摘要
【目的】研究自然环境条件下剪叶和油松毛虫不同取食刺激程度双重诱导对油松体内蛋白质表达的影响,揭示油松诱导防御反应的蛋白质基础。【方法】在河北省平泉县的油松-山杏混交林中,选取15年生左右生长势一致的油松,进行树上套笼饲养10头、30头油松毛虫和剪叶处理,以未处理油松为对照,采用双向电泳法测定油松体内蛋白质表达差异。【结果】通过蛋白质点表达量的计算,与对照相比,油松毛虫取食及剪叶刺激后,均有特异表达蛋白点、消失蛋白点、上调蛋白点及下调蛋白点,但不同处理间蛋白点的表达差异不同。对差异蛋白点进行质谱鉴定,数据库检索结果表明,油松毛虫取食后特异表达的蛋白点是叶绿体ATP合酶CF0β亚基、抗坏血酸过氧化物酶、噻唑生物合成酶、Lhcb5蛋白、ATP合酶β亚基以及蛋白类谷胱甘肽S-转移酶,下调表达的蛋白点是蛋白酶亚基和假定蛋白CL304Contig1_01;剪叶刺激后特异表达的蛋白点是tau蛋白类谷胱甘肽S-转移酶和Rubisco加氧酶,下调表达的蛋白点是磷酸甘油酸激酶Ⅰ和核酮糖-1,5-二磷酸羧化酶/加氧酶大亚基。进一步分析表明,取食刺激处理后,胁迫和防御相关蛋白高度表达,并且30头松毛虫取食后,与蛋白合成相关的蛋白质也特异表达;剪叶处理后,胁迫和防御相关蛋白也有特异表达,而且能量代谢相关蛋白下调。说明取食和创伤刺激可明显诱导油松防御有关蛋白的表达、光合作用的增强、蛋白质的合成和能量代谢的降低等防御反应。【结论】取食和剪叶刺激可明显诱导油松防御有关蛋白的高表达、促进光合作用的增强和蛋白质的合成,降低能量代谢等,这些变化应是构成油松防御反应的蛋白质基础。
[Objective]This study aimed at detecting the protein expression differences and the protein base of defense reaction in Chinese pine（Pinus tabulaeformis） induced by leaf-cutting and different number pine caterpillar（Dendrolimus tabulaeformis） feeding stimulation in the natural environment.[Method]15-year-old Chinese pines with the similar growth potential were selected as the research object.The sample trees were treated with feeding stimulation by 10 or 30 pine caterpillars laid on the branch or with leaf-cutting,and the untreated trees served as the control in a Pinus tabulaeformisAmeniaca sibirica mixed forest in Pingquan County,Hebei Province.The protein expression differences of the Chinese pine were studied by dimensional electrophoresis method.[Result]Through the calculation of protein point expression,compared with control,after feeding with 10 pine caterpillars the up-regulated protein spots were 9,down-regulated protein spots were 4,new increased protein spots were 20,and disappeared protein spots were 18.After feeding with 30 pine caterpillars the up-regulated protein spots were 5,down-regulated protein spots were 31,new increased protein spots were 23,and disappeared protein spots were 13.After leaf-cutting stimulation the up-regulated protein spots were 8,down-regulated protein spots were 9,new increased protein spots were 25,and disappeared protein spots were 17.Differential expression proteins were further identified by mass spectrometry.The database retrieval results showed that new increased proteins in the insect feeding trees were chloroplast ATP synthase CF0 β subunit,ascorbate peroxidase（APX）,thiazole biosynthetic enzyme,Lhcb5 protein,ATP synthase β subunit and glutathione S-transferase,and downregulated proteins were proteasome subunit and hypothesis protein CL304Contig1_ 01.In the trees simulated by leafcutting,new increased proteins were glutathione S-transferase and rubisco activase,down-regulated proteins were phosphoglycerate kinase Ⅰ and ribulose-1,5-bisphosphate carbox
出处
《林业科学》
EI
CAS
CSCD
北大核心
2016年第8期68-75,共8页
Scientia Silvae Sinicae
关键词
油松
油松毛虫
诱导防御反应
取食
剪叶
蛋白质表达差异
Pinus tabulaeformis
Dendrolimus tabulaeformis
induced defense responses
feeding stimulation
leafcutting stimulation
difference of protein expression
