Plant-mediated interactions between insects and a fungal plant pathogen and the role of plant chemical responses to infection

Diverse organisms simultaneously exploit plants in nature, but most studies do not examine multiple types of exploiters like phytophagous insects and fungal, bacterial, and viral plant pathogens. This study examined patterns of induction of antipathogenic peroxidase enzymes and phenolics after infection by the cucurbit scab fungus, Cladosporium cucumerinum, and then determined if induction mediated ecological effects on Colletotrichum orbiculare, another fungal pathogen, and two insect herbivores, spotted cucumber beetles, and melon aphids. Peroxidase induction occurred in inoculated, `local,' symptom-bearing leaves 3 days after inoculation, and in `systemic,' symptom-free leaves on the same plants 1 day later. Phenolics were elevated in systemic but not in local leaves 3 days after inoculation. Detached systemic leaves from plants inoculated with C. cucumerinum developed significantly fewer and smaller lesions after challenge with C. orbiculare. Spotted cucumber beetles did not show consistently significant preferences for infected versus control leaf disks in comparisons using local or systemic leaves, but trends differed significantly between leaf positions. In no-choice tests, beetles removed more leaf area from local but not from systemic infected leaves compared to control leaves, and melon aphid reproduction was enhanced on local infected leaves. In the field, cucumber beetle and melon aphid densities did not differ between infected and control plants. Antipathogenic plant chemical responses did not predict reduced herbivory by insects. Other changes in metabolism may explain the positive direction and spatially dependent nature of plant-mediated interactions between pathogens and insects in this system. Received: 28 September 1997 / Accepted: 9 February 1998     

The induction of soluble peroxidase activity in bean leaves by wind-induced mechanical perturbation

The induction of defense-related peroxidase (POD) activity in plants occurs in response to many biotic and abiotic stimuli. This controlled greenhouse study was an attempt to provide insight into the nature of the induction of soluble POD activity by noninjurious wind-induced mechanical perturbation (MP). In a time course study, exposure of common bean (Phaseolus vulgaris) seedlings to daily periods of fan-produced wind induced a significant and sustained increase in soluble POD activity in primary leaves of 7-9-d-old seedlings. In a wind-gradient study, wind-induced MP led to increases in soluble POD activity in leaves that were proportionally related to the wind speed experienced by individual seedlings. Wind-induced MP enhanced soluble POD activity to a degree similar to treatment with 5 mmol/L HgCl(2), a potent oxidizing elicitor of POD activity in plants. However, no further increases in POD activity were induced by HgCl(2) on plants that were preconditioned with wind-induced MP. Finally, short periods of brushing-induced MP enhanced soluble POD activity to the same degree as longer periods of wind-induced MS, suggesting a greater sensitivity to thigmic stimuli than to seismic stimuli in leaves of bean seedlings. This study illustrates the potential importance of wind and other mechanical stimuli as inducers of POD activity and interacting factors in the elicitation of POD activity by other environmental stimuli.     

The effects of <Emphasis Type="Italic">β</Emphasis>-amino-butyric acid on resistance of cucumber against root-knot nematode, <Emphasis Type="Italic">Meloidogyne javanica</Emphasis>

In this study, the effects of β-amino-butyric acid (BABA) on root-knot nematode(Meloidogyne javanica) infection of cucumber and accumulation of total phenolic compounds, hydrogen peroxide and activity of some enzymes related to plant defense mechanisms, i.e., guaiacol peroxidase (GPOX), polyphenol oxidase (PPO), catalase (CAT) in cucumber roots infected with nematode were investigated. Results of this study show that treating the cucumber seedlings with the above elicitor significantly reduces the nematode infection level (the nematode galls, number of egg masses per plant and number of eggs per individual egg mass) compared to control. Additionally, treatment of cucumber roots by BABA and BABA + nematode, significantly increased peroxidase, polyphenol oxidase and catalase activities in root tissues, 1 day after nematode inoculation in comparison to nematode inoculated plants as control and sterile water-treated plants. Enzyme activities reached to a maximum level at 4, 4 and 3 days after nematode inoculation, respectively. Additionally, the amount of H₂O₂, a product of oxidative stress, was significantly increased in the BABA and BABA + nematode treatments in comparison to control. Such increases have occurred in two phases and maximum levels of it were observed at 5 days after inoculation. Inoculation of cucumber plants by BABA also significantly increased accumulation of total phenol in comparison to control and maximum level of it was observed at 7 days after nematode inoculation. The results suggest that the inhibitory effect of BABA on the root-knot nematode (M. javanica) may be related to its ability to enhance defense responses in the cucumber roots.     

Peroxidase Activity in Leaves of Wheat Cultivars Differing in Resistance to Erysiphe graminis DC.

The peroxidase activities in leaves from resistant and susceptible cultivars of wheat infected and non-infected by Erysiphe graminis DC were studied. In non-infected wheat, soluble and ionic bound peroxidase activity level was found to be higher in the resistant cultivar than that in the one susceptible to Erysiphe graminis DC. After infecting wheat leaves with Erysiphe graminis DC a remarkable increase in the activity of soluble and ionic bound peroxidases was detected 5 days after inoculation only in the resistant cultivar. In the susceptible cultivar a high increase in the activity of the soluble and ionic bound peroxidases occurred only 15 days after inoculation. Using ion exchange chromatography four peroxidase fractions were obtained from infected susceptible and resistant cultivars as from non-infected ones. The fraction II in non-inoculated resistant cultivars was much higher than that in the susceptible one. This fraction increased after inoculation in both cases reaching a higher level in resistant cultivars. Fraction I was higher in the susceptible cultivar. Electrofocusing profiles of peroxidase from the susceptible and resistant cultivar differed from one another. New peroxidase bands after inoculation appeared only in the resistant cultivar.     

颈盲蝽取食对薇甘菊叶片营养物质和防御酶的影响

【目的】薇甘菊是世界最具危害性的入侵杂草之一,对我国生态环境和农业、林业生产造成了严重的危害。颈盲蝽是控制薇甘菊的一种潜在的重要天敌昆虫。本研究旨在探讨薇甘菊被颈盲蝽取食后,叶片防御相关酶系活性、营养物质和叶绿素含量的变化,阐明颈盲蝽取食对薇甘菊生理功能的影响,为利用颈盲蝽防控薇甘菊提供依据。【方法】从云南瑞丽野外采集薇甘菊的本地天敌昆虫颈盲蝽,测定了颈盲蝽取食前及取食12、24、48、96 h后,薇甘菊叶片中过氧化物酶(POD)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)活性,以及可溶性糖、可溶性蛋白及叶绿素含量,以取食前的植株作为对照。【结果】与对照相比,颈盲蝽取食12 h时,薇甘菊叶片中的POD、CAT活性升高,SOD活性降低;之后POD、SOD活性上升,CAT活性降低,取食48 h时,POD和SOD活性达到最高值,CAT活性达到最低值;取食96 h时,POD与SOD活性降低,但仍高于对照,CAT活性与取食48 h时相近。颈盲蝽取食后,薇甘菊叶片中的可溶性糖含量明显上升,取食96 h达到最高值;可溶性蛋白和叶绿素含量显著降低,在96 h达到最低值,分别低于对照39.30%和69.94%。【结论】颈盲蝽取食严重破坏了薇甘菊叶片的正常生理功能,并最终导致其叶片萎蔫和坏疽。     

球孢白僵菌对黄瓜代谢产物及西花蓟马取食行为的影响

球孢白僵菌Beauveria bassiana灌根处理黄瓜后,发现西花蓟马Frankliniella occidentalis对叶片危害面积显著低于对照组;为探究产生这一现象的原因,测定黄瓜叶片中防御酶系和次生代谢产物等含量变化,采用直流刺吸电位仪(direct current electrical penetration graph,DC-EPG)分析西花蓟马成虫取食黄瓜叶片的行为.球孢白僵菌对黄瓜灌根后叶片中过氧化氢酶(CAT)、过氧化物酶(POD)和超氧化物歧化酶(SOD)含量均呈上升趋势,其中CAT活性增加最高,增加率为37.26％;丙二醛MDA的含量呈下降趋势;可溶性蛋白、类黄酮的含量增加.DC-EPG显示西花蓟马在经球孢白僵菌处理的黄瓜上,取食时间和次数均低于对照组.结果表明,黄瓜植株经球孢白僵菌处理后防御酶物质增加,进而导致西花蓟马对叶片危害下降,利于对西花蓟马的防治.     

Induction of the activities of antioxidative enzymes and the levels of malondialdehyde in cucumber seedlings as a consequence of Bemisia tabaci (Hemiptera: Aleyrodidae) infestation

The activities of antioxidative enzymes and the concentration of malondialdehyde were assayed in cucumber leaves (Cucumis sativus L.) 0, 6, 12, 24, 48, 72, and 96 h after the cucumber seedlings were infested by Bemisia tabaci (Gennadius). The results indicated that the activities of antioxidative enzymes were increased after herbivore infestation, including superoxide dismutase (SOD), and peroxidase (POD), and the levels of malondialdehyde (MDA), which is a product of membrane lipid peroxidation in the leaves. The enzymes and MDA showed peaks of different activity levels at 24 and 48 h after the infestation. SOD activity reached the highest peak, 10.4% higher than control, at 24 h, POD activity reached the highest peak, 213.2% higher than control, at 6 h, catalase (CAT) activity was not statistically significant compared with the control, and MDA content reached the highest peak, 59.9% higher than control, at 48 h. The results suggested that the enhanced activities of antioxidative enzymes and MDA content may contribute to bioprotection of cucumber plants against B. tabaci infestation. Handling editor: Henryk Czosnek     

Xanthomonas oryzae pv. oryzae infection triggers accumulation of phenolics,defense-related enzymes and thaumatin-like proteins in rice leaves

Abstract

The effect of Xanthomonas oryzae pv. oryzae infection on induction of phenylalanine ammonia-lyase (PAL), peroxidase (PO), phenolics and thaumatin-like proteins (TLPs) in rice was studied. PAL activity increased significantly one day after inoculation with X. o. pv. oryzae and the maximum enzyme activity was observed two days after inoculation. The phenolic content in rice leaves increased significantly one day after inoculation and the maximum accumulation of phenols was observed two days after inoculation. Significant increase in peroxidase activity was observed in rice leaves one day after inoculation with X. o. pv. oryzae. Isozyme analysis indicated that three peroxidase isozymes (PO-1, PO-2 and PO-3) were induced after inoculation with X. o. pv. oryzae. Immunoblot analysis of protein extracts from control and pathogen inoculated rice plants revealed the induced accumulation of 16 and 24 kDa TLPs in rice leaves in response to X. o. pv. oryzae infection. TLP mRNA accumulation was induced strongly in rice leaves in response to infection by X. o. pv. oryzae.     

Changes in antioxidant enzyme activities in detached leaves of cucumber exposed to chilling

We studied changes in biochemical and physiological status, level of oxidative damage, and antioxidant enzyme activities in detached leaves of cucumber plants (Cucumis sativus L. cv. Pyunggangnaebyungsamchuk) that were exposed to a low temperature (4°C). Chlorophyll fluorescence (F_v/F_m) declined during the chilling treatment, but was slowly restored after the tissues were returned to 25°C. Likewise, the fluorescence quenching coefficient and relative water content decreased during the stress period, but then increased during recovery. In contrast, we detected a significant rise in protein and hydrogen peroxide contents in the chilled leaves, as well as higher activities for superoxide dismutase, ascorbate peroxidase, peroxidase, and glutathione reductase. However, the level of catalase decreased not only during chilling but also after 24 h of recovery. These results indicate that exposure to low temperatures acts as an oxidative stress. Moreover, we propose that a regulating mechanism exists in the detached cucumber leaves and contains an antioxidant defense system that induces active oxygen species, thereby alleviating the effects of chilling stress within 12 h.     

海滨滨麦叶片和根对不同厚度沙埋的生理响应差异分析

以烟台海岸抗风沙植物滨麦为研究材料,通过对不同厚度沙埋下其叶片和根部抗氧化酶活力(超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT))、丙二醛(MDA)含量和渗透调节物含量变化的分析,探讨了叶片和根部对沙埋生理响应的差异。试验按滨麦成株株高(约40 cm)对其进行了轻度沙埋(在株高1/4处)、中度沙埋(2/4处)和重度沙埋(3/4处)。在沙埋第6天,分别测定了不同厚度沙埋处理下,植株各段叶片和根抗氧化酶活力、MDA和渗透调节物含量。结果表明,轻度和中度沙埋均加速植株生长。与对照相比,经轻度、中度沙埋处理6 d,叶片平均MDA含量增加,在重度沙埋下降低。不同厚度沙埋6 d,叶片平均SOD活力和脯氨酸含量增加,而CAT活力、可溶性糖和可溶性蛋白质含量下降。但不同厚度沙埋均使沙上叶片MDA、脯氨酸、可溶性蛋白质含量和SOD和CAT活力增加,尤其是叶片顶部增加最为明显,使沙下叶片MDA、可溶性糖、可溶性蛋白质含量和CAT活力下降,导致同株沙上和沙下叶片MDA、脯氨酸、可溶性糖、可溶性蛋白质含量和SOD和CAT活力差异显著(P0.05)。与叶片相比,根中MDA、可溶性蛋白质含量和SOD和CAT活力较低,而POD活力和可溶性糖含量较高并与叶片差异显著(P0.05)。不同厚度沙埋6 d,滨麦根中MDA和可溶性蛋白质含量变化较小,可溶性糖含量和CAT、POD、SOD活力略有降低。研究表明,滨麦根和叶片对不同厚度沙埋的生理响应不同。沙埋直接作用于叶片并诱发叶内氧自由基积累,但叶片通过快速激活的抗氧化酶保护系统(CAT、SOD)维持氧自由基代谢平衡,以及渗透调节物(脯氨酸、可溶性糖)的积累维护细胞水分代谢平衡,并满足能量的需求和快速生长。但在不同厚度沙埋下,由于根系不受沙埋直接影响而生理变化较小,并且还维持较低的膜脂过氧化水平,这可能是根能维持正常的吸水输水功能并在沙埋处理过程中和沙埋后地上叶片快速生长摆脱沙埋的重要物质基础。     

Gene expression and activity of antioxidant enzymes in barley (Hordeum vulgare L.) under controlled severe drought

The objective of the present study was to determine the activity of antioxidant enzymes: superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT) and the expression of their genes in two barley genotypes under controlled severe drought. To fulfill this objective, 21-day-old barley plants of two genotypes: Rum and Yarmouk were exposed to controlled severe drought (25% field capacity) for 2, 9, and 16 days. The activity of SOD was significantly high in Rum genotype after 2 days of drought treatment. In Yarmouk genotype, the activity of APX was significantly high after 2 and 9 days of drought treatment. In Rum genotype, CAT2 was upregulated after 9 days of drought treatment and SOD and APX were upregulated after 16 days of drought treatment, whereas CAT2, SOD, and APX were upregulated in Yarmouk genotype after 2 days of drought treatment. The results indicate a unique pattern of activity and gene expression of the antioxidant enzymes in the two barley genotypes under controlled severe drought. Moreover, the data suggest that each genotype utilizes different molecular and biochemical responses under the same drought conditions.     

The Decreased Cold-Resistance of Chilling-Sensitive Plants Is Related to Suppressed CO2 Assimilation in Leaves and Sugar Accumulation in Roots

Tomato (Lycopersicon esculentum L., cv. Sibirskii skorospelyi) and cucumber (Cucumis sativus L., cv. Konkurent) plants were grown in a soil culture in a greenhouse at an average daily temperature of 20°C and ambient illumination until the development of five and eight true leaves, respectively. During the subsequent three days, some plants were kept in a climatic chamber at 6°C in the light, whereas other plants remained in a greenhouse (control). The cold-resistance of cucumber leaves and roots, as assayed from the electrolyte leakage, was reduced after cold exposure stronger than cold-resistance of tomato organs. The ratio photosynthesis/dark respiration was lower in cucumber than in tomato leaves at all measurement temperatures. The concentrations of sugars (sucrose + glucose + fructose) increased in chilled tomato roots but decreased in cucumber roots. Cold exposure changed the activities of various invertase forms (soluble and insoluble acidic and alkaline invertases). The total invertase activity and the ratio of mono- to disaccharides increased. The lower cucumber cold-resistance is related to the higher sensitivity of its photosynthetic apparatus to chilling and, as a consequence, insufficient root supply with sugars.     

Induced resistance against Alternaria brassicae blight of mustard through plant extracts

An experiment was conducted to study the effect of plant extracts on soluble sugar, soluble phenol and defence-related enzymes response against Alternaria blight in mustard crop. The efficacy of six selected plant extracts (5 and 10%) used as foliar sprays at 60 and 70 days after sowing and mustard leaves was used for investigation. The results indicate that soluble phenol and sugar content in mustard leaves significantly increases in response to spraying of Azadirachta indica seed kernel, Calotropis procera and A. indica leaf extracts. The soluble protein, viz. peroxidase, polyphenol oxidase and phenylalanine ammonia lyase content, was higher in mustard leaves sprayed with C. procera leaves extract, A. indica seed kernel and Allium sativum bulb extract.     

Changes in Peroxidases and Catalase Activity During in vitro Rooting

Enzyme changes in non-rooted (treated with Fe-EDTA) and rooted (treated with Fe-EDDHA) stems of rootstock GF-677 (Prunus amygdalus×P. persica) during adventitious root formation in vitro have been recorded. The first roots appeared approximately after 12 d on the rooting medium. By contrast to non-rooted stems, rooted stems showed a maximum of soluble peroxidase activity on the 9^th day, of ionically bound peroxidase to cell wall on the 6^th and 12^th day and of catalase on the 6^th and the 15^th day. A time course study of changes of soluble peroxidases isoenzymes showed that there was a band visible only in the rooted stems and also a new band appeared three days before the emergence of roots.     

Induction of enzymatic scavengers of active oxygen species in rice in response to infection by <Emphasis Type="Italic">Rhizoctonia solani</Emphasis>

Changes in the activities of peroxidase, ascorbate peroxidase, catalase and superoxide dismutase in rice in response to infection by Rhizoctonia solani were studied. A significant increase in peroxidase activity was observed in R. solani-inoculated rice leaf sheaths 1 day after inoculation and the maximum enzyme activity was recorded 3 days after inoculation at which period a 3-fold increase in peroxidase activity was observed compared to the untreated control. Three peroxidase isozymes viz., PO-4, PO-5 and PO-6 were induced in rice upon infection by R. solani. Ascorbate peroxidase and catalase activities significantly increased 1–2 days after inoculation and the maximum enzyme activities were recorded 5 days after inoculation. Superoxide dismutase activity increased significantly 2 days after inoculation and increased progressively, reaching four times the control value at 7 days after inoculation.     

The Activity of the Peroxidase System in the Course of Stress-Induced CAM Development

The activity of the peroxidase system in Mesembryanthemum crystallinum L. plants in relation to the shift from C₃ to CAM photosynthesis was studied. In detached leaves of the fourth and fifth stories treated with NaCl (0.3 M), a rapid (after 30 min) transient induction of the ionically bound peroxidase (the first maximum) was observed followed by a second weak increase in the enzyme activity (90 min after salt treatment). In the leaves of intact plants, which received a longer treatment with NaCl, a two-phase change in the enzyme activity was also observed. It was most pronounced at the early stages of the NaCl-induced plant shift from C₃ to CAM photosynthesis. In this case, in both detached and intact leaves of juvenile plants, the activity of soluble peroxidase was at a low steady-state level. The situation changed dramatically when M. crystallinum plants transitioned to the reproductive developmental phase and photosynthesis switched from C₃ to CAM. The time dependence of the activities of both peroxidase types, the soluble ones in particular, was characterized by marked diurnal oscillations (light–dark), which coincided with the fluctuations of the total titratable acidity. In this case, the activity of the soluble enzyme was several orders of magnitude higher than the activity of the ionically bound peroxidase, even though the optimum pH for both isoforms was similar (pH 5.0). Three acid isoforms of soluble peroxidases, which operated more actively when the cytoplasm had a higher acidity, were distinguished by isoelectrofocusing. Their activity increased under salinity. Alkaline and neutral components were predominant in more than 30 molecular forms of the soluble peroxidase detected. We concluded that the operation of the peroxidase system changed substantially when plants shifted from the juvenile to the reproductive state and switched from C₃ to CAM photosynthesis: the activity of stress-induced ionically bound peroxidase was drastically inhibited with a concurrent increase in the activity of soluble peroxidase and a change in the spectrum of its molecular forms.     

Responses of the sweetpotato whitefly, Bemisia tabaci, to the chinaberry tree (Melia azedarach L.) and its extracts

Host preference bioassays for adults of the sweetpotato whitefly were performed with leaves of the chinaberry tree Melia azedarach L., tomato, cucumber and bean. Fruit and leaf extracts of the chinaberry tree were tested against adults of the sweetpotato whitefly. Fruit extracts were tested against eggs, first and second instar nymphs, and pupae of the insect. Treatments included aqueous, methanol, and acetone fruit extracts of 200 mg ml^?1 and serial dilutions of 20.0 and 2.00 mg ml^?1, ether extract, the botanical insecticides Azatin® and Margosan® ‐O and the control, water or water with Triton®. Mortality data was collected at 6, 7, and 8 days after treatment of the eggs, nymphs and pupae, respectively. Results of the host preference bioassays indicated a significantly lower number of live insects on leaves of the chinaberry tree vs leaves of bean, cucumber, and tomato after 24 h. This indicates that M. azedarach is not a good host for the sweetpotato whitefly. Adults of the insect were significantly more repelled from tomato plants treated with the undiluted extracts when compared to the control after 72 h. There were significant differences in percent mortality of nymphal instars when exposed to the undiluted extracts compared to other extracts and the control. However, there was no significant effect of the fruit extracts on the egg and pupa instars. Thus M. azedarach extracts were found to be repellent to the whitefly adults, while the fruit extracts have shown a significant detrimental effect against early nymphal instars. In general, the methanol extracts were more active against B. tabaci than extracts with other solvents.     

Adsorption of peroxidase on Celite 545 directly from ammonium sulfate fractionated white radish (Raphanus sativus) proteins

This paper demonstrates the direct immobilization of peroxidase from ammonium sulfate fractionated white radish proteins on an inorganic support, Celite 545. The adsorbed peroxidase was crosslinked by using glutaraldehyde. The activity yield for white radish peroxidase was adsorbed on Celite 545 was 70% and this activity was decreased and remained 60% of the initial activity after crosslinking by glutaraldehyde. The pH and temperature-optima for both soluble and immobilized peroxidase was at pH 5.5 and 40°C. Immobilized peroxidase retained higher stability against heat and water-miscible organic solvents. In the presence of 5.0 mM mercuric chloride, immobilized white radish peroxidase retained 41% of its initial activity while the free enzyme lost 93% activity. Soluble enzyme lost 61% of its initial activity while immobilized peroxidase retained 86% of the original activity when exposed to 0.02 mM sodium azide for 1 h. The K_m values were 0.056 and 0.07 mM for free and immobilized enzyme, respectively. Immobilized white radish peroxidase exhibited lower V_max as compared to the soluble enzyme. Immobilized peroxidase preparation showed better storage stability as compared to its soluble counterpart.     

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.