苹果树腐烂病菌挥发性代谢产物的分离提取与致病性研究
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摘要
为了研究苹果树腐烂病菌的致病活性物质,本研究利用MS液体培养基培养苹果树腐烂病菌,采用普通蒸馏法用乙酸乙酯作溶剂,收集其培养滤液中的挥发性代谢产物,利用气相色谱质谱联用仪(GC-MS)分离各个成分,将各组分质谱图与数据库标准物质质谱图对比,初步进行结构定性,用苹果枝条进行致病活性测定。结果表明,苹果树腐烂病菌在发酵过程中产生的2种挥发性物质的结构分别为异戊醇和2-苯乙醇,这2种物质均对苹果树枝具有致病活性。
Valsa mali was cultured in MS liquid medium to explore its pathogenic active substances.Its volatile metabolites were collected by distillation method with ethyl acetate as absorbing solvent.Volatile constituents dissolved in ethyl acetate were separated and identified by GC-MS.To confirm the structure of each component,their mass spectra were compared with those of standard substances in the database,and their bioactivities were tested on apple branches.Two substances obtained from the culture filtrate were 3-methylbutan-1-ol and 2-phenylethan-1-ol respectively.Both of these substances showed pathogenic activity against apple branches.
Valsa mali was cultured in MS liquid medium to explore its pathogenic active substances.Its volatile metabolites were collected by distillation method with ethyl acetate as absorbing solvent.Volatile constituents dissolved in ethyl acetate were separated and identified by GC-MS.To confirm the structure of each component,their mass spectra were compared with those of standard substances in the database,and their bioactivities were tested on apple branches.Two substances obtained from the culture filtrate were 3-methylbutan-1-ol and 2-phenylethan-1-ol respectively.Both of these substances showed pathogenic activity against apple branches.
引文
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[19]黄丹,方春玉,尚志超,等.一株酯化酶霉菌的分离、鉴定及代谢产物特征[J].中国酿造,2010,(6):62-64.HUANG D,FANG C Y,CHU Z C,et al.Isolation and identification of esterase-produce mould and characterization of its metabolites[J].China Brewing,2010(6):62-64.(in Chinese)
[20]黄丹,方春玉,储玉龙,等.一株酯化酶细菌的分离、鉴定及代谢产物特征[J].四川理工学院学报:自然科学版,2010(3):321-323.HUANG D,FANG C Y,CHU Y L,et al.Isolation and characterization of bacteria producing esterifying enzyme[J].Journal of Sichuan University of Science&Engineering:Natural Science Edition,2010(3):321-323.(in Chinese)
[21]张静,周小婷,胡立盼,等.SPME-GC-MS测定不同品种牡丹花挥发性物质成分分析[J].西北林学院学报,2013,28(4):136-143.ZHANG J,ZHOU X T,HU L P,et al.SPME-GC-MS measurement of volatile in different peony varieties[J].Journal of Northwest Forestry University,2013,28(4):136-143.(in Chinese)
[2]刘福昌,李美娜,王永洤.苹果树腐烂病菌致病因素——果胶酶的初步探讨[J].中国果树,1980(4):45-48.
[3]王娟,马强,庄霞,等.苹果腐烂病病原菌分泌物中果胶酶的测定[J].内蒙古农业科技,2009(4):39-40.WANG J,MA Q,ZHUANG X,et al.Determination of pectinase in fungi sesretion of apple tree canker[J].Inner Mongolia Agricultural Science and Technology,2009(4):39-40.(in Chinese)
[4]王建华,柯希望,黄以超,等.苹果树腐烂病菌发酵液中根皮苷主要降解成分的分析[J].西北农林科技大学学报:自然科学版,2012(8):89-94.WANG J H,KE X W,HUANG Y C,et al.Component analysis of Valsa mali var.mali fermentation liquid degrading phlorizin[J].Journal of Northwest A&F University:Nat.Sci.Ed.,2012(8):89-94.(in Chinese)
[5]何媛媛,于哲,王海英,等.不同碳氮源对苹果腐烂病菌胞外果胶酶活性的影响[J].西北农林科技大学学报:自然科学版,2014(11):81-86.HE Y Y,YU Z,WANG H Y,et al.Effects of different carbon and nitrogen sources on activity of extracellular pectinase of Valsa mali var.mali[J].Journal of Northwest A&F University:Nat.Sci.Ed.,2014(11):81-86.(in Chinese)
[6]柯希望.苹果树腐烂病菌在树皮组织内的扩展观察[D].陕西杨陵:西北农林科技大学,2009.
[7]NATSUME H,SETO H,OTAKE N.Studies on apple canker disease.The necrotic toxins produced by Valsa ceratosperma[J].Agricultural&Biological Chemistry,1982,46(8):2101-2106.
[8]KOGANEZAWA H,SAKUMA T.Possible role of breakdown products of phloridzin in symptom development by Valsa ceratosperma[J].Annals of the Phytopathological Society of Japan,1982,48(4):521-528.
[9]TRAQUAIR J.Oxalic acid and calcium oxalate produced by Leucostoma cinctaand L.persooniiin culture and in peach bark tissues[J].Canadian Journal of Botany,1988,65(9):1952-1956.
[10]SVIRCEV A M,BIGGS A R,MILES N W.Isolation and partial purification of phytotoxins from liquid cultures[J].Canadian Journal of Botany,1991,69(9):1998-2003.
[11]王建华.苹果树腐烂病菌致病物质的初步研究[D].陕西杨陵:西北农林科技大学,2012.
[12]祝云飞,黄治国,邓杰,等.浓香型大曲中一株酵母菌的分离鉴定及其挥发性产物分析[J].四川理工学院学报:自然科学版,2016,29(1):7-11.
[13]白岩岩,贾薇,张劲松.樟芝发酵液挥发性物质提取方法及成分比较[J].食品科学,2013,34(24):163-167.BAI Y Y,JIA W,ZHANG J S.Comparison of different extraction methods for the analysis of volatile components of the fermentation broth of Taiwanofungus comphoratus[J].Food Science,2013,34(24):163-167.(in Chinese)
[14]王丽娟,刘苏萌,楚明忠.百合产香内生真菌的分离鉴定及其挥发性成分分析[J].食品工业,2013(9):203-205.WANG L J,LIU S M,CHU M Z.Isolation and identification endophytic fungi fromLilium Siberia and analysis of volatile components[J].The Food Industry,2013(9):203-205.(in Chinese)
[15]谷运璀,宦月琴,钱莉群,等.天然肉桂酸及其酯类的制备[J].香料香精化妆品,2013(Supp.1):36-38.GU Y C,HUAN Y Q,QIAN L Q,et al.The synthesis of natural cinnamic acid and its esters[J].Flavour Fragrance Cosmetics,2013(Supp.1):36-38.(in Chinese)
[16]晏慧君,王娟,陈敏,等.朋粉(Rosa chinensis‘Pallida')、大马士革蔷薇(R.damascene)、百叶蔷薇(R.centifolia)香气成分分析[J].云南农业大学学报:自然科学,2017,32(1):78-82.YAN H J,WANG J,CHEN M,et al.The analysis of scent compounds from Rosa chinensis‘Pallida',R.damascene and R.centifoliaby GC/MS[J].Journal of Yunnan Agricultural University:Natural Science,2017,32(1):78-82.(in Chinese)
[17]曹建宏,张振文,张莉,等.霞多丽干白葡萄酒香气成分的GC/MS分析[J].西北林学院学报,2006,21(4):132-134.CAO J H,ZHANG Z W,ZHANG L,et al.Analysis of roma components of chardonnay dry white wine by GC/MS[J].Journal of Northwest Forestry University,2006,21(4):132-134.(in Chinese)
[18]陈利军,王国君,田雪亮,等.产香真菌ZY-2菌株鉴定及其挥发性物质抑菌活性测定与组分分析[J].南方农业学报,2013,44(11):1818-1822.CHEN L J,WANG G J,TIAN X L,et al.Identification of an aroma-producing fungus ZY-2and its analysis on antifungal activity and chemical component of volatile compounds[J].Journal of Southern Agriculture,2013,44(11):1818-1822.(in Chinese)
[19]黄丹,方春玉,尚志超,等.一株酯化酶霉菌的分离、鉴定及代谢产物特征[J].中国酿造,2010,(6):62-64.HUANG D,FANG C Y,CHU Z C,et al.Isolation and identification of esterase-produce mould and characterization of its metabolites[J].China Brewing,2010(6):62-64.(in Chinese)
[20]黄丹,方春玉,储玉龙,等.一株酯化酶细菌的分离、鉴定及代谢产物特征[J].四川理工学院学报:自然科学版,2010(3):321-323.HUANG D,FANG C Y,CHU Y L,et al.Isolation and characterization of bacteria producing esterifying enzyme[J].Journal of Sichuan University of Science&Engineering:Natural Science Edition,2010(3):321-323.(in Chinese)
[21]张静,周小婷,胡立盼,等.SPME-GC-MS测定不同品种牡丹花挥发性物质成分分析[J].西北林学院学报,2013,28(4):136-143.ZHANG J,ZHOU X T,HU L P,et al.SPME-GC-MS measurement of volatile in different peony varieties[J].Journal of Northwest Forestry University,2013,28(4):136-143.(in Chinese)