|
|
|
| Identification and Expression Analysis of Siderophore Synthetic Gene Cluster of Setosphaeria turcica |
| YUE Hao-Feng1,3, ZHAO Tian-Yi2,3, LI Hai-Xiao2,3, YAN Qiu-Jing2,3, XU Ya-Di1,3, MENG Ya-Nan1,3, CAO Zhi-Yan2,3,*, LIU Ning2,3,*, DONG Jin-Gao2,3 |
1 School of Life Sciences, Hebei Agricultural University, Baoding 071001, China;
2 College of Plant Protection, Hebei Agricultural University, Baoding 071001, China;
3 State Key Laboratory of North China Crop Improvement and Regulation/Key Laboratory of Hebei Province for Plant Physiology and Molecular Pathology, Baoding 071000, China |
|
|
|
|
Abstract Iron is an essential element for microbial growth and plays a crucial role in processes such as oxygen metabolism, electron transfer, DNA and RNA synthesis. Microbes primarily rely on siderophores to absorb iron ions from their environment. Siderophores, as low molecular weight iron chelators secreted by microbes, assist in transporting iron ions within the organism. Fungi primarily use non-ribosomal peptide synthetase-encoded genes as the core of secondary metabolic biosynthetic gene clusters to synthesize siderophores . This study aimed to clarify the siderophore function and biosynthetic process of Setosphaeria turcica. The type of siderophores synthesized by S. turcica was identified using CAS (chrome azurol sulphonate), and its biosynthetic gene cluster was predicted using the AntiSMASH website. Transcriptional data from different developmental stages of S. turcica spores and qRT-PCR analysis of the expression levels of key genes in the siderophore biosynthetic gene cluster during pathogen infection were utilized. The results showed that S. turcica produces hydroxamate-type siderophores. One biosynthetic gene cluster responsible for hydroxamate-type siderophore production was identified in the genome. This cluster consisted of 12 genes with the core gene StNPS6 encoding non-ribosomal peptide synthetase. During spore germination and infection process of S. turcica, several genes in the cluster exhibited significant upregulation. The core gene StNPS6 had an expression level 5.52 times higher at 6 d post-infection compared to the early infection stage, and the auxiliary biosynthesis gene StrhbE showed a 24.12-fold increase in expression at 9 d post-infection. Additionally, the transport-related gene Stpmd1 increased significantly by 19.95-fold at 9 d post-infection. At the same time, it was observed that both iron-rich and iron-deficient conditions had a certain inhibitory effect on the growth of the pathogen. Moreover, under iron-rich conditions, melanin synthesis in the pathogen was hindered. The differences in iron content in the medium led to significant changes in the expression levels of multiple genes within the gene cluster, especially the transport-related protein gene Stpmd1, which showed a 73.28-fold increase in expression under iron-rich conditions. This study provides a foundation for understanding the potential role of siderophores in S.turcica.
|
|
Received: 04 July 2023
|
|
|
|
Corresponding Authors:
*caozhiyan@hebau.edu.cn; lning121@126.com
|
|
|
|
[1] 陈伟, 王小利, 付薇, 等. 2016. 黑麦草根际产铁载体细菌HMGY6B的筛选鉴定及对病原菌的拮抗作用[J]. 微生物学通报, 43(10): 2207-2215.
(Chen W, Wang X L, Fu W, et al.2016. Screening and identification of rhizometropic bacterium HMGY6B in ryegrass rhizosphere and its antagonism against pathogenic bacteria[J]. Microbiology Bulletin, 43(10): 2207-2215.)
[2] 王迪. 2014. 玉米大斑病菌T-DNA插入突变体库的构建及StNPS6基因的功能研究[D]. 硕士学位论文, 吉林大学, 导师: 晋齐鸣, pp. 46-54.
(Wang D.2014. Construction of T-DNA insertion mutant library and functional study of StNPS6 gene of maize large spot pathogen[D].Thesis for M.S., Jilin University, Supervisor: Jin Q M, pp. 46-54.)
[3] Bölker M, Basse C W, Schirawski J.2008. Ustilago maydis secondary metabolism-from genomics to biochemistry[J]. Fungal Genetics And Biology, 1: S88-S93.
[4] Chen L H, Lin C H, Chung K R.2013. A nonribosomal peptide synthetase mediates siderophore production and virulence in the citrus fungal pathogen Alternaria alternata[J]. Molecular Plant Pathology, 14(5): 497-505.
[5] Griffin D, Winkelmann G, Winge D.1994. Metal Ions in Fungi[M]. Boca Raton: CRC Press, USA, pp. 425-427.
[6] Gsaller F, Blatzer M, Abt B.2013. The first promoter for conditional gene expression in Acremonium chrysogenum: Iron starvation-inducible mir1[P] Journal Of Biotechnol, 163(1): 77-80.
[7] Haas H.2003. Molecular genetics of fungal siderophore biosynthesis and uptake: the role of siderophores in iron uptake and storage[J]. Applied Microbiology and Biotechnology, 62(4): 316-330.
[8] Haas H, Eisendle M, Turgeon B G.2008. Siderophores in fungal physiology and virulence[J]. Annual Review of Phytopathology, 46: 149-187.
[9] Hof C, Eisfeld K, Antelo L, et al.2009. Siderophore synthesis in Magnaporthe grisea is essential for vegetative growth, conidiation and resistance to oxidative stress[J]. Fungal Genetics and Biology, 46(4): 321-332.
[10] Hof C, Eisfeld K, Welzel K, et al.2007. Ferricrocin synthesis in Magnaporthe grisea and its role in pathogenicity in rice[J]. Molecular Plant Pathology, 8(2): 163-172.
[11] Jalal M A, Love S K, van der Helm D.1988. N alpha-dimethylcoprogens. Three novel trihydroxamate siderophores from pathogenic fungi[J]. Biology of Metals, 1(1): 4-8.
[12] Johnson L.2008. Iron and siderophores in fungal-host interactions[J]. Mycological Research , 112(Pt 2): 170-183.
[13] Khaldi N, Seifuddin F T, Turner G, et al.2010. SMURF: Genomic mapping of fungal secondary metabolite clusters[J]. Fungal Genetics and Biology, 47(9): 736-741.
[14] Lee B N, Kroken S, Chou D Y, et al.2005. Functional analysis of all nonribosomal peptide synthetases in Cochliobolus heterostrophus reveals a factor, NPS6, involved in virulence and resistance to oxidative stress[J]. Eukaryot Cell, 4(3): 545-555.
[15] Lemanceau P, Alabouvette C, Meyer J M J S U.1986. Production of fusarinine and iron assimilation by pathogenic and non-pathogenic Fusarium[J]. Iron, Siderophores, and Plant Diseases, 117: 251-259.
[16] Liu G, Greenshields D L, Sammynaiken R, et al.2007. Targeted alterations in iron homeostasis underlie plant defense responses[J]. Journal of Cell Science, 120(Pt4): 596-605.
[17] De Luca, N G, P M Wood.2000. Iron uptake by fungi: Contrasted mechanisms with internal or external reduction[J]. Advances in Microbial Physiology, 43: 39-74.
[18] Mei B, Budde A D, Leong S A.1993. sid1, a gene initiating siderophore biosynthesis in Ustilago maydis: Molecular characterization, regulation by iron, and role in phytopathogenicity[J]. Proceedings of the National Academy of Sciences of The USA, 90(3): 903-907.
[19] Meng Y, Zeng F, Hu J, et al.2022. Novel factors contributing to fungal pathogenicity at early stages of Setosphaeria turcica infection[J]. Molecular Plant Pathology, 23(1): 32-44.
[20] Miethke M, Marahiel M A.2007. Siderophore-based iron acquisition and pathogen control[J]. Microbiology and Molecular Biology Reviews, 71(3): 413-451.
[21] Narh Mensah D L, Wingfield B D, Coetzee M P A.2023. Nonribosomal peptide synthetase gene clusters and characteristics of predicted NRPS-dependent siderophore synthetases in Armillaria and other species in the Physalacriaceae[J]. Current Genetics, 69(1): 7-24.
[22] Oide S, Moeder W, Krasnoff S, et al.2006. NPS6, encoding a nonribosomal peptide synthetase involved in siderophore-mediated iron metabolism, is a conserved virulence determinant of plant pathogenic ascomycetes[J]. The Plant Cell, 18(10): 2836-2853.
[23] Plattner H J, Diekmann H.1994. Enzymology of siderophore biosynthesis in fungi: Metal Ions in Fungi[M]. Boca Raton: CRC Press, USA, pp. 99-116.
[24] Rondon M R, Ballering K S, Thomas M G.2004. Identification and analysis of a siderophore biosynthetic gene cluster from Agrobacterium tumefaciens C58[J]. Microbiology (Reading), 150(Pt 11): 3857-3866.
[25] Sánchez-Sanuy F, Mateluna-Cuadra R, Tomita K, et al.2022. Iron induces resistance against the rice blast fungus Magnaporthe oryzae through potentiation of immune responses[J]. Rice (New York), 15(1): 68.
[26] Silva M G, Schrank A, Bailão E F, et al.2011. The homeostasis of iron, copper, and zinc in paracoccidioides brasiliensis, cryptococcus neoformans var. Grubii, and cryptococcus gattii: A comparative analysis[J]. Frontiers Microbiology, 2: 49.
[27] Tremblay N, Hill P, Conway K R, et al.2016. The use of clustermine360 for the analysis of polyketide and nonribosomal peptide biosynthetic pathways[J]. Methods in Molecular Biology, 1401: 233-252.
[28] Wilson B R, Bogdan A R, Miyazawa M, et al.2016. Siderophores in iron metabolism: From mechanism to therapy potential[J]. Trends in Molecular Medicine, 22(12): 1077-1090.
[29] Winkelmann G.2003. Siderophore transport in fungi: Microbial transport systems[J]. Microbial Transport Systems, 21: 463-480. |
|
|
|
