A preliminary study of the antifungal activity and antagonism mechanisms of Trichoderma spp. against turfgrass pathogens

doi:10.11686/cyxb2020333

Abstract

Abstract:

Trichoderma spp.， is an antagonistic group of microorganisms widely found in nature and many species have important biocontrol value for use against plant pathogens. In order to evaluate Trichoderma spp. as a potential biocontrol agent for turfgrass disease， the antagonistic effects of four Trichoderma strains against 14 turfgrass pathogens were determined by agar confrontation culture and the mechanism of inhibition was studied by microscopic observation， the ‘cellophane method’ and bacteriostatic testing of crude extract. The results showed that tested Trichoderma spp. isolates had antagonism against all tested pathogens. Among the tested Trichoderma spp. isolates， an isolate designated SQ-1Q-18 had the greatest antibacterial efficacy. The antagonistic activity against Rhizoctonia zeae and Laetisaria fuciformis was scored as level I， with 100% inhibition rate； for Botrytis cinerea， Colletotrichum hainanese and Rhizoctonia solani the antagonistic activity was scored as level Ⅱ， with over 80% inhibition rate for the most potent Trichoderma isolate， and an average inhibition rate against these three pathogens of up to 72.8%. Further study of the inhibitory mechanism of the SQ-1Q-18 isolate on 14 pathogens was that Trichoderma surrounded the growth space of pathogens by generating a bacteriostatic circle， and covered and inserted into the pathogen colony and entwined with it to make the pathogen mycelium become thin， constricted and even broken. Trichoderma could also penetrate directly into a pathogen mycelium to absorb nutrients and cause mycelium cytolysis. In addition， the SQ-1Q-18 isolate could produce antagonistic compounds with inhibitory activity against the turfgrass pathogens. The antagonistic substances produced by the cellophane culture method had strong antagonistic effects against Sclerotinia homoeocarpa， Nigrospora sphaerica and L. fuciformis， and the relative inhibitory rate was as high as 91.77%， 89.80% and 79.59%， respectively. The inhibition rate of Trichoderma crude extract against S. homoeocarpa and Bipolaris peregianensis was 43.60% and 42.91%， respectively. Furthermore， the SQ-1Q-18 isolate was identified as Trichoderma harzianum by morphological characteristics and ITS-rDNA sequence. This study provides scientific data for the research and development of a biocontrol agent for efficient control of turfgrass fungal diseases.

Key words: Trichoderma spp., turfgrass pathogens, antibacterial activity, antagonistic mechanism, Trichoderma harzianum

Wu ZHANG, Jin-yu YANG, Xiang LU, Jin-mei LIN, Xue-li NIU. A preliminary study of the antifungal activity and antagonism mechanisms of Trichoderma spp. against turfgrass pathogens[J]. Acta Prataculturae Sinica, 2021, 30(9): 137-149.

Figures/Tables 9

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序号No.	菌株Isolates	菌株代号Isolate code
1	木贼镰刀菌Fusarium equiseti	FE
2	新月弯孢霉Curvularia lunata	CL
3	球黑孢霉Nigrospora sphaerica	NS
4	草坪草墨斑病菌Curvularia malina	CM
5	佩立金平脐蠕孢Bipolaris peregianensis	BP
6	草茎点霉Phoma herbarum	PH
7	立枯丝核菌Rhizoctonia solani	RS
8	地衣状伏革菌Laetisaria fuciformis	LF
9	地毯草炭疽菌Colletotrichum hainanese	CH
10	灰葡萄孢菌Botrytis cinerea	BC
11	草坪草币斑病病原菌Sclerotinia homoeocarpa	SH
12	禾顶囊壳禾谷变种Gaeumannomyces graminis var. graminis	GG
13	雀稗微座孢Microdochium paspali	MP
14	玉蜀黍丝核菌Rhizoctonia zeae	RZ

序号No.	菌株Isolates	菌株代号Isolate code
1	木贼镰刀菌Fusarium equiseti	FE
2	新月弯孢霉Curvularia lunata	CL
3	球黑孢霉Nigrospora sphaerica	NS
4	草坪草墨斑病菌Curvularia malina	CM
5	佩立金平脐蠕孢Bipolaris peregianensis	BP
6	草茎点霉Phoma herbarum	PH
7	立枯丝核菌Rhizoctonia solani	RS
8	地衣状伏革菌Laetisaria fuciformis	LF
9	地毯草炭疽菌Colletotrichum hainanese	CH
10	灰葡萄孢菌Botrytis cinerea	BC
11	草坪草币斑病病原菌Sclerotinia homoeocarpa	SH
12	禾顶囊壳禾谷变种Gaeumannomyces graminis var. graminis	GG
13	雀稗微座孢Microdochium paspali	MP
14	玉蜀黍丝核菌Rhizoctonia zeae	RZ

病原菌 Pathogens	对峙培养结果 Confrontation incubation results	拮抗等级 Antagonist level
地衣状伏革菌 L. fuciformis	当两菌菌落接触后地衣状伏革菌停止生长，3 d后木霉菌菌丝迅速覆盖地衣状伏革菌菌落表面，5 d后地衣状伏革菌整个菌落被长枝木霉菌菌丝完全覆盖，并产生大量孢子。When the two colonies contact， the growth of L. fuciformis stop， Trichoderma spp. mycelium quickly cover the surface of L. fuciformis colony after 3 days， and Trichoderma spp. mycelium completely covered the entire colony after 5 days， and produced a large number of conidia.	Ⅰ
玉蜀黍丝核菌 R. zeae	对峙培养2 d后两菌落开始接触，木霉菌菌丝开始覆盖玉蜀黍丝核菌菌落表面，5 d后玉蜀黍丝核菌整个菌落被木霉菌菌丝完全侵占，并产生大量孢子抑制其生长。After 2 days of culture， the two colonies began to contact， and Trichoderma spp. mycelium began to cover the surface of R. zeae colony. After 5 days， the entire colony of R. zeae was completely invaded by Trichoderma spp. mycelium， and a large number of conidia were produced to R. zeae its growth.	Ⅰ
草坪草墨斑病菌 C. malina	两菌菌落交接后，草坪草墨斑病菌停止生长，无明显的抑菌圈，但5 d后SQ-1Q-18木霉菌菌丝围绕草坪草墨斑病菌菌落周围生长，并逐渐向草坪草墨斑病菌菌落中心扩展及蔓延。After the two colonies contract， the growth of C. malina stopped and there was no obvious bacteriostatic circle. However， after 5 days， the mycelium of Trichohypha spp. isolate SQ-1Q-18 grew around the C. malina， and gradually expanded and spread to the center of the colony of C. malina.	Ⅱ
佩立金平脐蠕孢B. peregianensis	两菌落接触后，SQ-1Q-18木霉菌大面积侵占佩立金平脐蠕孢的生长空间，使佩立金平脐蠕孢的生长受到抑制，5 d后木霉菌菌丝将其包围，并在其边缘产生大量的分生孢子。After the two colonies contacted， the Trichohypha spp. isolate SQ-1Q-18 occupied a large area of the growth space of B. peregianensis， which inhibited the growth of B. peregianensis. After 5 days， Trichoderma spp. surrounded the colony of B. peregianensis and produced a large number of conidia at its edge.	Ⅱ
立枯丝核菌 R. solani	立枯丝核菌生长速度快，但菌丝少，2 d后两菌落开始接触，5 d后木霉菌落几乎蔓延整个平板，使立枯丝核菌的生长空间受到极大的限制，无明显的交界及抑菌带。The rate of growth of R. solani was fast， but the mycelium were sparse. After 2 days， the two colonies began to contact， and after 5 days， Trichoderma spp. colonies almost occupied over the entire plate， which greatly restricted the growth space of R. solani.	Ⅱ
地毯草炭疽菌C. hainanese	地毯草炭疽菌生长较慢，木霉菌生长较快，当两菌落菌丝接触时，木霉菌菌落已占据大部分生长空间，导致地毯草炭疽菌停止生长，被木霉菌菌落迅速包围并覆盖。C. hainanese mycelium grew slowly， while Trichoderma spp. grows faster. When the two colonies contact， Trichoderma spp. colonies have occupied most of the growth space， resulting in the stopped growth of C. hainanese mycelium and the colonies are quickly surrounded and covered by Trichoderma spp.	Ⅱ
灰葡萄孢菌 B. cinerea	当两菌菌落接触后，灰葡萄孢菌停止生长，3 d后木霉菌菌丝迅速覆盖灰葡萄孢菌菌落表面，5 d后木霉菌菌丝已将灰葡萄孢菌菌落覆盖了一半，并产生大量孢子。When the two colonies contacted， the growth of B. cinerea stopped， and Trichoderma spp. mycelium covered the surface of B. cinerea rapidly after 3 days， then Trichoderma spp. mycelium covered half of B. cinerea after 5 days， and produced a large number of conidia.	Ⅱ
禾顶囊壳禾谷变种G. graminis var. graminis	禾顶囊壳禾谷变种病原菌生长较慢，当两菌落菌丝接触时，木霉菌菌落已占据大部分生长空间，导致禾顶囊壳禾谷变种停止生长，被木霉菌菌落迅速覆盖。The growth rate of G. graminis var. graminis was relatively slow. When the two colonies contacted， Trichoderma spp. had occupied most of the growth space， leading to the stop of growth of G. graminis var. graminis and the rapid coverage Trichoderma spp.	Ⅱ
雀稗微座孢 M. paspali	两菌落开始接触后，木霉菌逐渐侵占雀稗微座孢的营养及生长空间，使雀稗微座孢无法继续生长，很快被木霉菌落覆盖。After the two colonies began to contact， Trichoderma spp. gradually invaded the nutrition and growth space of M. paspali， making it impossible to continue to grow and was soon covered by Trichoderma spp. colony.	Ⅱ
木贼镰刀菌 F. equiseti	两菌落接触后，木贼镰刀菌菌落停止生长，5 d后整个菌落被木霉菌包围，但没有被覆盖，并在两菌交接处产生大量分生孢子。After the two colonies began to contact， the growth of F. equiseti stopped. After 5 days growth， the entire colony was surrounded but not covered by Trichoderma spp.， and a large number of conidia were produced at the junction of the two colonies.	Ⅲ
草坪草币斑病病原菌S. homoeocarpa	当两菌落接触后，木霉菌菌丝开始向草坪草币斑病病原菌中心扩展和蔓延，使其生长受到抑制，无明显的抑菌带。5 d后，草坪草币斑病病原菌已被大面积覆盖。When the two colonies contacted， Trichoderma spp. mycelium began to expand and spread to the center of the colony S. homoeocarpa， which inhibited its growth without obvious bacteriostatic zone. After 5 days growth， the colony S. homoeocarpa had been extensively covered.	Ⅲ
新月弯孢霉 C. lunata	两菌落接触后，在接触边缘产生了棕色抑菌带，新月弯孢霉停止生长，抑菌带处木霉孢子较少，也无覆盖病原菌的趋势。After the two colonies contacted， there was a brown bacteriostatic zone at the contact edge， and the growth of C. lunata stopped， and there were few of Trichoderma spp. conidia in the bacteriostatic zone， and there was no tendency to cover the colony of C. lunata.	Ⅳ
球黑孢霉 N. sphaerica	两菌菌落交接后球黑孢霉停止生长，产生明显棕黄色的抑菌圈，且交接处球黑孢霉部分菌丝消解，使球黑孢霉菌丝不能向外扩展和生长，但木霉菌菌丝并没有将其覆盖。After the two colonies contacted， the growth of N. sphaerica stopped， resulting in an obvious brownish yellow inhibition zone. Moreover， part of mycelium of N. sphaerica were destroyed at the junction， making the mycelium of N. sphaerica unable to grow outward， but the mycelium of Trichoderma spp. did not cover colony of N. sphaerica.	Ⅳ
草茎点霉 P. herbarum	当两菌落接触后，SQ-1Q-18木霉大量侵占草茎点霉的生长空间，使草茎点霉的生长受到抑制，5 d后木霉菌菌丝将其包围，并在其边缘产生大量的分生孢子。When the two colonies contacted， Trichoderma spp. isolate SQ-1Q-18 invaded the growth space of P. herbarum， which inhibited its growth. After 5 days， Trichoderma spp. mycelium surrounded the colony of P. herbarum and produced a large number of conidia at its edge.	Ⅳ

病原菌 Pathogens	对峙培养结果 Confrontation incubation results	拮抗等级 Antagonist level
地衣状伏革菌 L. fuciformis	当两菌菌落接触后地衣状伏革菌停止生长，3 d后木霉菌菌丝迅速覆盖地衣状伏革菌菌落表面，5 d后地衣状伏革菌整个菌落被长枝木霉菌菌丝完全覆盖，并产生大量孢子。When the two colonies contact， the growth of L. fuciformis stop， Trichoderma spp. mycelium quickly cover the surface of L. fuciformis colony after 3 days， and Trichoderma spp. mycelium completely covered the entire colony after 5 days， and produced a large number of conidia.	Ⅰ
玉蜀黍丝核菌 R. zeae	对峙培养2 d后两菌落开始接触，木霉菌菌丝开始覆盖玉蜀黍丝核菌菌落表面，5 d后玉蜀黍丝核菌整个菌落被木霉菌菌丝完全侵占，并产生大量孢子抑制其生长。After 2 days of culture， the two colonies began to contact， and Trichoderma spp. mycelium began to cover the surface of R. zeae colony. After 5 days， the entire colony of R. zeae was completely invaded by Trichoderma spp. mycelium， and a large number of conidia were produced to R. zeae its growth.	Ⅰ
草坪草墨斑病菌 C. malina	两菌菌落交接后，草坪草墨斑病菌停止生长，无明显的抑菌圈，但5 d后SQ-1Q-18木霉菌菌丝围绕草坪草墨斑病菌菌落周围生长，并逐渐向草坪草墨斑病菌菌落中心扩展及蔓延。After the two colonies contract， the growth of C. malina stopped and there was no obvious bacteriostatic circle. However， after 5 days， the mycelium of Trichohypha spp. isolate SQ-1Q-18 grew around the C. malina， and gradually expanded and spread to the center of the colony of C. malina.	Ⅱ
佩立金平脐蠕孢B. peregianensis	两菌落接触后，SQ-1Q-18木霉菌大面积侵占佩立金平脐蠕孢的生长空间，使佩立金平脐蠕孢的生长受到抑制，5 d后木霉菌菌丝将其包围，并在其边缘产生大量的分生孢子。After the two colonies contacted， the Trichohypha spp. isolate SQ-1Q-18 occupied a large area of the growth space of B. peregianensis， which inhibited the growth of B. peregianensis. After 5 days， Trichoderma spp. surrounded the colony of B. peregianensis and produced a large number of conidia at its edge.	Ⅱ
立枯丝核菌 R. solani	立枯丝核菌生长速度快，但菌丝少，2 d后两菌落开始接触，5 d后木霉菌落几乎蔓延整个平板，使立枯丝核菌的生长空间受到极大的限制，无明显的交界及抑菌带。The rate of growth of R. solani was fast， but the mycelium were sparse. After 2 days， the two colonies began to contact， and after 5 days， Trichoderma spp. colonies almost occupied over the entire plate， which greatly restricted the growth space of R. solani.	Ⅱ
地毯草炭疽菌C. hainanese	地毯草炭疽菌生长较慢，木霉菌生长较快，当两菌落菌丝接触时，木霉菌菌落已占据大部分生长空间，导致地毯草炭疽菌停止生长，被木霉菌菌落迅速包围并覆盖。C. hainanese mycelium grew slowly， while Trichoderma spp. grows faster. When the two colonies contact， Trichoderma spp. colonies have occupied most of the growth space， resulting in the stopped growth of C. hainanese mycelium and the colonies are quickly surrounded and covered by Trichoderma spp.	Ⅱ
灰葡萄孢菌 B. cinerea	当两菌菌落接触后，灰葡萄孢菌停止生长，3 d后木霉菌菌丝迅速覆盖灰葡萄孢菌菌落表面，5 d后木霉菌菌丝已将灰葡萄孢菌菌落覆盖了一半，并产生大量孢子。When the two colonies contacted， the growth of B. cinerea stopped， and Trichoderma spp. mycelium covered the surface of B. cinerea rapidly after 3 days， then Trichoderma spp. mycelium covered half of B. cinerea after 5 days， and produced a large number of conidia.	Ⅱ
禾顶囊壳禾谷变种G. graminis var. graminis	禾顶囊壳禾谷变种病原菌生长较慢，当两菌落菌丝接触时，木霉菌菌落已占据大部分生长空间，导致禾顶囊壳禾谷变种停止生长，被木霉菌菌落迅速覆盖。The growth rate of G. graminis var. graminis was relatively slow. When the two colonies contacted， Trichoderma spp. had occupied most of the growth space， leading to the stop of growth of G. graminis var. graminis and the rapid coverage Trichoderma spp.	Ⅱ
雀稗微座孢 M. paspali	两菌落开始接触后，木霉菌逐渐侵占雀稗微座孢的营养及生长空间，使雀稗微座孢无法继续生长，很快被木霉菌落覆盖。After the two colonies began to contact， Trichoderma spp. gradually invaded the nutrition and growth space of M. paspali， making it impossible to continue to grow and was soon covered by Trichoderma spp. colony.	Ⅱ
木贼镰刀菌 F. equiseti	两菌落接触后，木贼镰刀菌菌落停止生长，5 d后整个菌落被木霉菌包围，但没有被覆盖，并在两菌交接处产生大量分生孢子。After the two colonies began to contact， the growth of F. equiseti stopped. After 5 days growth， the entire colony was surrounded but not covered by Trichoderma spp.， and a large number of conidia were produced at the junction of the two colonies.	Ⅲ
草坪草币斑病病原菌S. homoeocarpa	当两菌落接触后，木霉菌菌丝开始向草坪草币斑病病原菌中心扩展和蔓延，使其生长受到抑制，无明显的抑菌带。5 d后，草坪草币斑病病原菌已被大面积覆盖。When the two colonies contacted， Trichoderma spp. mycelium began to expand and spread to the center of the colony S. homoeocarpa， which inhibited its growth without obvious bacteriostatic zone. After 5 days growth， the colony S. homoeocarpa had been extensively covered.	Ⅲ
新月弯孢霉 C. lunata	两菌落接触后，在接触边缘产生了棕色抑菌带，新月弯孢霉停止生长，抑菌带处木霉孢子较少，也无覆盖病原菌的趋势。After the two colonies contacted， there was a brown bacteriostatic zone at the contact edge， and the growth of C. lunata stopped， and there were few of Trichoderma spp. conidia in the bacteriostatic zone， and there was no tendency to cover the colony of C. lunata.	Ⅳ
球黑孢霉 N. sphaerica	两菌菌落交接后球黑孢霉停止生长，产生明显棕黄色的抑菌圈，且交接处球黑孢霉部分菌丝消解，使球黑孢霉菌丝不能向外扩展和生长，但木霉菌菌丝并没有将其覆盖。After the two colonies contacted， the growth of N. sphaerica stopped， resulting in an obvious brownish yellow inhibition zone. Moreover， part of mycelium of N. sphaerica were destroyed at the junction， making the mycelium of N. sphaerica unable to grow outward， but the mycelium of Trichoderma spp. did not cover colony of N. sphaerica.	Ⅳ
草茎点霉 P. herbarum	当两菌落接触后，SQ-1Q-18木霉大量侵占草茎点霉的生长空间，使草茎点霉的生长受到抑制，5 d后木霉菌菌丝将其包围，并在其边缘产生大量的分生孢子。When the two colonies contacted， Trichoderma spp. isolate SQ-1Q-18 invaded the growth space of P. herbarum， which inhibited its growth. After 5 days， Trichoderma spp. mycelium surrounded the colony of P. herbarum and produced a large number of conidia at its edge.	Ⅳ

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