棉铃虫持续取食对棉花三种防御酶活性的作用

昆虫取食作为一种关键的生物胁迫因子对棉花防御机制产生了重要影响。植物对昆虫取食产生的防御响应,在昆虫与植物的生态关系中具有重要作用。为了明确棉铃虫Helicoverpa armigera取食与棉花防御性之间的动态互作关系,本文研究了棉铃虫持续取食下及停止取食后,棉花中3种防御相关酶活性变化的时间效应。在明确了棉花受损程度与棉铃虫取食时间关系的基础上,分别考察了棉铃虫持续取食2、6、12、18和24h,对棉花中苯丙氨酸解氨酶(PAL)、脂氧合酶(LOX)和多酚氧化酶(PPO)活性的影响。针对棉铃虫持续取食棉叶12h后停止取食,研究了去除虫害胁迫后0、6、12、24和36h,棉花体内PAL、LOX、PPO活性的变化。结果表明:在棉铃虫持续取食棉叶24h内,棉花中3种防御酶的活性响应有所不同,其中,棉铃虫持续取食2和6h对棉花体内PAL活性没有产生显著影响,而持续取食12h显著诱导了PAL活性,持续取食24和36h,均极显著诱导了PAL活性;棉铃虫持续取食2、6、12、和18h均显著诱导了棉花体内LOX活性,持续取食24h极显著诱导了LOX活性;棉铃虫持续取食6h极显著诱导了棉花体内PPO活性,持续取食24h显著诱导了PPO活性。棉铃虫取食12h后停止取食,在去除虫害除胁迫后0、6、12、24和36h,棉花体内PAL活性均显著升高;而LOX活性则呈现出先升高后恢复正常的现象;PPO活性开始无变化,但在胁迫去除后12和24h显著增高,到36h恢复正常。可见,棉花体内PAL、LOX和PPO活性对棉铃虫取食产生的防御响应,与其受虫害持续取食胁迫时间的增长呈正相关,随着取食时间和受危害的程度加大而升高。并且,在虫害胁迫去除后的一定时间内,棉花体内PAL、LOX和PPO活性依然会保持较高的活性状态,而同等程度机械损伤后的棉叶内PAL、LOX、PPO活性均没有发生显著性变化。说明棉花对于棉铃虫取食胁迫的防御与棉花生理生化性质的改变有关,且具有持续性。     

Defence‐Related Enzymes in Soybean Resistance to Target Spot

Target spot, caused by the fungus Corynespora cassiicola, has become a serious foliar disease in soybean production in the Brazilian Cerrado. Information in the literature regarding the biochemical defence responses of soybean to C. cassiicola infection is rare. Therefore, the objective of this study was to determine the biochemical features associated with soybean resistance to target spot. The activities of chitinases (CHI), β‐1‐3‐glucanases (GLU), phenylalanine ammonia‐lyases (PAL), peroxidases (POX), polyphenol oxidases (PPO) and lipoxygenases (LOX), as well as the concentrations of total soluble phenolics (TSP) and lignin‐thioglycolic acid (LTGA) derivatives, were determined in soybean leaves from both a resistant (FUNDACEP 59) and a susceptible (TMG 132) cultivar. The target spot severity, number of lesions per cm² of leaflet and area under the disease progress curve were significantly lower for plants from cv. FUNDACEP 59 compared to plants from cv. TMG 132. The GLU, CHI, PAL, POX and PPO activities and the concentration of LTGA derivatives increased significantly, whereas LOX activity decreased significantly on the leaves infected by C. cassiicola. Inoculated plants from cv. FUNDACEP 59 showed a higher PPO activity and concentrations of TSP and LTGA derivatives at 4 and 6 days after inoculation compared to plants from cv. TMG 132. In conclusion, the results of this study demonstrated that the defence‐related enzyme activities increased upon C. cassiicola infection, regardless of the basal level of resistance of the cultivar studied. The increases in PPO activity and concentrations of TSP and LTGA derivatives, but lower LOX activity, at early stages of C. cassiicola infection were highly associated with soybean resistance to target spot.     

Active Oxygen-producing and -scavenging Enzymes in Pepper Leaves Infected with Xanthomonas campestris pv. vesicatoria

Lipoxygenase (LOX), peroxidase (POX), superoxide dismutase (SOD) and catalase (CAT) activities were determined in pepper (Capsicum annuum) leaves infected with Xanthomonas campestris pv. uesicatoria, from 3 to 15 days after inoculation, before symptom appearance and during the development of the disease. Strong Stimulation of LOX and POX activities was observed in infected leaves at an advanced stage of the disease (12–15 days after inoculation), when bacterial water-soaked spots and a slight chlorosis of the inoculated leaf areas were evident and a decrease in chlorophyll content of infected tissue was detected. The infection also induced a significant increase in CAT activity at the day 9 and a decrease in SOD and CAT activities at the day 12. On the basis of the changes observed, uncontrolled production of active oxygen species at advanced stages of infection is hypothesized.     

6种防御酶调控植物体应答虫害胁迫机制的研究进展

植物防御体系应对虫害胁迫产生一系列防御性生理生化反应,其中防御酶活性呈现显著变化.本文综述了超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)、多酚氧化酶(PPO)、脂氧合酶(LOX)和苯丙氨酸解氨酶(PAL) 6种常见防御酶应对虫害胁迫的机制,解析了6种防御酶的作用机理及其异同.梳理了6种防御酶应对虫害胁迫而相互协调的程序,总结了植物体遭虫害胁迫之后防御酶活性的变化及其与防御酶基因的关联,提出了植物体防御酶机制研究中的重要问题并展望前景.     

Induced defense-related proteins in soybean (Glycine max L. Merrill) plants by Carnobacterium sp. SJ-5 upon challenge inoculation of Fusarium oxysporum

The aim of the present study was to analyze induced expression of defense-related proteins in the soybean plants by rhizobacterial stain Carnobacterium sp. SJ-5 upon challenge inoculation with Fusarium oxysporum. Determination of the enzymatic activity of the different defense-related enzymes, phenylalanine ammonia lyase (PAL), lipoxygenase (LOX), peroxidase (POD) and polyphenol oxidase (PPO) was performed in the major parts of Glycine max L. Merrill using spectrophotometric method. Native-polyacrylamide gel electrophoresis analysis of the POD and PPO was employed followed by activity staining to find out the isoforms of respective enzymes. Activities of the PAL, LOX, POD and PPO were found to be highest in the bacterized root tissue of the soybean plants challenged with F. oxysporum. Isoform analysis revealed that PPO1, PPO4 and POD2 isoforms were expressed at higher levels in bacterized soybean root tissues challenge inoculated with the pathogen. Conclusively it was found that bacterial strain Carnobacterium sp. SJ-5 protect soybean plants from wilt disease caused by F. oxysporum by elicitation of the defense-related enzymes.     

Biochemical alterations in leaves of resistant and susceptible cotton genotypes infected systemically by cotton leaf curl Burewala virus

Leaf curl disease caused by Cotton Leaf Curl Burewala virus (CLCuBuV) has been recognized as serious threat to cotton in Indian subcontinent. However, information about cotton–CLCuBuV interaction is still limited. In this study, the level of phenolic compounds, total soluble proteins, and malondialdehyde (MDA) and the activities of phenylalanine ammonia-lyase (PAL), peroxidase (POX), catalase (CAT), proteases, superoxide dismutase (SOD), and polyphenol oxidase (PPO) were studied in leaves of two susceptible (CIM-496 & NIAB-111) and two resistant (Ravi and Co Tiep Khac) cotton genotypes. Disease symptoms were mild in the resistant genotypes but were severe in highly susceptible genotypes. The results showed that phenolic compounds, proteins, PAL, POX, CAT, proteases, SOD, PPO, and MDA play an active role in disease resistance against CLCuBuV. The amount of total phenols, proteases, MDA, and PPO was significantly higher in leaves of CLCuBuV-inoculated plants of both resistant genotypes as in non-inoculated plants, and decreased in CLCuBuV-inoculated plants of both susceptible genotypes over their healthy plants. POX, protein content, SOD, and PAL activities showed lower values in resistant genotypes, while they decreased significantly in susceptible genotypes as compared to the noninoculated plants except PAL, which showed non-significant decrease. CAT was found to be increased in both susceptible and resistant genotypes with maximum percent increase in resistant genotype Ravi, as compared to non-inoculated plants. The results showed significantly higher concentrations of total phenols and higher activity of protease, MDA, SOD, and PPO in resistant genotype Ravi after infection with CLCuBuV, suggesting that there is a correlation between constitutive induced levels of these enzymes and plant resistance that could be considered as biochemical markers for studying plant-virus compatible and incompatible interactions.     

Effects of silicon on defense of wheat against oxidative stress under drought at different developmental stages

The effects of silicon application before sowing on the drought-induced oxidative stress and antioxidant defense in wheat (Triticum aestivum L.) were investigated. Drought stress was applied by withholding watering till sampling at booting or filling stage. Application of Si increased the water potential of drought-stressed plants at filling stage, whereas it did not at booting stage. The superoxide dismutase (SOD) activity was inhibited and peroxidase (POD) activity was enhanced by drought at booting stage, and no differences were observed due to the Si treatment. At filling stage, however, application of Si increased the SOD activity and decreased the POD activity of drought-stressed plants. The catalase (CAT) activity was slightly increased by drought only in the absence of Si and at booting stage. The activity of glutathione reductase (GR) was not greatly influenced. Application of Si did not change the contents of H₂O₂, total soluble protein and protein carbonyl of drought-stressed plants at booting stage, whereas at filling stage, it decreased the content of H₂O₂ and protein carbonyl and increased the content of total soluble protein. The content of thiobarbituric acid reactive substances (TBARS) and the activities of acid phospholipase (AP) and lipoxygenase (LOX) in drought-stressed plants were also decreased by application of Si at both stages.     

南美斑潜蝇为害对黄瓜体内4种防御酶活性的影响

植物对昆虫取食产生的防御反应,在昆虫与植物相互作用关系中起着重要的作用。为明确南美斑潜蝇Liriomyza huidobrensis(Blanchard)取食与植物防御之间的作用关系,本文分别测定了南美斑潜蝇幼虫持续为害1、3、5、7及9d后黄瓜叶片中苯丙氨酸解氨酶(PAL)、多酚氧化酶(PPO)、过氧化物酶(POD)及超氧化物歧化酶(SOD)活性的变化。结果表明：在南美斑潜蝇幼虫持续取食下,PAL、PPO、POD和SOD等4种酶活性显著上升;PAL、PPO和POD等3种酶活性随受害程度的加重而上升幅度加大,即重度受害>轻度受害>系统对照>健康对照,而SOD活性变化没有规律。在系统对照黄瓜叶片中,PAL、PPO、POD和SOD等4种酶活性最大值分别在第5、1、5和5d;在轻度受害黄瓜叶片中,PAL、PPO、POD和SOD等4种酶活性最大值分别在第5、9、5和9d;在重度受害黄瓜叶片中,PAL、PPO、POD和SOD等4种酶活性最大值分别在第7、7、5和5d。黄瓜叶片受害后,POD和PPO活性上升幅度较大,PAL和SOD活性上升幅度较小,说明POD和PPO对取食胁迫响应比PAL和SOD更灵敏。     

Induction of total phenolics and defence-related enzymes during β-aminobutyric acid-induced resistance in Brassica carinata against Alternaria blight

Exogenous foliar application of β-aminobutyric acid (BABA) led to a significant reduction in disease severity in Brassica carinata caused by Alternaria brassicae. To get a better insight about changes in defence-related enzymes like phenylalanine ammonia lyase (PAL) and polyphenol oxidase (PPO), isoform analysis of superoxide dismutase (SOD) and peroxidase (POX) were studied. BABA-treated plants showed a significant increase in PAL, PPO enzyme activities and total phenolic content in response to pathogen inoculation. However, isoform analysis of SOD and POX revealed no change in isoform number but a quantitative change in activity was observed in response to pathogen.     

Induction of defence-related enzymes in tomato (Solanum lycopersicum) plants treated with Bacillus subtilis CBR05 against Xanthomonas campestris pv. vesicatoria

Bacterial spot disease caused by Xanthomonas campestris pv. vesicatoria is one of the most important destructive diseases of tomato in many parts of the agricultural world. Therefore, the present study aims to determine the effects of Bacillus subtilis CBR05 inoculation on bacterial spot disease severity and the induction of defence-related enzymes response in tomato. Tomato leaves were evaluated to determine the activities of antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), and polyphenol oxidase (PPO)) and the content of malondialdehyde (MDA). A reduction in bacterial spot severity was observed in plants inoculated with B. subtilis, compared with those of uninoculated controls. A significant increase in SOD, CAT, POD, and PPO activities was observed in plants treated with B. subtilis after 24?h inoculation compared with non-inoculated pathogen control and mock-inoculated controls. Moreover, the MDA content was induced by pathogen infection, and its amount in B. subtilis inoculated plants was significantly lower than that in pathogen control. Our results suggest that early increases in antioxidant enzymes and the reduction in MDA content with B. subtilis inoculation may play a pivotal role in mitigating oxidative stress, thereby induced systemic resistance against bacterial spot disease in tomato.     

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.     

二斑叶螨危害对棉花应激防御酶活性的影响

探明二斑叶螨取食对棉花叶片内应激防御性酶活性的影响。在棉苗5-6叶期接种不同数量(10-40头)的二斑叶螨雌成虫,取食1-4 d后采摘不同处理的叶片,利用分光光度计测定过氧化氢酶(CAT)、过氧化物酶(POD)、多酚氧化酶(PPO)、脂氧合酶(LOX)和苯丙氨酸解氨酶(PAL)活性。结果表明二斑叶螨取食后,棉花叶片CAT活性显著升高,且随诱导时间的延长其活性变化呈先上升后下降的趋势,为害3 d时,10、20、H(每株接棉蚜跟叶螨各40头)头/叶的CAT活性达到最高峰,显著高于对照(P0.05);POD、PAL活性也随诱导时间的延长和螨口数量的增加呈上升趋势,为害4 d时达到最高峰;而PPO、LOX活性随着时间的增加呈现上升后下降趋势,且为害2 d时达到最高峰,仍显著高于对照(P0.05)。棉叶应激防御酶活性随着二斑叶螨为害时间与螨口密度的增加而增加,表明二斑叶螨的危害胁迫能诱导寄主植物产生一系列应激生化反应,不同的酶活性表现有所不同。     

Defence-related enzymatic response in cabbage to Xanthomonas campestris pv. campestris

Black rot of cabbage caused by Xanthomonas campestris pv. campestris is one of the most important diseases of crucifers worldwide. Expression of defence-related enzymes in cabbage in response to X. campestris pv. campestris was investigated in the current experiment. Among the defence-related enzymes (phynylalanine ammonia lyase, peroxidase, polyphenol oxidase, superoxide dismutase [SOD] and chitinase) and quantity of phenolic compounds studied in the present investigation, phenylalanine ammonia lyase (PAL), the key enzyme in the phenylpropanoid pathway was the first enzyme suppressed at three days after inoculation in X. campestris pv. campestris-cabbage system. Correlation analysis indicated that PAL and phenolic compounds are the two most important compounds determining the susceptibility of cabbage to X. campestris pv. campestris. Induction of peroxidase isoform-1 (Rf value: 0.059) and SOD isoform-1 (Rf value: 0.179) three days after pathogen inoculation implicated the role of these isozymes in susceptible cabbage – X. campestris pv. campestris interaction. This study demonstrates the susceptibility of cabbage to X. campestris pv. campestris is a result of declination of PAL and phenolic contents at biochemical level as a manifestation of increase in bacterial population at the cellular level within the host tissues.     

短枝木麻黄家系对青枯病的抗性评价与选择

为探讨短枝木麻黄(Casuarina equisetifolia)对青枯病(Ralstonia solanacearum)侵染后的生理生化响应,利用其生理生化响应指标与抗病性特征的相关性开展短枝木麻黄家系的抗病评价和选择。结果表明,26个短枝木麻黄家系褐梗小枝接种青枯病后,病情指数(DI)和相对病害强度(RDI)均存在极显著差异(P<0.01)。在接种青枯病后,家系间的超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、苯丙氨酸解氨酶(PAL)、多酚氧化酶(PPO)、总酚(TP)和类黄酮(Flav)差异均达显著或极显著水平,说明参试家系对青枯病抗性的变异丰富,具有较大的选择潜力。短枝木麻黄家系的病情症状与6个生理生化指标间均呈显著或极显著的负相关关系,说明青枯病侵染后防御酶活性或次生代谢物含量越高,木麻黄的病害程度越低。利用2个病情特征性状和6个生理生化指标对短枝木麻黄不同家系进行综合评价和选择,按照30%的入选率初步选出201、206、198、207、202、213、208和195号等8个抗病家系。这些抗病家系的RDI和DI比总体均值分别低25.94%和28.93%,SOD、CAT...     

Olive (Olea europaea L.) freezing tolerance related to antioxidant enzymes activity during cold acclimation and non acclimation

The changes in the antioxidant enzymes activity, total protein and proline content and their correlations with freezing tolerance (FT) (expressed as LT₅₀) were investigated at 11 different olive cultivars at cold-acclimation (CA, in February) and non-acclimation (NA, in August) stages. Leaf samples were collected from each cultivar and were divided into two groups. The first group was immediately frozen in liquid nitrogen for further biochemical analysis. The second ones was subjected to different freezing temperatures (?5, ?10, ?15 and ?20 °C) for 10 h, in order to determine their FT. The unfrozen control samples were kept at 4 °C. The results showed that Fishomi, Mission and Shengeh were the most freezing tolerant among other cultivars. In contrast, Zard, Manzanilla and Amigdalolia were the most sensitive ones. The cold acclimation enhanced the activities of superoxide dismutase (SOD), peroxidase (POD), ascorbate peroxidase (APX), catalase (CAT), polyphenol oxidase (PPO) and total protein content. However, proline content and phenylalanine ammonia-lyase (PAL) activity did not change or even decreased slightly at CA stage, compare to those samples at NA stage. It was found that LT₅₀ to be closely correlated to POD, CAT, and PPO activity at CA and NA stages. Overall, higher leaf POD, CAT, and PPO activity could be used as important selection criteria in screening tolerant olive cultivars for cold zone climatic.     

Fe2O3 nanoparticles induced biochemical responses and expression of genes involved in rosmarinic acid biosynthesis pathway in Moldavian balm under salinity stress

The effect of iron oxide nanoparticle (NP) at four concentrations (0, 30, 60 and 90 ppm) and salinity at three levels (0, 50 and 100 mM) were investigated on rosmarinic acid (RA) production in 5-week-old Moldavian balm (Dracocephalum moldavica L.) plants. Salinity and spraying iron oxide NPs significantly affected tyrosine (Tyr), phenylalanine (Phe) and proline (Pro) amino acids content, Phenylalanine Ammonia-Lyase (PAL), Tyrosine Aminotransferase (TAT) and Rosmarinic Acid Synthase (RAS) genes expression levels, RA content, Polyphenol Oxidase (PPO), PAL and Superoxide Dismutase (SOD) activities, malondialdehyde (MDA) content and DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity. PAL, TAT and RAS genes expression rate and content of RA were enhanced in Moldavian balm plants exposed by NaCl + NPs. The results of high performance liquid chromatography (HPLC) revealed that simultaneous application of 50 mM NaCl and 90 ppm NPs increases the RA content in leaf by 81.15% as compared to control plants. The Tyr and Phe contents decreased in Moldavian balm plants exposed to salt stress. Application of NPs had a positive effect on the content of these amino acids. Proline content increased under salinity stress and application of iron NPs induced a significant increase in the Pro content of leaf. The results revealed that PAL, PPO and SOD enzymes activities increased under salinity conditions. The highest activity of PPO and SOD was observed in 100 mM NaCl + 60 ppm NPs treatment. Simultaneous application of 100 mM NaCl + 90 ppm NPs increased the MDA content and DPPH radical scavenging activity compared to control plants. It can be concluded that the application of appropriate levels of NPs moderates the effect of salinity stress in D. moldavica L. and results in an increased amount of RA compared to control plants.     

Cerebroside mediated elicitation of defense response in chilli (Capsicum annuum L.) against Colletotrichum capsici infection

Abstract

Enhancing the host resistance by using naturally occurring elicitors derived from pathogenic organisms is emerging as an ecofriendly approach in plant disease management. Cerebrosides, categorized as glycosphingolipids, were isolated and partially purified from the wilt causing fungus (Fusarium oxysporum f. sp. lycopersici). Cerebroside treatment significantly reduced the anthracnose disease incidence under greenhouse conditions. Cerebroside elicitors were found to stimulate the early H₂O₂ accumulation followed by the production of plant defense-related enzymes such as Phenylalanine ammonia lyase (PAL), Peroxidase (POX), Polyphenol oxidase (PPO), and Lipoxygenase (LOX) when applied to chilli (Capsicum annuum L.) plants by spray treatment and also induced the accumulation of capsidiol. Defense-related enzyme activities were increased by the elicitor treatment and an high level in activity was maintained during the experimental period. Under greenhouse conditions, the cerebroside elicitors effectively protected chilli plants against infection by anthracnose causing organism, Colletotrichum capsici.     

Biological control of southern corn leaf blight by Trichoderma atroviride SG3403

Trichoderma atroviride SG3403 showed high biocontrol activity against southern corn leaf blight (SCLB; pathogen: Cochliobolus heterostrophus). T. atroviride SG3403 could cause death of C. heterostrophus race O hypha on plates. Spraying T. atroviride SG3403 conidia suspension over maize seedling leaves protected the corn from SCLB infection. Biocontrol effect lasted for 30 days in the field. Trichoderma strain was able to induce resistance response in corn leaves against pathogen infection. In corn leaves treated with T. atroviride SG3403, the enzyme activities of phenylalanine ammonia lyase (PAL) and superoxide dismutase (SOD) reached the highest at 24 h, enzyme activity of catalase (CAT) reached the highest at 36 h after inoculation of pathogen C. heterostrophus race O. RNA expression levels of Pal, Sod and Cat (which synthesis enzyme PAL, SOD and CAT) were also upregulated and corresponded to the enzyme activity at the same time point. Enzyme activities and corresponding genes expression induced by Trichoderma SG3403 was more obvious than that induced by pathogen only, which implies that T. atroviride SG3403 induced corn defense gene expression against pathogen infection. Thus, induced resistance mechanism was possibly involved in the biocontrol of SCLB by T. atroviride SG3403.