Development and evaluation of a technique for screening watercress (Rorippa nasturtium-aquaticum) for resistance to watercress yellow spot virus and crook-root fungus (Spongospora subterranea f. sp. nasturtii)

A technique was developed for screening large numbers of watercress plants for resistance to watercress yellow spot virus (WYSV) and the crook-root fungus. Plants were raised in modules containing sand in the glasshouse, transported and placed in experimental watercress beds, recovered after 5 or 6 weeks, examined visually for crook-root infection and tested by ELISA for infection by WYSV. High incidences of crook root (98.8%) and virus (88.9%) were obtained in a watercress line known to be susceptible to both pathogens. Evaluation of the technique using 10 different watercress lines showed that it was capable of revealing a range of responses from very susceptible to very resistant, with UK lines being most susceptible to both diseases. When grown on, the very resistant line was different morphologically from UK watercress and was identified as early winter-cress (Barbarea verna). Results showed an association between crook root scores and ELISA values, providing further circumstantial evidence for the close relationship between the two pathogens. The implications of the results for watercress resistance screening are discussed.     

Epidemiology of root rot caused by <Emphasis Type="Italic">Leptosphaeria maculans</Emphasis> in <Emphasis Type="Italic">Brassica napus</Emphasis> crops

The infection of above-ground tissues of Brassica napus by Leptosphaeria maculans is well understood. However, root infection (root rot) under field conditions, the development of root rot over time and its relationship to other disease symptoms caused by L. maculans has not been described. A survey of B. napus crops was conducted in Australia to investigate the incidence and severity of root rot. Additionally, the pathway of root infection was examined in field experiments. Root rot was present in 95% of the 127 crops surveyed. The severity and incidence of root rot was significantly correlated with that of crown canker; however, the strength of this relationship was dependent on the season. Root rot symptoms appeared before flowering and increased in severity during flowering and at maturity, a pattern similar to crown canker suggesting that the infection of the root is an extension of the crown canker phase of the L. maculans lifecycle. All isolates of L. maculans tested in glasshouse experiments caused root rot and crown canker in B. napus and Brassica juncea. In the field, the main pathway of root infection is via invasion of cotyledons or leaves by airborne ascospores, rather than from inoculum in the soil. Root rot was present in crops in fields that had never been sown to B. napus previously, in plants grown in fumigated fields, and in glasshouse-grown plants inoculated in the hypocotyl with L. maculans.     

棉花抗枯萎病品种苗期鉴定初步研究

通过棉花抗枯萎病品种苗期鉴定,可以反应出不同品种和品系的抗病性。温室系统观察结果,棉苗在1—2片真叶时即有症状出现,以感病品种发生较早。一般在4—5片真叶期,枯萎病发生甚烈。不同品种(系)反应的症状类型有所差异,但大多数属于三个类型:“网纹型”、“青枯型”、“黄叶型”。观察到“网纹”和“黄叶”属慢性类型;“青枯”属急性类型。从242个品种(系)材抖中,鉴定出苗期抗病的品种(系)23个,均属抗型(R)。发病指数0—10%有11系,11—20%有8系,21—30%有4系;其中有免疫类型1系(61—84),发病指数为0。苗期属抗病类型,移植到病圃后,成株期仍反应出较多抗病类型,其中苗期属高抗和免疫株系,成株期的抗性反应亦基本一致。     

Methods for Sampling and Assessment in Relation to the Spatial Pattern of Phoma Stem Canker (Leptosphaeria maculans) in Oilseed Rape

A sound assessment of phoma stem canker symptoms is needed to develop epidemiological, agronomical and physiological studies on the pathosystem. A specific analysis was therefore carried out to: (i) compare four methods of crown canker assessment; (ii) test the among and within assessor repeatability of one of the methods compared; (iii) characterise the spatial pattern of the disease; and (iv) define the sample size required to achieve a given level of disease assessment precision. The methods compared examined the symptoms with different procedures and graded the plants observed into six severity classes. A disease index (DI) summarised the severity distribution observed. Examination of crown cross-sections was the most precise method for assessing crown cankers. The method was repeatable, though an 'assessor effect' was apparent. The disease generally had a random pattern although significant spatial correlations were detected for four out of the fifteen plots studied at the scales examined. A relationship between the coefficient of variation of the DI and the sample size was established, evaluated with experimental field data and exemplified for typical severity distributions.     

Initial report of glufosinate and paraquat multiple resistance that evolved in a biotype of goosegrass (Eleusine indica) in Malaysia

Field and glasshouse studies have confirmed the presence of a glufosinate‐ and paraquat‐resistant goosegrass biotype that has infested a bitter gourd field in Air Kuning, Perak, Malaysia. Glufosinate and paraquat had been applied at least six times per year to the affected fields (originally a rubber plantation) for more than four consecutive years. Paraquat had been used since 1970 for weed control in the rubber plantation. An on‐site field trial revealed that the control of the goosegrass plants, measuring 20–35 cm in height, ranged from 20 to 35% 3 weeks after being treated with each herbicide at twice the recommended rate. Dose–response tests were conducted in the glasshouse, using seedlings at the three‐to‐four‐leaf stage that had been obtained from the plants that had received repeated exposure to these herbicides and a biotype with no history of any herbicide resistance. The comparison of the GR₅₀ (the herbicide rate that is required to reduce the shoot fresh weight by 50%) of the seedlings indicated that the resistant biotype of goosegrass is 3.4‐fold and 3.6‐fold more resistant than the susceptible biotype following treatment with glufosinate and paraquat, respectively. This study has demonstrated the world's first field‐evolved instance of multiple resistance in goosegrass to two non‐selective herbicides, glufosinate and paraquat.     

Screening tomato and cucurbit rootstocks for resistance to Verticillium dahliae*

Seedlings of tomato (19) and cucurbits (33), previously selected as rootstocks for commercial cultivars, were evaluated for their resistance to verticillium wilt under glasshouse conditions. Disease scoring was based on foliar symptoms, but in plants with no visual or ambiguous symptoms the presence of the pathogen was confirmed by isolations or PCR using specific DNA primers. Five of the cucurbit rootstocks showed resistance to infection, 11 were moderately infected, 11 were susceptible, while the remaining six were very susceptible. In general, Cucurbita pepo (courgette, pumpkin) and Lagenaria siceraria tolerated infection, Cucumis melo rootstocks were susceptible and Citrullus vulgaris was the most susceptible. When inoculated with race 1 of the pathogen, one of the tomato rootstocks was very resistant, three moderately resistant, nine tolerant, ten susceptible and one very susceptible. Pathogenicity tests with race 2 showed that none of the rootstocks exhibited high tolerance, but that there was some variation in susceptibility. Research is under way to evaluate the reaction of commercial cultivars to verticillium wilt when grafted onto resistant rootstocks.     

Examination of resistance to clubroot in accessions of Brassica oleracea using a glasshouse seedling test

A glasshouse test was elaborated for assessing large numbers of seedlings ofBrassica oleracea for resistance to clubroot, a disease caused by the fungusPlasmodiophora brassicae. The method offers good control of inoculum density per plant, and requires 6–7 weeks from sowing. The results from the glasshouse test correlated well with field test results. With this method, 71 accessions ofB. oleracea reported to carry resistance to clubroot, and one susceptible control cultivar were tested with a Dutch clubroot isolate. High levels of resistance were found in several accessions of cabbage, broccoli and curly kale. F₁-populations of resistant cabbage or curly kale × susceptible cabbage were fully susceptible, indicating recessive inheritance of resistance in all cases.     

Pesticide resistance in glasshouse whitefly (Trialeurodes vaporariorum west

A survey in SE England showed that seven out of 17 populations of glasshouse whitefly were resistant to DDT. Resistance to malathion occurred in 15 populations and to resmethrin in eight. Mecarbam and methiocarb gave good kills in screening tests but some insect growth regulators gave variable results. The synthetic pyrethroid permethrin was slightly more effective against a population of whitefly resistant to both DDT and malathion than against a susceptible population. Parthenogenetic females from a population susceptible to DDT and malathion produced almost entirely male progeny. An unsuccessful attempt was made to lower the resistance level of a DDT/malathion resistant strain by introducing these males to resistant females.     

Selection methods for determining resistance of carnation cultivars to Fusarium oxysporum f.sp. dianthi

The level of resistance of carnation ( Dianthus caryophyllus ) cultivars to wilt caused by Fusarium oxysporum f.sp. dianthi was compared in root-dip-inoculated plants grown in pots (filled with tuff or sandy soil) in a greenhouse and plants grown in a field where the soil was artificially infested with the fungus. In the field, wilt symptoms appeared first in susceptible and subsequently in resistant cultivars; none was immune. Variations in the level of resistance were expressed either by different percentages of wilted plants (i.e. disease incidence) or by delayed disease progress as compared to a susceptible cultivar. The range of disease severity in the field, ranked on a scale from 0 to 4, was highly and significantly correlated with the percentage of diseased plants. The greenhouse test was unreliable as a predictor of the degree of resistance observed in the field. Similar wilt levels in the greenhouse and the field were found only in susceptible cultivars.     

A Laboratory Assay for Phytophthora infestans Resistance in Various Solanum Species Reflects the Field Situation

Physiological and molecular research on resistance responses of Solanum tuberosum cultivars and partially resistant Solanum species to Phytophthora infestans requires a reliable resistance test that can be used in the laboratory. Laboratory tests performed on detached leaves and intact plants were compared with field tests for similarity of late blight reactions. Detached leaves from field-grown plants were as resistant as detached leaves from climate chamber-grown plants when challenged with P. infestans. However, detached leaves incubated in covered trays at high relative humidity were more susceptible than detached leaves kept in open trays or leaves on intact plants. The incubation conditions of detached leaves in covered trays rather than detachment itself appeared to affect the resistance expression. Detached leaves of some wild Solanum genotypes became partially infected, whereas intact plants were completely resistant when inoculated. Inoculation of leaves on intact plants, however, resulted in lower infection efficiencies. These limitations should be taken into account when choosing the appropriate inoculation method for specific purposes. For resistance screening, laboratory tests proved to be a good alternative for field tests. The ranking of resistance levels for twenty plant genotypes was similar under laboratory and field conditions.     

Assessment of resistance to Plasmodiophora brassicae in swedes

A method is described in which swede seedlings inoculated with standardized concentrations of resting spores of specific populations of Plasmodiophora brassicae can be evaluated for resistance in different environments. Similar ranking for resistance of four swede cultivars inoculated with one pathogen population was obtained from seedling tests in a glasshouse, from young plants in a polythene tunnel, and from mature plants in field trials. Differential resistance of the four cultivars to two pathogen populations evident in glasshouse seedling tests was more clearly demonstrated in the field where there was a highly significant cuitivar × pathogen population interaction (p<0 001) for both root fresh weight and mean disease category.
The inclusion of uninoculated control plants in field trials enabled a direct comparison of yield with that from inoculated plants. Cultivars were defined as resistant in terms of yield if they did not suffer any crop loss in comparison with uninoculated plants, even though some plants showed restricted gall development. The distribution of fresh weight to galls, roots and shoots could be used to characterize the relative resistance of cultivars; in the most susceptible cultivars there was rapid gall development but little increase in root or shoot fresh weight between 6 and 12 weeks after inoculation. Differential response was determined during the 6 weeks following inoculation; the implications of this observation are discussed in relation to growth stage and rate of development of host and pathogen.     

杨树水泡型溃疡病菌的致病性研究

 从杨树病斑上分离所得的水泡型溃疡病病原菌(Dothiorella gregaria Sacc.),经室内外的人工接种试验,均能诱致发病,产生与自然罹病相同的典型症状。室内人工接种保湿时间以10~14天较为适宜,接种最适温度为24℃~28℃,温度低于13℃不发病。该菌对阔叶树中的柳树、刺槐、核桃均有致病性。不同杨树种及品种对该菌的抗病性测定结果,白杨派的种及品种较抗病,其中尤以新疆杨最抗病;青杨派、黑杨派×青杨派、青杨派×黑杨派的派间杂种最易感病,其中尤以青杨、小青杨、北京杨感病最重;黑杨派的品种为中等感病。     

Effect of method of inoculation, inoculum density and seedling age at inoculation on the expression of resistance of cocoa (Theobroma cacao L.) to Verticillium dahliae Kleb.

Soil drench and stem puncture inoculation were compared as methods for selecting cocoa cultivars with resistance to Verticillium dahliae. Disease progress was more rapid and induced symptoms were more severe following stem puncture and, under glasshouse conditions, differences between cultivars were detected 15 days after inoculation. Moreover, using stem puncture, inoculum densities of 10⁴ conidia/ml were sufficient to differentiate resistant and susceptible cultivars, whereas with the soil drench method, inoculum densities of 10⁷ conidia/ml were necessary. Although a substantially higher proportion of plants were affected by stem puncture inoculation, the resistance of cultivar Pound-7 remained effective at high inoculum densities of 10⁸ conidia/ml. With either method, older seedlings were more susceptible to V. dahliae than younger ones. However, with stem puncture, 15-day-old seedlings were sufficiently susceptible for a valid disease assessment. In contrast, with soil inoculation, 60-day-old plants were required. In a nursery trial with 15-day-old seedlings, seven cocoa genotypes previously selected as resistant, moderately resistant or susceptible to Verticillium dahliae , on the basis of root inoculation, were ranked in the same order when stem punctured. Stem puncture inoculation of young seedlings is cost-effective in terms of time and space, and is therefore recommended for screening of cocoa for wilt resistance, especially in large-scale breeding programmes.     

Effect of Host Plant Resistance to Tomato yellow leaf curl virus (TYLCV) on Virus Acquisition and Transmission by Its Whitefly Vector

ABSTRACT The effect that Tomato yellow leaf curl virus (TYLCV)-infected resistant tomato plants may have on virus epidemiology was studied. Four tomato genotypes that exhibit different levels of viral resistance, ranging from fully susceptible to highly resistant, served as TYLCV-infected source plants. Viral acquisition and transmission rates by white-flies following feeding on the different source plants were evaluated. TYLCV transmission rate by whiteflies that had fed on infected source plants 21 days postinoculation (DPI), shortly after the appearance of TYLCV symptoms, was negatively correlated with the level of resistance displayed by the source plant. Therefore, the higher the resistance, the lower the transmission rate. In addition, TYLCV DNA accumulation was shown to be lower in the resistant source plants compared with the susceptible plants. Whitefly survival rate, following feeding on source plants 21 DPI, was similar for all the cultivars tested. Significant differences in whitefly survival were found, however, following feeding on the infected source plants at 35 DPI; here, whitefly survival rate increased with higher levels of resistance displayed by the source plant. At 35 DPI, the susceptible plants had developed severe TYLCV disease symptoms, and transmission rates from these plants were the lowest, presumably due to the poor condition of these plants. Transmission rates from source plants displaying a medium level of resistance level were highest, with rates declining following feeding on source plants displaying higher levels of TYLCV resistance. TYLCV DNA accumulation in whiteflies following feeding on infected source plants at both 21 and 35 DPI was directly correlated with viral DNA accumulation in source plants. Results show that, in essence, the higher the resistance expressed, the less suitable the plant was as a viral source. Consequently, following acquisition from a highly resistant plant, TYLCV transmission by whiteflies will be less efficient.     

Comparison of Resistance to Tomato Leaf Curl Virus (India) and Tomato Yellow Leaf Curl Virus (Israel) among Lycopersicon Wild Species, Breeding Lines and Hybrids

The objective of this study was to screen wild and domesticated tomatoes for resistance to Tomato yellow leaf curl virus, Israel (TYLCV-Is) and Tomato leaf curl virus from Bangalore isolate 4, India (ToLCV-[Ban4]) to find sources of resistance to both viruses. A total of 34 tomato genotypes resistant/tolerant to TYLCV-Is were screened for resistance to ToLCV-[Ban4] under glasshouse and field conditions at the University of Agricultural Sciences, Bangalore, India. Resistance was assessed by criteria like disease incidence, symptom severity and squash-blot hybridization. All the tomato genotypes inoculated with ToLCV-[Ban4] by the whitefly vector Bemisia tabaci (Gennadius) produced disease symptoms. In some plants of the lines 902 and 910, however, the virus was not detected by hybridization. The tomato genotypes susceptible to ToLCV-[Ban4] by whitefly-mediated inoculation were also found susceptible to the virus under field conditions. However, there were substantial differences between genotypes in disease incidence, spread, symptom severity and crop yield. Despite early disease incidence, many genotypes produced substantially higher yields than the local hybrid, Avinash-2. Sixteen tomato genotypes from India resistant/tolerant to ToLCV-[Ban4] were also tested for TYLCV-Is resistance at the Hebrew University of Jerusalem, Rehovot, Israel. Accessions of wild species, Lycopersicon hirsutum LA 1777 and PI 390659 were the best sources of resistance to both viruses. Lines 902 and 910, which were, resistant to TYLCV-Is were only tolerant to ToLCV-[Ban4] and accession Lycopersicon peruvianum CMV Sel. INRA, resistant to ToLCV-[Ban4], was only tolerant to TYLCV-Is. Implications of using the resistant lines in breeding programme is discussed.     

溃疡病菌对杨树几丁酶、β-N-乙酰氨基葡萄糖苷酶的诱导作用

 杨树溃疡病菌Dothiorella gregaria及其菌丝体提取物都能诱导杨树几丁酶、β-N-乙酰氨基葡萄糖苷酶的积累,但在诱导的速度和强度上,抗病品种和感病品种有明显的差异。抗病品种毛白杨这两种酶不仅积累的速度快,而且幅度也远大于感病品种北京杨。在D.gregaria感染实验中,毛白杨几丁酶和β-N-乙酰氨基葡萄糖苷酶活性升高的幅度分别是北京杨的4.7倍和3倍;在菌丝体提取物的诱导实验中,毛白杨这两种酶升高的幅度分别是北京杨的4.5倍和2.7倍。因此可以认为几丁酶和β-N-乙酰氨基葡萄糖苷酶与杨树对溃疡病的抗性反应有关,是其防卫系统的要素之一。     

Development of a rapid,accurate glasshouse bioassay for assessing fusarium wilt disease responses in cultivated Gossypium species

A rapid glasshouse‐based bioassay method to screen large numbers of cotton plants for responses to Fusarium oxysporum f. sp. vasinfectum (Fov) was developed. Different Fov inoculum concentrations and methods of inoculation were assessed using resistant and susceptible cotton cultivars. Cotton seeds were planted directly into Fov‐inoculated soil. Studies of seed germination, seedling establishment, seedling mortality and fusarium wilt symptoms (i.e. stunting, foliar symptoms and vascular browning) were performed to optimize the bioassay parameters. Growing seedlings in Fov‐inoculated soils at 5 × 10⁴ or 1 × 10⁵ CFU g^?1 soil, in individual seedling tubes with 12 h at 28–30°C and 12 h at 15–18°C, gave consistent results when assessing Fov disease responses 6 weeks after inoculation. When fusarium wilt resistance ranks (FWRRs) and vascular browning index (VBI) means of 18 Australian and other cotton cultivars from the Fov glasshouse bioassay were compared against their fusarium field performance ranks (F‐ranks), assessed on adult plants for cotton cultivar release, Pearson’s correlation was highly significant for both comparisons. The level of congruence between field and glasshouse data indicated that this protocol should be an effective tool for large‐scale screening for Fov‐resistance responses in diverse germplasm and breeding populations and for advancing genetic research to develop molecular markers for Fov resistance in cotton.     

Systemic growth of Leptosphaeria maculans from cotyledons to hypocotyls in oilseed rape: influence of number of infection sites, competitive growth and host polygenic resistance

The influence of competitive effects between two isolates, of the number of infection sites on cotyledons and of host polygenic resistance on the systemic growth of Leptosphaeria maculans , the cause of phoma stem canker in oilseed rape ( Brassica napus ), were investigated. Controlled-condition experiments were conducted with two oilseed rape doubled haploid lines, one susceptible and the other with a high level of polygenic resistance, inoculated via wounded cotyledons with conidial suspensions obtained from two isolates. Expression of cankers in plants was enhanced by exposing inoculated plants to low temperature (6°C) followed by warm temperature (20°C). The fungus was detected by PCR amplifications of three minisatellite markers in all stems with visible canker symptoms and also in the stems of 14 of the 59 plants without visible cankers on the hypocotyls. Disease severity increased with the number of infection sites on cotyledons: in one of the three replicate experiments, the mean external necrosis length on the hypocotyl ranged from 6·47 to 35·3 mm for one and eight infections sites on cotyledons, respectively. The probability of an isolate reaching the hypocotyl from inoculated cotyledons decreased with increasing competing inoculum load on cotyledons: for instance, for isolate A290v it decreased from 1 when inoculated alone to 0·28 when coinoculated with six drops of competing isolate P27d. Polygenic resistance significantly reduced disease incidence and severity. For instance, in one of the three replicate experiments, disease incidence ranged from more than 74% in susceptible plants to 16% in resistant ones, while mean external necrosis length was up to 35·3 and 6·5 mm on susceptible and on resistant plants, respectively. This study offers new possibilities for assessing levels of polygenic resistance to stem canker in B. napus and studying the aggressiveness of L. maculans isolates.     

Factors affecting the susceptibility of six cocoa clones to Phytophthora palmivora (Butl.) Butler bark canker in Trinidad

An assessment of canker incidence in six cocoa clones revealed IMC 67 to be least susceptible and TSH 1076 most susceptible among the clones tested (ICS 1, IMC 67, P 18, SCA 6, TSH 1076 and TSH 1188). These results were confirmed by greenhouse studies on budded plants using artificial inoculation. IMC 67 with the smallest canker lesion size was found to have the highest value (3.75 N/m²) for extra-xylary tissue hardness while TSH 1076 with the largest lesion size had the lowest value (1.9 N/m²). Bark moisture content determination also showed lowest values for IMC 67 and highest for TSH 1076. Bark hardness and moisture content were highly correlated with canker resistance.     

Susceptibility of Douglas fir (Pseudotsuga menziesii) to pitch canker, caused by Gibberella circinata (anamorph = Fusarium circinatum)

For better characterization of the risk of pitch canker (caused by Gibberella circinata , anamorph =  Fusarium circinatum ) to Douglas fir ( Pseudotsuga menziesii ), Californian isolates, selected exotic isolates, and ascospore progeny of a cross between wild-type Californian isolates were tested for aggressiveness to this host species. In addition, seedlings from representative provenances of P. menziesii in California were tested for susceptibility to pitch canker. The results revealed only minor differences between isolates, but differences in susceptibility between trees were often significant. The majority of the tested trees were relatively resistant as indicated by the development of only very short lesions, but some were clearly susceptible.