Biological control mechanisms of D-pinitol against powdery mildew in cucumber

D-pinitol is an effective agent for controlling powdery mildew (Podosphaera xanthii) in cucumber. In this study, we determined the mechanisms of D-pinitol in controlling powdery mildew in cucumber plants. We compared P. xanthii development on cucumber leaf surface treated with D-pinitol or water (2 mg ml⁻¹) at different time points after inoculation. The germinating conidia, hyphae, and conidiophores of the pathogen were severely damaged by D-pinitol at any time of application tested. The highest level of suppression of fungal development was obtained at 3 days after inoculation. The contents of chlorophyll, total phenolics, flavonoid, and gallic acid and its derivatives (GAD); the activities of phenylalanine ammonialyase (PAL), polyphenoloxidase (PPO), peroxidase (POX), and superoxide dismutase (SOD); and the expression of the genes encoding for PR-1, peroxidase (POX), lipoxygenase (LOX1), chitinase (Chit1) were higher in the cucumber leaves treated with D-pinitol and inoculated than in the leaves either treated with D-pinitol or inoculated with the pathogen. These results suggest that D-pinitol triggers several plant defense responses in cucumber leading to pathogen suppression and resistance to powdery mildew.     

寡聚糖诱导黄瓜对白粉病抗病反应的超微结构研究

 中科6号(2%氨基寡糖素)处理黄瓜植株叶片,5 d后接种白粉菌Sphaerotheca fuliginea(Schlecht.) Poll.,可诱导黄瓜Cucumis sativus产生对白粉病的抗病性,寄主细胞对病原菌的侵入产生了防卫反应结构和物质以及过敏性坏死反应。表现为寄主细胞壁加厚,染色加深,寄主细胞壁下产生多层次结构的乳突,在寄主细胞壁与质膜之间有黑色物质沉积;吸器外质膜皱褶,染色加深,吸器外基质中出现染色加深的颗粒状电子沉积物;寄主细胞质紊乱,细胞器解体,整个寄主细胞解体、坏死。     

Prior Inoculation with Non-pathogenic Fungi Induces Systemic Resistance to Powdery Mildew On Cucumber Plants

A spray inoculation of the first leaf of 2-leaf stage cucumber plants with a non-pathogenic isolate of Alternaria cucumarina or Cladosporium fulvum before a challenge inoculation with the pathogen Sphaerotheca fuliginea induced systemic resistance to powdery mildew on leaves 2–5. Systemic resistance was expressed by a significant (p < 0.05) reduction in the number of powdery mildew colonies produced on each leaf of the induced plants, as compared with water-sprayed plants. Systemic resistance was evident when a prior inoculation with each of the inducing fungi was administered 1, 3 or 6 days before the challenge inoculation with S. fuliginea. Increasing the inoculum concentration of A. cucumarina or C. fulvum enhanced the systemic protection and provided up to 71.6% or 80.0% reduction, respectively, in the number of colonies produced on upper leaves, relative to controls. Increasing the inoculum concentration of S. fuliginea used for challenge inoculation, increased the number of powdery mildew colonies produced on both induced and non-induced plants. Pre-treated plants, however, were still better protected than controls, indicating that the level of systemic protection was related to the S. fuliginea inoculum concentration. The induction of systemic resistance against powdery mildew by biotic agents, facilitates the development of a wide range of disease management tools.     

白粉寄生孢寄生黄瓜白粉菌的特性研究

采用白粉寄生孢（Ampelomyces quisqualis Ces.AQ）接种离体条件下赛璐酚上黄瓜白粉菌［Sphaerotheca fuliginea (Schlecht)Poll.］和活体条件下黄瓜白粉菌后,通过棉兰染色和显微观察分析,初步明确了白粉寄生孢的侵染寄生过程。白粉寄生孢分生孢子产生芽管可入侵黄瓜白粉菌的分生孢子、菌丝、分生孢子梗;有时黄瓜白粉菌串生的分生孢子、分生孢子梗可被2~3条白粉寄生孢的菌丝寄生,随着寄生过程的进一步发展,黄瓜白粉菌的分生孢子梗基部膨大成无色的椭圆形或球形,其上逐渐产生由黄色至褐色的白粉寄生孢的分生孢子器,器内含有大量的分生孢子。     

Resistance levels of cucumber to <Emphasis Type="Italic">Podosphaera xanthii</Emphasis> in a growth chamber are related to haustorial formation and hyphal branching frequency

To clarify relationships between powdery mildew resistance of cucumber and hyphal developmental state of the pathogen, haustorial formation and the hyphal branching frequency were compared among cucumber varieties that differ in resistance levels to powdery mildew pathogen Podosphaera xanthii. Cotyledons of four cultivars were inoculated with P. xanthii. By 2 days after inoculation, secondary haustoria had developed from the first hyphal cell that formed beside the conidium in susceptible cultivars. The fungus on susceptible cultivars also tended to have hyphal branches just after the hyphal cell producing haustoria.     

Potassium phosphate induces systemic protection in barley to powdery mildew infection

In laboratory tests, treatment of the first leaves of barley (Hordeum vulgare L cv Golden Promise) with potassium phosphate led to significant reduction in infection of the second leaves with the powdery mildew fungus Blumeria graminis f sp hordei Marchal, with a 25 mM treatment giving 89% reduction in infection. Although the optimal interval between phosphate treatment of the first leaves and mildew inoculation of the second leaves was 2 days, significant protection was still obtained if the interval was increased to 12 days. Protection against powdery mildew infection was not as effective when the potassium phosphate was applied as a seed treatment or root drench. Phosphate treatment of the first leaves led to significant increases in activities of phenylalanine ammonia lyase (PAL), peroxidase and lipoxygenase in second leaves. Enzyme activities, especially PAL and peroxidase, were increased further when second leaves of phosphate-treated plants were inoculated with powdery mildew. Phosphate treatment of the first leaves did not adversely affect plant growth and, in a field trial, 25 mM potassium phosphate provided 70% control of mildew and gave a small increase in grain yield.     

Saccharin-induced protection against powdery mildew in barley: effects on growth and phenylpropanoid metabolism

Treatment of barley ( Hordeum vulgare ) with 3 m m saccharin, applied as a foliar treatment to the first leaf or as a soil drench, provided significant control of powdery mildew ( Blumeria graminis f.sp. hordei ) on first and second leaves. This was unlikely to be the result of a direct effect of saccharin on the fungus, as application of the chemical to first leaves 2 h before inoculation did not affect conidial germination or formation of appressoria. Saccharin treatment had no significant effect on plant growth, except for a reduction in total leaf area in plants treated with a saccharin drench 14 days before inoculation with mildew. Phenylalanine ammonia-lyase activity was reduced significantly in second leaves 18 and 48 h after inoculation in plants treated with saccharin 14 days earlier. Peroxidase activity increased significantly in plants challenged with mildew within 6 days of saccharin application, although changes were not apparent until 48 h after pathogen challenge. On these occasions, treatment with saccharin resulted in a 33% increase in peroxidase activity compared with controls. In plants inoculated 10 or 14 days after saccharin application, cinnamyl alcohol dehydrogenase (CAD) activity increased prior to, and 18, 24 and 48 h after, inoculation of the barley plants with mildew. CAD activity increased approximately twofold compared with controls. However, in contrast to peroxidase, CAD activity was significantly higher in saccharin-treated plants prior to and after inoculation with powdery mildew, suggesting that saccharin primes CAD activity prior to pathogen challenge.     

Suppression of conidial germination and appressorial formation by silicate treatment in powdery mildew of strawberry

The mode of action of soluble silicon against strawberry powdery mildew (Sphaerotheca aphanis var. aphanis) was investigated in four experiments. First, silicon-treated leaves from plants grown with silicate (Si+) and control leaves were excised, inoculated with conidia, and subsequent germination and formation of appressoria in a petri dish was assessed after 24 h. The germination rate was 49.7% on Si+ leaves, and was 67.2% on control leaves (t-test, P < 0.01). Second, we soaked cellulose membranes in various solvents and then placed the membranes on 4% water agar, dusted the membranes with conidia, and examined after 12 h. No difference was apparent between any treatment and the control (distilled water). Third, strawberries growing hydroponically with additional silicon in the medium were inoculated with conidia, and leaves were observed with a scanning electron microscope 1–2 days after inoculation. Germ tubes and secondary hyphae were shorter and had fewer branches on Si+ leaves than on the control. Moreover, penetration appeared to be inhibited. Fourth, the cuticle was separated from leaves from plants grown as in the third experiment, placed on water agar, and dusted with conidia. Germination of conidia, observed with a light microscope, on Si+ leaves was suppressed markedly to 40%–60% of that of the control. These results suggested that soluble silicon induced physiological changes in the cuticle layer after absorption by the plant. In addition, soluble silicate reduced germination of conidia, formation of appressoria, and possibly the penetration of powdery mildew.     

Changes in carbohydrate composition of cucumber leaves during the development of powdery mildew infection

Within 3 days after inoculation, greater rates of decrease of hexoses, and of increases in inositol and α-glucans, were detected in leaves of cucumber leaves infected with Sphaerotheca fuliginea (now reclassified as Podosphaera xanthii ) than in healthy leaves, together with synthesis of fungal mannitol, arabitol and trehalose. With the onset of sporulation (5 days after inoculation, dai), glucose, polyols, trehalose and glycogen content, as well as fresh and dry weight of leaf tissue, increased more rapidly than in healthy plants, while sucrose, raffinose and starch declined. Starch, glycogen and other α-glucans, extracted from ethanol-insoluble tissue fractions of different water-solubility from healthy and infected leaves, were quantified after amyloglucosidase treatment. Infection-induced alterations in partitioning of photosynthate during a 14 h photoperiod at 7 dai, including a close correspondence in patterns of synthesis of glucose, polyols and glycogen, further clarified the process of diversion of host assimilate to the pathogen. The presence of glucose, but only traces of sucrose in spores, supported evidence from other powdery mildew infections for photosynthate being transferred to S. fuliginea as glucose.     

Evaluation of the systemic activity of simeconazole in comparison with that of other DMI fungicides

The systemic activity of simeconazole (RS-2-(4-fluorophenyl)-1-(1H-1,2,4-triazol-1-yl)-3-trimethylsilylpropan-2-ol) in plants was compared with those of eight other sterol demethylation inhibitor (DMI) fungicides. Simeconazole prevented the infection of Blumeria graminis (DC) Speer f sp hordei Marchal on barley leaves within a radius of several centimeters from the edge of local treatment on the leaves when the compound was separated from the leaves by glass coverslips. This reveals that simeconazole has prominent vapour-phase activity. Simeconazole showed excellent curative activity against barley powdery mildew when treated 1-3 days after inoculation. Furthermore, this indicates that simeconazole has notable translaminar activity because, when the compound was applied onto either the adaxial or abaxial leaf surface, it showed excellent efficacy against powdery mildew on the opposite side of the leaf surface of barley and cucumber. Simeconazole also showed excellent efficacy against barley powdery mildew by soil drench 24h after inoculation. This suggests that simeconazole was absorbed very quickly into the barley plant after treatment. The permeability of the compound through cuticular membranes prepared from tomato fruits was about 20% at 22 h after the treatment and was much superior to that of the other DMI fungicides tested.     

三个小麦农家品种的苗期抗白粉病遗传分析

[目的]对3份小麦农家品种‘矮秆芒麦’、‘红头麦’和‘大红头’进行苗期抗性的遗传分析,研究它们的抗白粉病遗传特点,为其在抗病育种中的有效利用提供依据.[方法]将这3份小麦农家品种分别与感病品种‘铭贤169’正、反杂交,获得了F1和F2代.利用白粉菌E09菌株,分别对这3份农家品种、感病亲本‘铭贤169’以及各自的F1和F2代植株进行抗性鉴定.调查统计的数据经卡方测验分析其符合度.[结果]这3份农家品种对白粉菌E09菌株的抗性均由1对隐性核基因控制.[结论]3份农家品种对石家庄本地区的混合白粉病菌表现出良好的抗性,并且对E09的抗性均由1对隐性基因控制.可以进一步对它们进行分子标记及定位研究,为其作为抗源在小麦抗白粉病育种中的应用奠定基础.     

Silicon enhances the accumulation of diterpenoid phytoalexins in rice: a potential mechanism for blast resistance

ABSTRACT Although several reports underscore the importance of silicon (Si) in controlling Magnaporthe grisea on rice, no study has associated this beneficial effect with specific mechanisms of host defense responses against this fungal attack. In this study, however, we provide evidence that higher levels of momilactone phytoalexins were found in leaf extracts from plants inoculated with M. grisea and amended with silicon (Si(+)) than in leaf extracts from inoculated plants not amended with silicon (Si(-) ) or noninoculated Si(+) and Si(-) plants. On this basis, the more efficient stimulation of the terpenoid pathway in Si(+) plants and, consequently, the increase in the levels of momilactones appears to be a factor contributing to enhanced rice resistance to blast. This may explain the lower level of blast severity observed on leaves of Si(+) plants at 96 h after inoculation with M. grisea. The results of this study strongly suggest that Si plays an active role in the resistance of rice to blast rather than the formation of a physical barrier to penetration by M. grisea.     

Powdery mildew of Arabidopsis thaliana: a pathosystem for exploring the role of silicon in plant–microbe interactions

Silicon (Si) has been used in agriculture to protect plants against disease for hundreds of years and its prophylactic effects in monocots and dicots are well documented. The mechanisms by which Si exerts its protective effects in planta, however, are uncertain and presently the subject of debate. In this study, we sought to determine if Arabidopsis thaliana could be used to clarify the role of Si in plant–pathogen interactions. Accordingly, X-ray microanalysis mapping, light microscopy, scanning and transmission electron microscope techniques were used to examine the leaves of Si− fed A. thaliana plants inoculated with the powdery mildew fungus, Erysiphe cichoracearum. The results of this study demonstrate for the first time, that A. thaliana is a species that absorbs Si and that the incidence of powdery mildew disease for Si− fed plants is significantly lower compared to control plants. In particular, treatment with Si appeared to induce the production of an electron-dense, fungitoxic substance that accumulated within and around the collapsed fungal haustoria of infected epidermal cells within the leaves of disease-resistant plants. These results with Arabidopsis corroborate recent observations in other non-related species and support the emerging theory that the mechanisms by which Si imparts resistance to plants are complex and are not entirely explained by the traditionally proposed role of Si as a reinforcer of mechanical resistance. Collectively, the findings of the present study have established the Arabidopsis thaliana-Erysiphe cichoracearum pathosystem as a valid model to investigate the role of Si in plant–microbe interactions.     

Effect of potassium silicate on epidemic components of powdery mildew on melon

The effects of silicon (Si) supplied in the form of potassium silicate (PS) were evaluated on epidemic components of powdery mildew of melon under greenhouse conditions. The PS was applied to the roots or to leaves. In the first experiment, epidemic components were evaluated after inoculation with Podosphaera xanthii. In the second experiment, the disease progress rate was evaluated on plants subjected to natural infection. The area under the disease progress curve was reduced by 65% and 73% in the foliar and root treatments, respectively, compared to control plants, as a consequence of reductions in infection efficiency, colony expansion rate, colony area, conidial production and disease progress rate. However, root application of PS was more effective than foliar application in reducing most of the epidemic components, except for infection efficiency. This can be explained by the high Si concentration in leaf tissues with root application, in contrast to the foliar treatment where Si was only deposited on the external leaf surfaces. The effects of PS reported in this study demonstrated that powdery mildew of melon can be controlled, and that the best results can be achieved when PS is supplied to the roots.     

Nitrogen-induced changes in colony density and spore production of Erysiphe graminis f.sp. hordei on seedlings of six spring barley cultivars

The influence of increasing nitrogen supply (30, 60, 120 and 240 mg N per pot) on susceptibility was studied on seedlings of six cultivars of spring barley inoculated with virulent isolates of powdery mildew. The colony density (CD) measured as colonies per cm² was significantly increased with increasing application of nitrogen on all cultivars, and a significant interaction was found between N and cultivar. The different reactions of the cultivars could not be ascribed to lack of N uptake. In general, increasing N application enhanced the sporulation capacity of colonies (CSC) irrespective of increased CD and the cumulative production of spores per cm² leaf (CSCM) increased strongly with N application in all cultivars. No interaction between N and cultivar was found for the latter component. The increase in CSCM closely corresponded with the increase in N content and fresh weight of uninoculated leaves. No interaction between N treatment and powdery mildew isolates was found for infection efficiency and spore production per colony, when tested on one cultivar. The N-induced changes in infection and sporulation can explain the main part of the increasing effect of N fertilization on powdery mildew development in the field. The results indicate that it may be possible to breed for or select barley cultivars with low N impact on powdery mildew development.     

Lithium chloride and cucumber powdery mildew infection

Lithium chloride solutions (1 mM), supplied to cucumber plants via the root system, conferred protection against powdery mildew infection of the leaves by Sphaerotheca fuliginea. The development of infection structures was significantly inhibited by this treatment. Effective concentrations of the lithium salt increased the growth of shoots but resulted in some reduction of root growth, The incorporation of lithium by different parts of plants was assessed during a 20-day treatment period, The possible mode of action of lithium on this host-pathogen system is discussed.     

黄瓜白粉病菌在不同抗性黄瓜材料上的侵染过程

白粉病是黄瓜生产中发生普遍,危害严重的主要病害之一。pm5.1和PM5.2是黄瓜上的2个白粉病抗性位点,本文对7份不同抗病基因型的黄瓜自交系进行了黄瓜白粉病抗性鉴定,并开展了黄瓜白粉病菌侵染过程的研究,对侵染后12、24、72 h的萌发率、菌丝形成率及菌落形成率等进行了分析。结果表明,当基因型为PM5.1PM5.1 pm5.2pm5.2时,黄瓜病情指数最高,表现为高感白粉病;当基因型为PM5.1PM5.1PM5.2 PM5.2和pm5.1pm5.1pm5.2pm5.2时,表现为中感白粉病;当基因型为pm5.1pm5.1PM5.2PM5.2时黄瓜自交系病情指数最低,表现为抗白粉病。分生孢子在抗、感黄瓜自交系植株叶片上均能萌发,但只能在感病黄瓜材料上完成整个无性生长周期,产生分生孢子。此外,黄瓜白粉病菌分生孢子在感病材料上的萌发率、菌丝形成率及菌落形成率均高于抗病材料。     

公主岭霉素对草莓的防病诱抗和促生效应

为明确公主岭霉素在草莓生产中的防病诱抗效应,于大田采用五点取样法调查公主岭霉素对草莓白粉病的防控效果及对植株生长性状的影响,并于室内采用生测法测定其对草莓叶片防御酶活性和根际土壤酶活性的影响。结果显示,以公主岭霉素水浸提液100倍稀释液进行灌根并协同叶面喷雾对草莓白粉病的预防效果为71.90%;单独叶面喷雾对草莓白粉病的治疗效果为69.90%;灌根协同叶面喷雾预防与叶面喷雾治疗的综合防控处理对草莓白粉病的防治效果可达85.37%。公主岭霉素水浸提液100倍稀释液灌根后草莓幼苗的株高、叶柄长、叶长、叶宽、叶数、径冠等方面均优于清水对照;叶面喷施可提升叶片中多酚氧化酶和超氧化物歧化酶的活性,喷雾后1 d内即较清水对照显著提高;5 d后过氧化氢酶活性显著高于清水对照。灌根后根际土壤中蔗糖酶的活性在用药后15 d内均显著高于清水对照。表明合理施用公主岭霉素可实现对草莓白粉病的有效防控,并诱导提升草莓植株的抗病性,改善草莓生长环境,促进其生长。     

苦瓜白粉病病原菌和生理小种的鉴定及苦瓜对白粉病的抗性遗传分析

为明确海南省苦瓜白粉病的病原菌、生理小种及苦瓜对白粉病的抗性遗传规律,结合形态学鉴定和分子鉴定解析白粉病菌及生理小种种类,通过显微镜观察白粉病菌侵染过程,并应用主基因+多基因混合遗传模型分析法探讨苦瓜对白粉病的主要抗性遗传规律。结果表明:采集自海南省6个市(县)的苦瓜白粉病病原菌均为单囊壳白粉菌Sphaerotheca fuliginea,属生理小种2F,该菌在侵染苦瓜叶片时有4个关键时期:接种后4 h为分生孢子萌发高峰期,8 h为附着孢形成高峰期,16～24 h为次生菌丝形成高峰期,5 d为分生孢子梗形成高峰期。将其接种于苦瓜抗、感品系,对白粉病的抗性符合2对加性-显性-上位性主基因+加性-显性多基因模型,主基因和多基因共同控制苦瓜对白粉病的抗性,其中以主基因遗传为主,且会受到环境变异的影响。根据苦瓜抗性遗传规律,F2代主基因遗传率最高,受环境影响最小,在苦瓜的白粉病抗性育种中,以早期世代F2代作为有效选择世代。研究表明白粉病菌侵染叶片的前2 d是白粉病防治的最佳时期,所以在白粉病易发的物候期,可将防治时间提前1～2 d。     

Suppression of induced resistance in cucumber through disruption of the flavonoid pathway

ABSTRACT In this study, cucumber plants (Cucumis sativus) expressing induced resistance against powdery mildew (caused by Podosphaera xanthii) were infiltrated with inhibitors of cinnamate 4-hydroxylase, 4-coumarate:CoA ligase (4CL), and chalcone synthase (CHS) to evaluate the role of flavonoid phytoalexin production in induced disease resistance. Light and transmission electron microscopy demonstrated ultrastructural changes in inhibited plants, and biochemical analyses determined levels of CHS and beta-glucosidase enzyme activity and 4CL protein accumulation. Our results showed that elicited plants displayed a high level of induced resistance. In contrast, down regulation of CHS, a key enzyme of the flavonoid pathway, resulted in nearly complete suppression of induced resistance, and microscopy confirmed the development of healthy fungal haustoria within these plants. Inhibition of 4CL ligase, an enzyme largely responsible for channeling phenylpropanoid metabolites into the lignin pathway, had little effect on induced disease resistance. Biochemical analyses revealed similar levels of 4CL protein accumulation for all treatments, suggesting no alterations of nontargeted functions within inhibited plants. Collectively, the results of this study support the idea that induced resistance in cucumber is largely correlated with rapid de novo biosynthesis of flavonoid phytoalexin compounds.