茉莉酸甲酯对高温胁迫、生物蚕食无花果幼苗的影响

以波姬红无花果为试材,40℃高温胁迫经茉莉酸甲酯处理的无花果幼苗,分析与抗逆相关的超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)、茉莉酸合成路径中脂氧合酶(LOX)活性变化及丙二醛、脯氨酸、多酚含量,同时,采用黑绒金龟蚕食伤害经茉莉酸甲酯处理的无花果幼苗,分析与生物胁迫相关的多酚氧化酶(PPO)、苯丙氨酸解氨酶(PAL)活性变化及单宁、胰蛋白酶抑制素含量。结果表明,高温胁迫经50μmol/L茉莉酸甲酯处理的无花果幼苗,其SOD、POD、CAT活性与未用茉莉酸甲酯处理(对照)相比有显著增加,脯氨酸、多酚含量提高,丙二醛含量降低;生物蚕食经50μmol/L茉莉酸甲酯处理的幼苗叶片,其PPO、PAL活性增加,单宁、胰蛋白酶抑制素含量提高。经茉莉酸甲酯处理的无花果幼苗,高温胁迫、生物蚕食能通过增加SOD、POD、CAT活性抵御非生物逆境的胁迫,通过提高PPO、PAL活性增强对生物胁迫的抗性。     

喷施外源茉莉酸类化合物对红松防御酶及蛋白酶抑制剂的影响

冷杉梢斑螟(Dioryctria abietella)和赤松梢斑螟(D.sylvestrella)常常伴随发生,前者危害球果,后者在球果和主梢间转移危害.为了研究外源茉莉酸类化合物对红松(Pinus koraiensis诱导防御的影响,以不同浓度茉莉酸甲酯、茉莉酮喷雾处理红松球果和主梢.采集健康、虫害和喷雾处理的红松球果、主梢和侧枝,用紫外分光光度法测定采样部位防御酶及蛋白酶抑制剂活性,以冷杉梢斑螟和赤松梢斑螟幼虫危害的球果、赤松梢斑螟幼虫取食的主梢为虫害对照.结果 表明:喷施不同浓度茉莉酸类化合物使球果和主梢超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、苯丙氨酸解氨酶(PAL)和多酚氧化酶(PPO)活性显著升高,原位诱导与系统诱导的结果无显著差异.POD活性仅在处理后1d高于对照.喷施茉莉酮诱导的防御酶活性更接近幼虫取食.喷施不同浓度茉莉酸类化合物使球果和主梢胰蛋白酶抑制剂活性分别在处理后的1～60 d和1～30 d显著增加.系统诱导主梢胰凝乳蛋白酶抑制剂活性显著高于原位诱导.茉莉酮诱导的2种蛋白酶抑制剂活性更接近幼虫取食.可见,茉莉酸类化合物在提高红松防御能力方面具有重要的价值.     

茉莉酸对棉花单宁含量和抗虫相关酶活性的诱导效应

以植物生长调节物茉莉酸(Jasmonic acid,JA)为诱导子,以常规棉为研究对象,探讨了外源茉莉酸对棉花幼苗单宁和蛋白酶抑制素以及其它抗虫相关酶活性诱导的浓度依赖性和持久性,讨论了棉花抗虫相关物质的抗虫效果.结果表明,0.01、0.1和1.0 mmol/L茉莉酸都能在2周内诱导棉花单宁和胰蛋白酶抑制素(Proteinase inhibitors,PIs)含量增加,诱导多酚氧化酶(Polyphenol oxidase,PPO)、苯丙氨酸解氨酶(Phenylalanine ammonia-lyase,PAL)、过氧化物酶(Peroxidase,POD)和过氧化氢酶(Catalase,CAT)活性升高.对3种浓度茉莉酸的诱导效应进行分析表明,0.1 mmol/L茉莉酸对于诱导PIs、PPO、POD和CAT最有效,0.1和1.0 mmol/L茉莉酸对于诱导棉花单宁和苯丙氨酸解氨酶等效,二者的诱导效应均高于0.01 mmol/L.对茉莉酸诱导抗性的持久性进行分析表明,最佳诱导效应发生的时间各不相同:POD活性在JA处理后第1天最高,随后呈下降趋势,PIs和单宁含量分别在JA处理后第7天和第14天达最大值;JA处理后第1天和第7天的PPO活性无明显差异,但明显高于第14天;JA处理后第7天和第14天的PAL活性无明显差异,但明显高于第1天;JA处理后第1、7和14天棉花叶片的CAT活性均无明显差异.以上结果表明,茉莉酸可通过增加棉叶单宁和PIs含量、提高棉叶PAL、PPO、POD和CAT活性等增强棉花幼苗的抗虫性.     

落叶松诱导抗性对舞毒蛾生长发育的影响

以喷施茉莉酸甲酯、茉莉酮、舞毒蛾幼虫取食和松毛虫幼虫取食4种方法处理落叶松幼苗,研究了不同诱导方法对舞毒蛾各龄期幼虫体质量、存活率、蛹质量及成虫产卵量的影响.结果表明:舞毒蛾取食不同诱导处理的落叶松后,其幼虫体质量显著下降,存活率降低,蛹质量减轻,产卵量减少.不同诱导因子对舞毒蛾幼虫生长发育的影响存在差异.其中,取食舞毒蛾诱导落叶松的舞毒蛾各项生长指标,低于取食落叶松毛虫诱导落叶松的舞毒蛾.喷施茉莉酸甲酯的落叶松对舞毒蛾幼虫生长发育影响最大,4龄后幼虫体质量、蛹质量和成虫产卵量也显著低于其它处理的幼虫.说明外源化合物和昆虫取食均能诱导落叶松的防御反应,且茉莉酸甲酯的诱导效果最好.因此,茉莉酸甲酯作为一种外源植物激素,能够有效诱导落叶松产生抗性,可以用于害虫的综合治理.     

植物内源茉莉酸类生长物质研究进展

植物体内的茉莉酸类化合物（茉莉素）是一类新确认的植物内源生长物质，高浓度茉莉酸或茉莉酸甲酯抑制植生长，低浓度则促进植物生长。茉莉酸或茉莉酸甲酯还能调节植物体保护性酶以及次生代谢酶类的活性，促进基因表达，诱导蛋白质及次生物质的合成，从而提高植物抗逆能力。     

茉莉酸甲酯采前处理对花生成熟种子黄曲霉抗性的影响

以湛秋48和粤油5号花生(Arachis hypogaea L.)为试验材料，在荚果发育初期用茉莉酸甲酯处理，再进行干旱处理，于干旱处理两周后接种黄曲霉(Aspergillus flavus)，收获时测定花生种子的黄曲霉毒素Bl含量、蛋白酶抑制剂、脂肪氧化酶的活性．结果表明，茉莉酸甲酯能减少干旱情况下花生成熟种子黄曲霉毒素Bl的含量，诱导蛋白酶抑制剂的形成，诱导脂肪氧化酶活性升高．     

茉莉酸信号转导途径介导番茄对棉铃虫的抗性

以过表达前系统素转基因番茄(35S∷prosys)、茉莉酸合成突变体番茄(spr2)和野生型番茄(WT)为试验材料,研究了其被棉铃虫取食后叶片保护酶活性和防御反应基因表达情况,以及对棉铃虫抗性的影响.结果表明:棉铃虫取食不同基因型番茄叶片后,多酚氧化酶、过氧化物酶、超氧化物歧化酶和脂氧合酶活性在35S∷prosys植株中最高,在WT植株中次之,在spr2植株中最低.荧光定量PCR检测表明,棉铃虫取食不同基因型番茄叶片后,脂氧合酶基因、丙二烯氧化物环化酶基因和蛋白酶抑制剂基因的转录水平在35S∷prosys植株中最高,在WT植株中次之,而在spr2植株中这些抗性基因的表达未受到虫害的诱导.生物测定表明,取食spr2的棉铃虫体重是取食WT和35S∷prosys棉铃虫体重的2.17和2.96倍.可见,茉莉酸信号途径介导了番茄对棉铃虫的抗性.     

外源茉莉酸对菜豆和番茄幼苗3种防御酶的诱导效应

为明确外源茉莉酸（JA）对菜豆和番茄化学防御的诱导作用,以鲁阳地豆菜豆和红珍珠番茄为材料,研究4种浓度（0.001、0.01、0.1和1 mmol/L）JA喷施2种植物幼苗后叶片多酚氧化酶(PPO)、过氧化物酶(POD) 和脂氧合酶(LOX)的活性变化。结果表明,JA处理可以提高菜豆和番茄叶片3种防御酶的活性,活性提高与诱导的JA浓度和诱导时间有关。0.001 mmol/L JA对菜豆叶片3种防御酶诱导作用不明显,0.1 mmol/L JA诱导效果均显著。4种JA浓度对番茄叶片POD和PPO均具有显著的诱导作用,但0.001和1 mmol/L JA对LOX诱导效果不明显。0.1 mmol/L JA处理72 h后对番茄叶片LOX诱导作用最强,但处理24 h后对番茄叶片POD和PPO以及菜豆叶片3种防御酶诱导作用最强。     

大豆植株挥发物组分变化与大豆食心虫为害的关系

检测了经茉莉酸甲酯和水杨酸甲酯诱导处理后大豆植株挥发物组分的相对含量,并调查了诱导处理小区的大豆食心虫为害情况。结果表明:对照大豆叶片挥发物检测到8种,豆荚挥发物15种;茉莉酸甲酯诱导后叶片和豆荚挥发物中的酯类和萜类组分相对含量明显增加;水杨酸甲酯诱导后大豆叶片挥发物组分含量与对照差异不大,豆荚中烷类组分含量增加。2种诱导剂处理小区内的大豆食心虫成虫数量、卵荚率、幼虫入荚率和虫食率明显低于对照,且差异显著。茉莉酸甲酯处理区效果好于水杨酸甲酯处理区。     

外源茉莉酸甲酯对柳杉幼苗内含物及柳杉毛虫幼虫食物利用的影响

茉莉酸信号通路在柳杉响应柳杉毛虫取食过程中具有重要作用。为了评价外源茉莉酸甲酯(MeJA)对柳杉幼苗抗虫性的诱导性能,对1年生柳杉幼苗喷施3种浓度(0.001、0.01、0.1 mmol·L^-1)的MeJA,分别于处理4、8、12、24、48 h后采集柳杉针叶,测定其营养物质(可溶性糖和可溶性蛋白)、次生物质(单宁、黄酮和总酚)的质量分数和防御酶(多酚氧化酶和苯丙氨酸解氨酶)活性,以及柳杉毛虫5龄幼虫对3种处理柳杉幼苗的食物利用率。结果表明：与对照相比,经不同浓度外源MeJA处理后,柳杉幼苗体内可溶性糖和可溶性蛋白质量分数总体上呈降低趋势,单宁、黄酮和总酚质量分数有不同程度的增大,多酚氧化酶和苯丙氨酸解氨酶活性均升高;柳杉毛虫的食物利用率显著降低。这表明外源MeJA可通过抑制2种营养物质和促进3种植物次生物质的合成增强植株的抗虫性。本研究可为柳杉的抗虫性研究提供新思路。     

Effect of Exogenous MJA Treatment of Tea Plants on the Growth of Geometrid Larvae

The effect of tea plant Camellia sinensis induced by exogenous methyl jasmonate （MJA） on lipoxygenase （LOX）, polyphenol oxidase （PPO） and proteinase inhibitor （PI） activity in the leaves of tea plants, as well as the growth and midgut proteinase activity of the geometrid Ectropis obliqua larvae were studied. MJA significantly induced LOX, PPO and PI activity in leaves of tea plants. When geometrid larvae have fed on leaves of tea plants treated with MJA, the activities of the high alkaline trypsin-like enzyme and chymotrypsin-like enzyme in their midgut were significantly inhibited, but the activities of the low alkaline trypsin-like enzyme in their midgut were unaffected, leading to imbalance between different types of proteinase activity in the midgut of the larvae and in turn, the growth were inhibited. These chains of response may be an important mechanism of the direct resistance induced by MJA-treatment of tea plant on geometrid larvae.     

稀土对茶树不同部位叶片中酶活性的影响

以安徽农业大学茶业系品种园的安农早抗 1号为材料 ,稀土处理后测定不同部位叶片中酶活性 ,并与对照相比较 ,结果表明 :稀土处理的茶树谷氨酸脱氢酶、过氧化物酶、多酚氧化酶及肽酶在第 1叶位的活性、总活性均得到提高 ;而在第 4叶及老叶部位酶活性与对照相比未呈现规律性。第 1叶位中酶活性的提高 ,有利于茶树经济性状的改善 ,这可能是茶园喷施稀土肥料的增产提质机理之一。     

Response dynamics of three defense related enzymes in cotton leaves to the interactive stress of Helicoverpa armigera (Hübner) herbivory and omethoate application

In order to explore the response dynamics of the activities of defense related enzymes in cotton leaves towards the interactive stress of Helicoverpa armigera herbivory and omethoate application, the activities of phenylalanine ammonia-lyase (PAL), lipoxygenase (LOX), and polyphenol oxidase (PPO) were examined from 6 to 126 h after cotton leaves were treated 12 h of H. armigera herbivory, and then sprayed with 800 mg L–1 omethoate. The results showed that the changes in the activities of PAL, LOX and PPO that occured under the interactive stress of H. armigera herbivory and omethoate application reflected the interactive effects of the two stresses on cotton defense. The similarity between the response dynamics of PAL, LOX, and PPO activities in cotton leaves under the interactive stress and that under H. armigera herbivory treatment alone showed that the induction of H. armigera herbivory on the activities of PAL, LOX and PPO in cotton leaves played a leading role in the interactive effects, and the effect of omethoate application played only a minor role. A joint factor analysis was performed according to a method which has been used to analyze the joint toxicity of pesticides; this analysis sought to clarify if there was a synergistic, antagonistic, or additive effect on PAL, LOX, and PPO activity in cotton leaves resulting from the interactive H. armigera herbivory and omethoate treatment. In the interactive effect on the response of PAL activity in cotton leaves, antagonistic effects of the omethoate application towards H. armigera herbivory were observed at 6 and 12 h. Synergistic effects were then observed at 18 and 30 h. Antagonistic effects were observed from 54 to 78 h and synergistic effects were finally observed at 126 h. The correlation between H. armigera herbivory and omethoate application in the interactive effect on cotton defense responses of LOX activity also fluctuated from synergism to antagonism during the time course. In the interactive effect on PPO activity, only antagonism was observed between H. armigera herbivory and omethoate application. In the interactive stress of H. armigera herbivory and omethoate application on cotton defense responses, omethoate affected the defense responses of cotton to H. armigera herbivory by producing antagonistic and synergistic effects. These results will be useful to understand the relationship between host plant and herbivorous pest.     

松油烯-4-醇对粘虫几种代谢酶及酚氧化酶的影响

【目的】探讨杀虫植物砂地柏Sabina vulgaris精油中主要杀虫成分松油烯-4-醇的作用机理。【方法】以点滴法处理粘虫5龄幼虫后,测定兴奋期、痉挛期、麻痹期及复苏期等4个中毒阶段试虫体内4种代谢酶及酚氧化酶（PPO）的活性。【结果】松油烯-4-醇仅在麻痹期对酸性磷酸酯酶（ACP）具有一定的抑制作用,而在其它时期均可明显激活ACP及碱性磷酸酯酶（AKP）;对谷胱甘肽S转移酶（GSTs）则表现出先抑制后恢复的趋势;可显著抑制细胞色素P450 O-脱甲基活性,各中毒阶段酶抑制率分别为26.27%、46.03%、80.24%和90.22%;离体和活体条件下,均可显著抑制PPO活性。【结论】松油烯-4-醇对粘虫代谢酶系细胞色素P450、GSTs及PPO的抑制与其毒杀活性有关,而中毒试虫的复苏可能与GSTs活性的恢复有关。该化合物可开发为新型农药或增效剂,在害虫抗性治理中具有潜在的应用价值。     

转Bt基因杨树对美国白蛾幼虫中肠解毒酶及乙酰胆碱酯酶的影响

以转Bt基因欧洲黑杨叶片饲喂4-5龄美国白蛾幼虫，对其中肠几种重要解毒酶及乙酰胆碱酯酶的活性进行了测定。结果表明，中肠多功能氧化酶受到强烈抑制，饲喂4h后，多功能氧化酶的活性比对照降低50.89%，48h后，该酶活性仅为对照的1/6，酯酶及羟酸酯酶的活性也受到明显抑制，饲喂48h后，上述两种酶的活性分别比对照降低73.58%和56.55%，谷胱甘肽S-转移酶变化比较复杂，饲喂24h后其活性出现峰值；48h后与对照持平，中肠乙酰胆碱酯酶活性无明显变化，转基因叶片对幼虫中肠解毒酶活性的抑制可能是其毒杀害虫的作用机制之一。     

Effects of Terpinen-4-ol on Four Metabolic Enzymes and Polyphenol Oxidase （PPO） in Mythimna separta Walker

To study insecticidal mechanism of terpinen-4-ol, a main insecticidal composition in the essential oil of Sabina vulgaris, the 5th instar larvae of Mythimna separta, were investigated with terpinen-4-ol by topical application. The activities of phosphatase, glutathione S-transferase （GSTs）, cytochrome P450 （P450）, and polyphenol oxidase （PPO） of tested insects were determined in all poisoning stages, including exciting stage, convulsing stage, paralysis stage, and recover stage. The result showed that the activities of both acid phosphatase （ACP） and alkaline phosphatase （AKP） in treated insects were induced by terpinen-4-ol, but ACP was inhibited in paralysis stage. The activities of GSTs were inhibited in exciting stage, convulsing stage, and paralysis stage, but gradually recovered in recover stage. O-demethylase activity of cytochrome P450 was inhibited by terpinen-4-ol, and the inhibition rate in all poisoning stages were 26.27, 46.03, 80.24, and 90.22%, respectively. PPO activities were strongly inhibited by terpinen-4-ol both in vitro and in vivo. In conclusion, the activities of P450, GSTs, and PPO could have relation with toxicity of terpinen-4-ol against larvae of the Mythimna separta, but recover stage of the poisoning insects might be related to GSTs induced. As a new insecticide or synergist, terpinen- 4-ol has a potential value in field of insecticide resistance management.     

Effects of Terpinen-4-ol on Four Metabolic Enzymes and Polyphenol Oxidase (PPO) in Mythimna separta Walker

To study insecticidal mechanism of terpinen-4-ol, a main insecticidal composition in the essential oil of Sabina vulgaris, the 5th instar larvae of Mythimna separta, were investigated with terpinen-4-ol by topical application. The activities of phosphatase, glutathione S-transferase (GSTs), cytochrome P450 (P450), and polyphenol oxidase (PPO) of tested insects were determined in all poisoning stages, including exciting stage, convulsing stage, paralysis stage, and recover stage. The result showed that the activities of both acid phosphatase (ACP) and alkaline phosphatase (AKP) in treated insects were induced by terpinen-4-ol, but ACP was inhibited in paralysis stage. The activities of GSTs were inhibited in exciting stage, convulsing stage, and paralysis stage, but gradually recovered in recover stage. O-demethylase activity of cytochrome P450 was inhibited by terpinen-4-ol, and the inhibition rate in all poisoning stages were 26.27, 46.03, 80.24, and 90.22%, respectively. PPO activities were strongly inhibited by terpinen-4-ol both in vitro and in vivo. In conclusion, the activities of P450, GSTs, and PPO could have relation with toxicity of terpinen-4-ol against larvae of the Mythimna separta, but recover stage of the poisoning insects might be related to GSTs induced. As a new insecticide or synergist, terpinen- 4-ol has a potential value in field of insecticide resistance management.     

转双抗三倍体毛白杨对舞毒蛾幼虫中肠解毒酶的影响

[目的]为转基因植物的合理利用提供理论依据。[方法]以转Bt基因和慈姑蛋白酶抑制剂基因三倍体毛白杨叶片喂4～5龄舞毒蛾幼虫,对其中肠几种重要解毒酶的活性进行了测定。[结果]幼虫中肠酯酶的比活力变化是先升高后降低,24 h后达到高峰,而在48 h后,比活力明显下降;谷胱甘肽-S-转移酶的比活力是一直在升高。取食抗虫杨Pb1、Pb33和Pb47叶片48 h后的舞毒蛾幼虫中肠酯酶的比活力分别为取食CK幼虫酯酶比活力的62.9%、70.7%和74.5%;24 h后的幼虫中肠谷胱甘肽-S-转移酶的比活力分别比取食CK幼虫谷胱甘肽-S-转移酶比活力下降了25.7%、27.7%和17.9%。[结论]导致幼虫中肠解毒酶活性的改变可能是转基因叶片毒杀害虫的作用机制之一。     

棉花多酚氧化酶基因GhPPO1的克隆及在棉铃虫取食诱导反应中的作用

【目的】克隆棉花多酚氧化酶基因（GhPPO1）全长cDNA序列,分析其序列特征,并研究其及多酚氧化酶系对棉铃虫（Helicoverpa armigera）的取食诱导反应,明确其在棉花防御棉铃虫取食中的作用。【方法】根据笔者研究组前期从棉花SSH文库中获得的对昆虫取食诱导响应明显的棉花PPO基因序列片段,设计5′RACE特异引物,进行5′端RACE反应,测序后进行序列拼接。拼接序列在NCBI棉花dbEST数据库中,用Blastn程序进行同源检索,获得一条EST（GenBank登录号：DR461072.1）与测序结果有410 bp重复。用DNAstar进行组装、电子延伸,获得一条棉花PPO基因序列,命名为GhPPO1。在GhPPO1序列两端设计引物进行全长验证,同时设计引物,以棉花基因组DNA为模板,进行PCR扩增测序,验证GhPPO1是否含有内含子。采用BLASTX在NCBI数据库中对验证后的GhPPO1序列进行同源序列分析;ClustalW软件进行多重序列比对;MEGA 4.0软件构建系统进化树;同时对推测的编码区氨基酸序列进行功能位点及理化性质的预测与分析,所用软件包括ANTHEPRO5.0、ExPASy、InterProscan等。利用实时荧光定量PCR方法测定机械损伤、棉铃虫取食和口腔分泌物处理后,棉花叶片中GhPPO1的mRNA表达量;并通过分光光度法测定棉花叶片中PPO酶系的活性变化趋势。【结果】 GhPPO1的cDNA序列全长为2 022 bp,推测该基因编码区（ORF）为1 797 bp,编码598个氨基酸,5′UTR 含102 bp,3′UTR含123 bp,预测其等电点为6.11,蛋白分子量约67.18 kD,无内含子。GhPPO1编码区氨基酸序列包含CuA和CuB结合位点、3个N-糖基化位点、8个N-豆蔻酰化位点、6个蛋白激酶C磷酸化位点、8个酪蛋白激酶Ⅱ磷酸化位点、1个依赖于cAMP和cGMP的蛋白激酶磷酸化位点及1个酰胺化位点。GhPPO1的CuA和CuB离子结合区,有PPO蛋白关键氨基酸组氨酸和半胱氨酸。GhPPO1蛋白与其他植物的PPO蛋白序列最大相似度几乎均在50%以上,与其他植物PPO蛋白进化关系相对较远。机械损伤棉花叶片后,GhPPO1表达量呈现出先升高后下降的趋势,至6 h表达量最高,为无处理对照的3.29倍;1龄末期棉铃虫幼虫取食棉花后,叶片中该基因的表达量呈现先升高后下降,然后又升高的趋势,至12 h表达量最高,为无处理对照的6.04倍;棉铃虫取食3 h和6 h后的表达量低于机械损伤处理后相同时间的表达量。棉铃虫取食和机械损伤棉花叶片后,PPO酶活性均呈现出升高趋势,且机械损伤处理酶活性在各时间段均高于棉铃虫幼虫取食处理。与未做任何处理的正常叶片相比,机械损伤和清水共同处理的叶片GhPPO1表达量及PPO酶活性显著升高,而机械损伤和棉铃虫幼虫口腔分泌物共同处理的叶片GhPPO1表达量及PPO酶活性无显著变化,表明口腔分泌物对GhPPO1表达量及PPO酶活性有抑制作用。【结论】获得了GhPPO1的全长cDNA序列,证明GhPPO1是棉花防御棉铃虫取食的关键PPO基因,推测棉铃虫口腔分泌物中存在抑制棉花PPO酶活性升高物质,该物质在棉铃虫适应寄主植物产生的防御反应中发挥作用。