[논문]Endophytic Trichoderma gamsii YIM PH30019: a promising biocontrol agent with hyperosmolar, mycoparasitism, and antagonistic activities of induced volatile organic compounds on root-rot pathogenic fungi of Panax notoginseng

Chen, Jin-Lian; Sun, Shi-Zhong; Miao, Cui-Ping; Wu, Kai; Chen, You-Wei; Xu, Li-Hua; Guan, Hui-Lin; Zhao, Li-Xing

doi:10.1016/j.jgr.2015.09.006

[국내논문] Endophytic Trichoderma gamsii YIM PH30019: a promising biocontrol agent with hyperosmolar, mycoparasitism, and antagonistic activities of induced volatile organic compounds on root-rot pathogenic fungi of Panax notoginseng 원문보기

Journal of ginseng research = 高麗人參學會誌, v.40 no.4, 2016년, pp.315 - 324

Chen, Jin-Lian (School of Energy and Environment Science, Yunnan Normal University) , Sun, Shi-Zhong (School of Energy and Environment Science, Yunnan Normal University) , Miao, Cui-Ping (Key Laboratory of Microbial Diversity in South West China of Ministry of Education and Laboratory for Conservation and Utilization of Bio-Resources, Yunnan Institute of Microbiology, Yunnan University) , Wu, Kai (School of Energy and Environment Science, Yunnan Normal University) , Chen, You-Wei (Key Laboratory of Microbial Diversity in South West China of Ministry of Education and Laboratory for Conservation and Utilization of Bio-Resources, Yunnan Institute of Microbiology, Yunnan University) , Xu, Li-Hua (Key Laboratory of Microbial Diversity in South West China of Ministry of Education and Laboratory for Conservation and Utilization of Bio-Resources, Yunnan Institute of Microbiology, Yunnan University) , Guan, Hui-Lin (School of Energy and Environment Science, Yunnan Normal University) , Zhao, Li-Xing (Key Laboratory of Microbial Diversity in South West China of Ministry of Education and Laboratory for Conservation)

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Background: Biocontrol agents are regarded as promising and environmental friendly approaches as agrochemicals for phytodiseases that cause serious environmental and health problems. Trichoderma species have been widely used in suppression of soil-borne pathogens. In this study, an endophytic fungus, Trichoderma gamsii YIM PH30019, from healthy Panax notoginseng root was investigated for its biocontrol potential. Methods: In vitro detached healthy roots, and pot and field experiments were used to investigate the pathogenicity and biocontrol efficacy of T. gamsii YIM PH30019 to the host plant. The antagonistic mechanisms against test phytopathogens were analyzed using dual culture, scanning electron microscopy, and volatile organic compounds (VOCs). Tolerance to chemical fertilizers was also tested in a series of concentrations. Results: The results indicated that T. gamsii YIM PH30019 was nonpathogenic to the host, presented appreciable biocontrol efficacy, and could tolerate chemical fertilizer concentrations of up to 20%. T. gamsii YIM PH30019 displayed antagonistic activities against the pathogenic fungi of P. notoginseng via production of VOCs. On the basis of gas chromatography-mass spectrometry, VOCs were identified as dimethyl disulfide, dibenzofuran, methanethiol, ketones, etc., which are effective ingredients for antagonistic activity. T. gamsii YIM PH30019 was able to improve the seedlings' emergence and protect P. notoginseng plants from soil-borne disease in the continuous cropping field tests. Conclusion: The results suggest that the endophytic fungus T. gamsii YIM PH30019 may have a good potential as a biological control agent against notoginseng phytodiseases and can provide a clue to further illuminate the interactions between Trichoderma and phytopathogens.

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2) Colony diameter was an average value of three replicates.
3) No growth was observed in our tests.
1) T. gamsii YIM PH30019 was inoculated in the DCWB medium.
2) T. gamsii YIM PH30019 was inoculated in the above medium without deactivated cell wall.

제안 방법

Antagonistic activities caused by VOCs were tested on the deactivated cell wall agar medium (DCWA) (15.0 g deactivated cell walls, 6.9 g NaH2PO4, 2.0 g KH2PO4, 1.4 g (NH4)2SO4, 1.0 g peptone, 0.3 g MgSO4·7H2O, 0.3 g urea, and 15.0 g agar in 1 L distilled water, pH not adjusted), and VOCs were collected by culturing T. gamsii YIM PH30019 in the deactivated cell wall broth medium (DCWB).
notoginseng, an endophytic fungus with mycoparasitic activity was obtained from a healthy notoginseng root. In this study, experiments were carried out to estimate: (1) the pathogenicity of Trichoderma gamsii YIM PH30019 to its host plant; (2) biocontrol efficacy to phytodiseases in the continuous field experiments; (3) tolerance capacity to chemical fertilizers; (4) mycoparasitism and induced VOCs antagonistic activities against phytopathogens; (5) VOCs identified with gas chromatography-mass spectrometry (GCMS).
gamsii YIM PH30019, three chemical fertilizers were selected—ammonium chloride, potassium nitrate, and ammonium dihydrogen phosphate—and mixed at a ratio of 1:1:1 (w/w/w). A 6-mm plug of actively growing T. gamsii YIM PH30019 was placed on the center of a petri dish containing 25 mL PDA medium amended with chemical fertilizers at the following concentrations (w/v): 0%, 2%, 4%, 6%, 8%, 10%, 12%, 14%, 16%, 18%, 20%, 22%, 24%, 26%, 28%, and 30%. Each concentration was repeated thrice.
gamsii antagonism of VOCs induced by pathogenic fungi in the process of biocontrol has been poorly studied. In this study, deactivated cell walls of pathogenic fungi could induce T. gamsii YIM PH30019 to produce different VOC profiles, including dimethyl disulfide, dibenzofuran, methanethiol, and ketones. It is noteworthy that dimethyl disulfide is reported to have favorable antagonistic activity against insects and pathogens associated with many important plants and crops [48,49].
Disease coverage of type 0, 1, 2, 3, and 4 lesions for each P. notoginseng root was evaluated following indicators [27] with modification, where 0 ¼ no lesions, 1 = one to several lesions (roots blacking < 25%), 2 = extensive lesions or several entire roots necrotic (25-50% roots blacking), 3 = lesions on roots and darkening of crown (50-75% root blacking), 4 = extensive darkening of crown (75-100% roots blacking).

대상 데이터

The treatment without inoculation with YIM PH30019 was set as control. Six replications were used for treatment and control. The pots were incubated at 25°C for 2 mo, and P.
YIM PH30019, pathogenic fungi, and the mixture of YIM PH30019 and test pathogenic fungus were inoculated with a healthy root in a pot containing 400 g soil autoclaved at 121°C for 60 min. Four replicates were set for each treatment. The pots were kept in a shed similar to the field-planting condition.
notoginseng harvested in the previous year were used. The experimental field is located at Xiangshuilong Village, a traditional notoginseng cultivation center in Wenshan, Yunnan. Rice bran was mixed with equal mass water, and autoclaved at 121°C, for 60 min.

성능/효과

5). The highest inhibition rate appeared on the 4^th d (Fig. 6), reaching 45.0%, 37.5%, 38.6%, and 46.3% for E. nigrum, S. lignicola, P. herbarum, and F. flocciferum, respectively.
Nonchemical methods that effectively control plant diseases are highly desirable. The results give us insight that the deactivated cell walls of pathogenic fungi could be an effective activator for T. gamsii YIM PH30019 to produce more VOCs that inhibit the growth and metabolism of pathogenic fungi. This interesting phenomenon warrants further investigation in ongoing studies.

참고문헌 (53)

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[국내논문] Endophytic Trichoderma gamsii YIM PH30019: a promising biocontrol agent with hyperosmolar, mycoparasitism, and antagonistic activities of induced volatile organic compounds on root-rot pathogenic fungi of Panax notoginseng 원문보기

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[국내논문] Endophytic Trichoderma gamsii YIM PH30019: a promising biocontrol agent with hyperosmolar, mycoparasitism, and antagonistic activities of induced volatile organic compounds on root-rot pathogenic fungi of Panax notoginseng 원문보기

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