环链棒束孢Isaria cateinannulata发酵液的化学成分分析
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摘要
环链棒束孢广泛存在于自然界,并且在植物病虫害防治中起重要作用,目前相关的研究多集中于其发酵条件、生物学特性及杀虫毒力的研究,对该菌发酵液中的化学物质研究甚少。本文依次用石油醚和乙酸乙酯对环链棒束孢发酵液进行分级萃取,借助气相色谱-质谱(GC-MS)联用技术和超高效液相色谱串联四级杆飞行时间质谱法(UPLC/Q-TOF-MS)鉴定分析石油醚萃取物、乙酸乙酯萃取物和萃余物中的化学物质组成。共鉴定69个化合物的结构,其主要化学成分为酚类、酯类、烷烃、烯烃、卤代烃和酮类非极性/弱极性的小分子化合物等,其中邻苯二甲酸二丁酯、角鲨烯、环戊烯丙菊酯物质、瓜叶菊素Ⅰ等物质具有特殊的作用。发酵液提取物中还含有多种小分子化学物质,应结合多种色谱分离手段和波普学手段对代谢产物进行分离、纯化和结构鉴定,为其次生代谢产物的广泛利用提供数据支撑。
Isaria cateinannulata widely exists in nature and plays an important role in the control of plant disease and pest. At present, most of the related studies focus on the fermentation conditions, biological characteristics and insecticidal toxicity, while little research has been done on the chemical substances in the fungal fermentation broth. In this study, petroleum ether and ethyl acetate were used to extract the fermentation broth of Isaria cateinannulata. The chemical compositions of petroleum ether extract, ethyl acetate extract and raffinate of the fungus were identified and analyzed by gas chromatography-mass spectrometry(GC-MS) technology and high performance liquid chromatography tandem four-stage time-of-flight mass spectrometry(UPLC/Q-TOF-MS). A total of 69 compounds were identified, and the main chemical constituents were small molecular compounds such as phenols, esters, alkanes, olefins, halogenated hydrocarbons and ketones with non-polar/weak polarity. Among them, dibutyl phthalate, squalene,terallethrin and cinerinⅠhave special active effects. The fermentation broth extract also contained many kinds of other small molecular chemicals, and these metabolites yet await separation, purification and identification by using various chromatographic and spectroscopic methods.
Isaria cateinannulata widely exists in nature and plays an important role in the control of plant disease and pest. At present, most of the related studies focus on the fermentation conditions, biological characteristics and insecticidal toxicity, while little research has been done on the chemical substances in the fungal fermentation broth. In this study, petroleum ether and ethyl acetate were used to extract the fermentation broth of Isaria cateinannulata. The chemical compositions of petroleum ether extract, ethyl acetate extract and raffinate of the fungus were identified and analyzed by gas chromatography-mass spectrometry(GC-MS) technology and high performance liquid chromatography tandem four-stage time-of-flight mass spectrometry(UPLC/Q-TOF-MS). A total of 69 compounds were identified, and the main chemical constituents were small molecular compounds such as phenols, esters, alkanes, olefins, halogenated hydrocarbons and ketones with non-polar/weak polarity. Among them, dibutyl phthalate, squalene,terallethrin and cinerinⅠhave special active effects. The fermentation broth extract also contained many kinds of other small molecular chemicals, and these metabolites yet await separation, purification and identification by using various chromatographic and spectroscopic methods.
引文
Duan W,2007.Research on separation natural pyrethrin composition and immunological detection metheod of Chrysanthemum cineraraefolium.Master Thesis,Chongqing University,Chongqing.17-29(in Chinese)
Han YF,Zhang YW,Liang JD,Liang ZQ,2008.Preliminary studies on the pathogenicity of some Paecilomyces isolates to Plutella xylostella.Plant Protection,2008(2):46-49(in Chinese)
He J,Kang JC,Lei BX,Liu XY,Wen TC,2010.Identification of four entomogenous fungi and preliminary evaluation on their insecticidal activities to Plutella xylostella L.Journal of Plant Protection,37(4):341-346(in Chinese)
Li FB,Wang CY,Yao JF,Fang LJ,2011.Biological control of Cenopalpus lineola in Mountain Huangshan scenic spot by Paecilomyces cateniannulatus.Journal of Northeast Forestry University,39(7):77-78(in Chinese)
Li KL,Bao JY,Lu RL,Zhang LW,Zhang DL,Hu FL,2012.Analysis of volatile components of three entomogenous fungi by SDE-GC-MS.Mycosystema,31(1):92-101(in Chinese)
Li P,Mi SQ,Feng YF,Feng SW,Zhao Z,2017.Study on the effects of dibutyl phthalate on physiological activity in Caenorhabditis elegans.Science and Technology of Food Industry,38(6):129-131+136(in Chinese)
Li P,Mi SQ,Zhao Z,2016.Study on the oxidative damage effects of dibutyl phthalate on Caenorhabditis elegans.Science and Technology of Food Industry,37(24):149-152(in Chinese)
Liang ZQ,1981a.Two new species of Paecilomyces from insects.Acta Microbiologica Sinica,21(1):31-34+132(in Chinese)
Liang ZQ,1981b.Entomogenous fungi from the pests of tea-plants.Acta Phytopathologica Sinica,11(4):9-16(in Chinese)
Liu CY,Ma MH,Jin GF,Geng F,Wang QL,Sun SG,2015.Research process on squalene and bioactivities.Chinese Journal of Food Science,15(5):147-156(in Chinese)
Lu L,2011.Study on biological characteristics and some active metabolite of Cordyceps japonica.Master Thesis,Ocean University of China,Qingdao.57-59(in Chinese)
Luangsa-Ard JJ,Hywel-Jones NL,Manoch L,Samson RA,2005.On the relationships of Paecilomyces sect.Isarioidea species.Mycological Research,109(5):581-589
LüJ,Ruan XM,Sheng ZY,Xu HT,Li YQ,2003.Comparison between solid phase micro-extraction and simultaneous distillation and extraction methods for analysis of aroma components in tobacco flavor.Tobacco Science,2003(2):25-28(in Chinese)
Mitsuhashi W,Shimazu M,Hashimoto H,1992.Control of Epinotia granitalis(Lepidoptera:Tortricidae)with Paecilomyces spp.On cotton bands wrapped on the trunks of Cryptomeria japonica.Applied Entomology and Zoology,27(2):295-296(in Chinese)
Shimazu M,2001.Paecilomyces cateniannulatus Liang,commonly found,but an unrecorded entomogenous fungus in Japan.Applied Entomology and Zoology,36(3):283-288
Vél?z H,Glassbrook NJ,Daub ME,2007.Mannitol metabolism in the phytopathogenic fungus Alternaria alternata.Fungal Genetics and Biology,44(4):258-268
Wang DF,Yang G,Wang QS,Zeng MS,Wu GY,2014.Identification of two Isaria isolates and bioassay of their pathogenicity against tea tortrix Homona coffearia and smaller tea tortrix Adoxophyes honmai.Acta Phytophylacica Sinica,41(5):531-539(in Chinese)
Wang ZM,Zhou B,2003.Effect of pyrethrin on 3rd instar larva of Plutella xylostella at different temperature.Journal of Zhangzhou Institute of Technology,2003(2):120-121(in Chinese)
Wetmore DR,Joseloff E,Pilewski J,2010.Metabolomic profiling reveals biochemical pathways and biomarkers associated with pathogenesis in cystic fibrosis cells.Journal of Biological Chemistry,285(40):30516-30522
Yao T,Liang ZQ,Mo MH,Wang CY,2006.Nematicidal activity of Paecilomyces isolates from Yunnan Province.Chinese Journal of Biological Control,2006(3):226-229(in Chinese)
You FM,2010.Analysis method for four novel pyrethroids effective components in mosquito-repellent incense.Agrochemicals,49(11):819-821(in Chinese)
Zhang DL,Wang XD,Lu RL,Li KL,Hu FL,2011.Influence of liquid or solid culture conditions on the volatile components of mycelia of Isaria cateinannulata.Acta Microbiologica Sinica,51(12):1616-1624(in Chinese)
Zhang L,2013.Study on secondary metablites from endophytic fungi Pestalotiopsis karstenii and Isaria cateinannulata and their bioactivities.Master Thesis,Hebei University,Baoding.28-32(in Chinese)
Zhang XN,Jin DC,Zou X,Wang SC,Qu JJ,Guo JJ,2014.Screening of highly virulent strain of Isaria cateniannulate against Tetranychus urticae and its effect to Euseius nicholsi.Journal of Environmental Entomology,36(3):372-380(in Chinese)
Zhang XW,Li X,Li B,Yang QF,Xu MN,Cao XY,Chen ZS,Wang C,2009.Effects of dibutyl phthalate on procreant ability and its genotoxity in male Drosophila melanogaster.Journal of Fourth Military Medical University,30(1):62-65(in Chinese)
段伟,2007.天然除虫菊有效成分的分离及其免疫学检测方法研究.重庆大学硕士论文,重庆.17-29
韩燕峰,张延威,梁建东,梁宗琦,2008.一些拟青霉菌株对小菜蛾致病性的初步研究.植物保护,2008(2):46-49
何劲,康冀川,雷帮星,刘心怡,文庭池,2008.四株虫生真菌的鉴定及其对小菜蛾的室内毒力.植物保护学报,37(4):341-346
李丰伯,汪传友,姚剑飞,方乐金,2011.环链拟青霉防治黄山风景区细纹新须螨.东北林业大学学报,39(7):77-78
李康乐,包佳源,陆瑞利,张龙娃,张德龙,胡丰林,2012.SDE-GC-MS法分析三种虫生真菌菌丝中挥发性成分.菌物学报,31(1):92-101
李萍,米生权,赵卓,2016.塑化剂邻苯二甲酸二丁酯对秀丽隐杆线虫的氧化损伤作用研究.食品工业科技,37(24):149-152
李萍,米生权,冯亚芳,冯斯文,赵卓,2017.塑化剂邻苯二甲酸二丁酯对秀丽隐杆线虫生理活性影响研究.食品工业科技,38(6):129-131+136
梁宗琦,1981a.拟青霉属的两个新种.微生物学报,21(1):31-34
梁宗琦,1981b.茶树害虫的虫生真菌.植物病理学报,11(4):9-16
刘纯友,马美湖,靳国锋,耿放,王庆玲,孙术国,2015.角鲨烯及其生物活性研究进展.中国食品学报,15(5):147-156
鲁璐,2011.日本冬虫夏草(Cordyceps japonica)的生物学及活性产物分析.中国海洋大学硕士论文,青岛.57-59
吕健,阮晓明,盛志艺,徐海涛,李炎强,2003.固相微萃取与同时蒸馏萃取法分析香精成分比较.烟草科技,2003(2):25-28
王定锋,杨广,王庆森,曾明森,吴光远,2014.两株棒束孢菌的鉴定及其对茶卷叶蛾和茶小卷叶蛾的致病力.植物保护学报,41(5):531-539
王铮敏,周斌,2003.不同温度下除虫菊酯对小菜蛾3龄幼虫的药效试验.漳州职业大学学报,2003(2):120-121
姚婷,梁宗琦,莫明和,王春燕,2006.拟青霉云南菌株的杀线虫作用.中国生物防治,2006(3):226-229
游飞明,2010.蚊香中4种新型菊酯类有效成分分析方法.农药,49(11):819-821
张德龙,王小董,陆瑞利,李康乐,胡丰林,2011.液体或固体培养对环链棒束孢(Isaria cateinannulata)菌丝体中挥发性成分组成的影响.微生物学报,51(12):1616-1624
张磊,2013.内生真菌Peristalsis Kirsten与Isaria cateinannulata次级代谢产物及其生物活性研究.河北大学硕士论文,保定.28-32
张晓娜,金道超,邹晓,王思潮,瞿娇娇,郭建军,2014.杀二斑叶螨高毒力环链棒束孢菌株的筛选及其对尼氏真绥螨的影响.环境昆虫学报,36(3):372-380
张欣文,李响,李冰,杨巧凤,徐曼妮,曹响元,陈治松,王承,2009.邻苯二甲酸二丁酯对雄性果蝇生殖能力影响及其遗传毒性.第四军医大学学报,30(1):62-65
Han YF,Zhang YW,Liang JD,Liang ZQ,2008.Preliminary studies on the pathogenicity of some Paecilomyces isolates to Plutella xylostella.Plant Protection,2008(2):46-49(in Chinese)
He J,Kang JC,Lei BX,Liu XY,Wen TC,2010.Identification of four entomogenous fungi and preliminary evaluation on their insecticidal activities to Plutella xylostella L.Journal of Plant Protection,37(4):341-346(in Chinese)
Li FB,Wang CY,Yao JF,Fang LJ,2011.Biological control of Cenopalpus lineola in Mountain Huangshan scenic spot by Paecilomyces cateniannulatus.Journal of Northeast Forestry University,39(7):77-78(in Chinese)
Li KL,Bao JY,Lu RL,Zhang LW,Zhang DL,Hu FL,2012.Analysis of volatile components of three entomogenous fungi by SDE-GC-MS.Mycosystema,31(1):92-101(in Chinese)
Li P,Mi SQ,Feng YF,Feng SW,Zhao Z,2017.Study on the effects of dibutyl phthalate on physiological activity in Caenorhabditis elegans.Science and Technology of Food Industry,38(6):129-131+136(in Chinese)
Li P,Mi SQ,Zhao Z,2016.Study on the oxidative damage effects of dibutyl phthalate on Caenorhabditis elegans.Science and Technology of Food Industry,37(24):149-152(in Chinese)
Liang ZQ,1981a.Two new species of Paecilomyces from insects.Acta Microbiologica Sinica,21(1):31-34+132(in Chinese)
Liang ZQ,1981b.Entomogenous fungi from the pests of tea-plants.Acta Phytopathologica Sinica,11(4):9-16(in Chinese)
Liu CY,Ma MH,Jin GF,Geng F,Wang QL,Sun SG,2015.Research process on squalene and bioactivities.Chinese Journal of Food Science,15(5):147-156(in Chinese)
Lu L,2011.Study on biological characteristics and some active metabolite of Cordyceps japonica.Master Thesis,Ocean University of China,Qingdao.57-59(in Chinese)
Luangsa-Ard JJ,Hywel-Jones NL,Manoch L,Samson RA,2005.On the relationships of Paecilomyces sect.Isarioidea species.Mycological Research,109(5):581-589
LüJ,Ruan XM,Sheng ZY,Xu HT,Li YQ,2003.Comparison between solid phase micro-extraction and simultaneous distillation and extraction methods for analysis of aroma components in tobacco flavor.Tobacco Science,2003(2):25-28(in Chinese)
Mitsuhashi W,Shimazu M,Hashimoto H,1992.Control of Epinotia granitalis(Lepidoptera:Tortricidae)with Paecilomyces spp.On cotton bands wrapped on the trunks of Cryptomeria japonica.Applied Entomology and Zoology,27(2):295-296(in Chinese)
Shimazu M,2001.Paecilomyces cateniannulatus Liang,commonly found,but an unrecorded entomogenous fungus in Japan.Applied Entomology and Zoology,36(3):283-288
Vél?z H,Glassbrook NJ,Daub ME,2007.Mannitol metabolism in the phytopathogenic fungus Alternaria alternata.Fungal Genetics and Biology,44(4):258-268
Wang DF,Yang G,Wang QS,Zeng MS,Wu GY,2014.Identification of two Isaria isolates and bioassay of their pathogenicity against tea tortrix Homona coffearia and smaller tea tortrix Adoxophyes honmai.Acta Phytophylacica Sinica,41(5):531-539(in Chinese)
Wang ZM,Zhou B,2003.Effect of pyrethrin on 3rd instar larva of Plutella xylostella at different temperature.Journal of Zhangzhou Institute of Technology,2003(2):120-121(in Chinese)
Wetmore DR,Joseloff E,Pilewski J,2010.Metabolomic profiling reveals biochemical pathways and biomarkers associated with pathogenesis in cystic fibrosis cells.Journal of Biological Chemistry,285(40):30516-30522
Yao T,Liang ZQ,Mo MH,Wang CY,2006.Nematicidal activity of Paecilomyces isolates from Yunnan Province.Chinese Journal of Biological Control,2006(3):226-229(in Chinese)
You FM,2010.Analysis method for four novel pyrethroids effective components in mosquito-repellent incense.Agrochemicals,49(11):819-821(in Chinese)
Zhang DL,Wang XD,Lu RL,Li KL,Hu FL,2011.Influence of liquid or solid culture conditions on the volatile components of mycelia of Isaria cateinannulata.Acta Microbiologica Sinica,51(12):1616-1624(in Chinese)
Zhang L,2013.Study on secondary metablites from endophytic fungi Pestalotiopsis karstenii and Isaria cateinannulata and their bioactivities.Master Thesis,Hebei University,Baoding.28-32(in Chinese)
Zhang XN,Jin DC,Zou X,Wang SC,Qu JJ,Guo JJ,2014.Screening of highly virulent strain of Isaria cateniannulate against Tetranychus urticae and its effect to Euseius nicholsi.Journal of Environmental Entomology,36(3):372-380(in Chinese)
Zhang XW,Li X,Li B,Yang QF,Xu MN,Cao XY,Chen ZS,Wang C,2009.Effects of dibutyl phthalate on procreant ability and its genotoxity in male Drosophila melanogaster.Journal of Fourth Military Medical University,30(1):62-65(in Chinese)
段伟,2007.天然除虫菊有效成分的分离及其免疫学检测方法研究.重庆大学硕士论文,重庆.17-29
韩燕峰,张延威,梁建东,梁宗琦,2008.一些拟青霉菌株对小菜蛾致病性的初步研究.植物保护,2008(2):46-49
何劲,康冀川,雷帮星,刘心怡,文庭池,2008.四株虫生真菌的鉴定及其对小菜蛾的室内毒力.植物保护学报,37(4):341-346
李丰伯,汪传友,姚剑飞,方乐金,2011.环链拟青霉防治黄山风景区细纹新须螨.东北林业大学学报,39(7):77-78
李康乐,包佳源,陆瑞利,张龙娃,张德龙,胡丰林,2012.SDE-GC-MS法分析三种虫生真菌菌丝中挥发性成分.菌物学报,31(1):92-101
李萍,米生权,赵卓,2016.塑化剂邻苯二甲酸二丁酯对秀丽隐杆线虫的氧化损伤作用研究.食品工业科技,37(24):149-152
李萍,米生权,冯亚芳,冯斯文,赵卓,2017.塑化剂邻苯二甲酸二丁酯对秀丽隐杆线虫生理活性影响研究.食品工业科技,38(6):129-131+136
梁宗琦,1981a.拟青霉属的两个新种.微生物学报,21(1):31-34
梁宗琦,1981b.茶树害虫的虫生真菌.植物病理学报,11(4):9-16
刘纯友,马美湖,靳国锋,耿放,王庆玲,孙术国,2015.角鲨烯及其生物活性研究进展.中国食品学报,15(5):147-156
鲁璐,2011.日本冬虫夏草(Cordyceps japonica)的生物学及活性产物分析.中国海洋大学硕士论文,青岛.57-59
吕健,阮晓明,盛志艺,徐海涛,李炎强,2003.固相微萃取与同时蒸馏萃取法分析香精成分比较.烟草科技,2003(2):25-28
王定锋,杨广,王庆森,曾明森,吴光远,2014.两株棒束孢菌的鉴定及其对茶卷叶蛾和茶小卷叶蛾的致病力.植物保护学报,41(5):531-539
王铮敏,周斌,2003.不同温度下除虫菊酯对小菜蛾3龄幼虫的药效试验.漳州职业大学学报,2003(2):120-121
姚婷,梁宗琦,莫明和,王春燕,2006.拟青霉云南菌株的杀线虫作用.中国生物防治,2006(3):226-229
游飞明,2010.蚊香中4种新型菊酯类有效成分分析方法.农药,49(11):819-821
张德龙,王小董,陆瑞利,李康乐,胡丰林,2011.液体或固体培养对环链棒束孢(Isaria cateinannulata)菌丝体中挥发性成分组成的影响.微生物学报,51(12):1616-1624
张磊,2013.内生真菌Peristalsis Kirsten与Isaria cateinannulata次级代谢产物及其生物活性研究.河北大学硕士论文,保定.28-32
张晓娜,金道超,邹晓,王思潮,瞿娇娇,郭建军,2014.杀二斑叶螨高毒力环链棒束孢菌株的筛选及其对尼氏真绥螨的影响.环境昆虫学报,36(3):372-380
张欣文,李响,李冰,杨巧凤,徐曼妮,曹响元,陈治松,王承,2009.邻苯二甲酸二丁酯对雄性果蝇生殖能力影响及其遗传毒性.第四军医大学学报,30(1):62-65