A Novel Protein Com1 Is Required for Normal Conidium Morphology and Full Virulence in <i>Magnaporthe oryzae</i>

Yang, Jun; Zhao, Xiao-Yan; Sun, Jing; Kang, Zhensheng; Ding, Shengli; Xu, Jin‐Rong; Peng, You-Liang

doi:10.1094/mpmi-23-1-0112

Cited by 115 publications

(129 citation statements)

References 34 publications

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“…The two flanking sequences were cloned into pKNH (Yang et al, 2010) as the deletion vector pKNH-ALG3, which was linearized by NotI digestion and transformed into protoplasts of P131. Protoplasts were isolated and transformed with the polyethylene glycol/ CaCl 2 approach as described (Yang et al, 2010). For selecting hygromycin-resistant or neomycin-resistant transformants, CM plates were supplemented with 250 mg/mL hygromycin B (Roche) or 400 mg/mL neomycin (Ameresco).…”

Section: Gene Disruption and Complementation Of Alg3mentioning

confidence: 99%

“…For complementation assays, the ALG3 gene containing a 1.5-kb native promoter fragment and a 0.5-kb terminator region was amplified with primers ALG3CF/ALG3CR and cloned into pKN (Yang et al, 2010). The resulting construct pKN-ALG3 was digested with NotI and transformed into the Dalg3 mutant.…”

Section: Gene Disruption and Complementation Of Alg3mentioning

confidence: 99%

“…After inoculation, microscopy observations were performed at 2, 12, 18, 24, 30, and 36 hpi. Rice (Oryza sativa cv LTH) seedlings at the fifth leaf stage and 8-d-old barley seedlings (cv E9) were sprayed with conidia suspension of 10 5 conidia/mL and incubated as described previously (Yang et al, 2010). Lesion formation was examined at 5 d after inoculation.…”

Section: Virulence Test and Infection Process Observationmentioning

confidence: 99%

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N-Glycosylation of Effector Proteins by an α-1,3-Mannosyltransferase Is Required for the Rice Blast Fungus to Evade Host Innate Immunity

et al. 2014

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Plant pathogenic fungi deploy secreted effectors to suppress plant immunity responses. These effectors operate either in the apoplast or within host cells, so they are putatively glycosylated, but the posttranslational regulation of their activities has not been explored. In this study, the ASPARAGINE-LINKED GLYCOSYLATION3 (ALG3)-mediated N-glycosylation of the effector, Secreted LysM Protein1 (Slp1), was found to be essential for its activity in the rice blast fungus Magnaporthe oryzae. ALG3 encodes an a-1,3-mannosyltransferase for protein N-glycosylation. Deletion of ALG3 resulted in the arrest of secondary infection hyphae and a significant reduction in virulence. We observed that Dalg3 mutants induced massive production of reactive oxygen species in host cells, in a similar manner to Dslp1 mutants, which is a key factor responsible for arresting infection hyphae of the mutants. Slp1 sequesters chitin oligosaccharides to avoid their recognition by the rice (Oryza sativa) chitin elicitor binding protein CEBiP and the induction of innate immune responses, including reactive oxygen species production. We demonstrate that Slp1 has three N-glycosylation sites and that simultaneous Alg3-mediated N-glycosylation of each site is required to maintain protein stability and the chitin binding activity of Slp1, which are essential for its effector function. These results indicate that Alg3-mediated N-glycosylation of Slp1 is required to evade host innate immunity.

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Section: Gene Disruption and Complementation Of Alg3mentioning

confidence: 99%

Section: Gene Disruption and Complementation Of Alg3mentioning

confidence: 99%

Section: Virulence Test and Infection Process Observationmentioning

confidence: 99%

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N-Glycosylation of Effector Proteins by an α-1,3-Mannosyltransferase Is Required for the Rice Blast Fungus to Evade Host Innate Immunity

et al. 2014

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“…The resulting PCR products were sequentially cloned into the XhoI-KpnI (upstream sequence) and SpeIEcoRI (downstream sequence) sites of pKNH (Yang et al 2010). After linearization with NotI, pKOC5 was transformed into strain P131.…”

Section: Generation Of the Mocrc1 Null Mutantsmentioning

confidence: 99%

“…The MoCRC1-eGFP fusion vector pKGCRC1 was generated by cloning MoCRC1 along with its 1.3-kb upstream sequence, which had been amplified with the primer pair crc1gfpf/crc1gfpr (Table 2), into pKNTG (Yang et al 2010). After linearization, pKGCRC1 was transformed into the DMocrc1 null mutant.…”

Section: Complementationmentioning

confidence: 99%

A carnitine–acylcarnitine carrier protein, MoCrc1, is essential for pathogenicity in Magnaporthe oryzae

et al. 2012

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The rice blast fungus Magnaporthe oryzae forms a specialized infection structure called an appressorium to breach the host-plant epidermis for successful infection. In this study, a mutant defective in appressorial penetration was isolated by a mutagenesis approach, in which an exogenous DNA fragment was found to be inserted into the first exon of MoCRC1. This gene encodes a putative carnitine-acylcarnitine carrier protein that is widely conserved among eukaryotic organisms. Deletion of MoCRC1 severely reduces appressorium turgor generation, appressorial penetration, and development of infection hyphae. The null mutant of MoCRC1 lost pathogenicity on intact and abraded host leaves. MoCRC1 was also found to be required for growth on minimal medium containing sodium acetate or olive oil. Moreover, the transformed MoCrc1-eGFP fusion protein was expressed throughout the infection process. Our results suggest that the carnitine-acylcarnitine carrier protein plays vital roles in appressorium-mediated infection and is essential for pathogenesis of M. oryzae and perhaps other phytopathogenic fungi.

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A conidiation-related gene is highly expressed at the resting urediospore stage inPuccinia striiformisf. sp.tritici

Guo

Duan

Zhang

et al. 2012

J. Basic Microbiol.

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The production and germination of asexual spores in a diverse group of fungi play a crucial role in their infection cycles. These processes are regulated by a set of genes, namely, conidiation-related genes, involved in the production, morphological characteristics, and differentiation of conidia. In this study, we identified and characterized the PsCon1 gene, which is the first conidiation-related gene identified in Puccinia striiformis f. sp. tritici (Pst). Sequence analysis revealed that PsCON1 has two conserved conidiation-specific protein 6 domains. Single nucleotide polymorphisms and insertion/deletion variations were detected in the coding region of PsCon1 among five Pst races. Quantitative RT-PCR assays revealed that PsCon1 was expressed at the highest level in resting urediospores of Pst, and gradually decreased after germination and infection. However, at 312 hpi, at the stage of forming large amounts of urediospores on leaves, the amount of PsCon1 mRNA was sharply increased but only 0.1-fold that of resting urediospores. Subcellular localization assays indicated PsCon1 heterologously expressed in Fusarium graminearum was located in the cytoplasm of conidia. The results suggest that PsCon1 may play a role in formation or survival of Pst urediospores.

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A Novel Protein Com1 Is Required for Normal Conidium Morphology and Full Virulence in Magnaporthe oryzae

Cited by 115 publications

References 34 publications

N-Glycosylation of Effector Proteins by an α-1,3-Mannosyltransferase Is Required for the Rice Blast Fungus to Evade Host Innate Immunity

N-Glycosylation of Effector Proteins by an α-1,3-Mannosyltransferase Is Required for the Rice Blast Fungus to Evade Host Innate Immunity

A carnitine–acylcarnitine carrier protein, MoCrc1, is essential for pathogenicity in Magnaporthe oryzae

A conidiation-related gene is highly expressed at the resting urediospore stage inPuccinia striiformisf. sp.tritici

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