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A noncanonical poly(A) RNA polymerase gene affects morphology in Phoma medicaginis

Kihyuck Choi1, Stephen M Marek2

  • 1Department of Entomology and Plant Pathology, Oklahoma State University, Stillwater, OK, USA; Department of Applied Bioscience, Dong-A University, Busan, Republic of Korea.

Fungal Genetics and Biology : FG & B
|November 21, 2017
PubMed

Insights

Phoma medicaginis, a plant pathogen, has a noncanonical poly(A) RNA polymerase (ncPAP) gene, Pmncpap1, crucial for its growth. Disrupting this gene causes sensitivity to hydroxyurea, impacting fungal development.

Area of Science:

  • Plant Pathology
  • Mycology
  • Molecular Biology

Background:

  • Phoma medicaginis causes spring black stem and leaf spot disease in alfalfa and medics.
  • The fungus is amenable to Agrobacterium mediated transformation (ATMT), enabling genetic studies.
  • Random T-DNA integration during ATMT can generate useful mutants for gene function analysis.

Purpose of the Study:

  • To investigate the function of a putative noncanonical poly(A) RNA polymerase gene (Pmncpap1) in Phoma medicaginis.
  • To characterize the role of Pmncpap1 in fungal morphology and response to stress.
  • To establish Pmncpap1 as the first noncanonical poly(A) RNA polymerase examined in filamentous fungi.

Main Methods:

  • Generated a T-DNA tagged mutant (P265) via ATMT, observing altered pycnidia and hyphal growth.
  • Identified T-DNA disruption in the Pmncpap1 gene and confirmed its interaction with ribonucleotide reductase (RNR) in yeast.
  • Created targeted deletion mutants (ΔPmncpap1) and performed gene rescue experiments using a nourseothricin-resistant plasmid.

Main Results:

  • The P265 mutant exhibited smaller pycnidia and increased aerial hyphae compared to wild-type P. medicaginis.
  • Mutants showed sensitivity to hydroxyurea (HU), an RNR inhibitor, similar to yeast mutants.
  • ΔPmncpap1 mutants displayed comparable morphological defects to P265, and gene rescue partially restored wild-type characteristics.

Conclusions:

  • Pmncpap1 plays a significant role in the morphology and development of Phoma medicaginis.
  • The Pmncpap1 gene is essential for resistance to RNR inhibition, highlighting its importance in fungal stress response.
  • This study provides the first examination of a noncanonical poly(A) RNA polymerase in a filamentous fungus, opening new avenues for research.
Keywords:
ATMTCid13Homologous recombinationNon-canonical poly (A) RNA polymerasePhoma medicaginisPmncPAP1PycnidiaRNRSpring black stemTrf4Trf5

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