The Aspergillus flavus hacA Gene in the Unfolded Protein Response Pathway Is a Candidate Target for Host-Induced Gene Silencing

被引:0
作者
Chang, Perng-Kuang [1 ]
机构
[1] ARS, Southern Reg Res Ctr, USDA, 1100 Allen Toussaint Blvd, New Orleans, LA 70124 USA
关键词
<italic>Aspergillus flavus</italic>; aflatoxin; unfolded protein response; bZip; CRISPR; host-induced gene silencing; RNAi; sclerotia; secondary metabolism; MESSENGER-RNA; AFLATOXIN BIOSYNTHESIS; SECONDARY METABOLISM; TRANSGENIC MAIZE; TRANSCRIPTION; IDENTIFICATION; EXPRESSION; GROWTH; FUNGI; NIGER;
D O I
10.3390/jof10100719
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Fungal HacA/Hac1 transcription factors play a crucial role in regulating the unfolded protein response (UPR). The UPR helps cells to maintain endoplasmic reticulum (ER) protein homeostasis, which is critical for growth, development, and virulence. The Aspergillus flavus hacA gene encodes a domain rich in basic and acidic amino acids (Bsc) and a basic leucine zipper (bZip) domain, and features a non-conventional intron (Nt20). In this study, CRISPR/Cas9 was utilized to dissect the Bsc-coding, bZip-coding, and Nt20 sequences to elucidate the relationship between genotype and phenotype. In the Bsc and bZip experimental sets, all observed mutations in both coding sequences were in frame, suggesting that out-of-frame mutations are lethal. The survival rate of transformants in the Nt20 experiment set was low, at approximately 7%. Mutations in the intron primarily consisted of out-of-frame insertions and deletions. In addition to the wild-type-like conidial morphology, the mutants exhibited varied colony morphologies, including sclerotial, mixed (conidial and sclerotial), and mycelial morphologies. An ER stress test using dithiothreitol revealed that the sclerotial and mycelial mutants were much more sensitive than the conidial mutants. Additionally, the mycelial mutants were unable to produce aflatoxin but still produced aspergillic acid and kojic acid. RNAi experiments targeting the region encompassing Bsc and bZip indicated that transformant survival rates generally decreased, with a small number of transformants displaying phenotypic changes. Defects in the hacA gene at the DNA and transcript levels affected the survival, growth, and development of A. flavus. Thus, this gene may serve as a promising target for future host-induced gene-silencing strategies aimed at controlling infection and reducing aflatoxin contamination in crops.
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页数:17
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  • [1] The molecular mechanism and functional diversity of UPR signaling sensor IRE1
    Bashir, Samirul
    Banday, Mariam
    Qadri, Ozaira
    Bashir, Arif
    Hilal, Nazia
    Nida-i-Fatima
    Rader, Stephen
    Fazili, Khalid Majid
    [J]. LIFE SCIENCES, 2021, 265
  • [2] Coordination of secondary metabolism and development in fungi: the velvet family of regulatory proteins
    Bayram, Oezguer
    Braus, Gerhard H.
    [J]. FEMS MICROBIOLOGY REVIEWS, 2012, 36 (01) : 1 - 24
  • [3] HacA Governs Virulence Traits and Adaptive Stress Responses in Trichophyton rubrum
    Bitencourt, Tamires A.
    Lang, Elza A. S.
    Sanches, Pablo R.
    Peres, Nalu T. A.
    Oliveira, Vanderci M.
    Fachin, Ana Lucia
    Rossi, Antonio
    Martinez-Rossi, Nilce M.
    [J]. FRONTIERS IN MICROBIOLOGY, 2020, 11
  • [4] MEDIUM FOR RAPID IDENTIFICATION AND ENUMERATION OF ASPERGILLUS-FLAVUS AND RELATED ORGANISMS
    BOTHAST, RJ
    FENNELL, DI
    [J]. MYCOLOGIA, 1974, 66 (02) : 365 - 369
  • [5] Relationship between secondary metabolism and fungal development
    Calvo, AM
    Wilson, RA
    Bok, JW
    Keller, NP
    [J]. MICROBIOLOGY AND MOLECULAR BIOLOGY REVIEWS, 2002, 66 (03) : 447 - +
  • [6] Genome-wide expression analysis upon constitutive activation of the HacA bZIP transcription factor in Aspergillus niger reveals a coordinated cellular response to counteract ER stress
    Carvalho, Neuza D. S. P.
    Jorgensen, Thomas R.
    Arentshorst, Mark
    Nitsche, Benjamin M.
    van den Hondel, Cees A. M. J. J.
    Archer, David B.
    Ram, Arthur F. J.
    [J]. BMC GENOMICS, 2012, 13
  • [7] Expanding the ku70 toolbox for filamentous fungi: establishment of complementation vectors and recipient strains for advanced gene analyses
    Carvalho, Neuza D. S. P.
    Arentshorst, Mark
    Kwon, Min Jin
    Meyer, Vera
    Ram, Arthur F. J.
    [J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2010, 87 (04) : 1463 - 1473
  • [8] Creating large chromosomal segment deletions in Aspergillus flavus by a dual CRISPR/Cas9 system: Deletion of gene clusters for production of aflatoxin, cyclopiazonic acid, and ustiloxin B
    Chang, Perng-Kuang
    [J]. FUNGAL GENETICS AND BIOLOGY, 2024, 170
  • [9] A Simple CRISPR/Cas9 System for Efficiently Targeting Genes of Aspergillus Section Flavi Species, Aspergillus nidulans, Aspergillus fumigatus, Aspergillus terreus, and Aspergillus niger
    Chang, Perng-Kuang
    [J]. MICROBIOLOGY SPECTRUM, 2023, 11 (01):
  • [10] Aspergillus flavus GPI-anchored protein-encoding ecm33 has a role in growth, development, aflatoxin biosynthesis, and maize infection
    Chang, Perng-Kuang
    Zhang, Qi
    Scharfenstein, Leslie
    Mack, Brian
    Yoshimi, Akira
    Miyazawa, Ken
    Abe, Keietsu
    [J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2018, 102 (12) : 5209 - 5220