Bcl-2 proteins link programmed cell death with growth and morphogenetic adaptations in the fungal plant pathogen Colletotrichum gloeosporioides

Barhoom, Sima; Sharon, Amir

doi:10.1016/j.fgb.2006.06.007

Cited by 44 publications

(45 citation statements)

References 55 publications

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“…Genomic DNA was isolated according to the method of Barhoom and Sharon (2). Plasmid DNA was isolated using the GenElute plasmid miniprep kit (Sigma).…”

Section: Methodsmentioning

confidence: 99%

Regulation of Pathogenic Spore Germination by CgRac1 in the Fungal Plant Pathogen Colletotrichum gloeosporioides

Nesher

Minz

Kokkelink³

et al. 2011

Eukaryot Cell

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Colletotrichum gloeosporioides is a facultative plant pathogen: it can live as a saprophyte on dead organic matter or as a pathogen on a host plant. Different patterns of conidial germination have been recognized under saprophytic and pathogenic conditions, which also determine later development. Here we describe the role of CgRac1 in regulating pathogenic germination. The hallmark of pathogenic germination is unilateral formation of a single germ tube following the first cell division. However, transgenic strains expressing a constitutively active CgRac1 (CA-CgRac1) displayed simultaneous formation of two germ tubes, with nuclei continuing to divide in both cells after the first cell division. CA-CgRac1 also caused various other abnormalities, including difficulties in establishing and maintaining cell polarity, reduced conidial and hyphal adhesion, and formation of immature appressoria. Consequently, CA-CgRac1 isolates were completely nonpathogenic. Localization studies with cyan fluorescent protein (CFP)-CgRac1 fusion protein showed that the CgRac1 protein is abundant in conidia and in hyphal tips. Although the CFP signal was equally distributed in both cells of a germinating conidium, reactive oxygen species accumulated only in the cell that produced a germ tube, indicating that CgRac1 was active only in the germinating cell. Collectively, our results show that CgRac1 is a major regulator of asymmetric development and that it is involved in the regulation of both morphogenesis and nuclear division. Modification of CgRac1 activity disrupts the morphogenetic program and prevents fungal infection.

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“…Genomic DNA was isolated according to the method of Barhoom and Sharon (2). Plasmid DNA was isolated using the GenElute plasmid miniprep kit (Sigma).…”

Section: Methodsmentioning

confidence: 99%

Regulation of Pathogenic Spore Germination by CgRac1 in the Fungal Plant Pathogen Colletotrichum gloeosporioides

Nesher

Minz

Kokkelink³

et al. 2011

Eukaryot Cell

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“…The 700-bp fragment was cloned into SacI and SpeI sites of the resulting plasmid. The two fragments in opposite orientations were released by digestion with NcoI and BamHI and cloned between the GPDA promoter and TRPC terminator in pKsh52-1 (4).…”

Section: Methodsmentioning

confidence: 99%

“…Usually, the fungus will penetrate the plant several hours after appressorium formation, and further development will occur inside the plant tissues (3,4). Extensive hyphal growth on the plant surface was not observed in the wild-type strain 24 h postinoculation.…”

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confidence: 99%

Functional Characterization of CgCTR2, a Putative Vacuole Copper Transporter That Is Involved in Germination and Pathogenicity inColletotrichum gloeosporioides

et al. 2008

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Copper is a cofactor and transition metal involved in redox reactions that are essential in all eukaryotes. Here, we report that a vacuolar copper transporter that is highly expressed in resting spores is involved in germination and pathogenicity in the plant pathogen Colletotrichum gloeosporioides. A screen of C. gloeosporioides transformants obtained by means of a promoterless green fluorescent protein (GFP) construct led to the identification of transformant N159 in which GFP signal was observed in spores. The transforming vector was inserted 70 bp upstream of a putative gene with homology to the Saccharomyces cerevisiae vacuolar copper transporter gene CTR2. The C. gloeosporioides CTR2 (CgCTR2) gene fully complemented growth defects of yeast ctr2⌬ mutants, and a CgCTR2-cyan fluorescent protein (CFP) fusion protein accumulated in vacuole membranes, confirming the function of the protein as a vacuolar copper transporter. Expression analysis indicated that CgCTR2 transcript is abundant in resting conidia and during germination in rich medium and downregulated during "pathogenic" germination and the early stages of plant infection. CgCTR2 overexpression and silencing mutants were generated and characterized. The Cgctr2 mutants had markedly reduced Cu superoxide dismutase (SOD) activity, suggesting that CgCTR2 is important in providing copper to copper-dependent cytosolic activities. The Cgctr2-silenced mutants had increased sensitivity to H 2 O 2 and reduced germination rates. The mutants were also less virulent to plants, but they did not display any defects in appressorium formation and penetration efficiency. An external copper supply compensated for the hypersensitivity to H 2 O 2 but not for the germination and pathogenicity defects of the mutants. Similarly, overexpression of CgCTR2 enhanced resistance to H 2 O 2 but had no effect on germination or pathogenicity. Our results show that copper is necessary for optimal germination and pathogenicity and that CgCTR2 is involved in regulating cellular copper balance during these processes.

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“…PCR conditions followed the methods reported by the respective sources for each primer set. For amplifi cation of the SOD2 region of C. gloeosporioides, SODglo2-F (5′-CAG ATC ATG GAG CTG CAC CA-3′) and -R (5′-TAG TAC GCG TGC TCG GAC AT-3′) primers were newly designed in this study based on the sequence of C. gloeosporioides reported by Barhoom and Sharon (2007). The cycle parameters with these primers were an initial denaturation at 95°C for 4 min; followed by 30 cycles of denaturation at 95°C for 30 s, annealing at 58°C for 30 s, extension at 72°C for 2 min, and a fi nal extension for 5 min at 72°C.…”

Section: Cultural Characters and Inoculation Testmentioning

confidence: 99%

Colletotrichum echinochloae, a new species on Japanese barnyard millet (Echinochloa utilis)

Moriwaki

Tsukiboshi

2009

Mycoscience

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Bcl-2 proteins link programmed cell death with growth and morphogenetic adaptations in the fungal plant pathogen Colletotrichum gloeosporioides

Cited by 44 publications

References 55 publications

Regulation of Pathogenic Spore Germination by CgRac1 in the Fungal Plant Pathogen Colletotrichum gloeosporioides

Regulation of Pathogenic Spore Germination by CgRac1 in the Fungal Plant Pathogen Colletotrichum gloeosporioides

Functional Characterization of CgCTR2, a Putative Vacuole Copper Transporter That Is Involved in Germination and Pathogenicity inColletotrichum gloeosporioides

Colletotrichum echinochloae, a new species on Japanese barnyard millet (Echinochloa utilis)

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