A recessive resistance gene of alfalfa to alfalfa mosaic virus and impact of high ambient temperature on virus resistance

Twenty plants of alfalfa cv. Beaver were screened for resistance to alfalfa mosaic virus (AIMV) severe strain A-515. ELISA screening on both inoculated and apical leaves at fixed temperature (20°C day, 16°C night) suggested the following three types of clonal response to AIMV infection: extremely resistant, AIMV not normally detected from either inoculated or apical leaves; resistant, AIMV detected from inoculated leaves only and did not spread systemically; susceptible, AIMV detected from both inoculated and apical leaves. When plants in the second category were maintained at high temperature (30°C), AIMV was detected from inoculated and apical leaves 6 and 12 days after inoculation, but was not detectable in the apical leaves thereafter. Plants in the first category remained extremely resistant at all temperatures tested. The results of comparative tests using progeny plants of extremely resistant, resistant and susceptible plants, and of their hybrids, suggested that the resistance to AIMV A-515 was controlled by a temperature-dependent recessive gene.     

Effect of alfalfa mosaic virus isolates on forage yield of lucerne (Medicago sativa) in Britain

The effect of alfalfa mosaic virus (A1MV) on lucerne forage yield was investigated in experimental plots in which all plants were indexed for freedom from seed-borne A1MV. Plants which were artificially inoculated with A1MV were tested for successful infection prior to transfer to the plots. Inoculated plots yielded 14-8-22-8% (fresh weight) and 15.0 18.1% (dry weight) less than uninoculated plots. Isolate A1MV-I reduced the yield of five lucerne cultivars and yield was decreased by several A1MV isolates in cv. Vertus. Yield decreases tended to be greatest at the first harvest and progressively smaller at the second and third. Despite aphicide treatments and a cereal barrier crop, ELISA indexing showed natural infection developed in control plots. Most known infected plants failed to produce symptoms and yield decreases were associated with predominantly symptomless infection.     

豇豆退绿斑驳病毒(CCMV)突变体的混合侵染研究

 本研究采用PCR点突变的方法,对豇豆退绿斑驳病毒(CCMV) RNA3的cDNA克隆进行突变,突变体TP7是在外壳蛋白基因之前突变形成一个独特的Sal I酶切点;TP8是在3a基因之前突变形成一个独特的Bam H I酶切点;AG 1是在外壳蛋白基因之后突变形成一个独特的Not I酶切点。TP7、TP8、AG1体外转录产物分别与CCMV RNA1、RNA2的c DNA克隆的体外转录产物混和后接种豇豆,结果表明,接种后第6 d TP7植株首先表现出症状,9d后TP8、AG1和野生型CCMV接种的植株表现出症状,TP7的症状发展最快。
采用TP7、TP8、AG1接种20 d的毒源植株,按TP7+TP8、TP7+AG1、TP8+AG1等量混合汁液接种豇豆,接种20 d,通过RT-PCR分析接种植物,在TP7+TP8和TP7+AG1混合侵染中,TP7在混合侵染中占优势,在TP8+AG1的混合侵染中,TP8和AG1的种群十分接近。     

香蕉束顶病毒(BBTV)侵染对寄主内源激素的影响

 本试验用ELISA方法测定了香蕉感染束顶病毒(BBTV)后植株体内的3种内源激素,赤霉素(GA₃)、玉米素类(iPAs)和脱落酸(ABA)的变化。结果表明,感株的GA₃水平在侵染过程中虽有微弱增加,但含量和增长速度显著低于健株对照;iPAs在接种BBTV第14天后明显下降,并维持较低水平;ABA在BBTV侵染后被大量诱导增加并不断积累,在接种后第35天测定含量最高,为对照的3.34倍。试验还同时检测了BBTV在侵染过程中的运转。用间接ELISA测定的接种叶和顶叶的BBTV含量显示:接种21天后BBTV在接种叶和顶叶中还大量增殖,呈系统性分布。但寄主的症状在接种35天后才逐渐在顶叶表现。以上结果表明:香蕉束顶病的症状表现似乎主要与病株中的内源激素的失调有关,而与BBTV在体内的运转并不直接相关     

叶霉菌非亲和小种对番茄系统抗性的诱导及植株内水杨酸动态

 番茄叶霉菌小种4是番茄Cf5品系的非亲和小种,接种Cf5植株第3叶后,经不同诱导间隔期以亲和小种5接种第3叶和第4叶,15 d后检查叶霉病发病情况。试验表明,在诱导间隔期为3 d和5 d时,小种4诱导接种的第3叶和未经诱导接种的上位第4叶发病面积比不接种或接种小种5的对照显著降低,以5 d间隔期处理效果最好。上述2个叶位的发病分别比对照降低90%和85%。小种4接种第3叶后该叶位和上部未接种第4叶内水杨酸含量迅速增加,以接种后3 d含量最高,分别达4.02 μg/g鲜重和3.21μg/g鲜重,比对照分别高2倍和1.8倍。接种后5 d内始终保持较高水平。接种8 d后逐渐下降,但仍高于对照。水杨酸含量的增加早于抗性表现,因而可能在该系统的抗性诱导中起作用。     

Spread and interaction of Pepino mosaic virus (PepMV) and Pythium aphanidermatum in a closed nutrient solution recirculation system: effects on tomato growth and yield

Pepino mosaic virus (PepMV) was shown to be efficiently transmitted between tomato plants grown in a closed recirculating hydroponic system. PepMV was detected in all plant parts after transmission via contaminated nutrient solution using ELISA, immunocapture RT‐PCR, RT‐PCR, electron microscopy, and by inoculation to indicator plants. Detection of PepMV in nutrient solution was only possible after concentration by ultracentrifugation followed by RT‐PCR. Roots tested positive for PepMV 1–3 weeks after inoculation, and subsequently a rapid spread from the roots into the young leaves and developing fruits was found within 1 week. PepMV was only occasionally detected in the older leaves. None of the infected plants showed any symptoms on fruits, leaves or other organs. Pre‐infection of roots of tomato cv. Hildares with Pythium aphanidermatum significantly delayed PepMV root infections. When mechanically inoculated with PepMV at the 2–4 leaf stage, yield loss was observed in all plants. However, only plants of cv. Castle Rock recorded significant yield losses when infected via contaminated nutrient solution. Yield losses induced by infection with PepMV and/or P. aphanidermatum ranged from 0·4 up to 40% depending on experimental conditions.     

4种化学物质对南瓜抗白粉菌产生过敏性反应影响的研究

研究了4种化学物质对南瓜抗白粉病产生过敏性反应的影响,结果表明H2O2+接菌处理可使南瓜叶片中过敏性细胞数量增加,且随着浓度的升高过敏性细胞数量逐渐增多,经20mmol/L H2O2 + 接菌处理的南瓜叶片,在96h时产生的过敏性细胞比对照多81个/cm2; 而活性氧清除剂过氧化氢酶、抗坏血酸和谷胱甘肽3种化学物质处理+接菌南瓜叶片后,叶片中过敏性细胞的数量显著减少,且浓度越高,过敏性细胞数量越少,2000U/mL过氧化氢酶+接菌、20mmol/L抗坏血酸+接菌和40mg/L谷胱甘肽 + 接菌处理南瓜叶片,96h时过敏性细胞数量分别比对照 + 接菌少79、95和90个/cm2; 接菌10d后,20mmol/L H2O2 + 接菌处理的南瓜叶片的病情指数比对照 + 接菌低14.1,2000U/mL过氧化氢酶 + 接菌、20mmol/L抗坏血酸 + 接菌和40mg/L谷胱甘肽 + 接菌处理的南瓜叶片的病情指数分别比对照高7.47、4.95和6.83。     

Induction of systemic resistance to rust in Vicia faba by phosphate and EDTA: effects of calcium

Resistance in the upper leaves of broad bean ( Vicia faba ) plants to infection by the rust fungus, Uromyces viciae-fabae, was increased following treatment of the lower leaves with 10 m m potassium phosphate or 5 m m EDTA. Increasing the interval between treatment of the lower leaves and inoculation of the upper leaves had little effect on rust infection. Thus rust infection was reduced by 50 and 34% if the upper leaves were inoculated 1 day after treatment of the lower leaves with potassium phosphate or EDTA, respectively, while there was a 75% reduction in infection if the interval between treatment and inoculation was increased to 12 days. Application of calcium nitrate (10 m m ) after the phosphate or EDTA treatments prevented the induction of systemic resistance. Calcium nitrate applied alone to the lower leaves had no significant effect on rust infection of the upper leaves.     

Effect of Figaren, an Antiviral Protein from Cucumis figarei, on Cucumber mosaic virus Infection

Local lesion formation on cowpea leaves was more than 50% inhibited by treatment with a 23 kDa RNase-like glycoprotein from Cucumis figarei, figaren, from 24 hr before to 1 hr after inoculation with Cucumber mosaic virus (CMV). CMV accumulation detected by ELISA in tobacco leaves treated with figaren 6 or 0 hr before inoculation with CMV was suppressed. When upper leaves of tobacco plants were treated with figaren and inoculated 10 min later with CMV, mosaic symptoms were delayed for 5–7 days on most of the tobacco plants, and some plants remained asymptomatic. From fluorescence in situ hybridization, infection sites were present in figaren-treated cowpea or melon leaves after inoculation with CMV, though the sites were reduced in number and size compared with those in water-treated control leaves. The amount of CMV RNAs and CMV antigen in melon protoplasts inoculated with CMV and subsequently incubated with figaren similarly increased with time as did that in the control. ELISA and local lesion assays indicated that CMV infection on the upper surfaces of the leaves of tobacco, melon, cowpea and C. amaranticolor whose lower surfaces had been treated with figaren 5–10 min before CMV inoculation was almost completely inhibited. Figaren did not inhibit CMV infection on the opposite untreated leaf halves of melon, cowpea and C. amaranticolor, whereas it almost completely inhibited CMV infection on the untreated halves of leaves of tobacco. CMV infection was not inhibited in the untreated upper or lower leaves of the four plants. These data suggest that figaren does not completely prevent CMV invasion but does inhibit the initial infection processes. It may also induce localized acquired resistance in host plants. Received 10 October 2000/ Accepted in revised form 6 February 2001     

Some properties of alfalfa mosaic virus isolated from Buddleia davidii in the UK

Alfalfa mosaic virus (AMV) and cucumber mosaic virus (CMV) were isolated from clones of Buddleia davidii collected at Long Ashton Research Station for assessment of horticultural characteristics. The AMV isolate investigated in this study (AMV-B) infected 18 species and cultivars of herbaceous test plants. The in vitro properties of AMV-B were determined. The virus was purified by permeation chromatography on controlled-pore glass beads and an antiserum prepared. AMV-B appeared to be serologically indistinguishable from AMV isolate 15/64. The sizes and shapes of the virus particles were those of classical AMV components. The capsid protein was a single polypeptide of Mr 24 375. Buddleia seedlings inoculated with AMV-B showed only mild chlorosis and slight distortion of leaves.     

Effects of leaf wetness duration, relative humidity, light and dark on infection and sporulation by Didymella rabiei on chickpea

No infection occurred at less than 95% relative humidity (r.h.) when chickpea plants were dried after inoculation with conidia of Didymella rabiei. Infection was significant when the dry leaves were exposed to 98% r.h. for 48 h. When inoculated plants were subjected to different leaf wetness periods, some infection occurred with 4 h wetness, and disease severity increased with wetness duration according to an exponential asymptote, with a maximum value after about 18 h. Germination of conidia and germ tube penetration increased linearly with increasing wetness periods when recorded 42 h after inoculation. With a 24-h wetness period, germination of conidia was first observed 12 h after inoculation and increased linearly with time up to 52 h (end of the experiment). Dry periods immediately after inoculation, followed by 24-h leaf wetness, reduced disease severity; as the dry period increased the severity decreased. Disease severity increased with increasing periods of darkness after inoculation. The number of pycnidia and the production of conidia on infected leaves increased only slightly with high r.h. (either in the light or in the dark), but large increases occurred over an 8-day period when the leaves were kept wet.     

一种新的90 kD胞外蛋白激发子诱导烟草系统获得抗性研究

 就棉疫病菌(Phytophthora boehmeriae Saw.)培养滤液中提纯获得的90 kD胞外蛋白激发子诱发烟草系统获得抗性进行了研究。以10 nmol/L激发子溶液注射处理Samsun NN烟草叶片24 h后,在处理叶片及其上、下各2片叶片接种TMV,结果是处理叶及其上、下各2片叶片上的枯斑数显著少于对照,诱抗防效达40.9%~53.1%;接种TMV7d后处理叶片的枯斑平均直径为1.23mm,显著小于对照叶片上的枯斑直径2.97mm,但是处理叶片的上、下叶片上的枯斑平均直径与对照没有显著差别。以10 nmol/L激发子溶液注射处理Samsun NN烟草叶片分别立即接种或于1、2、4、7、15 d接种TMV,结果证明该激发子诱导烟草对TMV的抗性以处理后第1d至第4 d接种为较好,防效达30.2%~5 0.4%;处理后立即接种和第7d接种TMV诱抗防效仅4%左右,处理叶片上的枯斑数与对照没有显著差别,表明较强的诱导抗性可持续时间<7d。用不同浓度激发子处理烟叶,所测定的0.5~100 nmol/L各浓度均可显著地诱发烟草对TMV产生获得抗性,诱导抗性效果为34.9%~5 8.2%,诱导抗性效果随浓度的降低呈下降趋势。以10 nmol/L激发子溶液注射处理W38烟草叶片,2 d后分别注射接种烟草野火病菌(Pseudomonas syringae pv.tabaci)菌液或喷雾接种烟草赤星病菌(Alternaria alternata)分生孢子,结果是,注射接种烟草野火病菌5d后处理叶片及其上、下叶片上的野火病病斑均显著小于对照;处理叶片上的赤星病病斑数及病斑面积明显小于对照,表明该激发子可诱导烟草对野火病和赤星病产生抗性。上述结果表明,90kD蛋白激发子诱导烟草产生的获得抗性是一种典型的系统获得抗性,该系统获得抗性对病原菌具广谱抗性。     

Systemic infection of sorghum and corn by conidia ofSclerospora Sorghi

Conidia ofSclerospora sorghi, obtained from either systemically-infected or local-lesion-infected leaves of sorghum (cv. Vidan), were capable of inducing typical downy mildew systemic infection, including oospore formation, in sorghum and corn hybrids. Very young inoculated seedlings displayed chlorotic systemic symptoms already on the first leaf, and often died at fourth-leaf stage. Systemic infection was induced by conidia on sorghum 1–14 days old at inoculation. Incidence of infection was much higher and symptoms less delayed when the shoot rather than coleorhizas of young sorghum and corn seedlings were inoculated; in two-week-old sorghum with three leaves, inoculation of the coleoptile or of the base of the second and third blades resulted in systemic infection; with coleoptile inoculation partial leaf chlorosis was delayed until the fourth-or fifth-leaf stage, showing that penetration without symptoms had occurred as far as the meristematic tissues of young leaves still within the leaf tube. Conidial inoculation of young sorghum tillers sprouting after cutting down healthy mother shoots resulted in systemic infection. Conidial inoculum is deemed to be the probable major means for systemic infection of corn and sorghum sown in fields in which oospores are not present; inoculation of new tillers of forage sorghum by conidia from infected plants in a neighboring field can explain the rise in numbers of plants systemically stricken. Two sweet corn hybrids — one considered resistant in the field, the other very susceptible — proved equally susceptible when inoculated with conidia at 5 days of age.     

Response of Hard Red Winter Wheat to Soilborne wheat mosaic virus Using Novel Inoculation Methods

ABSTRACT Soilborne wheat mosaic virus (SBWMV) is an agronomically important pathogen of wheat that is transmitted by the soilborne plasmodiophorid vector Polymyxa graminis. In the laboratory, attempts to generate SBWMV-infected plants are often hampered by poor infectivity of the virus. To analyze the mechanism for virus resistance in wheat cultivars, we developed novel inoculation techniques. A new technique for foliar inoculation of SBWMV was developed that eliminated wound-induced necrosis normally associated with rub inoculating virus to wheat leaves. This new technique is important because we can now uniformly inoculate plants in the laboratory for studies of host resistance mechanisms in the inoculated leaf. Additionally, wheat plants were grown hydroponically in seed germination pouches and their roots were inoculated with SBWMV either by placing P. graminis-infested root material in the pouch or by mechanically inoculating the roots with purified virus. The susceptibility of one SBWMV susceptible and three field resistant wheat cultivars were analyzed following inoculation of plants using these novel inoculation techniques or the conventional inoculation technique of growing plants in P. graminis-infested soil. The results presented in this study suggest that virus resistance in wheat likely functions in the roots to block virus infection.     

Pathogenesis of Alfalfa mosaic virus in Soybean (Glycine max) and Expression of Chimeric Rabies Peptide in Virus-Infected Soybean Plants

ABSTRACT Infection of soybean (Glycine max) plants inoculated with particles of Alfalfa mosaic virus (AlMV) isolate 425 at 12 days after germination was monitored throughout the life cycle of the plant (vegetative growth, flowering, seed formation, and seed maturation) by western blot analysis of tissue samples. At 8 to 10 days after inoculation, the upper uninoculated leaves showed symptoms of virus infection and accumulation of viral coat protein (CP). Virus CP was detectable in leaves, stem, roots, seedpods, and seed coat up to 45 days postinoculation (dpi), but only in the seedpod and seed coat at 65 dpi. No virus accumulation was detected in embryos and cotyledons at any time during infection, and no seed transmission of virus was observed. Soybean plants inoculated with recombinant AlMV passaged from upper uninoculated leaves of infected plants showed accumulation of full-length chimeric AlMV CP containing rabies antigen in systemically infected leaves and seed coat. These results suggest the potential usefulness of plants and plant viruses as vehicles for producing proteins of biomedical importance in a safe and inexpensive manner. Moreover, even the soybean seed coat, treated as waste tissue during conventional processing for oil and other products, may be utilized for the expression of value-added proteins.     

Antioxidant,ethylene and membrane leakage responses to powdery mildew infection of near-isogenic barley lines with various types of resistance

Leaves of powdery mildew-susceptible barley (Hordeum vulgare cv. Ingrid) and related near-isogenic lines bearing various resistance genes (Mla12, Mlg or mlo5) were inoculated with Blumeria graminis f. sp. hordei race A6. Fungal attack induced several-fold increases in ethylene emission and electrolyte leakage in leaves of susceptible Ingrid beginning 3 days after inoculation. Activities of peroxidase, superoxide dismutase, glutathione S-transferase, ascorbate peroxidase and glutathione reductase enzymes were induced markedly in susceptible leaves 5–7 days after inoculation. Similar, but less pronounced pathogen-induced changes were detected in inoculated leaves of Mla-type resistant plants that show hypersensitive cell death upon inoculation, and, to an even lesser extent, in the Mlg and mlo lines, where no visible symptoms accompanied the incompatible interaction. Glutathione content increased only in susceptible barley 7 days after inoculation. Catalase activity, total ascorbate content and redox state were not influenced by inoculation in any of the genotypes. The activity of dehydroascorbate reductase was significantly reduced 3–5 days after inoculation in the susceptible parental plants and after 5 days in Mla and Mlg lines, while it was stable in the mlo barley. Slightly elevated levels of H₂O₂ were observed in the inoculated resistant plants. In contrast, H₂O₂ content decreased in the susceptible line 7 days after pathogen attack. These data indicate that high levels of antioxidants are involved in the compatible interaction of susceptible barley and powdery mildew by protecting the pathogen from oxidative damage.     

Verticillium dahliae Modifies the Concentrations of Proline, Soluble Sugars, Starch, Soluble Protein and Abscisic Acid in Pepper Plants

The objective of this research was to study the levels of some organic solutes, such as total protein, total soluble sugars, starch and proline in leaves, as well as abscisic acid concentration in xylem of pepper plants inoculated with Verticillium dahliae Kleb. Healthy and inoculated plants were always kept well watered. Measurements were made at time intervals after inoculation. Leaf water potential rapidly decreased as a consequence of fungal infection. However, relative water content in leaves only changed significantly from day 20 after inoculation, and such decreases coincided with a sharp build up of proline and total soluble sugars in leaves. Starch and protein levels, as well as abscisic acid concentration in xylem, declined in healthy and inoculated peppers as they became older. However, such decreases were more pronounced in infected plants, especially soon after inoculation. Results suggest that proline and total soluble sugars accumulation could be sensors of the damage caused by the fungal infection.     

Effects of foliar- and root-applied silicon on the enhancement of induced resistance to powdery mildew in Cucumis sativus

Two cucumber ( Cucumis sativus ) cultivars differing in their resistance to powdery mildew, Ningfeng No. 3 (susceptible) and Jinchun No. 4 (resistant), were used to study the effects of foliar- and root-applied silicon on resistance to infection by Podosphaera xanthii (syn. Sphaerotheca fuliginea ) and the production of pathogenesis-related proteins (PRs). The results indicated that inoculation with P. xanthii significantly suppressed subsequent infection by powdery mildew compared with noninoculation, regardless of Si application. Root-applied Si significantly suppressed powdery mildew, the disease index being lower in Si-supplied than in Si-deprived plants, regardless of inoculation treatment. The resistant cultivar had a more constant lower disease index than the susceptible cultivar, irrespective of inoculation or Si treatment. Moreover, with root-applied Si, activities of PRs (for example peroxidase, polyphenoloxidase and chitinase) were significantly enhanced in inoculated lower leaves or noninoculated upper leaves in inoculated plants of both cultivars. Root-applied Si significantly decreased the activity of phenylalanine ammonia-lyase in inoculated leaves, but increased it in noninoculated upper leaves. However, Si treatment failed to change significantly the activity of PRs in plants without fungal attack. Compared to the control (no Si), foliar-applied Si had no effects either on the suppression of subsequent infection by P. xanthii or on the activity of PRs, irrespective of inoculation. Based on the findings in this study and previous reports, it was concluded that foliar-applied Si can effectively control infections by P. xanthii only via the physical barrier of Si deposited on leaf surfaces, and/or osmotic effect of the silicate applied, but cannot enhance systemic acquired resistance induced by inoculation, while continuously root-applied Si can enhance defence resistance in response to infection by P. xanthii in cucumber.     

Involvement of lipoxygenase,lipoxygenase pathway volatiles,and lipid peroxidation during the hypersensitive reaction of pepper leaves to Xanthomonas campestris pv. vesicatoria

Lipoxygenase activity and protein, production of lipid-derived volatiles, and lipid peroxidation levels were determined in pepper (Capsicum annuum L., cv. Early Calwonder-10R) leaves during the hypersensitive reaction induced by avirulent race 2 of Xanthomonas campestris pv. vesicatoria. Lipoxygenase activity increased during the collapse phase of the hypersensitive reaction (8 to 12 h after inoculation), and an increase in electrolyte leakage occurred. However, Western blot analysis revealed that lipoxygenase proteins decreased during the same period. When only one longitudinal half of a pepper leaf was inoculated with the avirulent bacterium race, a significant increase in lipoxygenase activity was observed in both inoculated and noninoculated leaf halves, 10 h after inoculation. In addition, lipoxygenase protein decreased in inoculated leaf halves, but remained unchanged in noninoculated ones. The evolution of some volatile compounds derived from the lipoxygenase pathway [(E,E)-2,4-hexadienal, 1-hexanol, 3-hexen-1-ol, 2,4-hexadienal and 2,4-eptadienal] and carotenoid degradation (α- and β-ionone) increased in the incompatible interaction during the collapse phase of the hypersensitive reaction. The level of the oxidative index (A₂₃₅/A₂₀₅) of leaf lipid extracts, determined to estimate lipid peroxidation, significantly increased in the advanced stage of the hypersensitive reaction. Furthermore, determination of the oxidative index in neutral lipid, glycolipid and phospholipid fractions showed that the oxidative index was significantly increased only in the glycolipid fraction. Lipoxygenase activity and protein, electrolyte leakage, volatiles and lipid peroxidation were not changed in pepper leaves inoculated with the virulent race 1 of X. campestris pv.vesicatoria during the time interval considered (2–12 h after inoculations). The hypothesis that a lipoxygenase with chloroplastic location is induced in the incompatible interaction, and which is responsible for the increase in lipid peroxidation is advanced.