Susceptibility of Rubus Species and Cultivars to Blackberry Leaf Rust (Phragmidium violaceum) and its Control by Fungicides

Rubus cultivars were compared with R. procerus for their susceptibility to Phragmidium violaceum, a pathogen first recorded in Victoria in February 1984, by inoculation of detached leaflets in the laboratory. The cultivar ‘Thornless Evergreen’ was highly susceptible. Cultivars ‘Thornfree’, ‘Dirksen Thornless’ and ‘Smoothstem’ were moderately susceptible whilst cvs ‘Silvan’ and ‘Lawtonberry’ were resistant. Cultivars ‘Cherokee’, ‘Cheyenne’, ‘Comanche’, ‘Marionberry’, ‘Boysenberry’, ‘Youngberry’, and ‘Olallie’ were highly resistant. ‘Loganberry’ and the raspberry cv. ‘Malling Landmark’ were immune. Seven fungicides were evaluated for their effect on P. violaceum. Mancozeb and triforine were the most effective against spore germination in vitro. RH 3866, triforine, propiconazole and mancozeb were the most effective in protective tests on detached leaflets of R. procerus, while RH 3866, propiconazole and flutriafol were the most effective in similar, curative tests. Phytotoxicity on detached leaflets was severe at high concentrations of triadimefon and propiconazole, and moderate to slight at similar concentrations of RH 3866 and copper hydroxide.     

番茄抗黄化曲叶病毒育种研究进展

本文分别对近年番茄抗黄化曲叶病毒的传统育种、分子辅助育种、基因工程育种进展进行了综述.栽培番茄均不抗番茄黄化曲叶病毒,所以传统育种采用从野生近缘种中筛选抗性材料,以其为亲本与栽培番茄进行杂交来获得抗性;野生近缘种中的抗性位点Ty-1、Ty-2和Ty-3及一些QTLs先后被定位,也筛选出了可鉴定Ty-1基因的SSR-47标记及鉴定Ty-3的SCAR标记;通过转基因技术获得抗性是研究热点之一,目前转入番茄后表现出抗性的序列有TYLCV病毒的CP基因、尺EP基因的部分序列或反义序列、不编码的保守序列以及源于白粉虱的GroEL基因.同时讨论了今后的主要发展方向.     

Efficacy of Fungicides for Control of Phytophthora Leaf Blight of Taro

Six fungicides were evaluated under greenhouse, laboratory,and dryland field conditions for control of Phytophthora leafblight of taro incited by P. colocasiae. Five separate criteriawere utilized to evaluate these fungicides: fungicidal activityin vitro; and fungicidal activity in vivo under conditions ofsimulated dew, simulated rainfall, greenhouse, and dryland fieldenvironments. In in vitro tests zoospores were killed at thefollowing concentrations: Dithane M-45, 5 ppm; Difolatan, 9ppm; Polyram, 65 ppm; Tribasic Copper Sulphate, 145 ppm; Benlate,210 ppm; and Dyrene, 260 ppm. Excellent control was obtainedwith Difolatan; good control with Dithane M-45 and Polyram;and poor control with Benlate, Tribasic Copper Sulphate, andDyrene. Results of in vivo tests correlated with those of thein vitro tests. Difolatan, Benlate, and Dyrene were the mostphytotoxic while Tribasic Copper Sulphate, Polyram, and DithaneM-45 were the least phytotoxic.     

Ability of Moderately Halophilic Bacteria to Control Grey Mould Disease on Tomato Fruits

Tomato is one of the leading crops in Tunisia in terms of weight consumed (20 kg/per person/year). Preserving the quality of the fruit from field to consumer is essential to successful marketing. Grey mould rot induced by Botrytis cinerea is an important cause of postharvest loss depending on season and handling practices. We describe here the ability of halotolerant to moderately halophilic bacteria isolated from different Tunisian Sebkhas (hypersaline soils) to protect fresh‐market tomato fruits from B. cinerea. The tomatoes tested were at two different stages of ripening, (i) mature‐green and (ii) red. Six strains significantly reduced growth of the pathogens from 67% to 87%. The effectiveness of these antagonists was also confirmed on green tomatoes; in which the fruit rot protection rate ranged from 74% to 100%. The antagonists were characterized by morphological, biochemical and physiological tests as well as 16S rDNA sequencing. The halotolerant effective isolates were identified as belonging to one of the species Bacillus subtilis (M1‐20, J9) or B. licheniformis (J24). One effective moderately halophilic isolate (M2‐26) was identified as Planococcus rifietoensis. These strains are a source of hydrolytic enzymes such as chitinases, proteases, laminarinases, amylases, lipases and cellulases. For comparison, 12 halotolerant or moderately halophilic strains obtained from DSM culture collection were also evaluated for their antifungal activity against B. cinerea on tomato fruits. The most effective strains were Halomonas subglaciescola, Halobacillus litoralis, Marinococcus halophilus, Salinococcus roseus, Halovibrio variabilis and Halobacillus halophilus with a percentage of grey mould rot reduction ranging from 71% to 97%. Inoculation of mature‐green tomatoes by the bacterial antagonist of Halobacillus trueperi resulted in no disease development. Our results indicate that the use of halotolerant to halophilic micro‐organisms should be helpful in reducing grey mould disease of stored tomatoes.     

Electrolyte Leakage, Lipoxygenase, and Lipid Peroxidation Induced in Tomato Leaf Tissue by Specific and Nonspecific Elicitors from Cladosporium fulvum

Glycoprotein nonspecific elicitor (NSE) and a specific elicitor preparation from intercellular fluids (SE) of tomato (Lycopersicon esculentum Mill. cv Bonny Best or Potentate) infected with race 2.4.5 of Cladosporium fulvum Cooke [syn. Fulvia fulva (Cooke) Ciferri] were injected into cv Sonatine (resistant to race 2.4.5) to compare electrolyte leakage, lipoxygenase activity, and lipid peroxidation induced in response to these elicitors. Increased electrolyte leakage was induced by NSE or SE; the leakage due to NSE but not to SE was inhibited by the nonsteroidal antiinflammatory drug (NSAID) piroxicam. Under normal photoperiod conditions, higher levels of lipoxygenase activity were detected 6 hours after injection with either elicitor. This activity peaked by 12 hours with both elicitors and declined to control levels by 24 hours when visible necrosis could be detected. Both NSE and SE-induced lipoxygenase was inhibited by piroxicam in vitro. Lipid peroxidation in elicitor-treated tissue was also assayed at 6, 12, and 24 hours after injection using the TBA test for malonaldehyde. Increased peroxidation was detected in response to NSE or SE at 12 hours with similar values obtained at 24 hours. With plants incubated in the dark, lipoxygenase, and lipid peroxidation were similarly induced in SE-injected tissue whereas necrosis induction by SE was light dependent.     

Biocontrol of Postharvest Grey Mould on Tomato by Yeasts

The fungal pathogen Botrytis cinerea causes severe rots on tomato fruit during storage and shelf life. Biological control of postharvest diseases of fruit may be an effective alternative to chemical control. Yeasts are particularly suitable for postharvest use, proving to be highly effective in reducing the incidence of fungal pathogens. Yeast fungi isolated from the surface of solanaceous plants were evaluated for their activity in reducing the postharvest decay of tomato caused by B. cinerea. Of 300 isolates, 14 strains of Rhodotorula rubra and Candida pelliculosa were found to be strongly antagonistic to the pathogen in vitro and were selected for further storage experiment. The antagonists were evaluated for their effect on the biological control of postharvest grey mould. Artificially wounded fruits were treated by means of a novel technique: small sterile discs of filter paper imbibed separately in suspensions of each yeast and the pathogen were superposed onto each wound. After 1‐week, 11 isolates were significantly effective in reducing the diameter of lesions by more than 60% compared to the control treated with B. cinerea alone. Total protection was obtained with the strain 231 of R. rubra on fruits challenged with pathogen spores. To our knowledge, R. rubra and C. pelliculosa have not been described as biocontrol agents against grey mould caused by B. cinerea. Our data demonstrate that the application of antagonistic yeasts represents a promising and environmentally friendly alternative to fungicide treatments to control postharvest grey mould of tomato.     

上海地区番茄黄化曲叶病毒病的鉴定及嫁接接种法研究

番茄黄化曲叶病毒(tomato yellow leafcurl virus,TYLCV)是一种由烟粉虱(Bemisia tabaci)和嫁接传播的双生病毒,在热带、亚热带地区给番茄生产造成严重威胁.根据番茄黄化曲叶病毒的保守序列设计一对引物,运用PCR技术从上海地区的感病番茄中扩增出一条575bp的特异带,而健康植株无此带.测序表明该序列与番茄黄化曲叶病毒具有极高的同源性(97%～99%).将健康接穗嫁接到感染番茄黄化曲叶病毒的番茄砧木上,间隔15 d和30 d,分别提取接穗的DNA,并用PCR法检测病毒,发现嫁接15 d后在部分接穗中检测到TYLCV病毒,嫁接30 d后在所有的接穗中均检测到病毒,因此,嫁接法可以作为番茄黄化曲叶病毒病的接种鉴定方法.     

Reduced Development of Grey Mould (Botrytis cinerea) in Bean and Tomato Plants by Calcium Nutrition

Fertilization of bean plants grown in perlite with 1 and 3 mM CaCl₂ or Ca(NO₃)₂ reduced severity of grey mould as compared with control plants or plants fertilized with 5 mM of the compounds. Fertilization with Ca(NO₃)₂ reduced severity leaf grey mould and fruit ghost spots of tomato plants grown in perlite by 70 and 45%, respectively. The rate of decrease varied with the position of the fruits on the plants. Leaves from plants treated with calcium or otherwise [KNO₃, (NH₄)₂SO₄] produced less ethylene than leaves of nontreated plants. Rate of growth of B. cinerea was lower on growth medium prepared from washings from leaves of calcium fertilized plants than from leaves from other treatments. The fertilizer combination Ca(H₂PO₄)₂+ CaSO₄ (1 and 3 g/kg soil) applied once to tomato plants grown in soil reduced severity of leaf grey mould by 80 % (significant at P = 0.05) but 1–3 g CaSO₄/kg soil only tended to reduce disease severity (30–40 %, not significant) as compared with the control. The compounds CaCl₂ and Ca(NO₃)₂ increased significantly ( P = 0.05) the growth of B. cinerea on synthetic medium when applied at rates of 1 0–10.0 mM whereas reduction of growth was observed with 0.1 mM of the compounds and of CaSO₄.     

Secondary Metabolism and Biotrophic Lifestyle in the Tomato Pathogen Cladosporium fulvum

Cladosporium fulvum is a biotrophic fungal pathogen that causes leaf mould of tomato. Analysis of its genome suggested a high potential for production of secondary metabolites (SM), which might be harmful to plants and animals. Here, we have analysed in detail the predicted SM gene clusters of C. fulvum employing phylogenetic and comparative genomic approaches. Expression of the SM core genes was measured by RT-qrtPCR and produced SMs were determined by LC-MS and NMR analyses. The genome of C. fulvum contains six gene clusters that are conserved in other fungal species, which have undergone rearrangements and gene losses associated with the presence of transposable elements. Although being a biotroph, C. fulvum has the potential to produce elsinochrome and cercosporin toxins. However, the corresponding core genes are not expressed during infection of tomato. Only two core genes, PKS6 and NPS9, show high expression in planta, but both are significantly down regulated during colonization of the mesophyll tissue. In vitro SM profiling detected only one major compound that was identified as cladofulvin. PKS6 is likely involved in the production of this pigment because it is the only core gene significantly expressed under these conditions. Cladofulvin does not cause necrosis on Solanaceae plants and does not show any antimicrobial activity. In contrast to other biotrophic fungi that have a reduced SM production capacity, our studies on C. fulvum suggest that down-regulation of SM biosynthetic pathways might represent another mechanism associated with a biotrophic lifestyle.     

Effect of Ethephon on Protein Degradation and the Accumulation of ;Pathogenesis-Related' (PR) Proteins in Tomato Leaf Discs

The effect of ethephon (2-chloroetylphosphonic acid) on the degradation of proteins and on the induction of Lycopersicon esculentum pathogenesis-related (PR) proteins was studied in tomato leaf discs. The rate of ribulose, -1,5-bisphosphate carboxylase/oxygenase (Rubisco) degradation was maximal in discs after 48 hours of incubation with 1 millimolar ethephon, leading to complete disappearance of Rubisco after 96 hours. This effect was correlated with an increase in PR protein synthesis and the induction of the previously reported alkaline proteolytic enzyme PR-P69 (P Vera, V Conejero [1988] Plant Physiol 87: 58-63). In vivo pulse-chase experiments demonstrated that ethephon not only affected Rubisco content but that of many other ³⁵S-labeled proteins as well, indicating that ethylene activates a general and nonspecific mechanism of protein degradation. This effect was partially inhibited in vivo by the action of pCMB, a selective inhibitor of cysteine-proteinases such as P69. These data reinforce the hypothesis that P69 and perhaps other PR proteins are involved in the mechanism of accelerated protein degradation activated by ethylene.     

Seasonal Dynamics of Black Leaf Mould (Pseudocercospora fuligena) on Greenhouse‐Grown Fresh Market Tomatoes

Black leaf mould (BLM), caused by Pseudocercospora fuligena, is a major fungal foliar disease of greenhouse‐grown tomatoes in the humid tropics. Quantifying the disease and yield loss from seasonal plantings will help mitigate the heavy reliance on frequent sprays of curative fungicides. In this study, severity of BLM was investigated in sequential plantings of tomatoes in two 50‐mesh BioNet greenhouses in central Thailand from June 2005 to January 2007. Each planting consisted of a block of six plants that received two applications of mancozeb and another block of six plants with no fungicide application at all. Two BLM peak epidemic periods were identified, that is, from plantings that were started in August–September and in January. On average, a yield loss of 30.6% was recorded from the two peak epidemic periods based on comparison of fungicide‐sprayed and non‐sprayed plants. Two sprays of mancozeb resulted in 71.6% reduction in disease severity during these peak epidemic periods. Mean disease severity (DS*) was highly correlated with a favourability index of relative humidity (RH), which was quantified on a scale of 0 (<85%, non‐favourable) to 1 (≥85%, extremely favourable). A three‐parameter logistic function explained the data well (R² = 0.81, P < 0.0001). Marketable yield (MY) was positively correlated with maximum plant height (PH_max) but negatively correlated with DS*. In addition, MY was higher from plantings during October to December. It was predicted well (R² = 0.60; P < 0.0001) using the model MY = (a + b × PH_max) × (1‐c × DS*), which combined both PH_max and DS*. Using this model, a reduction of 1.05% in marketable yield was predicted for each 1% increase in mean disease severity. The outcomes of this study implicated the need for management of RH and critical relevance of protecting tomato plants against BLM when they are grown during the peak epidemic periods.     

Effects of Leaf Age and Position on the Shoot Axis on Potassium Absorption by Tomato Leaf Slices

The effect of leaf age on K (⁸⁶Rb) influx into tomato (Lycopersiconesculentum Mill.) leaf lamina slices was determined for leaves5, 9 and 13 counting acropetally. Potassium influx rates expressedon a leaf fresh weight basis declined rapidly during leaf elongationat external KCI concentrations between 0.5 and 20.0 mM. In fullyexpanded leaves, K influx rates declined more slowly with age.The onset of senescence in mature leaves did not result in alarge loss in K uptake capability. Leaf position on the shootaxis and the stage of whole plant development had little influenceon K influx into leaf cells. It is suggested that the rapiddecrease in K influx in growing leaves is related to a dilutionin the concentration of K transporter sites resulting from anincrease in cell volume and weight. Lycopersicon esculentum Mill, tomato, free space, potassium, influx rate, ion uptake, leaf slices, leaf age leaf ontogeny     

Effect of Leaf Age on Potassium Efflux and Net Flux in Tomato Leaf Slices

The effects on leaf age on K (⁸⁶Rb) efflux, influx and net fluxinto lamina slices from leaf 7 on a tomato plant (Lycopersiconesculentum Mill.) were determined. The ontogenetic trend inK efflux was dependent on the external K concentration. At externalKCI concentrations between 0.5 and 10.0 mM, K efflux rates increasedduring leaf elongation. Only a small increase in efflux occurredin mature leaves with increasing age. It is suggested that thetonoplast retains its structural integrity through the initialstages of leaf senescence. In fully expanded leaves, a zeronet K flux (a balance between influx and efflux) was achievedat external KCI concentrations between 1.0 and 3.5 mM. The Kcontent of lamina slices from leaves 5 and 13 remained constantwhen bathed in a solution containing 2 to 3 mM K. It is suggestedthat the decline in K concentration in mature tomato leaf tissueis due to a decline in leaf free space K concentrations below1 to 3 mM which would result in a net efflux out of leaf cells. Lycopersicon esculentum Mill., tor ato, free space, ion fluxes, leaf age, leaf ontogeny, potassium     

Tomato Yellow Leaf Curl Virus Infection in a Monocotyledonous Weed (Eleusine indica)

Tomato yellow leaf curl virus (TYLCV) is one of the most important plant viruses belonging to the genus Begomovirus of the family Geminiviridae. To identify natural weed hosts that could act as reservoirs of TYLCV, 100 samples were collected at a TYLCV-affected tomato farm in Iksan from 2013 to 2014. The sample weeds were identified as belonging to 40 species from 18 families. TYLCV was detected in 57 samples belonging to 28 species through polymerase chain reaction using root samples including five species (Eleusine indica, Digitaria ciliaris, Echinochloa crus-galli, Panicum dichotomiflorum, and Setaria faberi) from the family Poaceae. Whitefly Bemisia tabaci-mediated TYLCV transmission from TYLCV-infected E. indica plants to healthy tomatoes was confirmed, and inoculated tomatoes showed typical symptoms, such as leaf curling and yellowing. In addition, TYLCV was detected in leaf and root samples of E. indica plants inoculated by both whitefly-mediated transmission using TYLCV-viruliferous whitefly and agro-inoculation using a TYLCV infectious clone. The majority of mastreviruses infect monocotyledonous plants, but there have also been reports of mastreviruses that can infect dicotyledonous plants, such as the chickpea chlorotic dwarf virus. No exception was reported among begomoviruses known as infecting dicots only. This is the first report of TYLCV as a member of the genus Begomovirus infecting monocotyledonous plants.     

番茄黄化曲叶病毒对番茄叶片光合特性和叶绿体超微结构的影响

以番茄为试验材料,研究番茄黄化曲叶病毒(TYLCV)侵染对植株叶片叶绿素含量、净光合速率、气孔导度、胞间CO2浓度和叶绿体超微结构的影响.结果表明:TYLCV侵染番茄后,叶片叶绿素a、b以及总叶绿素含量分别下降50.2%、24.19%和43.84%,叶片净光合速率和气孔导度分别下降43.28%、27.07%,胞间CO2浓度增加13.04%.与健康叶片相比,叶绿体变形,叶绿体基质片层大部分消解,基粒结构消失,叶绿体外膜和内膜剥离,质壁分离和细胞膜内陷,细胞器消解.研究表明,TYLCV侵染破坏了番茄叶片的叶绿体结构,严重影响番茄叶片的光合作用.