茉莉酸甲酯对马尾松萜类合成酶活性及其细胞化学定位的影响
|
摘要
马尾松(Pinus massoniana)是我国南方生态建设与造林用材的主要树种,为了揭示马尾松抗虫机理尤其是诱导抗虫性的分子机制,该研究以马尾松幼苗为材料,通过外源喷施茉利酸甲酯(Me JA),分析了处理与对照间植株针叶显微结构、萜类合成酶活性及其细胞化学定位的变化。结果表明:在0.2 mmol·L~(-1)Me JA处理下马尾松植株松针中萜类物质,尤其是单萜、二萜的相对含量增加,马尾松毛虫拒食性明显,诱导抗性增强。显微观测中,针叶叶肉细胞内树脂道分泌物增加,叶绿体数目减少,但叶绿体体积增大,叶绿体片层结构增加。Me JA处理4周后,针叶中萜类合成酶活性增加,通过电镜酶细胞化学观察,膜系统尤其是叶绿体膜上萜类合成酶活性定位明显增强。这说明Me JA诱导的马尾松诱导抗性可能与改变的叶绿体结构及绿色质体萜类合成酶活性密切相关。
Pinus massoniana is one of the most important tree species for afforestation in South China,and it is of great economic value and ecological benefit. This study aimed to reveal the cytological basis in relation to induced resistance during the biosynthesis of terpenoids in P. massoniana,and provide the guide for the further research on molecular mechanism of induced insect resistance in P. massoniana. In the illumination incubator with strictly controlled conditions of light and temperature,variations of microscopic structure,terpenoid synthetase activity and its cytochemical localization of needles were investigated in P. massoniana seedlings treated by exogenous jasmonates. The results showed that there were increases in the relative contents of terpenoids,especially,in those of monoterpenoids and diterpenoids,and the antifeedant of Dendrolimus punctatus against needles in Pinus massoniana seedlings was increased following the treatment of 0.2 mmol·L~(-1) Me JA,which indicated that the induced resistance of P. massoniana were strengthened. For the variations of anatomical structure,secretions of resin canal were enhanced,the number of chloroplast was reduced,the volume of it was increased,and the thylakoid of it was enhanced in the needles of P. massoniana seedlings. The activity of terpenoid synthetase was increased after four weeks of Me JA treatment. The localization of terpenoid synthetase activity was remarkably increased from membrane system,especially,from chloroplast membrane in the needles of P. massoniana seedlings using the method for electron microscopic cytochemistry of enzyme. This implies that the induced resistance is closely related to the changed chloroplast structure and terpenoid synthetase activity in green plastids in P. massaniana exposed to Me JA treatment.
Pinus massoniana is one of the most important tree species for afforestation in South China,and it is of great economic value and ecological benefit. This study aimed to reveal the cytological basis in relation to induced resistance during the biosynthesis of terpenoids in P. massoniana,and provide the guide for the further research on molecular mechanism of induced insect resistance in P. massoniana. In the illumination incubator with strictly controlled conditions of light and temperature,variations of microscopic structure,terpenoid synthetase activity and its cytochemical localization of needles were investigated in P. massoniana seedlings treated by exogenous jasmonates. The results showed that there were increases in the relative contents of terpenoids,especially,in those of monoterpenoids and diterpenoids,and the antifeedant of Dendrolimus punctatus against needles in Pinus massoniana seedlings was increased following the treatment of 0.2 mmol·L~(-1) Me JA,which indicated that the induced resistance of P. massoniana were strengthened. For the variations of anatomical structure,secretions of resin canal were enhanced,the number of chloroplast was reduced,the volume of it was increased,and the thylakoid of it was enhanced in the needles of P. massoniana seedlings. The activity of terpenoid synthetase was increased after four weeks of Me JA treatment. The localization of terpenoid synthetase activity was remarkably increased from membrane system,especially,from chloroplast membrane in the needles of P. massoniana seedlings using the method for electron microscopic cytochemistry of enzyme. This implies that the induced resistance is closely related to the changed chloroplast structure and terpenoid synthetase activity in green plastids in P. massaniana exposed to Me JA treatment.
引文
ANANIEVA K,ANANIEW ED,MISHEV K,et al,2007.Methyl jas-monate is a more effective senescence-promoting factor in Cucurbita pepo(zucchini)cotyledons when compared with darkness at the early stage of senescence[J].JPlant Physiol,164:1179-1187.
ARIGONI D,SAGNER S,LATZEL C,et al,1997.Terpenoid biosynthesis from 1-deoxy-D-xylulose in higher plants by intramolecular skeletal rearrangement[J].Proc Natl Acad Sci(USA),94:10600-10605.
BARTRAM S,JUX A,GLEIXNER G,et al,2006.Dynamic pathway allocation in early terpenoid biosynthesis of stressinduced lima bean leaves[J].Phyto Chem,67:1661-1672.
BYUN-MCKAY A,GODARD K,TOUDEFALLAH M,et al,2006.Wound induced terpene synthase gene expression insitka spruce that exhibit resistance or susceptibility to attack by the white pine weevil[J].Plant Physiol,140(3):1009-1021.
CREELMAN RA,MULLET JE,1997.Biosynthesis and action of jasmonate in plants[J].Ann Rev Plant Physiol Plant Mol Biol,48:355-381.
DING GJ,ZHOU ZC,WANG ZR,et al,2006.Cultivation and utilization of pulpwood stand for Pinus massoniana[M].Beijing:China Forestry Press:1-10.[丁贵杰,周志春,王章荣,等,2006.马尾松纸浆用材林培育与利用[M].北京:中国林业出版社:1-10.]
EISENREICH W,ROHDICH F,BACHER A,2010.Deoxyxylulose phosphate pathway to terpenoids[J].Trends Plant Sci,6:78-84.]
ERBILGIN N,COLGAN LJ,2012.Differential effects of plant ontogeny and damage type on phloem and foliage monoterpenes in jack pine(Pinus banksiana)[J].Tree Physiol,32(8):946-957.
FAHN A,BENAYOUN J,1976.Ultrastructure of resin ducts in Pinus halepensis development,possible sites of resin synthesis,and mode of its elimination from the protoplast[J].Ann Bot,40:857-863.
F LDT J,MARTIN M,MILLER M,2003.Traumatic resin defense in Norway spruce(Picea abies):methyl jasmonate-induced terpene synthase gene expression and c DNA cloning and functional characterization of(+)-3-carene synthase[J].Plant Mol Biol,151(1):119-133.
FENG HZ,WANG L,YAO YS,et al,2013.Analysis of volatile compounds released from cotton,induced by Aphis gossypii and methyl jasmonate[J].Chin J Appl Entomol,50(3):742-748.[冯宏祖,王兰,姚永生,等,2013.棉蚜和茉莉酸甲酯诱导棉花挥发物组分分析[J].应用昆虫学报,50(3):742-748.]
GE F,LI ZY,XIE YP,et al,2002.Comparison for the rules of insect-induced resistance of main pine trees in China[J].JBeijing For Univ,24(3):61-65.[戈峰,李镇宇,谢映平,等,2002.我国主要松树诱导抗虫性的一些规律比较[J].北京林业大学学报,24(3):61-65.]
GONG Z,LI DM,ZHANG Z,2010.Research progress on terpene synthase enzymes of conifer trees[J].Sci Silv Sin,46(1):123-130.[龚治,李典谟,张真,2010.针叶树萜类合成酶研究进展[J].林业科学,46(1):123-130.]
HU SB,FENG YB,WEI CL,et al,1988.Resistance of Dendrolimus punctatus-resistant plants in Pinus massoniana[J].Guangxi Plant Prot,(4):47-48.[胡少波,冯远斌,韦成礼,等,1988.马尾松抗松毛虫植株的抗性研究[J].广西植保,(4):47-48.]
HU YJ,2010.Study on communication between healthy and pest-fed Pinus massoniana and differential expression proteins induced by Me JA[D].Beijing:Beijing Forestry University.[胡永建,2010.虫害马尾松邻株信号通讯及茉莉酸甲酯诱导的蛋白表达差异研究[D].北京:北京林业大学.]
HU YJ,REN Q,LI ZY,et al,2008.Rapid changes in volatile compounds and endogenousmethyl jasmonates from the adjacent healthy branch needles of damaged Pinus massioniana[J].Acta Ecol Sin,28(11):5331-5337.[胡永建,任琴,李镇宇,等,2008.虫害马尾松(Pinus massoniana Lamb.)邻枝针叶挥发物及其内源茉莉酸甲酯的快速变化[J].生态学报,28(11):5331-5337.]
HUNG KT,KAO CH,2004.Nitric oxide acts as an antioxidant and delays methyl jasmonate-induced senescence of rice leaves[J].J Plant Physiol,161:43-52.
KASAHARA H,HANADA A,KUZUYAMA T,et al,2002.Contribution of the mevalonate and methylerythritol phosphate pathways to the biosynthesis of gibberellins in Arabidopsis[J].J Biol Chem,77:45188-45194.
KENNETH JC,1987.Initiation of terpenoid synthesis in osmophores of Stanhopea anfracta(Orchidaceae):A cytochemical study[J].Am J Bot,74(9):1332-1338.
LI AM,2004.Histological and cytological studies on the development of resin canals in Pinus massoniana[D].Guangzhou:South China Agricultural University.[李爱民,2004.马尾松树脂道发育的组织学和细胞学研究[D].广州:华南农业大学.]
LI ZY,CHENH S,YUAN XH,et al,1998.Induced chemical defensesof Pinus tabulaeformis Carr.to Dendrolimus spectabil is Butler[J].Sci Silv Sin,34(2):43-50.[李镇宇,陈华盛,袁小环,等,1998.油松对赤松毛虫的诱导化学防御[J].林业科学,34(2):43-50.]
LIANG SJ,WU H,2010.Energy dispersive x-ray spectroscopy in cytochemistry analysis of key enzyme during terpenoid biosynthesis pathway[C].First International Conference on Cellular,Molecular Biology,Biophysics and Bioengineering(CMBB):18-21.[梁社坚,吴鸿,2010.萜类物质合成途径中关键酶细胞化学定位的X射线能谱分析[C].第一届细胞、分子生物、生物物理与生物工程国际会议:18-21.]
MITHFER A,BOLAND W,2012.Plant defense against herbivores:chemical aspects[J].Ann Rev Plant Biol,63:431-450.
Research Group of Dendrolimus Punctatus Resistance in Pinus massoniana,1990.Resistance mechanism of Dendrolimus punctatus-resistant plants in Pinus massoniana[J].Sci Silv Sin,26(2):133-141.[马尾松抗松毛虫抗性研究组,1990.马尾松抗松毛虫植株的抗性机制研究[J].林业科学,26(2):133-141.]
THOLL D,2006.Terpene synthases and the regulation,diversity and biological roles of terpene metabolism[J].Curr Opin Plant Biol,9(3):297-304.
WANG LC,REN Q,XU ZC,et al,2008.Effects of methyl jasmonate on Pinus massoniana needle terpenes volatiles and the growth of Dendrolimus punctatus[J].J Beijing For Univ,30(1):79-84.[王立春,任琴,许志春,等,2008.茉莉酸甲酯对马尾松松针萜烯类挥发物及马尾松毛虫生长发育的影响[J].北京林业大学学报,30(1):79-84.]
WANG LJ,FANG X,YANG CQ,et al,2013.Secondary metabolism and regulation of plants[J].Chin Sci(Vit),43(12):1030-1046.[王凌健,方欣,杨长青,等,2013.植物萜类次生代谢及其调控[J].中国科学:生命科学,43(12):1030-1046.]
YUE YC,FAN YP,2011.Research progress on terpene synthase enzymes and metabolic regulation of plants[J].Acta Hortic Sin,38(2):379-388.[岳跃冲,范燕萍,2011.植物萜类合成酶及其代谢调控的研究进展[J].园艺学报,38(2):379-388.]
ZHANG KH,CHEN CP,LIU XP,et al,2002.Influences of induced resistance of pine on growth and development of the larvae of pine caterpillar[J].Entomol Knowl,39(4):292-296.[章康华,陈春平,刘兴平,等,2002.马尾松诱导抗性对马尾松毛虫生长发育的影响[J].昆虫知识,39(4):292-296.]
ZHANG QJ,ZHANG AH,SU JB,et al,2012.Research progress on allelopathic effect of terpenoids in plants[J].Ecol Environ,21(1):187-193.[张秋菊,张爱华,孙晶波,等,2012.植物体中萜类物质化感作用的研究进展[J].生态环境学报,21(1):187-193.]
ZHENG GC,GU ZP,1987.Biology micro technology[M].2nd ed.Beijing:Higher Education Press:327-365.[郑国锠,谷祝平,1987.生物显微技术[M].第二版.北京:高等教育出版社:327-365.]
ARIGONI D,SAGNER S,LATZEL C,et al,1997.Terpenoid biosynthesis from 1-deoxy-D-xylulose in higher plants by intramolecular skeletal rearrangement[J].Proc Natl Acad Sci(USA),94:10600-10605.
BARTRAM S,JUX A,GLEIXNER G,et al,2006.Dynamic pathway allocation in early terpenoid biosynthesis of stressinduced lima bean leaves[J].Phyto Chem,67:1661-1672.
BYUN-MCKAY A,GODARD K,TOUDEFALLAH M,et al,2006.Wound induced terpene synthase gene expression insitka spruce that exhibit resistance or susceptibility to attack by the white pine weevil[J].Plant Physiol,140(3):1009-1021.
CREELMAN RA,MULLET JE,1997.Biosynthesis and action of jasmonate in plants[J].Ann Rev Plant Physiol Plant Mol Biol,48:355-381.
DING GJ,ZHOU ZC,WANG ZR,et al,2006.Cultivation and utilization of pulpwood stand for Pinus massoniana[M].Beijing:China Forestry Press:1-10.[丁贵杰,周志春,王章荣,等,2006.马尾松纸浆用材林培育与利用[M].北京:中国林业出版社:1-10.]
EISENREICH W,ROHDICH F,BACHER A,2010.Deoxyxylulose phosphate pathway to terpenoids[J].Trends Plant Sci,6:78-84.]
ERBILGIN N,COLGAN LJ,2012.Differential effects of plant ontogeny and damage type on phloem and foliage monoterpenes in jack pine(Pinus banksiana)[J].Tree Physiol,32(8):946-957.
FAHN A,BENAYOUN J,1976.Ultrastructure of resin ducts in Pinus halepensis development,possible sites of resin synthesis,and mode of its elimination from the protoplast[J].Ann Bot,40:857-863.
F LDT J,MARTIN M,MILLER M,2003.Traumatic resin defense in Norway spruce(Picea abies):methyl jasmonate-induced terpene synthase gene expression and c DNA cloning and functional characterization of(+)-3-carene synthase[J].Plant Mol Biol,151(1):119-133.
FENG HZ,WANG L,YAO YS,et al,2013.Analysis of volatile compounds released from cotton,induced by Aphis gossypii and methyl jasmonate[J].Chin J Appl Entomol,50(3):742-748.[冯宏祖,王兰,姚永生,等,2013.棉蚜和茉莉酸甲酯诱导棉花挥发物组分分析[J].应用昆虫学报,50(3):742-748.]
GE F,LI ZY,XIE YP,et al,2002.Comparison for the rules of insect-induced resistance of main pine trees in China[J].JBeijing For Univ,24(3):61-65.[戈峰,李镇宇,谢映平,等,2002.我国主要松树诱导抗虫性的一些规律比较[J].北京林业大学学报,24(3):61-65.]
GONG Z,LI DM,ZHANG Z,2010.Research progress on terpene synthase enzymes of conifer trees[J].Sci Silv Sin,46(1):123-130.[龚治,李典谟,张真,2010.针叶树萜类合成酶研究进展[J].林业科学,46(1):123-130.]
HU SB,FENG YB,WEI CL,et al,1988.Resistance of Dendrolimus punctatus-resistant plants in Pinus massoniana[J].Guangxi Plant Prot,(4):47-48.[胡少波,冯远斌,韦成礼,等,1988.马尾松抗松毛虫植株的抗性研究[J].广西植保,(4):47-48.]
HU YJ,2010.Study on communication between healthy and pest-fed Pinus massoniana and differential expression proteins induced by Me JA[D].Beijing:Beijing Forestry University.[胡永建,2010.虫害马尾松邻株信号通讯及茉莉酸甲酯诱导的蛋白表达差异研究[D].北京:北京林业大学.]
HU YJ,REN Q,LI ZY,et al,2008.Rapid changes in volatile compounds and endogenousmethyl jasmonates from the adjacent healthy branch needles of damaged Pinus massioniana[J].Acta Ecol Sin,28(11):5331-5337.[胡永建,任琴,李镇宇,等,2008.虫害马尾松(Pinus massoniana Lamb.)邻枝针叶挥发物及其内源茉莉酸甲酯的快速变化[J].生态学报,28(11):5331-5337.]
HUNG KT,KAO CH,2004.Nitric oxide acts as an antioxidant and delays methyl jasmonate-induced senescence of rice leaves[J].J Plant Physiol,161:43-52.
KASAHARA H,HANADA A,KUZUYAMA T,et al,2002.Contribution of the mevalonate and methylerythritol phosphate pathways to the biosynthesis of gibberellins in Arabidopsis[J].J Biol Chem,77:45188-45194.
KENNETH JC,1987.Initiation of terpenoid synthesis in osmophores of Stanhopea anfracta(Orchidaceae):A cytochemical study[J].Am J Bot,74(9):1332-1338.
LI AM,2004.Histological and cytological studies on the development of resin canals in Pinus massoniana[D].Guangzhou:South China Agricultural University.[李爱民,2004.马尾松树脂道发育的组织学和细胞学研究[D].广州:华南农业大学.]
LI ZY,CHENH S,YUAN XH,et al,1998.Induced chemical defensesof Pinus tabulaeformis Carr.to Dendrolimus spectabil is Butler[J].Sci Silv Sin,34(2):43-50.[李镇宇,陈华盛,袁小环,等,1998.油松对赤松毛虫的诱导化学防御[J].林业科学,34(2):43-50.]
LIANG SJ,WU H,2010.Energy dispersive x-ray spectroscopy in cytochemistry analysis of key enzyme during terpenoid biosynthesis pathway[C].First International Conference on Cellular,Molecular Biology,Biophysics and Bioengineering(CMBB):18-21.[梁社坚,吴鸿,2010.萜类物质合成途径中关键酶细胞化学定位的X射线能谱分析[C].第一届细胞、分子生物、生物物理与生物工程国际会议:18-21.]
MITHFER A,BOLAND W,2012.Plant defense against herbivores:chemical aspects[J].Ann Rev Plant Biol,63:431-450.
Research Group of Dendrolimus Punctatus Resistance in Pinus massoniana,1990.Resistance mechanism of Dendrolimus punctatus-resistant plants in Pinus massoniana[J].Sci Silv Sin,26(2):133-141.[马尾松抗松毛虫抗性研究组,1990.马尾松抗松毛虫植株的抗性机制研究[J].林业科学,26(2):133-141.]
THOLL D,2006.Terpene synthases and the regulation,diversity and biological roles of terpene metabolism[J].Curr Opin Plant Biol,9(3):297-304.
WANG LC,REN Q,XU ZC,et al,2008.Effects of methyl jasmonate on Pinus massoniana needle terpenes volatiles and the growth of Dendrolimus punctatus[J].J Beijing For Univ,30(1):79-84.[王立春,任琴,许志春,等,2008.茉莉酸甲酯对马尾松松针萜烯类挥发物及马尾松毛虫生长发育的影响[J].北京林业大学学报,30(1):79-84.]
WANG LJ,FANG X,YANG CQ,et al,2013.Secondary metabolism and regulation of plants[J].Chin Sci(Vit),43(12):1030-1046.[王凌健,方欣,杨长青,等,2013.植物萜类次生代谢及其调控[J].中国科学:生命科学,43(12):1030-1046.]
YUE YC,FAN YP,2011.Research progress on terpene synthase enzymes and metabolic regulation of plants[J].Acta Hortic Sin,38(2):379-388.[岳跃冲,范燕萍,2011.植物萜类合成酶及其代谢调控的研究进展[J].园艺学报,38(2):379-388.]
ZHANG KH,CHEN CP,LIU XP,et al,2002.Influences of induced resistance of pine on growth and development of the larvae of pine caterpillar[J].Entomol Knowl,39(4):292-296.[章康华,陈春平,刘兴平,等,2002.马尾松诱导抗性对马尾松毛虫生长发育的影响[J].昆虫知识,39(4):292-296.]
ZHANG QJ,ZHANG AH,SU JB,et al,2012.Research progress on allelopathic effect of terpenoids in plants[J].Ecol Environ,21(1):187-193.[张秋菊,张爱华,孙晶波,等,2012.植物体中萜类物质化感作用的研究进展[J].生态环境学报,21(1):187-193.]
ZHENG GC,GU ZP,1987.Biology micro technology[M].2nd ed.Beijing:Higher Education Press:327-365.[郑国锠,谷祝平,1987.生物显微技术[M].第二版.北京:高等教育出版社:327-365.]