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Haijuan Fan,Zhihua Liu,Rongshu Zhang,Na Wang,Kai Dou,Gulijimila Mijiti,Guiping Diao,Zhiying Wang 한국미생물학회 2014 The journal of microbiology Vol.52 No.2
The subtilisin-like serine protease gene ThSS45 has beencloned from Trichoderma harzianum ACCC30371. Its codingregion is 1302 bp in length, encoding 433 amino acids,with a predicted protein molecular weight of 44.9 kDa andpI of 5.91. ThSS45 was shown by RT-qPCR analysis to bedifferentially transcribed in response to eight different treatments. The transcription of ThSS45 was up-regulated whengrown in mineral medium, under carbon starvation, andnitrogen starvation, and in the presence of 1% root powder,1% stem powder, and 1% leaf powder derived from Populusdavidiana × P. bolleana (Shanxin poplar) aseptic seedlings. The highest increase in transcription approached 3.5-foldthat of the control at 6 h under induction with 1% poplarroot powder. The transcription of ThSS45 was also slightlyup-regulated by 1% Alternaria alternata cell wall and 5% A. alternata fermentation liquid. Moreover, the analyses of codingand promoter regions of ThSS45 homologs indicatedthat serine protease may be involved in both mycoparasitismand antibiotic secretion. ThSS45 was cloned into the pGEX-4T-2 vector and then expressed in Escherichia coli BL21. Therecombinant protein, with an expected molecular weight ofapproximately 69 kDa, was then purified. When transformantBL21-ss was induced with 1 mM IPTG for 6 h, thepurified protease activity reached a peak of 18.25 U/ml atpH 7.0 and 40°C. In antifungal assays the purified proteaseobviously inhibited the growth of A. alternata mycelia.
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Yue-Feng Wang,Xue-Yue Hou,Chuan-Ying Jiang,Tong-Tong Zhai,Rui Miao,Jun-Jie Deng,Zhi-Hong Yao,Rongshu Zhang 한국원예학회 2021 Horticulture, Environment, and Biotechnology Vol.62 No.2
Trichoderma spp. are widely applied, eco-friendly mycofungicides and plant growth promoters. Native Trichoderma strainsare likely to have more productive, stable biocontrol and biofertilizer activities since they are well adapted to the local environment. In this study, we isolated a native Trichoderma strain ‘Th62’ from the rhizosphere soil of wild Chelidonium majusplants in Harbin, China (126.6341°E, 45.7242°N). The isolated Trichoderma strain was identifi ed as a T. harzianum strainvia morphological observation and molecular methods based on the rDNA internal transcribed spacer region and elongationfactor-1α gene sequences. Signifi cant antagonistic activities of Th62 against fi ve soil-borne fungal phytopathogens,Fusarium oxysporum , Sclerotinia sclerotiorum , Alternaria alternata , Cytospora chrysosperma, and Rhizoctonia solani ,were confi rmed by dual-culture assays. Furthermore, the crude fermentation products of Th62 also displayed antifungalactivities against these fi ve pathogens. To evaluate the function of Th62 as a biofertilizer, we subsequently applied Th62on cockscomb ( Celosia cristata L), a plant species with both ornamental and medicinal values, by inoculation with Th62conidia at diff erent concentrations, 1 × 10 10 cfu mL −1 , 1 × 10 11 cfu mL −1 , and 1 × 10 12 cfu mL −1 . The benefi cial eff ects ofTh62 were evaluated by measuring the growth and photosynthetic traits of the inoculated cockscomb plants, and the resultsdemonstrated that Th62 signifi cantly improved the photosynthetic effi ciency, photosynthetic capacity, and the adaptabilityto intense light of the inoculated cockscomb plants compared to the controls. Consistently, Th62 inoculation signifi cantlyimproved the growth and fl ower yield of cockscomb. We presented a positive case of isolating and applying native microbialresources on local plantation practices.
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