. Department of Neurosurgery, The Fourth Affiliated Hospital of Harbin Medical University, Harbin, 150001, China

    Chao Wang

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  1. Kai Zhao
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  2. Ye Xiao
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  3. Chao Wang
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  4. Dan Liu
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  5. Yue Zhang
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  6. Xin Wang
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  7. Xiuliang Li
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  8. Tao Jin
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Corresponding authors

Correspondence to Kai Zhao, Xin Wang or Tao Jin.

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Kai Zhao, Ye Xiao and Chao Wang contributed equally to this study.

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Zhao, K., Xiao, Y., Wang, C. et al. Screening of taxol biosynthesis-related genes in taxol produced from Nodulisporium sylviforme HDF-68 by mRNA differential display. Ann Microbiol 64, 1633–1642 (2014). https://doi.org/10.1007/s13213-014-0807-1

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  • DOI: https://doi.org/10.1007/s13213-014-0807-1

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