Effects of feeding Spodoptera littoralis on lima bean leaves: IV. Diurnal and nocturnal damage differentially initiate plant volatile emission

Continuous mechanical damage initiates the rhythmic emission of volatiles in lima bean (Phaseolus lunatus) leaves; the emission resembles that induced by herbivore damage. The effect of diurnal versus nocturnal damage on the initiation of plant defense responses was investigated using MecWorm, a robotic device designed to reproduce tissue damage caused by herbivore attack. Lima bean leaves that were damaged by MecWorm during the photophase emitted maximal levels of beta-ocimene and (Z)-3-hexenyl acetate in the late photophase. Leaves damaged during the dark phase responded with the nocturnal emission of (Z)-3-hexenyl acetate, but with only low amounts of beta-ocimene; this emission was followed by an emission burst directly after the onset of light. In the presence of (13)CO(2), this light-dependent synthesis of beta-ocimene resulted in incorporation of 75% to 85% of (13)C, demonstrating that biosynthesis of beta-ocimene is almost exclusively fueled by the photosynthetic fixation of CO(2) along the plastidial 2-C-methyl-D-erythritol 4-P pathway. Jasmonic acid (JA) accumulated locally in direct response to the damage and led to immediate up-regulation of the P. lunatus beta-ocimene synthase gene (PlOS) independent of the phase, that is, light or dark. Nocturnal damage caused significantly higher concentrations of JA (approximately 2-3 times) along with enhanced expression levels of PlOS. Transgenic Arabidopsis thaliana transformed with PlOS promoter :: beta-glucuronidase fusion constructs confirmed expression of the enzyme at the wounded sites. In summary, damage-dependent JA levels directly control the expression level of PlOS, regardless of light or dark conditions, and photosynthesis is the major source for the early precursors of the 2-C-methyl-D-erythritol 4-P pathway.     

Effects of feeding Spodoptera littoralis on lima bean leaves. III. Membrane depolarization and involvement of hydrogen peroxide

In response to herbivore (Spodoptera littoralis) attack, lima bean (Phaseolus lunatus) leaves produced hydrogen peroxide (H(2)O(2)) in concentrations that were higher when compared to mechanically damaged (MD) leaves. Cellular and subcellular localization analyses revealed that H(2)O(2) was mainly localized in MD and herbivore-wounded (HW) zones and spread throughout the veins and tissues. Preferentially, H(2)O(2) was found in cell walls of spongy and mesophyll cells facing intercellular spaces, even though confocal laser scanning microscopy analyses also revealed the presence of H(2)O(2) in mitochondria/peroxisomes. Increased gene and enzyme activations of superoxide dismutase after HW were in agreement with confocal laser scanning microscopy data. After MD, additional application of H(2)O(2) prompted a transient transmembrane potential (V(m)) depolarization, with a V(m) depolarization rate that was higher when compared to HW leaves. In transgenic soybean (Glycine max) suspension cells expressing the Ca(2+)-sensing aequorin system, increasing amounts of added H(2)O(2) correlated with a higher cytosolic calcium ([Ca(2+)](cyt)) concentration. In MD and HW leaves, H(2)O(2) also triggered the increase of [Ca(2+)](cyt), but MD-elicited [Ca(2+)](cyt) increase was more pronounced when compared to HW leaves after addition of exogenous H(2)O(2). The results clearly indicate that V(m) depolarization caused by HW makes the membrane potential more positive and reduces the ability of lima bean leaves to react to signaling molecules.     

Effects of feeding Spodoptera littoralis on lima bean leaves. I. Membrane potentials, intracellular calcium variations, oral secretions, and regurgitate components

Membrane potentials (V(m)) and intracellular calcium variations were studied in Lima bean (Phaseolus lunatus) leaves when the Mediterranean climbing cutworm (Spodoptera littoralis) was attacking the plants. In addition to the effect of the feeding insect the impact of several N-acyl Glns (volicitin, N-palmitoyl-Gln, N-linolenoyl-Gln) from the larval oral secretion was studied. The results showed that the early events upon herbivore attack were: a) a strong V(m) depolarization at the bite zone and an isotropic wave of V(m) depolarization spreading throughout the entire attacked leaf; b) a V(m) depolarization observed for the regurgitant but not with volicitin [N-(17-hydroxy-linolenoyl)-Gln] alone; c) an enhanced influx of Ca(2+) at the very edge of the bite, which is halved, if the Ca(2+) channel blocker Verapamil is used. Furthermore, the dose-dependence effects of N-acyl Gln conjugates-triggered influx of Ca(2+) studied in transgenic aequorin-expressing soybean (Glycine max) cells, showed: a) a concentration-dependent influx of Ca(2+); b) a configuration-independent effect concerning the stereochemistry of the amino acid moiety; c) a slightly reduced influx of Ca(2+) after modification of the fatty acid backbone by functionalization with oxygen and; d) a comparable effect with the detergent SDS. Finally, the herbivore wounding causes a response in the plant cells that cannot be mimicked by mechanical wounding. The involvement of Ca(2+) in signaling after herbivore wounding is discussed.     

西花蓟马取食与机械损伤对菜豆叶片抗氧化系统的影响

本文研究了西花蓟马Frankliniella occidentalis(Pergande)取食和机械损伤诱导对菜豆抗氧化系统的影响,比较了不同损伤形式诱导的抗氧化酶活力和抗氧化物质含量的变化差异。结果表明,西花蓟马取食和机械损伤均可诱导菜豆叶片内过氧化物酶(POD)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)3种抗氧化酶活性有不同程度的升高,但两种处理诱导的抗氧化酶活性变化规律不完全相同,各种酶活性达到最高点时间不同,西花蓟马取食对抗氧化酶活力的诱导作用大于机械损伤的。两种处理诱导下的类胡萝卜素的含量变化不大,但不论西花蓟马取食还是机械损伤均导致菜豆叶片内类黄酮和总酚含量整体呈下降趋势,西花蓟马的取食诱导的下降幅度大于机械损伤的。因此,西花蓟马取食诱导明显高于机械损伤对菜豆抗氧化系统的影响。     

Effects of extracellular ATP on local and systemic responses of bean (Phaseolus vulgaris L) leaves to wounding

Wounding increased the extracellular Adenosine 5?-triphosphate (eATP) level of kidney bean leaves. Treatment with wounding or exogenous ATP increased the hydrogen peroxide (H₂O₂) content, activities of catalase and polyphenol oxidase, and malondialdehyde content in both the treated and systemic leaves. Pre-treatment with ATP-degrading enzyme, apyrase, to the wounded leaves reduced the wound-induced local and systemic increases in H₂O₂ content, activities of catalase and polyphenol oxidase, and malondialdehyde content. Application of dimethylthiourea (DMTU) and diphenylene iodonium (DPI) to the wounded and ATP-treated leaves, respectively, reduced the wound- and ATP-induced local and systemic increases in H₂O₂ content, activities of catalase and polyphenol oxidase, and malondialdehyde content. Moreover, the wound- and ATP-induced systemic increases of these physiological parameters were suppressed when DMTU or DPI applied to leaf petiole of the wounded and ATP-treated leaves. These results suggest that eATP at wounded sites could mediate the wound-induced local and systemic responses by H₂O₂-dependent signal transduction.     

蓟马取食、机械损伤以及外源水杨酸甲酯和茉莉酸对菜豆叶片防御酶活性的影响

【目的】探讨菜豆对昆虫取食防御反应的生化机制。【方法】研究了西花蓟马Frankliniella occidentalis取食、机械损伤以及外源水杨酸甲酯（MeSA）和茉莉酸（JA）处理后菜豆叶片防御酶活性的变化。【结果】西花蓟马取食、机械损伤及MeSA和JA处理均能明显提高过氧化物酶（POD）的活性,前2种处理POD活性在72 h上升到最高峰,而后2种处理则在48 h达到最高峰。蛋白酶抑制剂（PI）活性在西花蓟马取食后升高最明显。JA途径关键酶脂氧合酶（LOX）和多酚氧化酶（PPO）的活性在西花蓟马取食、机械损伤和JA诱导处理均升高,但外源MeSA诱导处理则不能诱导它们的活性(P>0.05)。SA途径的关键酶苯丙氨酸解氨酶（PAL）在西花蓟马取食和机械损伤后均有一个先升高后下降的过程,外源MeSA诱导只在24 h引起PAL活性升高,其余时间下和对照没有明显的区别,外源JA诱导未能引起PAL活性的显著变化(P>0.05)。西花蓟马取食、JA和MeSA诱导以及机械损伤均能诱导β-1,3 葡聚糖酶（PR-2）活性上升(P<0.05)。【结论】结果说明,不同处理可诱导菜豆植株产生明显的防御反应,但酶活性的变化与处理方式和处理时间有关。     

The glutathione-deficient mutant pad2-1 accumulates lower amounts of glucosinolates and is more susceptible to the insect herbivore Spodoptera littoralis

Plants often respond to pathogen or insect attack by inducing the synthesis of toxic compounds such as phytoalexins and glucosinolates (GS). The Arabidopsis mutant pad2-1 has reduced levels of the phytoalexin camalexin and is known for its increased susceptibility to fungal and bacterial pathogens. We found that pad2-1 is also more susceptible to the generalist insect Spodoptera littoralis but not to the specialist Pieris brassicae . The PAD2 gene encodes a γ-glutamylcysteine synthetase that is involved in glutathione (GSH) synthesis, and consequently the pad2-1 mutant contains about 20% of the GSH found in wild-type plants. Lower GSH levels of pad2-1 were correlated with reduced accumulation of the two major indole and aliphatic GSs of Arabidopsis, indolyl-3-methyl-GS and 4-methylsulfinylbutyl-GS, in response to insect feeding. This effect was specific to GSH, was not complemented by treatment of pad2-1 with the strong reducing agent dithiothreitol, and was not observed with the ascorbate-deficient mutant vtc1-1 . In contrast to the jasmonate-insensitive mutant coi1-1 , expression of insect-regulated and GS biosynthesis genes was not affected in pad2-1 . Our data suggest a crucial role for GSH in GS biosynthesis and insect resistance.     

Effects of water stress on plant volatile emission and insect oviposition preference in an agroecosystem

1. Drought has become more common and severe in many parts of the world due to climate change. The effect of water stress on insect oviposition preference that is key determinant for their fitness has received less attention.
2. Here, we examined how water stress may affect oviposition selection of Ostrinia furnacalis for maize plants in the greenhouse, and tested difference in volatile compounds emitted from treated maize, and electronantennogram and bioassay responses of O. furnacalis to the volatile profiles in maize plants.
3. Ostrinia furnacalis were more prone to lay eggs on the well-watered maize. Most plant volatile compounds differed significantly among the three water treatments, including increased emissions of β-caryophyllene, (E)-2-hexenal, and linalool, and decreased emission of (Z)-3-hexen-1-ol when subjected to increasing intensity of drought.
4. Varied volatile profiles of maize may drive oviposition decision of O. furnacalis, because O. furnacalis showed a clear oviposition preference for (Z)-3-hexen-1-ol, while not for β-caryophyllene, (E)-2-hexenal, and linalool, at the concentration of 1000 ppm.
5. This study advances understanding of drought effects on plant–insect interactions through volatile profiles. Our finding calls for attention to oviposition selection for insect pest management in agricultural settings, especially in regions under changing precipitation patterns.

     

Ion channel-forming alamethicin is a potent elicitor of volatile biosynthesis and tendril coiling. Cross talk between jasmonate and salicylate signaling in lima bean

Alamethicin (ALA), a voltage-gated, ion channel-forming peptide mixture from Trichoderma viride, is a potent elicitor of the biosynthesis of volatile compounds in lima bean (Phaseolus lunatus). Unlike elicitation with jasmonic acid or herbivore damage, the blend of substances emitted comprises only the two homoterpenes, 4,11-dimethylnona-1,3,7-triene and 4,8,12-trimethyltrideca-1,3,7,11-tetraene, and methyl salicylate. Inhibition of octadecanoid signaling by aristolochic acid and phenidone as well as mass spectrometric analysis of endogenous jasmonate demonstrate that ALA induces the biosynthesis of volatile compounds principally via the octadecanoid-signaling pathway (20-fold increase of jasmonic acid). ALA also up-regulates salicylate biosynthesis, and the time course of the production of endogenous salicylate correlates well with the appearance of the methyl ester in the gas phase. The massive up-regulation of the SA-pathway (90-fold) interferes with steps in the biosynthetic pathway downstream of 12-oxophytodienoic acid and thereby reduces the pattern of emitted volatiles to compounds previously shown to be induced by early octadecanoids. ALA also induces tendril coiling in various species like Pisum, Lathyrus, and Bryonia, but the response appears to be independent from octadecanoid biosynthesis, because inhibitors of lipoxygenase and phospholipase A(2) do not prevent the coiling reaction.     

抗虫辣椒叶片对斜纹夜蛾幼虫相关生理生化指标的影响

为明确朝天椒(品种：BLTY2)叶片饲喂的斜纹夜蛾Spodoptera litura幼虫生长发育受到抑制的关键原因,本研究以人工饲料及牛角椒（品种：FXBX）叶片饲喂的斜纹夜蛾幼虫为对照,通过比较3种食料饲喂后斜纹夜蛾幼虫相关生理生化指标的变化,分析BLTY 2辣椒叶片饲喂所致的斜纹夜蛾幼虫生长发育异常的原因。结果表明,朝天椒辣椒叶片饲喂的斜纹夜蛾幼虫生长发育速率滞后于牛角椒辣椒叶片及人工饲料饲喂的斜纹夜蛾幼虫,且每头幼虫的发育时长均显著延长（P<0.05）。饲喂后第1天、第3天、第4天、第5天、第6天和第7天,饲喂BLTY 2辣椒叶片的斜纹夜蛾幼虫体内保幼激素（JH）含量均显著高于其他2种食料饲喂的斜纹夜蛾幼虫（P<0.05）。饲喂后第3天、第4天、第5天和第7天,饲喂BLTY 2辣椒叶片的斜纹夜蛾幼虫体内蜕皮激素（Ecd）含量均显著高于其他2种食料饲喂的斜纹夜蛾幼虫（P<0.05）。3种食料饲喂后斜纹夜蛾幼虫体内的几丁质酶（CHT）活性变化差异较大,而2种辣椒叶片饲喂后斜纹夜蛾幼虫体内的组织蛋白酶（CTS）活性变化趋势相似,且不同于人工饲料饲喂的斜纹夜蛾幼虫体内的CTS活性变化趋势。由此可见,BLTY 2辣椒主要干扰斜纹夜蛾幼虫体内JH与Ecd的含量抑制其生长发育。本研究结果可为筛选具有抗虫活性的植物次生代谢物质奠定基础。     

绿盲蝽取食与机械损伤对棉花叶片内防御性酶活性的影响

为探明绿盲蝽Apolygus lucorum (Mayer-Dür)取食和机械损伤对不同抗性棉花叶片内主要防御酶活性的影响以及防御酶与棉花抗绿盲蝽性的关系,以棉花3个不同抗性品系为材料,室内条件下测定绿盲蝽取食和机械损伤处理后棉叶中苯丙氨酸解氨酶(PAL)、过氧化物酶(POD)和多酚氧化酶(PPO)的活性.结果表明:对...     

Effects of juvenile hormone I and precocene I & II on Microplitis rufiventris when administered via its host, Spodoptera littoralis

We investigated the effect of juvenile hormoneI (JHI) and precocene I & II (PI & PII) onthe endo-developmental stages, reproductiveperformance and longevity of Microplitisrufiventris Kok. after topical treatment ofhost larvae of Spodoptera littoralis(Boisd.). Host larvae were treated with each ofthe test compounds by a single dose 2–4 h afterparasitization or 1 day prior to parasitoidemergence, or by repeated daily applications 2–4 h after parasitization to parasitoid emergence.JHI-treatments increased parasitoiddevelopmental time, induced developmentalabnormalities and inhibition of parasitoidegression. Interestingly, one of the causal factorsthat inhibited the parasitoid egression was themoulting of some parasitoid larvae into asupernumerary instar. Applications of PI or PIIto parasitized S. littoralis larvae didnot affect parasitoid developmental time, butsome developmental abnormalities were noted.Melanin formation in host's blood was greaterin PI-treatments than in PII-treatments.PII-treatments arrested development of somefirst instar parasitoid larvae typical of JHIexcess. Adult female wasps that emerged fromJHI-treated hosts lived for significantlylonger periods than control females, but didnot produce more offspring. The longevity andreproductive capacity of females whichdeveloped in hosts treated by precocenes werereduced compared with the females emerged fromcontrol hosts. Even the small number ofadult wasps from PII-treated hosts which livedfor the same period as control wasps, showed asignificant reduction in their reproductiveperformance. Repeated daily applications of thetest compounds reduced wasp fecundity andlongevity to a significantly greater degreethan the single applications, except for a lowdose of JHI (1 µg). The study demonstratesthat growth disrupters, such as JHs andprecocenes have a delayed developmental effect onM. rufiventris and points out the need tobe aware of these effects in implementingcontrol strategies of S. littoralis innatural habitats.     

Bioassaying an insect virus on leaves. II. The influence of certain factors associated with the larvae and the leaves

The importance of certain factors which act through the larvae or through the leaves when a virus suspension is bioassayed by applying a known volume in droplets to a leaf of standard size has been investigated. The most critical was the age of the second-instar larvae. In 24 hr their resistance to infection increased significantly. On the other hand, starving the test larvae for 6 hr to assist randomization of factors that influenced the area of treated leaf consumed did not produce a measurable effect on mortality. A 2°C increase in the temperature reduced the incubation period of the disease but produced no effect on the final kill due to the virus. The practical implications of these results are discussed in relation to the bioassay procedure.     

樟树幼苗机械损伤叶片对挥发性有机化合物及叶绿素荧光参数的影响

为探讨植物在机械损伤后C₆-C₁₀醛类化合物的释放机理, 及C₆-C₁₀醛类化合物对叶片光系统II (PSII)的影响, 以樟树(Cinnamomum camphora)为材料, 采用动态顶空气体循环法和热脱附/气相色谱/质谱联用技术(TDS-GC-MS), 对樟树幼苗叶片损伤后释放的C₆-C₁₀醛类化合物进行采集与分析, 并同步测定了脂氧合酶活性和损伤叶片的叶绿素荧光动力学参数。结果表明: 樟树幼苗叶片损伤后, 其挥发性有机化合物中己醛、庚醛、辛醛、壬醛和癸醛的释放量比损伤前分别增加了2.47、0.99、1.34、0.91和28.38倍(p < 0.01); 同时新增4种醛类化合物, 分别是: 2-己烯醛、2,4-己二烯醛、(E)-2-辛烯醛和(E)-2-壬烯醛。脂氧合酶活性比损伤前增加1.2倍(p < 0.01)。PSII单位反应中心复合体吸收的能量和被核心捕获的能量分别比损伤前下降12.8%和9.8% (p < 0.01)。单位面积反应中心的数量、电子传递量子产额、捕获激子能导致电子传递效率和叶片性能指数分别比损伤前增加23.3%、24.4%、22.6%和82.7% (p < 0.01)。损伤24 h后, 醛类化合物的种类、释放量、脂氧合酶活性及叶片叶绿素荧光动力学参数基本恢复到损伤前水平。说明机械损伤使PSII供体侧受损、脂氧合酶活性升高, 致使C₆-C₁₀醛类化合物大量释放, 樟树幼苗通过增加单位面积反应中心的数量来提高光合效率应对胁迫。     

Hylobius abietis L. feeding on the novel host Pinus brutia Ten. increases emission of volatile organic compounds

Plants respond to feeding by herbivorous insects by producing volatile organic chemicals, which mediate interactions between herbivores and plants. Yet, few studies investigated whether such plant responses to herbivory differ between historical host and novel plants. Here, we investigated whether herbivory by the pine weevil Hylobius abietis causes a release of volatile organic chemicals from a novel tree Pinus brutia and compared the relative amounts of volatiles released from herbivore's historical hosts and P. brutia. We collected volatiles emitted from P. brutia seedlings that were either subjected to feeding by H. abietis or no feeding. Our results indicated that feeding increased emission of volatile compounds, composed of monoterpenes and sesquiterpenes, and that the emission was several fold higher in the damaged seedlings than in undamaged seedlings. In particular, emission of monoterpenes and sesquiterpenes increased by 4.4‐and 10‐fold in the damaged plants, respectively. Strikingly, individual monoterpenes and sesquiterpenes showed much greater dissimilarity between damaged and undamaged seedlings. Furthermore, several minor monoterpenes showed negative relationships with the weevil gnawed area. We discussed these results with the results of previous studies focused on historical host plants of H. abietis and hypothesized the ecological relevance and importance of our results pertaining relevance to the plant–herbivory interactions.     

Intermediate steps in cellular iron uptake from transferrin. II. A cytoplasmic pool of iron is released from cultured cells via temperature-dependent mechanical wounding

Summary A previous study described a cytoplasmic, transferrin (Tf)-free, iron (Fe) pool that was detected only when cells were mechanically detached from the culture substratum at 4°C, after initial incubation with⁵⁹Fe-¹²⁵I-Tf at 37°C (Richardson and Baker, 1992a). The release of this internalized⁵⁹Fe could be markedly reduced if the cells were treated with proteases or incubated at 37°C prior to detachment. The present study was designed to characterize this Fe pool and understand the mechanism of its release. The results show that cellular⁵⁹Fe release increased linearly as a function of preincubation time with⁵⁹Fe-Tf subsequent to mechanical detachment at 4°C using a spatula. These data suggest that the⁵⁹Fe released was largely composed of end product(s) and was not an “intermediate Fe pool.” When the Fe(II) chelator, dipyridyl (DP), was incubated with⁵⁹Fe-Tf and the cells, it prevented the accumulation of⁵⁹Fe that was released following mechanical detachment at 4°C. Other chelators had much less effect on the proportion of⁵⁹Fe released. Examination of the⁵⁹Fe released showed that after a 4-h preincubation with⁵⁹Fe-Tf, approximately 50% of the⁵⁹Fe was present in ferritin. These data indicate that mechanical detachment of cells at 4°C resulted in membrane disruptions that allow the release of high M, molecules. Moreover, electron microscopy studies showed that detachment of cells from the substratum at 4°C resulted in pronounced membrane damage. In contrast, when cells were detached at 37°C, or at 4°C after treatment with pronase, membrane damage was minimal or not apparent. These results may imply that temperature-dependent processes prevent the release of intracellular contents on membrane wounding, or alternatively, prevent wounding at 37°C. The evidence also indicates that caution is required when interpreting data from expriments where cells have been mechanically detached at 4°C.     

Effect of cold-storage of bean leaves on photosynthetic reactions of isolated chloroplasts. Inability to donate electrons to photosystem II and relation to manganese content

1.

1. Chloroplasts from bean leaves which have been aged in darkness at o °C do not carry out Hill reactions. They reduce NADP with reduced 2,6-dichlorophenolindophenol (DCIP) in a reaction that is insensitive to 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU). They reduce NADP with hydroquinone, p-phenylenediamine or benzidine, and reduce DCIP with hydroxylamine, 1,4-diphenylsemicarbazide, 1,5-diphenylcarbohydrazide or manganous ion in DCMU-sensitive reactions.     

两种不同种植方式对龙葵叶片虫食状的影响研究

昆虫与植被之间相互关系的深入研究,有助于更好的理解生态系统结构与功能之间的关系以及生态系统生物多样性维持机理。本文通过对集中种植和分块种植下两种不同种植方式龙葵叶片虫食状进行调查分析,以期对龙葵的栽培和养护提供理论基础和技术建议。研究结果表明:龙葵叶片中共识别出11种虫食状类型,其中,集中种植方式有10种,分块种植方式有11种,各种虫食状类型出现频率在0.4%-24.7%;集中种植样地的龙葵叶片虫食状种类数、Shannon-Wiener指数和Pielou均匀度指数均低于分块种植;分块种植的龙葵叶片受到植食性昆虫的伤害频率较高,但是集中种植的龙葵叶片受到的伤害程度却明显高于分块种植方式。因此,种植龙葵应尽量避免大规模集中种植方式,不同生态系统边界之间边缘效应的尺度和强弱的关系是未来研究的重点和核心问题。