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Volume 169, Issue 4

T-DNA insertion mutagenesis in results in identification of an enolase gene mutant impaired in secretion of organic acids and phosphate solubilization Open Access

Abstract

strain P6 is a phosphate-solubilizing fungus isolated from farmland in Guangdong Province, China. To gain better insights into the phosphate solubilization mechanisms of strain P6, a T-DNA insertion population containing approximately 4500 transformants was generated by -mediated transformation. The transformation procedure was optimized by using a Hybond N membrane for co-cultivation of and . A mutant impaired in phosphate solubilization (named MT27) was obtained from the T-DNA insertion population. Thermal asymmetric interlaced PCR was then used to identify the nucleotide sequences flanking the T-DNA insertion site. The T-DNA in MT27 was inserted into the fourth exon of an enolase gene, which shows 90.8 % nucleotide identity with enolase mRNA from . Amino acid sequence homology analysis indicated that the enolase is well conserved among filamentous fungi and . Complementation tests with the MT27 mutant confirmed that the enolase gene is involved in phosphate solubilization. Analysis of organic acids in culture supernatants indicated reduced levels of oxalic acid and lactic acid for the MT27 mutant compared to the parent strain P6 or the complementation strain. In conclusion, we suggest that the identified enolase gene of is involved in production of specific organic acids, which, when secreted, act as phosphate solubilizing agents.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): enolase , phosphate solubilization and T-DNA insertion
Funding
This study was supported by the:
  • Guangdong Provincial Key Laboratory of Radionuclides Pollution Control and Resources (Award 2020PlantKF06)
    • Principle Award Recipient: XiaogeHan
  • Guangzhou Science and Technology Plan Projects People's Livelihood Special Fund (Award 202005000002)
    • Principle Award Recipient: NotApplicable
  • Guangzhou Science and Technology Innovation Center (Award 202206010058)
    • Principle Award Recipient: JuntaoZhang
  • Guangzhou Municipal Science and Technology Plan Project (Award 202102020517)
    • Principle Award Recipient: XuChangchao
© 2023 The Authors
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License. The Microbiology Society waived the open access fees for this article.
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2023-04-17
2024-05-22
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http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.001325
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T-DNA insertion mutagenesis in Penicillium brocae results in identification of an enolase gene mutant impaired in secretion of organic acids and phosphate solubilization
Microbiology 169, 001325 (2023); https://doi.org/10.1099/mic.0.001325
/content/journal/micro/10.1099/mic.0.001325
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