Basal expression of abscisic acid inducing immunity against Karnal bunt of wheat

In order to understand the intricate mechanism of differential immunity against Karnal bunt (KB), basal levels of carotenoids, abscisic acid (ABA), total protease and protease inhibitor were determined in resistant and susceptible genotypes during transition from vegetative to developing stages of wheat spikes. The lower levels of carotenoid precursor of ABA in resistant genotype than in susceptible genotype could be explained by more inter-conversion of carotenoids into ABA pool which was evident from the results of determining ABA by enzyme-linked sorbent assay. The ABA was significantly higher in resistant genotype at all stages than in susceptible genotype, while a sharp increase was observed at S2 stage. The activity of total protease was higher at initial stages of resistant genotype and gradually declined in later stages after anthesis. In contrast to the protease activity, a reverse trend was observed for the levels of cysteine protease inhibitor, suggesting a negative correlation with each other. Level of cysteine protease inhibitor was observed to be three-folds higher at S2 stage in the resistant genotype than in the susceptible genotype. The results provided the clue for the involvement of ABA-dependent pathways in upregulation of cystatin that leads to induction of differential immunity against the KB pathogen.     

Expression and in silico characterization of Phenylalanine ammonium lyase against karnal bunt (Tilletia indica) in wheat (Triticum aestivum)

To investigate the lignifications process and its physiological significance under Karnal Bunt (KB), the changes in enzymes responsible for lignifications likes, phenylalanine ammonia lyase (PAL), were determined in resistant (HD-29) and susceptible genotype (WH-542) of wheat during different developmental stages. The PAL gene was cloned and sequenced. The expression of PAL gene was measured by means of semi-quantitative RT-PCR. The enzyme was expressed constitutively in both the susceptible and resistant genotype. However, the activity was higher in all the developmental stages of resistant genotype, indicating that this genotype has a significant higher basal level of these enzymes as compared to the susceptible line and could be used as marker(s) to define KB resistance. The activity of PAL was significantly higher in WSv stage (Z=16). Structural comparisons based on alignments of all the protein sequences using the clustal W program and searches for conserved motifs using the MEME program have revealed broad conservation of main motifs characteristic of the plant PAL. MSA and phylogenetic analyses of different plants PAL demonstrate that all PAL cluster divided in to two main cluster. The PAL also possesses a specific consensus sequences [GS]- [STG]-[LIVM]-[STG]-[SAC]-S-G-[DH]-L-x-[PN]-L-[SA]-x(2,3)-[SAGVTL]. The pathway might be associated with the enhancement of structural defense barrier due to lignifications of cell wall as evident from the enhanced synthesis of lignin in all the stages of resistant genotype. Our results clearly indicate the possible role of enzymes of Phenyl propanoid pathway metabolism provides genotype and stage dependant structural barrier resistance in wheat against KB.     

Hormonal Balance in Wheat Infected with Helminthosporium sativum

Using immunoenzyme assay, we studied the levels of phytohormones in resistant and susceptible varieties of wheat infected with Helminthosporium sativum. The infection of wheat with this pathogen leads to an increase in cytokinins in plants of the resistant variety in the presence of stable IAA/ABA ratio. The level of ABA increases in the infected plants of the susceptible variety. The role of hormones in the mechanisms of plant resistance to fungal infections is discussed.     

Induction of resistance in host against the infection of leaf blight pathogen (Alternaria palandui) in onion (Allium cepa var aggregatum)

The Pseudomonas fluorescens isolate Pfl was found to inhibit the growth of pathogen Alternaria palandui, in vitro. In the present study, foliar application of a talc-based formulation of Pfl significantly reduced the incidence of leaf blight of onion, caused by A. palandui. Induction of defense-related proteins viz., chitinase, beta-1,3 glucanase, peroxidase (PO) and polyphenol oxidase (PPO) by application of Pfl, was studied against A. palandui infection in resistant (IHR 56) and susceptible (MDUI) onion cultivars. Chitinase in both cultivars, with or without challenge-inoculation of A. palandui revealed changes in the isoform pattern. The Native-PAGE of PO showed induction of PO2 isoform in both the cultivars, in response to inoculation of pathogen. Isoform analysis of PPO also exhibited induction in the Pfl-treated plants challenged with pathogen. Similarly, the activity of beta-1,3-glucanase was greatly induced in Pfl-treated plants, challenged with pathogen as compared to controls. Thus, the P. fluorescens-treated plants showed significant increase in the levels of the defense enzymes, in comparison to the plants challenged with the pathogen.     

Central Role of Salicylic Acid in Resistance of Wheat Against <Emphasis Type="Italic">Fusarium graminearum</Emphasis>

Head blight caused by Fusarium graminearum (F. graminearum) is one of the major threats to wheat and barley around the world. The importance of this disease is due to a reduction in both grain yield and quality in infected plants. Currently, there is limited knowledge about the physiological mechanisms involved in plant resistance against this pathogen. To reveal the physiological mechanisms underlying the resistance to F. graminearum, spikes of resistant (Sumai3) and susceptible (Falat) wheat cultivars were analyzed 4 days after inoculation, as the first symptoms of pathogen infection appeared. F. graminearum inoculation resulted in a greater induction level and activity of salicylic acid (SA), callose, phenolic compounds, peroxidase, phenylalanine ammonia lyase (PAL), and polyphenol oxidase in resistant versus susceptible cultivars. Soil drench application to spikes of SA, 24 h before inoculation with F. graminearum alleviated Fusarium head blight symptoms in both resistant and susceptible cultivars. SA treated plants showed a significant increment in hydrogen peroxide (H₂O₂) production, lipid peroxidation, SA, and callose content. SA-induced H₂O₂ level seems to be related to increased superoxide dismutase and decreased catalase activities. In addition, real-time quantitative PCR analysis showed that SA pretreatment induced expression of PAL genes in both infected and non-infected head tissues of the susceptible and resistant cultivars. Our data showed that soil drench application of SA activates antioxidant defense responses and may subsequently induce systemic acquired resistance, which may contribute to the resistance against F. graminearum. These results provide novel insights about the physiological and molecular role of SA in plant resistance against hemi-biotrophic pathogen infection.     

小麦对赤霉病抗性不同品种的SOD活性

本研究对9个赤霉病抗性不同小麦品种采用赤霉病菌分生孢子悬浮液以单花针注法进行了田间和温室抗病性鉴定;测定了各品种的胚性愈伤组织和盛花期麦穗分别经赤霉病菌毒素和分生孢子接种前后SOD活性的变化。结果表明,各品种SOD活性与其对赤霉病抗性呈极显著的正相关。接种后寄主的SOD活性均有提高,抗病品种比感病品种提高幅度大,且有新的同工酶带出现。抗病品种望水白比感病品种Alondra“S”多出两条SOD同工酶谱带。SOD在小麦抗赤霉病上可能起积极作用,其活性有可能作为鉴定小麦抗赤霉病的一种生理生化指标。     

Role of salicylic acid and oxidative burst in expression of resistance to Alternaria blight

The accumulation of salicylic acid and H₂O₂ during pathogenic infection of mustard plants with Alternaria brassicae spores was investigated to understand the role of these two defense compounds in the expression of resistance. Comparisons were made between a susceptible Brassica juncea variety RH30 and a Brassica carinata variety HC1, which is known to be resistant. An oxidative burst was detected as in situ accumulation of H₂O₂, in both the Brassica spp. after pathogen application. However, H₂O₂ generation was extracellular in the resistant variety and both extra- and intracellular in the susceptible variety. Endogenous levels of SA increased over 2.5-fold in the resistant variety HC1 in response to pathogen application and this increase was observed only in conjugated SA levels. Pathogen application also led to an increase in the antioxidant enzymes, guaiacol-dependent peroxidase (GDP) and superoxide dismutase (SOD) in HC1. Exogenous SA application to leaves led to over threefold increase in the free and conjugated SA levels in both varieties. Pathogen application to the SA pretreated plants led to over 10-fold increase in endogenous SA levels in both varieties as compared to the levels in controls and this correlated with a decrease in disease symptoms in both species. SA appeared to regulate defense responses in Brassica spp. in a concentration-dependent manner. While 2.7-fold increase in endogenous SA levels (as seen in HC1) led to an induction of antioxidant enzymes, over 10-fold increases in endogenous SA levels (as seen after exogenous SA application in both varieties) brought about no induction of the antioxidant enzymes, probably because SA itself served as an antioxidant.     

Herbivore- and Elicitor- Induced Resistance in Groundnut to Asian armyworm,Spodoptera litura (Fab.) (Lepidoptera: Noctuidae)

Induced defense was studied in three groundnut genotypes ICGV 86699 (resistant), NCAc 343 (resistant) and TMV 2 (susceptible) in response to Spodoptera litura infestation and jasmonic acid (JA) application. The activity of the oxidative enzymes [peroxidase (POD) and polyphenol oxidase (PPO)] and the amounts other host plant defense components [total phenols, hydrogen peroxide (H2O2), malondialdehyde (MDA), and protein content] were recorded at 24, 48, 72 and 96 h in JA pretreated (one day before) plants and infested with S. litura, and JA application and simultaneous infestation with S. litura to understand the defense response of groundnut genotypes against S. litura damage. Data on plant damage, larval survival and larval weights were also recorded. There was a rapid increase in the activities of POD and PPO and in the quantities of total phenols, H2O2, MDA and protein content in the JA pretreated + S. litura infested plants. All the three genotypes showed quick response to JA application and S. litura infestation by increasing the defensive compounds. Among all the genotypes, higher induction was recorded in ICGV 86699 in most of the parameters. Reduced plant damage, low larval survival and larval weights were observed in JA pretreated plants. It suggests that pretreatment with elicitors, such as JA could provide more opportunity for plant defense against herbivores.     

斜纹夜蛾幼虫诱导的油菜抗虫性及其与茉莉酸信号途径的关系

植物在受植食性昆虫为害时能产生防御反应,并且植物的茉莉酸信号转导途径在这一过程中发挥着重要作用。然而, 迄今为止对于油菜Brassica campestris的诱导防御反应很少有研究报道。为此, 本实验通过测定油菜内茉莉酸和胰蛋白酶抑制剂含量研究了油菜在斜纹夜蛾Spodoptera litura幼虫为害后的抗虫性和胰蛋白酶抑制剂含量的变化,并分析这些变化在油菜诱导抗虫性与茉莉酸信号转导途径中的关系。结果表明：斜纹夜蛾幼虫取食能导致油菜体内茉莉酸和胰蛋白酶抑制剂含量系统性上升,外用茉莉酸甲酯处理也能系统性增加油菜的胰蛋白酶抑制剂含量,并且取食茉莉酸甲酯处理或斜纹夜蛾幼虫取食过的叶片能显著降低斜纹夜蛾幼虫的体重,两者的体重分别为对照植株上的67.5%和60.2%。机械损伤加斜纹夜蛾幼虫口腔分泌物处理能引起处理叶中茉莉酸和胰蛋白酶抑制剂含量的增加,但其诱导效果与机械损伤加水没有显著差异,并且两者明显低于虫害的诱导效果,两种处理的茉莉酸和胰蛋白酶抑制剂含量分别为虫害诱导的68.4%和24.4%及62.9%和36.9%;多次连续机械损伤的诱导效果与一次损伤的没有明显差异。结果说明斜纹夜蛾幼虫诱导的油菜抗虫性与茉莉酸信号转导途径有关,而其激活油菜抗虫反应的机理则可能与其特定的取食行为相关。     

Cereal host interactions with Russian wheat aphid: A review

Abstract

A lack of understanding the interaction between the Russian wheat aphid (RWA) and its host plant is a limitation in developing effective strategies for controlling the aphid. It is generally assumed that the interaction between aphid and plant is similar to that between plant and pathogen; that is, an elicitor from the insect is recognized by a protein from the host plant and a cascade of signal transduction events follows. However, evidence suggests that RWA feeding is eliciting both the SA- and JA/ethylene-dependent signaling pathways by mimicking aspects of both pathogen and herbivorous insect attacks. Results further suggest that phenotypic symptoms after RWA feeding are under regulation via two independent reactions, namely an immediate response (i.e., leaf rolling) and a downstream event (i.e., chlorosis). These defense responses enable a resistant host plant to defend itself and overcome the stress response, while their susceptible counterparts die. The processes involved in the onset of the defense response are discussed, and mechanisms enabling resistant plants to overcome the stress associated with the feeding process are presented as a working model for RWA-cereal host interaction. Knowledge of genes involved in wheat's defense responses against the RWA and an understanding of their functions may provide additional strategies for developing broad-spectrum resistance in plants.     

Enhanced proteolysis leads to pre-mature cell death under the influence of elicitor like mycelial components from karnal bunt (tilletia indica) pathogen in wheat callus cultures

Calli raised from mature embryos of susceptible wheat cultivar WH 542 were used in the present study as in vitro bioassay system to study the influence of disease determinant(s) of Karnal bunt (Tilletia indica), a semi-biotrophic fungal pathogen of wheat. Influence of elicitor and conditioned medium (CM) prepared from fungal cultures of T. indica was investigated on induction of programmed cell death (PCD). Induction of PCD was observed as hypersensitive response (HR) in terms of browning at localized regions of callus cultures and induction of proteolytic enzyme(s). Elicitor treated calli showed higher induction of protease activity than untreated and CM-treated cultures, which showed not much change in the activity. It was further substantiated by gel protease assay and activation of caspase-3 like protein(s) in callus cultures that clearly suggested the presence of signaling molecule(s) in the fungal elicitor preparation rather than in conditioned medium. This study further demonstrated that only elicitor preparation possesses such molecule(s), which might be cell wall bound components, rather than secretory in nature as CM was unable to induce PCD in wheat callus cultivars.     

小麦品种抗麦红吸浆虫鉴定与抗性分析

利用一套较完整的田间自然感虫鉴定法,对183份小麦品种进行了抗麦红吸浆虫鉴定.鉴定结果表明,小麦不同品种对麦红吸浆虫的抗性存在着显著差异:高抗、中抗、低抗、感虫、高感的小麦品种,分别占参试品种的24.59%、16.94%、18.58%、14.21%、25.68%.不同抗性类型的小麦品种其穗被害率、粒被害率、估计损失率、抗性指数均有较大的差异,其中表现高抗类型45份,中抗类型31份.这些品种既可作为麦红吸浆虫发生区控制麦红吸浆虫危害的主推品种和后备品种,也可作为亲本材料提供给育种单位利用.