Carbamoyl phosphate synthetase subunit Cpa1 interacting with Dut1, controls development, arginine biosynthesis, and pathogenicity of Colletotrichum gloeosporioides

Tan, Qingqun; Zhao, Xuanzhu; He, Haiyong; Zhang, Junxiang; Yi, Tuyong

doi:10.1016/j.funbio.2020.10.009

Cited by 11 publications

(10 citation statements)

References 30 publications

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“…Our growth assay showed attenuated growth of ΔMocpa1 deletion mutants on MM medium, with the growth defects being rescued upon addition of exogenous arginine, clearly demonstrating the importance of MoCpa1 in arginine biosynthesis. This results were in agreement with a recently reported study on the role of CPA1 in glutamine biosynthesis in fungal pathogen Colletotrichum gloeosporioides (Tan et al, 2020).…”

Section: Discussionsupporting

confidence: 94%

“…In rice blast fungus, it was reported that deletion of carbamoyl phosphate synthase arginine specific large subunit MoCpa2, leads to attenuated virulence (Xinyu . Moreover, a recently reported study, showed that loss of CPA1 in Colletotrichum gloeosporioides resulted in complete loss of virulence (Tan et al, 2020). Similarly, the ΔMocpa1 mutant was nonpathogenic on barley and rice leaves.…”

Section: Discussionmentioning

confidence: 88%

“…Previous study showed that amino acid biosynthesis is important for the fungal infection cycle (Xin Liu et al, 2019;Orasch et al, 2019;Tan, Zhao, He, Zhang, & Yi, 2020) . In M. oryzae, arginine biosynthesis catalyzed by MoArg1, MoArg5, MoArg6, MoArg7, and MoCpa2 have been shown to be indispensable for the development and pathogenicity of M. oryzae (Xinyu Liu, Cai, et al, 2016;Y.…”

Section: Discussionmentioning

confidence: 99%

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MoCpa1-mediated arginine biosynthesis is crucial for fungal growth, conidiation, and plant infection ofMagnaporthe oryzae

Aron

Wang

Mabeche

et al. 2020

Preprint

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Arginine is an important amino acid involved in processes such as cell signal transduction, protein synthesis, and sexual reproduction. To understand the biological roles of arginine biosynthesis in pathogenic fungi, we used Cpa1, the carbamoyl phosphate synthase arginine-specific small chain subunit in Saccharomyces cerevisiae as a query to identify its ortholog in Magnaporthe oryzae genome database and named it MoCpa1. MoCpa1 is a 471-amino acid protein containing the CPSase_sm_chain domain and the GATase domain. MoCpa1 transcripts were highly expressed at the conidiation, early-infection, and late-infection stages of the fungus. Targeted deletion of MoCPA1 gene resulted in the ΔMocpa1 mutant exhibiting arginine auxotrophy on MM, confirming its role in de novo arginine biosynthesis. The ΔMocpa1 mutant presented significantly decreased sporulation with some of its conidia being defective in morphology. Furthermore, the ΔMocpa1 mutant was nonpathogenic on rice and barley leaves, which was a result of defects in appressorium-mediated penetration and restricted invasive hyphal growth within host cells. Addition of exogenous arginine partially rescued conidiation and pathogenicity defects on the barley and rice leaves, while introduction of MoCPA1 gene in ΔMocpa1 mutant fully complemented the lost phenotype. Further confocal microscopy examination revealed that MoCpa1 is localized in the mitochondria. In summary, our results demonstrate that MoCpa1-mediated arginine biosynthesis is crucial for fungal development, conidiation, appressorium formation and infection-related morphogenesis in M. oryzae, thus serving as an attractive target for mitigating obstinate fungal plant pathogens.

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Section: Discussionsupporting

confidence: 94%

Section: Discussionmentioning

confidence: 88%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

MoCpa1-mediated arginine biosynthesis is crucial for fungal growth, conidiation, and plant infection ofMagnaporthe oryzae

Aron

Wang

Mabeche

et al. 2020

Preprint

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show abstract

“…Previous studies showed that amino acid biosynthesis is important for growth and the fungal infection (Liu et al 2019;Orasch et al 2019;Tan et al 2020). In M. oryzae, it has been reported that deletion of genes involved in arginine biosynthesis resulted in mutant cells requiring exogenous arginine for growth on MM medium (Liu et al 2016;Zhang et al 2015).…”

Section: Discussionmentioning

confidence: 99%

MoCpa1-mediated arginine biosynthesis is crucial for fungal growth, conidiation, and plant infection of Magnaporthe oryzae

Aron

Wang

Mabeche

et al. 2021

Appl Microbiol Biotechnol

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Arginine is an important amino acid involved in processes such as cell signal transduction, protein synthesis, and sexual reproduction. To understand the biological roles of arginine biosynthesis in pathogenic fungi, we used Cpa1, the carbamoyl phosphate synthase arginine-specific small chain subunit in Saccharomyces cerevisiae as a query to identify its ortholog in the Magnaporthe oryzae genome and named it MoCpa1. MoCpa1 is a 471-amino acid protein containing a CPSase_sm_chain domain and a GATase domain. MoCpa1 transcripts were highly expressed at the conidiation, early-infection, and late-infection stages of the fungus. Targeted deletion of the MoCPA1 gene resulted in a ΔMocpa1 mutant exhibiting arginine auxotrophy on minimum culture medium (MM), confirming its role in de novo arginine biosynthesis. The ΔMocpa1 mutant presented significantly decreased sporulation with some of its conidia being defective in morphology. Furthermore, the ΔMocpa1 mutant was nonpathogenic on rice and barley leaves, which was a result of defects in appressorium-mediated penetration and restricted invasive hyphal growth within host cells. Addition of exogenous arginine partially rescued conidiation and pathogenicity defects on the barley and rice leaves, while introduction of the MoCPA1 gene into the ΔMocpa1 mutant fully complemented the lost phenotype. Further confocal microscopy examination revealed that MoCpa1 is localized in the mitochondria. In summary, our results demonstrate that MoCpa1-mediated arginine biosynthesis is crucial for fungal development, conidiation, appressorium formation, and infection-related morphogenesis in M. oryzae, thus serving as an attractive target for mitigating obstinate fungal plant pathogens. Key points• MoCpa1 is important for aerial hyphal growth and arginine biosynthesis.• MoCpa1 is pivotal for conidial morphogenesis and appressorium formation.• MoCpa1 is crucial for full virulence in M. oryzae.

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“…To analyse the integration site of T-DNA in S58 mutant, genomic DNA was attained through hiTAIL PCR products and then sequencing was performed. The position of the mutant S58 was located at 399,910 bp which is a part of the first exon of a hypothetical-gene CPA1 (3,999,310-4,000,751 bp) located on contig 39 ( Tan et al, 2021 ). It contained two exon and one intron.…”

Section: Resultsmentioning

confidence: 99%

Carbamoyl Phosphate Synthase Subunit CgCPS1 Is Necessary for Virulence and to Regulate Stress Tolerance in Colletotrichum gloeosporioides

et al. 2021

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Glomerella leaf spot (GLS) is a severe infectious disease of apple whose infective area is growing gradually and thus poses a huge economic threat to the world. Different species of Colletotrichum including Colletotrichum gloeosporioides are responsible for GLS. For efficient GLS control, it is important to understand the mechanism by which the cruciferous crops and C. gloeosporioides interact. Arginine is among one of the several types of amino acids, which plays crucial role in biochemical and physiological functions of fungi. The arginine biosynthesis pathway involved in virulence among plant pathogenic fungi is poorly understood. In this study, CgCPS1 gene encoding carbamoyl phosphate synthase involved in arginine biosynthesis has been identified and inactivated experimentally. To assess the effects of CgCPS1 , we knocked out CgCPS1 in C. gloeosporioides and evaluated its effects on virulence and stress tolerance. The results showed that deletion of CgCPS1 resulted in loss of pathogenicity. The Δcgcps1 mutants showed slow growth rate, defects in appressorium formation and failed to develop lesions on apple leaves and fruits leading to loss of virulence while complementation strain (CgCPS1-C) fully restored its pathogenicity. Furthermore, mutant strains showed extreme sensitivity to high osmotic stress displaying that CgCPS1 plays a vital role in stress response. These findings suggest that CgCPS1 is major factor that mediates pathogenicity in C. gloeosporioides by encoding carbamoyl phosphate that is involved in arginine biosynthesis and conferring virulence in C. gloeosporioides .

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Carbamoyl phosphate synthetase subunit Cpa1 interacting with Dut1, controls development, arginine biosynthesis, and pathogenicity of Colletotrichum gloeosporioides

Cited by 11 publications

References 30 publications

MoCpa1-mediated arginine biosynthesis is crucial for fungal growth, conidiation, and plant infection ofMagnaporthe oryzae

MoCpa1-mediated arginine biosynthesis is crucial for fungal growth, conidiation, and plant infection ofMagnaporthe oryzae

MoCpa1-mediated arginine biosynthesis is crucial for fungal growth, conidiation, and plant infection of Magnaporthe oryzae

Carbamoyl Phosphate Synthase Subunit CgCPS1 Is Necessary for Virulence and to Regulate Stress Tolerance in Colletotrichum gloeosporioides

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