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- Functional Analysis of a Subtilisin-like Serine Protease Gene from Biocontrol Fungus Trichoderma harzianum
- Haijuan Fan , Zhihua Liu , Rongshu Zhang , Na Wang , Kai Dou , Gulijimila Mijiti , Guiping Diao , Zhiying Wang
- J. Microbiol. 2014;52(2):129-138. Published online February 1, 2014
- DOI: https://doi.org/10.1007/s12275-014-3308-9
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- 20 Crossref
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Abstract
- The subtilisin-like serine protease gene ThSS45 has been cloned from Trichoderma harzianum ACCC30371. Its coding region is 1302 bp in length, encoding 433 amino acids, with a predicted protein molecular weight of 44.9 kDa and pI of 5.91. ThSS45 was shown by RT-qPCR analysis to be differentially transcribed in response to eight different treatments. The transcription of ThSS45 was up-regulated when grown in mineral medium, under carbon starvation, and nitrogen starvation, and in the presence of 1% root powder, 1% stem powder, and 1% leaf powder derived from Populus davidiana × P. bolleana (Shanxin poplar) aseptic seedlings. The highest increase in transcription approached 3.5-fold that of the control at 6 h under induction with 1% poplar root powder. The transcription of ThSS45 was also slightly up-regulated by 1% Alternaria alternata cell wall and 5% A. alternata fermentation liquid. Moreover, the analyses of coding and promoter regions of ThSS45 homologs indicated that serine protease may be involved in both mycoparasitism and antibiotic secretion. ThSS45 was cloned into the pGEX-4T-2 vector and then expressed in Escherichia coli BL21. The recombinant protein, with an expected molecular weight of approximately 69 kDa, was then purified. When transformant BL21-ss was induced with 1 mM IPTG for 6 h, the purified protease activity reached a peak of 18.25 U/ml at pH 7.0 and 40°C. In antifungal assays the purified protease obviously inhibited the growth of A. alternata mycelia.
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Citations
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- The role of Trichoderma koningii and Trichoderma harzianum in mitigating the combined stresses motivated by Sclerotiniasclerotiorum and salinity in common bean (Phaseolusvulgaris)
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Ying Sun, Na Yang, Sirui Li, Fei Chen, Yijing Xie, Canming Tang, Monica Höfte
Journal of Experimental Botany.2024; 75(11): 3500. CrossRef - Purification and Identification of the Nematicidal Activity of S1 Family Trypsin-Like Serine Protease (PRA1) from Trichoderma longibrachiatum T6 Through Prokaryotic Expression and Biological Function Assays
Nan Ma, Hang Lv, Solomon Boamah, Shuwu Zhang, Bingliang Xu
Genes.2024; 15(11): 1437. CrossRef - Genome and transcriptome sequencing of Trichoderma harzianum T4, an important biocontrol fungus of Rhizoctonia solani, reveals genes related to mycoparasitism
Yaping Wang, Jian Wang, Xiaochong Zhu, Wei Wang
Canadian Journal of Microbiology.2024; 70(3): 86. CrossRef - Strain improvement of Trichoderma harzianum for enhanced biocontrol capacity: Strategies and prospects
Ziyang Xiao, Qinqin Zhao, Wei Li, Liwei Gao, Guodong Liu
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Tree Physiology.2022; 42(12): 2627. CrossRef - Extracellular proteins of Trichoderma and their role in plant health
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Wenjing Yu, Gulijimila Mijiti, Ying Huang, Haijuan Fan, Yucheng Wang, Zhihua Liu
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Journal of Plant Biochemistry and Biotechnology.2018; 27(3): 351. CrossRef - Subtilisin-like serine protease gene TghSS42 from Trichoderma ghanense ACCC 30153 was successfully expressed in Escherichia coli and recombinant protease rTghSS42 exhibited antifungal ability to five phytopathogens
HUIFANG ZHANG, NA WANG, YUCHENG WANG, JINJIE WANG, HONG ZHENG, ZHIHUA LIU
Biocontrol Science.2017; 22(3): 145. CrossRef - A novel organic solvent- and detergent-stable serine alkaline protease from Trametes cingulata strain CTM10101
Maroua Omrane Benmrad, Emna Moujehed, Mouna Ben Elhoul, Nadia Zaraî Jaouadi, Sondes Mechri, Hatem Rekik, Sidali Kourdali, Mohamed El Hattab, Abdelmalek Badis, Sami Sayadi, Samir Bejar, Bassem Jaouadi
International Journal of Biological Macromolecules.2016; 91: 961. CrossRef - Differential Response of Extracellular Proteases of Trichoderma Harzianum Against Fungal Phytopathogens
Vivek Sharma, Richa Salwan, Prem N. Sharma
Current Microbiology.2016; 73(3): 419. CrossRef - Fungal proteins and genes associated with biocontrol mechanisms of soil-borne pathogens: a review
Yohann Daguerre, Katarzyna Siegel, Véronique Edel-Hermann, Christian Steinberg
Fungal Biology Reviews.2014; 28(4): 97. CrossRef
- The role of Trichoderma koningii and Trichoderma harzianum in mitigating the combined stresses motivated by Sclerotiniasclerotiorum and salinity in common bean (Phaseolusvulgaris)
- Cloning, Annotation and Expression Analysis of Mycoparasitism-Related Genes in Trichoderma harzianum 88
- Lin Yao , Qian Yang , Jinzhu Song , Chong Tan , Changhong Guo , Li Wang , Lianhai Qu , Yun Wang
- J. Microbiol. 2013;51(2):174-182. Published online April 27, 2013
- DOI: https://doi.org/10.1007/s12275-013-2545-7
- 53 View
- 0 Download
- 9 Scopus
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Abstract
- Trichoderma harzianum 88, a filamentous soil fungus, is an effective biocontrol agent against several plant pathogens. High-throughput sequencing was used here to study the mycoparasitism mechanisms of T. harzianum 88. Plate confrontation tests of T. harzianum 88 against plant pathogens were conducted, and a cDNA library was constructed from T. harzianum 88 mycelia in the presence of plant pathogen cell walls. Randomly selected transcripts from the cDNA library were compared with eukaryotic plant and fungal genomes. Of the 1,386 transcripts sequenced, the most abundant Gene Ontology (GO) classification group was “physiological process”. Differential expression of 19 genes was confirmed by real-time RT-PCR at different mycoparasitism stages against plant pathogens. Gene expression analysis revealed the transcription of various genes involved in mycoparasitism of T. harzianum 88. Our study provides helpful insights into the mechanisms of T. harzianum 88-plant pathogen interactions.
- Construction of a Streptomyces lydicus A01 Transformant with a chit42 Gene from Trichoderma harzianum P1 and Evaluation of Its Biocontrol Activity against Botrytis cinerea
- Qiong Wu , Linquan Bai , Weicheng Liu , Yingying Li , Caige Lu , Yaqian Li , Kehe Fu , Chuanjin Yu , Jie Chen
- J. Microbiol. 2013;51(2):166-173. Published online April 27, 2013
- DOI: https://doi.org/10.1007/s12275-013-2321-8
- 51 View
- 0 Download
- 16 Scopus
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Abstract
- Streptomyces lydicus A01 and Trichoderma harzianum P1 are potential biocontrol agents of fungal diseases in plants. S. lydicus A01 produces natamycin to bind the ergosterol of the fungal cell membrane and inhibits the growth of Botrytis cinerea. T. harzianum P1, on the other hand, features high chitinase activity and decomposes the chitin in the cell wall of B. cinerea. To obtain the synergistic biocontrol effects of chitinase and natamycin on Botrytis cinerea, this study transformed the chit42 gene from T. harzianum P1 to S. lydicus A01. The conjugal transformant (CT) of S. lydicus A01 with the chit42 gene was detected using polymerase chain reaction (PCR). Associated chitinase activity and natamycin production were examined using the 3, 5-dinitrosalicylic acid (DNS) method and ultraviolet spectrophotometry, respectively. The S. lydicus A01-chit42 CT showed substantially higher chitinase activity and natamycin production than its wild type strain (WT). Consequently, the biocontrol effects of S. lydicus A01-chit42 CT on B. cinerea, including inhibition to spore germination and mycelial growth, were highly improved compared with those of the WT. Our research indicates that the biocontrol effect of Streptomyces can be highly improved by transforming the exogenous resistance gene, i.e. chit42 from Trichoderma, which not only enhances the production of antibiotics, but also provides a supplementary function by degrading the cell walls of the pathogens.
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