The relationship between hypersensitive tissue and resistance in wheat seedlings infected with Puccinia graminis tritici

Data obtained during the first 120 hr. of the infection process in several resistant and susceptible varieties of wheat infected with Puccinia graminis tritici did not show any consistent relationship between the growth of the rust colony and the rate of increase of host tissue showing hypersensitive necrosis. Furthermore, no apparent relationship existed between the area of the rust colony at any given time after inoculation, and the amount of hypersensitive necrotic tissue per unit area of the leaf colonized. Thus, it is suggested that hypersensitive necrosis of the host tissue is a consequence and not the cause of resistance of wheat to infection by P. graminis tritici.     

Effect of Host Plant Resistance on Haustorium Formation in Cereal Rust Fungi

Haustoria of Puccinia triticina (wheat leaf rust fungus) and P. hordei (barley leaf rust fungus) were isolated from susceptible and partially resistant wheat lines, and susceptible, hypersensitive and partially resistant barley lines. Haustoria were counted and measured. The size of haustoria was similar in the partially resistant and susceptible genotypes but haustoria were smaller in the hypersensitive barley line L94+Pa7. The number of haustoria was reduced in both partially and hypersensitive lines when compared with susceptible ones. Therefore it seems that the reduction in the number of haustoria is a consequence of the resistance that can be attributable either to early abortion of infection units or reduced colony growth. The reduction of the number of haustoria was more pronounced in the adult plant stage.     

Selection, biological properties and fitness of resistance-breaking strains of Tomato spotted wilt virus in pepper

In glasshouse tests, infective sap from plants infected with 17 different isolates of Tomato spotted wilt virus (TSWV) from four Australian states was inoculated to three Capsicum chinense accessions (PI 152225, PI 159236 and C00943) carrying single genes that confer hypersensitive resistance to TSWV. The normal response to inoculation was development of necrotic (hypersensitive) local lesions in inoculated leaves without systemic invasion, but 3/1386 infected plants also developed systemic susceptible reactions in addition to hypersensitive ones. Similarly when two isolates were inoculated to C. chinense backcross progeny plants, 1/72 developed systemic susceptible reactions in addition to localised hypersensitive ones. Using cultures from the four plants with susceptible reactions and following three to five further cycles of serial subculture in TSWV‐resistant C. chinense plants, four isolates were obtained that gave systemic susceptible type reactions in the three TSWV‐resistant accessions, and in TSWV‐resistant cultivated pepper (C. annuum). When three of these isolates were inoculated to tomato (Lycopersicon esculentum) breeding lines with single gene resistance to TSWV, resistance was not overcome. Similarly, none of the four isolates overcame partial resistance to TSWV in Lactuca virosa. When TSWV isolates were inoculated to tomato breeding lines carrying partial resistance from L. chilense, systemic infection developed which was sometimes followed by ‘recovery’. After four successive cycles of serial passage in susceptible cultivated pepper of a mixed culture of a resistance‐breaking isolate with the non resistance‐breaking isolate from which it came, the resistance‐breaking isolate remained competitive as both were still found. However, when the same resistance‐ breaking isolate was cultured alone, evidence of partial reversion to wild‐type behaviour was eventually obtained after five but not four cycles of long term serial subculture in susceptible pepper, as by then the culture had become a mixture of both types of strain. This work suggests that resistance‐breaking strains of TSWV that overcome single gene hypersensitive resistance in pepper are relatively stable. The findings have important implications for situations where resistant pepper cultivars are deployed widely in the field without taking other control measures as part of an integrated TSWV management strategy.     

Isolation of Race-and Cultivar-Specific Elicitors from Intercellular Fluids of Compatible Interactions of Pseudomonas syringae pv. pisi and Pea

Intercellular fluids obtained by an in vacuo infiltration technique from compatible race-cultivar interactions of five races of Pseudomonas syringae pv. pisi and pea induced extensive light brown necrotic (hypersensitive type of) lesions in resistant but not susceptible cultivars. In susceptible cultivars the intercellular fluids induced extetensive water soaking symptoms. The intercellular fluids elicited intermediate reactions in pea cultivars of moderate resistance. The intensity of the light brown necrotic reactions in pea appears to be positively correlated with the degree of resistance.     

Induction of systemic resistance in different varieties of Solanum tuberosum by pure and crude elicitor treatment

A 10 kD elicitor protein (infestin) produced by Phytopthora infestans was purified and its efficacy for induction of systemic resistance in resistant and susceptible varieties of Solanum tuberosum was studied. Culture filtrates from P. infestans with and without purified elicitor (infestin) were used as elicitors to understand the effect of purified elicitor (infestin) on development of systemic resistance. Culture filtrate and purified elicitor (infestin) were found to induce hypersensitive reaction on the leaves of resistant varieties, but not on susceptible varieties after 48 h. Culture filtrate devoid of purified elicitor (infestin) did not induce any necrotic spots even on resistant variety. Purified elicitor (infestin) was found to induce glucose oxidase, NADPH oxidase, superoxide dismutase, glutathione reductase, catalase and peroxidase enzymes in resistant S. tuberosum plants, however the induction of these enzymes was low in susceptible varieties. The oxidative enzymes were found to induce earlier than antioxidative enzymes and there was negative correlation between these two groups of enzymes. Levels of salicylic acid, phenylalanine ammonia lyase (PAL), beta-1, 3 glucanase and chitinase activities were also found higher in resistant than in susceptible varieties. It was observed that purified elicitor (infestin) was superior to crude culture filtrate, but was not capable of inducing systemic resistance in susceptible varieties.     

Behavioral and physiological responses of susceptible and resistant diamondback moth larvae to Bacillus thuringiensis

To determine whether field-selected resistance of diamondback moth (Plutella xylostella L.) (Lepidoptera: Plutellidae) to Bacillus thuringiensis is based on behavioral or physiological adaptation, we measured mortality, consumption, and movement of larvae from a susceptible and a resistant colony when placed on untreated and B. thuringiensis treated cabbage. Colonies did not differ in mortality, consumption, or movement on untreated cabbage. However, for a given amount of consumption of treated cabbage, resistant larvae had lower mortality than susceptible larvae, demonstrating that resistance had a physiological basis. The movement patterns could not account for the differences between colonies in survival. Resistant larvae did not avoid B. thuringiensis more than did susceptible larvae. Thus, we found no evidence for behavioral resistance.     

The behaviour of Bretnia lactucae on cultivars of Lactuca sativa and on other composites

The spores of an isolate of Bremia lactucae obtained from lettuce germinated equally well on Cellophane and on the surfaces of all plant species inoculated. The fungus penetrated the epidermal cells of both host and non-host plants but extensively invaded only certain lettuce cultivars. Many of the species and cultivars inoculated showed no macroscopic symptoms but exhibited a hypersensitive response to invasion that was limited to a few cells and only visible microscopically. However, the hypersensitive response was not always so limited and in some species, fungal development and associated host cell necrosis was more extensive and visible macroscopically. In such instances, limited sporulation occasionally occurred. On some lettuce cultivars symptoms developed more slowly than on fully susceptible ones and less seedlings became diseased. This was due to the heterogeneity of the inoculum with respect to its ability to overcome certain race specific resistance genes. Such cultivars reacted hypersensitively to the majority, but not all, of the fungal propagules applied and the observed effect was thus due to a lower effective inoculum concentration. Without due care, both these types of ‘intermediate’ reaction could easily be misinterpreted as race non-specific resistance particularly under field conditions. These observations emphasized the problems of categorizing cultivars and species rigidly as resistant or susceptible when inoculum could be heterogenous or when resistance reactions allowed some macroscopic symptom development.     

Studies on the nature of resistance in tomato plants to Verticillium albo-atrum

Extracts of healthy resistant and of healthy susceptible plants of tomato had the same effect on growth of Verticillium albo-atrum in vitro. Tracheal saps from resistant and from susceptible plants showed no difference in their effect on spore germination and mycelial growth of V. albo-atrum. Cuttings from resistant plants, inoculated with V. albo-atrum and fed with low concentrations of casamino acids, or glucose, at first wilted more than controls but soon recovered. Continuous treatment with dilute ethanol solutions for 2 weeks induced marked wilting in inoculated cuttings of resistant plants: treatment for shorter periods caused less severe symptoms, from which cuttings recovered slowly. Metabolic inhibitors did not break resistance of cuttings, but the pathogen survived longer in cuttings treated with sodium diethyldithiocarbamate, salicylaldoxime or 8-hydroxyquinoline than in controls. When one end of segments of stems of resistant plants was inoculated with the pathogen, and 48 h later the uninoculated end was placed near a colony of V. albo-atrum on agar, growth of the fungus colony towards the stem segment was sometimes inhibited. There was no such inhibition when segments from susceptible plants were used. Both tracheal sap and diffusates from segments of inoculated resistant plants supported less growth of germ tubes of V. albo-atrum than sap and diffusates from uninoculated plants. These differences were not obtained with the susceptible variety and production of fungitoxic substances in resistant plants after infection is inferred.     

Antixenotic resistance of subterranean clover cotyledons to redlegged earth mite, Halotydeus destructor

The short-term response of redlegged earth mite, Halotydeus destructor (Tucker) (Acarina: Penthaleidae) to cotyledons of different varieties of subterranean clover (Trifolium subterraneum subsp. subterraneum L.) was assessed by means of paired choice tests, and no-choice tests. H. destructor had lower numbers and fed less on detached cotyledons of resistant than susceptible varieties, yielding a correlation between the numbers of mites and feeding damage to the cotyledons during a three hour test period. For a number of resistant and susceptible varieties, there was a negative correlation between cotyledon deterrence in the three hour choice test and feeding damage to seedling after a two week period. Since the response of the mites to different subclover varieties occurred within three hours, it is concluded that the resistance is based on antixenosis.No evidence was obtained for antifeedant activity in organic solvent extracts from the variety DGI007 (resistant) in comparison with those from the variety Dalkeith (susceptible). Water soluble compounds from DGI007 cotyledons were preferred by mites, in feeding tests in terms of numbers, over those from Dalkeith (susceptible). Squeezed sap from the cotyledons of both varieties showed the same effects on mites as 5% glucose and were more phagostimulatory than water extracts. Mechanically damaged cotyledons of Dalkeith and DGI007 attracted more mites than the undamaged counterparts. The toughness of cotyledons in 17 varieties of T. subterraneum subsp. subterraneum was measured with a manual penetrometer. Results showed a negative correlation between toughness values and mite feeding damage scores (r²=0.752) for all varieties except S3615D (resistant). This implies a likely involvement of epidermal toughness as a contributor in the antixenotic resistance of these varieties. Other mechanisms may be involved in the resistance of S3615D.     

Phytophthora Root Rot of Pepper Influence of Host Genotype and Pathogen Strain on the Inoculum Density-Disease Severity Relationships

The relationship between inoculum density and mortality or infection was studied for various pepper varieties (Capsicum annuum L.) inoculated with zoospores of two P. capsici isolates. The inoculum concentrations required for 50% mortality (LD 50) varied greatly between pepper varieties and P. capsici isolates: with one isolate, LD 50 was 40 zoospores/ml for a susceptible variety and reached 4,380 to 97,300 zoospores/ml for resistant varieties. For another isolate, LD 50 for the, same varieties ranged from 26 to 800 zoospores/ml. Comparisons between LD 50 and inoculum doses required for 50 % Infection (ID 50) also revealed differences between varieties but not between isolates. After multiple infection correction, regression slopes of infections/inoculum concentration were low for resistant varieties (0.28 to 0.50) but higher for susceptible varieties (0.72 to 0.94), indicating strong competition between spores for infection of resistant plants, but not for infection of susceptible plants. This analysis provided many criteria which can be used to differentiate susceptible from resistant varieties and to evaluate with precision the resistance level of the different resistance genitors used in our breeding program.     

Deletion of a large genomic segment in tobacco varieties that are resistant to potato virus Y (PVY)

Recessive alleles (va, va ¹ , va ² , etc) of the tobacco Va locus confer resistance to potato virus Y (PVY). To elucidate the mechanism underlying this resistance, we attempted to identify randomly amplified polymorphic (RAPD) markers that reveal polymorphism between two nearly isogenic lines (NILs) that differ in their susceptibility to PVY. Using each of 500 primers and 800 pairs of primers, we identified over 100 RAPD fragments that differed between the NILs. We applied these RAPD primers or primer combinations to an F₂ population obtained from a cross between the susceptible line BY4 and the resistant va ² -bearing NIL, F55. It was found that only 10 RAPD markers were polymorphic between resistant and susceptible plants. Unexpectedly, these markers were all linked to Va. All 10 RAPD markers were present in all 8 susceptible varieties tested. At least one RAPD marker was not detected in 8 out of 10 resistant varieties. Southern analysis revealed that the sequences of markers were not present in the genomes of resistant varieties, and the markers were found in individually distinct positions on the chromosomes of susceptible tobacco varieties. These results strongly suggest that the resistance conferred by va is due to deletions at the Va locus governing susceptibility to PVY. Received: 20 May 1999 / Accepted: 17 August 1999     

The response of tomato seedling roots to infection by Verticillium albo-atrum or Fusarium oxysporum f. sp. lycopersici

The response of seedling roots of near-isogenic tomato varieties to infection by Verticillium albo-atrum or Fusarium oxysporum f. sp. lycopersici was investigated. Studies of the infection of seedling roots not artificially damaged indicated that there was an extra-vascular expression of resistance towards V. albo-atrum but not to F. oxysporum. Roots of resistant tomato seedlings infected by V. albo-atrum contained the fungus in the epidermis and outer cortex while susceptible roots became heavily colonised. Observations made by transmission electron microscopy showed that the fungus appeared to be abnormal in growth and appearance in the epidermal and cortical cells of resistant seedling roots but normal in susceptible roots. Two preformed antifungal terpenoids were detected in seedling roots in greater amounts in resistant that in susceptible varieties. The possible mechanisms of seedling root resistance to vascular wilts are discussed.     

cDNAs generated from individual epidermal cells reveal that differential gene expression predicting subsequent resistance or susceptibility to rust fungal infection occurs prior to the fungus entering the cell lumen

As the cowpea rust fungus penetrates the wall of a cowpea epidermal cell, resistant and susceptible plants exhibit different ultrastructural and cytochemical changes within the epidermal protoplast. To examine plant gene expression at this stage of infection, cytoplasm was extracted from individual inoculated or uninoculated epidermal cells before the fungal penetration peg reached the cell lumen. Initial differential colony hybridization screening of an expressed sequence tag library constructed from globally amplified cDNAs generated from the inoculated resistant cells resulted in 80 clones (out of 835) with a differential hybridization pattern. Further slot-blot screening and screening of the amplified cDNAs generated from inoculated or uninoculated, resistant or susceptible cells revealed 28 separate genes, mostly with no matching sequences in the databases, that were up-regulated in response to the growth of the fungus through the wall of resistant or susceptible cells. Five genes, including those coding for beta- and alpha-tubulin, were found to be down-regulated specifically in inoculated, susceptible cells, and five were specifically up-regulated in inoculated, resistant cells, including a PR-10 homolog and a phenylalanine ammonia-lyase gene. Probing the amplified cDNAs from each cell type for the expression of cell death-related genes revealed that an LLS1 homolog (vuLLS1), cloned from cowpea, was up-regulated by infection in both resistant and susceptible cells and that a homolog of HSR203J was differentially up-regulated in resistant cells. These data show that changes in gene expression predicting the subsequent expression of susceptibility or hypersensitive resistance to fungal infection occur prior to the fungus entering the cell lumen.     

Blockage of stylet tips as the mechanism of resistance to virus transmission by Aphis gossypii in melon lines bearing the Vat gene

Aphis gossypii is the main virus vector in muskmelon crops. The melon gene Vat confers resistance to non‐persistent virus transmission by this aphid. The mechanism of this resistance is not well understood, but no relationship has been detected between resistance and the probing behaviour of aphids on resistant plants. Results presented here suggest that temporary blockage of aphid stylet tips preventing virus particle release may explain the resistance conferred by Vat gene. We performed experiments in which viruliferous aphids were allowed to probe different sequences of resistant (Vat‐bearing) and/or susceptible melon plants. The results demonstrated that A. gossypii inoculates Cucumber mosaic virus (CMV) efficiently in susceptible plants having previously probed resistant plants, showing that the resistance mechanism is reversible. Furthermore, the infection rate obtained for susceptible plants was the same (25%) regardless of whether the transmitting aphid had come directly from the CMV source or had subsequently probed on resistant plants. This result suggests that virus is not lost from stylet to plant during probing of resistant plants, supporting the temporary blockage hypothesis. We also found that the ability of Myzus persicae to transmit CMV is noticeably reduced after probing on resistant plants, providing evidence that this aphid species also responds to the presence of the Vat gene. Finally, we also found that in probes immediately after virus acquisition M. persicae inoculates resistant plants with CMV more efficiently than susceptible plants, perhaps because the Vat gene product induces increased salivation by this aphid.     

Sources and inheritance of resistance to downy mildew of the pea, Peronospora pisi

Six hundred and one lines from the John Innes Pisum germplasm collection were surveyed for resistance to downy mildew (Peronospora pisi). Potential sources of resistance were identified in forty-seven lines. Using the inoculation methods described resistant varieties/lines showed no evidence of infection. Isolates from recent outbreaks in the United Kingdom when screened against a representative test array of resistant and susceptible lines showed no evidence for a race structure in Peronospora pisi, although differences were found in overall virulence. The inheritance of resistance was studied in F₂ and F₃ families. Under the test conditions adopted the results obtained suggest that resistance may either be determined by a single dominant gene or by two recessive genes, but the lack of concordance between F₂ and F₃ segregation patterns was a disturbing feature despite careful control of experimental conditions. This, coupled with difficulties in obtaining large F₃ families presents considerable problems in interpretation. It is proposed that inbred lines of JI 411 Cobri and JI 399 Cennia be adopted as standards.     

甘薯品种抗黑斑病鉴定及其遗传趋势

对2745份甘薯品种及育种材料的黑斑病抗性鉴定结果表明,高抗型占7．7％,抗病型占17．7％,中抗型占23.2％,感病型占27．2％,高感型占24．2％。对898份材料及其亲本的抗性分析表明,不同抗性组合后代中均可分离出高抗至高感类型的材料,杂交后代的抗性强弱随双亲抗性水平的增加而提高。相关分析显示,甘薯品种抗黑斑病性与其它主要经济性状间没有相关关系。采用品种阃杂交育种技术先后育成了一批高产、优质的抗病型优良品种。