棘孢木霉活性代谢产物的初步分离及其抑菌与促生长功能
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摘要
为探究棘孢木霉CBS 433.97(Trichoderma asperellum CBS 433.97)的活性代谢产物在促进植物生长与抑制病原菌等方面的功能,本研究初步分离了棘孢木霉代谢产物,并分析了其活性成分对尖孢镰刀菌MC-1和CS-5的抑菌效果,同时研究了棘孢木霉代谢产物中的活性成分对小麦的株高、根长和发芽率的影响。用不同的萃取剂萃取棘孢木霉发酵液中的活性成分,正丁醇萃取相获得的粗提物对MC-1与CS-5的抑菌效果最好。进一步对正丁醇萃取相获得的粗提物进行薄层层析,获得了4个组分TA_1,TA_2,TA_3和TA_4,4个组分对MC-1与CS-5均无明显抑制作用,但组分TA_3在浓度为1 g·L~(-1)时,对小麦生长具有明显的促进作用。由此可见,棘孢木霉CBS 433.97代谢产物中具有能够抑制病原菌的生长、促进植物生长发育的活性成分,具有较好的开发潜力。
To explore the functions of active metabolites of Trichoderma asperellum CBS 433.97 on promoting plants growth and inhibitting phytopathogen, the metabolites were separated preliminarily, and the inhibitions of active components on Fusarium oxysporum MC-1 and Fusarium oxysporum CS-5, as well as the effects of the active components from T. asperellum CBS 433.97 metabolites on plant height, root length and germination rate of wheat were analyzed. The active components of the fermentation broth from T. asperellum CBS 433.97 were extracted with different ectracting agent, the crude extracts obtained by the n-butanol extraction phase had the best inhibitory effect on MC-1 and CS-5. The crude extracts from the n-butanol phase were further extracted by TLC, and four components, TA_1, TA_2, TA_3 and TA_4, were obtained. The four components had no significant inhibitory effect on MC-1 and CS-5.However, component TA_3 of 1 g·L~(-1) significantly promoted germination and growth of wheat. The results showed that the metabolites of T. asperellum CBS 433.97 possessed active components that could inhibit phytopathogen, as well as promote growth and development of plants. Therefore, this Trichoderma strain has good development potential.
To explore the functions of active metabolites of Trichoderma asperellum CBS 433.97 on promoting plants growth and inhibitting phytopathogen, the metabolites were separated preliminarily, and the inhibitions of active components on Fusarium oxysporum MC-1 and Fusarium oxysporum CS-5, as well as the effects of the active components from T. asperellum CBS 433.97 metabolites on plant height, root length and germination rate of wheat were analyzed. The active components of the fermentation broth from T. asperellum CBS 433.97 were extracted with different ectracting agent, the crude extracts obtained by the n-butanol extraction phase had the best inhibitory effect on MC-1 and CS-5. The crude extracts from the n-butanol phase were further extracted by TLC, and four components, TA_1, TA_2, TA_3 and TA_4, were obtained. The four components had no significant inhibitory effect on MC-1 and CS-5.However, component TA_3 of 1 g·L~(-1) significantly promoted germination and growth of wheat. The results showed that the metabolites of T. asperellum CBS 433.97 possessed active components that could inhibit phytopathogen, as well as promote growth and development of plants. Therefore, this Trichoderma strain has good development potential.
引文
[1]NKMAKCI R,ERAT M,ERDOGAN,et al.The influence of plant growth promoting rhizobacteria on growth and enzyme activities in wheat and spinach plants[J].Journal of plant nutrition and soil science,2007,170(2):288-295.
[2]El-HASAN A,SCHONE J,HOGLINGER B,et al.Assessment of the antifungal activity of selected biocontrol agents and their secondary metabolites against Fusarium graminearum[J].European journal of plant pathology,2018,150(1):91-103.
[3]胡云平.微生物农业产业技术研究进展及发展前景[J].河南农业,2018,467(15):49-50.
[4]赵嘉宁.农业微生物资源在国家经济及生态环境建设中的重要作用[J].江西农业,2018(12):87.
[5]FIEDLER H P,BRUNTNER C,BULL A T,et al.Marine actinomycetes as a source of novel secondary metabolites[J].Antonie van leeuwenhoek,2005,87(1):37-42.
[6]郝泽婷,郝晓慧,阎婧,等.淡紫拟青霉次级代谢产物的研究进展[J].生物技术通报,2018,34(10):11-17.
[7]HWANG K S,KIM H U,CHARUANTI P,et al.Systems biology and biotechnology of Streptomyces species for the production of secondary metabolites[J].Biotechnology advances,2014,32(2):255-268.
[8]VINALE F,SIVASITHAMPARAM K,GHISALBERTI E L,et al.Trichoderma secondary metabolites active on plants and fungal pathogens[J].The open mycology journal,2014,8(1):127-139.
[9]CONTRERAS-CORNEJO H A,MACIAS-RODRIGUEZ L,DEL-VAL E,et al.Ecological functions of Trichoderma spp.and their secondary metabolites in the rhizosphere-interactions with plants[J].FEMS microbiology ecology,2016,92(4):36.
[10]JOSHI D,SINGH P,SINGH A K,et al.Antifungal potential of metabolites from Trichoderma sp.against Colletotrichum falcatum Went causing red rot of sugarcanes[J].Sugar tech,2016,18(5):529-536.
[11]CHEN S C,ZHAO H J,WANG Z H,et al.Trichoderma harzianum-induced resistance against Fusarium oxysporum involves regulation of nuclear DNA content,cell viability and cell cycle-related genes expression in cucumber roots[J].European journal of plant pathology,2016,147(1):43-53.
[12]VINALE F,FLEMATTI G,SIVASITHAMPARAM K,et al.Harzianic acid,an antifungal and plant growth promoting metabolite from Trichoderma[J].Journal of natural products,2009,72(11):2032-2035.
[13]KESWANI C,MISHRA S,SARMA B K,et al.Unraveling the efficient applications of secondary metabolites of various Trichoderma spp[J].Applied microbiology and biotechnology,2014,98(2):533-544.
[14]崔振玲,王洁,胡忠义.液体培养基比色法快速检测结核分枝杆菌耐药性[J].中国抗生素杂志,2006,31(9):555-558.
[15]张量.防治辣椒疫病的木霉拮抗化合物筛选及其生防制剂制备[D].杭州:浙江大学,2015.
[2]El-HASAN A,SCHONE J,HOGLINGER B,et al.Assessment of the antifungal activity of selected biocontrol agents and their secondary metabolites against Fusarium graminearum[J].European journal of plant pathology,2018,150(1):91-103.
[3]胡云平.微生物农业产业技术研究进展及发展前景[J].河南农业,2018,467(15):49-50.
[4]赵嘉宁.农业微生物资源在国家经济及生态环境建设中的重要作用[J].江西农业,2018(12):87.
[5]FIEDLER H P,BRUNTNER C,BULL A T,et al.Marine actinomycetes as a source of novel secondary metabolites[J].Antonie van leeuwenhoek,2005,87(1):37-42.
[6]郝泽婷,郝晓慧,阎婧,等.淡紫拟青霉次级代谢产物的研究进展[J].生物技术通报,2018,34(10):11-17.
[7]HWANG K S,KIM H U,CHARUANTI P,et al.Systems biology and biotechnology of Streptomyces species for the production of secondary metabolites[J].Biotechnology advances,2014,32(2):255-268.
[8]VINALE F,SIVASITHAMPARAM K,GHISALBERTI E L,et al.Trichoderma secondary metabolites active on plants and fungal pathogens[J].The open mycology journal,2014,8(1):127-139.
[9]CONTRERAS-CORNEJO H A,MACIAS-RODRIGUEZ L,DEL-VAL E,et al.Ecological functions of Trichoderma spp.and their secondary metabolites in the rhizosphere-interactions with plants[J].FEMS microbiology ecology,2016,92(4):36.
[10]JOSHI D,SINGH P,SINGH A K,et al.Antifungal potential of metabolites from Trichoderma sp.against Colletotrichum falcatum Went causing red rot of sugarcanes[J].Sugar tech,2016,18(5):529-536.
[11]CHEN S C,ZHAO H J,WANG Z H,et al.Trichoderma harzianum-induced resistance against Fusarium oxysporum involves regulation of nuclear DNA content,cell viability and cell cycle-related genes expression in cucumber roots[J].European journal of plant pathology,2016,147(1):43-53.
[12]VINALE F,FLEMATTI G,SIVASITHAMPARAM K,et al.Harzianic acid,an antifungal and plant growth promoting metabolite from Trichoderma[J].Journal of natural products,2009,72(11):2032-2035.
[13]KESWANI C,MISHRA S,SARMA B K,et al.Unraveling the efficient applications of secondary metabolites of various Trichoderma spp[J].Applied microbiology and biotechnology,2014,98(2):533-544.
[14]崔振玲,王洁,胡忠义.液体培养基比色法快速检测结核分枝杆菌耐药性[J].中国抗生素杂志,2006,31(9):555-558.
[15]张量.防治辣椒疫病的木霉拮抗化合物筛选及其生防制剂制备[D].杭州:浙江大学,2015.