An efficient genetic transformation system for Chinese medicine fungus Tolypocladium ophioglossoides
Journal of Microbiological Methods, ISSN: 0167-7012, Vol: 176, Page: 106032
2020
- 6Citations
- 16Captures
- 1Mentions
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Metrics Details
- Citations6
- Citation Indexes6
- CrossRef5
- Captures16
- Readers16
- 16
- Mentions1
- References1
- Wikipedia1
Article Description
Tolypocladium ophioglossoides is a rare and valuable fungus extensively used in Chinese medicine for relieving postmenopausal syndrome in women yet its bioactive molecules are unknown. To explore its molecular mechanisms, we have developed a reliable Agrobacterium -mediated transformation system using the selective marker: the chlorimuron ethyl-resistance gene sur. For this purpose, we firstly constructed a T-DNA binary vector system and then improved the transformation efficiency by optimizing conditional parameters including the Agrobacterium tumefaciens concentration, the conidia number of T. ophioglossoides, the co-culture time and the concentration of acetosyringone. Furthermore, we have knocked-out the ku70 gene,which is a key gene in non-homologous end joining (NHEJ) DNA repair pathway,and the effect of the length of the homologous arms (HA) on the genetic transformation efficacy was also examined, which increased by 60% when HA was about 3 kb in length. Our results suggest that the genetic transformation system is efficient and feasible for the truffle-parasite fungus T. ophioglossoides, which can further be used in large-scale experiments for characterization of genes of interest in future work.
Bibliographic Details
http://www.sciencedirect.com/science/article/pii/S016770122030748X; http://dx.doi.org/10.1016/j.mimet.2020.106032; http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=85089545147&origin=inward; http://www.ncbi.nlm.nih.gov/pubmed/32805368; https://linkinghub.elsevier.com/retrieve/pii/S016770122030748X; https://dx.doi.org/10.1016/j.mimet.2020.106032
Elsevier BV
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