Draft Genome Sequence of Mycoparasite
            <i>Clonostachys rosea</i>
            Strain 67-1

Sun, Zhan‐Bin; Sun, Man-Hong; Li, Shidong

doi:10.1128/genomea.00546-15

Cited by 31 publications

(34 citation statements)

References 14 publications

(12 reference statements)

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“…The gpdA and trpC fragments were amplified using the primers qdz F/ qdz R and zzz F/ zzz R, respectively, with a concentration of 10 μM. The open reading frame (ORF) of Chi67 - 1 was amplified from the genomic DNA of C. rosea 67-1 using the CF and CR primers (Sun et al 2015c; Table 1). …”

Section: Methodsmentioning

confidence: 99%

Transformation of the endochitinase gene Chi67-1 in Clonostachys rosea 67-1 increases its biocontrol activity against Sclerotinia sclerotiorum

et al. 2017

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Clonostachys rosea is a promising biocontrol fungus active against various plant fungal pathogens. In this study, the endochitinase-encoding gene Chi67-1, the expression of which is sharply upregulated in C. rosea 67-1 when induced by sclerotia, was transformed into the original isolate by protoplast transformation, and transformants were screened against Sclerotinia rot of soybean. The transformation efficiency was approximately 50 transformants per 1 × 107 protoplasts, and 68 stably heritable recombinants were assayed. The parasitic rates of 32.4% of the tested strains increased by more than 50% compared to 43.3% of the wild type strain in 16 h, and the Rc4-4 transformant showed a parasitic rate of 100% in 16 h. The control efficiencies of the selected efficient transformants to soybean Sclerotinia stem rot were evaluated in pots in the greenhouse, and the results revealed that Rc4-4 achieved the highest efficiency of 81.4%, which was 31.7% and 28.7% higher than the control achieved by the wide type and the pesticide carbendazim, respectively. Furthermore, the expression level of Chi67-1 was 107-fold higher in Rc4-4 than in the wild type, and accordingly, the chitinase activity of the recombinant increased by 140%. The results lay a foundation for the development of efficient genetically engineered strains of C. rosea.

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Section: Methodsmentioning

confidence: 99%

Transformation of the endochitinase gene Chi67-1 in Clonostachys rosea 67-1 increases its biocontrol activity against Sclerotinia sclerotiorum

et al. 2017

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“…The gene tub was also used in the quantification of gene expressions in C. rosea when confronting with pathogenic fungi and plant nematodes (Mamarabadi et al., ; Zou, Tu, Liu, Tao, & Zhang, ). In our previous study, we found that EF1 was expressed most stably in C. rosea 67‐1 during parasitism on Sclerotinia sclerotiorum sclerotia, and we obtained a reliable expression profile of mycoparasitism‐related genes by using the reference gene EF1 (Sun, Li et al., ; Sun, Sun et al., ). In this study, analyses using geNorm, BestKeeper, and NormFinder showed that the expression of EF1 was not satisfactory during chlamydospore formation in C. rosea .…”

Section: Discussionmentioning

confidence: 94%

“…On the basis of the transcriptome data for C. rosea 67‐1 chlamydospore formation, a total of nine candidates were selected and cloned from the genome of C. rosea (Sun, Li et al., ; Sun, Sun et al., ), namely, ACT , EF1 , GAPDH , HIS , RPP , SSD , TBP , UBQ , and UCE (GenBank accession numbers: KP274072, KP274074‐KP274077, KX686112‐KX686115). The primers for the reference genes used for RT‐qPCR were designed using Primer Premier 6 (Table ), and their specificity was assessed with the following PCR program: 94°C for 3 min, followed by 30 cycles at 94°C for 30 s, 55°C for 30 s, and 72°C for 20 s; final extension at 72°C for 10 min.…”

Section: Methodsmentioning

confidence: 99%

“…However, no reference gene is absolutely consistent in different organisms and their developmental stages; instead, the expression level of a reference gene is stable only under specific conditions. β ‐tubulin was used to normalize N‐acety1‐β‐D‐glucosaminidase gene in C. rosea when confronting with Fusarium culmorum (Mamarabadi, Jensen, & Lübeck, ), however, it was not available in mycoparasitic process (Sun, Li, & Sun, ; Sun, Sun, & Li, ). In Neurospora crassa , the expression of act was not affected by temperature, but it was inhibited under all‐dark conditions (Cusick et al., ).…”

Section: Introductionmentioning

confidence: 99%

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Identification of suitable reference genes during the formation of chlamydospores in Clonostachys rosea 67‐1

Zhang

Sun

et al. 2017

MicrobiologyOpen

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Clonostachys rosea is a potential biocontrol fungus that can produce highly resistant chlamydospores under specific conditions. To investigate the genes related to chlamydospore formation, we identified reliable reference genes for quantification of gene expression in C. rosea 67‐1 during sporulation. In this study, nine reference genes, actin (ACT), elongation factor 1 (EF1), glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH), histone (HIS), RNA polymerase II CTD phosphatase Fcp1 (RPP), succinate‐semialdehyde dehydrogenase (SSD), TATA‐binding protein (TBP), ubiquitin (UBQ), and ubiquitin‐conjugating enzyme (UCE), were selected and cloned from 67‐1, and their expression stability during chlamydospore formation was determined using reverse transcription quantitative PCR and assessed using the software geNorm, NormFinder and BestKeeper. The Ct values of the candidates ranged from 19.9 to 29.7, among which HIS,ACT and SSD exhibited high expression levels. The statistical analysis showed that ACT and SSD were most stably expressed, while UBQ and GAPDH showed relatively large variations under different culture conditions. Calculation of pairwise variation value indicated that two reference genes were required for precise quantification. Finally, ACT and SSD were selected to normalize gene expression during chlamydospore production in C. rosea 67‐1. To the best of our knowledge, this is the first report of SSD as a reference gene. This study will facilitate the accurate quantification of differentially expressed genes during the generation of chlamydospores and contribute to the investigation of the molecular mechanism underlying chlamydospore formation in C. rosea.

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“…All clean data were uploaded to the NCBI SRA repository with the accession numbers SRR4017358, 4017394−4017397, 4017399−4017403, 4017405, and 4017407. The average number of clean reads of the samples was 45,805,098, and each sample was mapped to the genome of C. rosea 67‐1 (Sun et al., ) with a map rate above 92.42%. The average number of expressed genes during all sampling time points was 14,755 (Table ).…”

Section: Resultsmentioning

confidence: 99%

Identification of genes related to chlamydospore formation in Clonostachys rosea 67‐1

et al. 2018

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Chlamydospores are specific structures that are of great significance to the commercialization of fungal biopesticides. To explore the genes associated with chlamydospore formation, a biocontrol fungus Clonostachys rosea 67-1 that is capable of producing resistant spores under particular conditions was investigated by transcriptome sequencing and analysis. A total of 549,661,174 clean reads were obtained, and a series of differentially expressed genes potentially involved in fungal chlamydospore formation were identified. At 36 hr, 67 and 117 genes were up- and downregulated in C. rosea during chlamydospore production, compared with the control for conidiation, and 53 and 24 genes were up- and downregulated at 72 hr. GO classification suggested that the differentially expressed genes were related to cellular component, biological process, and molecular function categories. A total of 188 metabolism pathways were linked to chlamydospore production by KEGG analysis. Sixteen differentially expressed genes were verified by reverse transcription quantitative PCR, and the expression profiles were consistent with the transcriptome data. To the best of our knowledge, it is the first report on the genes associated with chlamydospore formation in C. rosea. The results provide insight into the molecular mechanisms underlying C. rosea sporulation, which will assist the development of fungal biocontrol agents.

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Draft Genome Sequence of Mycoparasite Clonostachys rosea Strain 67-1

Cited by 31 publications

References 14 publications

Transformation of the endochitinase gene Chi67-1 in Clonostachys rosea 67-1 increases its biocontrol activity against Sclerotinia sclerotiorum

Transformation of the endochitinase gene Chi67-1 in Clonostachys rosea 67-1 increases its biocontrol activity against Sclerotinia sclerotiorum

Identification of suitable reference genes during the formation of chlamydospores in Clonostachys rosea 67‐1

Identification of genes related to chlamydospore formation in Clonostachys rosea 67‐1

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