Screening techniques and sources of resistance against parasitic weeds in grain legumes

Summary A number of parasitic plants have become weeds, posing severe constraints to major crops including grain legumes. Breeding for resistance is acknowledged as the major component of an integrated control strategy. However, resistance against most parasitic weeds is difficult to access, scarce, of complex nature and of low heritability, making breeding for resistance a difficult task. As an exception, resistance against Striga gesnerioides based on a single gene has been identified in cowpea and widely exploited in breeding. In other crops, only moderate to low levels of incomplete resistance of complex inheritance against Orobanche species has been identified. This has made selection more difficult and has slowed down the breeding process, but the quantitative resistance resulting from tedious selection procedures has resulted in the release of cultivars with useful levels of incomplete resistance. Resistance is a multicomponent event, being the result of a battery of escape factors or resistance mechanisms acting at different levels of the infection process. Understanding these will help to detect existing genetic diversity for mechanisms that hamper infection. The combination of different resistance mechanisms into a single cultivar will provide durable resistance in the field. This can be facilitated by the use of in vitro screening methods that allow highly heritable resistance components to be identified, together with adoption of marker-assisted selection techniques.     

Resistance of cowpea and cereals to the parasitic angiospermStriga

Summary Striga species are parasitic angiosperms that attack many crops grown by subsistence farmers in sub-Saharan Africa and India. Control of the parasite is difficult and genetically resistant crops are the most feasible and appropriate solution. In cowpea, complete resistance toStriga gesnerioides has been identified. Breeding for resistance in sorghum has identified varieties with good resistance toS. asiatica in Africa and India. One variety was also resistant toS. hermonthica in W. Africa. No such resistance toStriga has been found in maize or millets.Resistant varieties have usually been sought by screening germplasm in fields naturally infested withStriga. However, laboratory techniques have also been developed, including anin vitro growth system used to screen cowpeas for resistance toS. gesnerioides. Two new sources of resistance in cowpea have been identified using the system. The technique has also been used to investigate the mechanisms of resistance in this crop. Two mechanisms have been characterised, both were expressed after penetration of cowpea roots by the parasite.The resistance of some sorghum varieties toStriga is controlled by recessive genes. In cowpea, resistance toStriga is controlled by single dominant genes. The genes for resistance are currently being transferred to cowpea varieties which are high yielding or adapted to local agronomic conditions. OneStriga resistant cowpea variety, Suvita-2, is already being grown widely by farmers in Mali. Reports of breakdown of resistance in cowpea toStriga have not yet been confirmed, but a wider genetic base to the resistance is essential to ensure durability ofStriga resistance.Abbreviations ICRISAT International Crops Research Institute for the Semi-Arid Tropics - IITA International Institute of Tropical Agriculture - LARS Long Ashton Research Station - SAFGRAD Semi-Arid Food Grain Research and Development     

Screening for resistance against rhopalosiphum padi (L.). I. avena species and breeding lines

Summary Thirty-two accessions of Avena species and breeding lines were evaluated for resistance, primarily antibiosis, to the aphid Rhopalosiphum padi (L.) in greenhouse and growth room tests. The highest levels of resistance were found in A. barbata and in the perennial species A. macrostachya. One breeding line, Obee, was also found to have interesting resistance characters. Resistance in A. macrostachya is discussed in relation to perenniality. A different screening method for rapid testing of large collections of varieties is evaluated.     

Sources of resistance to Cephalosporium gramineum in Triticum and Agropyron species

Summary Resistance to the soil-borne pathogen Cephalosporium gramineum was evaluated in Agropyron elongatum, A. intermedium. A. intermedium var. trichophorum, an Agrotriticum, and eight species of Triticum. Only A. elongatum and A. intermedium showed high levels of resistance. Agrotriticum (56 chromosomes) was resistant too. High resistance to C. gramineum is available, but its utilization will probably require the use of chromosome substitution techniques to transfer the resistance into an agronomically useful wheat.     

Somaclonal variation for resistance to Verticillium dahliae in potato (Solanum tuberosum L.) plants regenerated from callus

Summary Plant tissue culture is recognized as an important tool to generate useful genetic variability for crop improvement. Regenerated plants from callus induced from stem explants of Solanum tuberosum cv Désirée were assessed by in vitro selection, for resistance to Verticillium dahliae. This fungus is the causal agent of Verticillium wilt, a serious vascular wilt disease both in crops and wild species.The rate of in vitro multiplication by single node cuttings was used as a parameter of screening in two selection cycles with different concentrations of V. dahliae filtrate. One resistant clone was selected and then evaluated by inoculation in the growth chamber. Induced damage, and morphological traits (dry weight, leaf area and tuber production) were estimated. The selected clone was comparable to the resistant control, cv Kondor.The results suggest that genetic variation induced in tissue culture cound be utilized to generate disease resistance.     

Selection parameters for resistance to Fusarium oxysporum f. sp. cubense race 1 and race 4 on diploid banana (Musa acuminata Colla)

Summary Shoot tip cultures from banana clones susceptible and resistant to Fusarium oxysporum f. sp. cubense (FOC) race 1 and race 4 were grown in vitro in the presence of different concentrations of fusaric acid and fungal crude filtrates or inoculated with a conidial suspension of FOC to assess correlation between in vivo and in vitro behaviour. Explants were susceptible to both filtrate and fusaric acid irrespective to their known field resistance/susceptibility response. No clear linkage between in vivo and in vitro behaviour was observed and our results suggest that the use of crude filtrate or non-host specific toxin (fusaric acid) in a screening programme for selecting a novel resistant genotype of Musa to FOC is not feasible. When peroxidase activity was used as a parameter to discriminate between sesceptibility and tolerance, results were in good agreement with field response of host plant to pathogens. Early enzymatic activity increased in the incompatible host-pathogen interaction but not in the compatible interaction.Abbreviations IBA Indolebutyric acid - 2iP 6-dimethylallylamino-purine - VCG Vegetative Compatibility Group - FOC Fusarium oxysporum f. sp. cubense - IEF isoelectrofocusing     

In vitro screening of strawberry plants for cold resistance

Formation of embryo autonomy of strawberry, plant regeneration fro membryo components, plant freezing conditions in vitro and the possibility to differentiate objectively genotypes by freezing them in vitro and in vivo were studied to create strawberry screening technology in vitro for cold resistance. It was established that autonomy of strawberry embryos manifests itself not earlier than on 14–16^th day after pollination and full autonomy is reached on 20–22^nd day. Plants regenerated from 26 days old embryos grew most intensively. At the highest rate strawberry plants regenerated from an isolated embryo axis on MS medium without phytohormones, and from rescued cotyledons x on the medium with 1.0 BA and 0.5 NAA. The temperature interval, at which genotypes differentiated according to cold resistance in vitro, was -8 to 12 ^°C. Differentiation of strawberry genotypes according to this character conformed to their differentiation in vivo, provided hardening proceeded not less than 21 days. The correlation between cold resistance in vitro and in vivo reached 0.93. Domination of cold resistance manifested itself in strawberry seedlings from various crossing combinations. This revised version was published online in August 2006 with corrections to the Cover Date.     

Resistance to stripe rust in Triticum turgidum,T. tauschii and their synthetic hexaploids

Resistance to stripe rust (caused by Puccinia striiformis Westend.) of 34 Triticum turgidum L. var.durum, 278 T. tauschii, and 267 synthetic hexaploid wheats (T. turgidum x T. tauschii) was evaluated at the seedling stage in the greenhouse and at the adult-plant stage at two field locations. Mexican pathotype 14E14 was used in all studies. Seedling resistance, expressed as low infection type, was present in all three species. One hundred and twenty-eight (46%) accessions of T. tauschii, 8 (23%) of T. turgidum and 31 (12%) of synthetic hexaploid wheats were highly resistant as seedlings. In the field tests, resistance was evaluated by estimating area under disease progress curve (AUDPC). Synthetic hexaploid wheats showed a wide range of variability for disease responses in both greenhouse and field tests, indicating the presence of a number of genes for resistance. In general, genotypes with seedling resistance were also found to be resistant as adult plants. Genotypes, which were susceptible or intermediate as seedlings but resistant as adult plants, were present in both T. turgidum and the synthetic hexaploids. Resistances from either T. turgidum or T. tauschii or both were identified in the synthetic hexaploids in this study. These new sources of resistance could be incorporated into cultivated hexaploid wheats to increase the existing gene pool of resistance to stripe rust.     

Stability of bacterial leaf spot resistance in peach regenerants under in vitro,greenhouse and field conditions

Summary Phenotypic stability of bacterial leaf spot resistance in peach (Prunus persica (L.) Batsch) regenerants, either selected at the cellular level for insensitivity to a toxic culture filtrate of Xanthomonas campestris pv. pruni or screened at the whole plant level for resistance to X. campestris pv. pruni, was investigated. A detached-leaf bioassay was used to evaluate the original regenerants again after three years in the greenhouse and also after a two to three year cycle of tissue culture propagation. Peach trees derived through micropropagation from the original regenerants were also evaluated after one to three years growth in the field. Although leaf spot resistance was retained in some regenerants over time in the greenhouse, following in vitro propagation, and under field conditions, resistance was either lost or not expressed in others. Regenerants # 19-1 and #156-6, derived from embryo callus of bacterial spot susceptible Sunhigh, were significantly more resistant than Sunhigh. High levels of resistance were exhibited in greenhouse plants and field-grown trees of regenerant #122-1, derived from embryo callus of moderately resistant Redhaven. This research provides additional evidence that selecting or screening for somaclonal variants with disease resistance is a feasible approach to obtaining peach trees with increased levels of bacterial spot resistance.Abbreviations TC Tissue-Cultured - TF Toxic culture Filtrate     

Potential of a Fusarium eumartii culture filtrate on the screening for wilting resistance in potato

Summary Fusarium solani f.sp. eumartii Carp. Snyder and Hansen (Fusarium eumartii) is a soil inhabitant that induces the so-called Potato Wilt and Stem End Rot disease. Prior to wilting, the pathogen induces peculiar small bronze spots on the leaflets. Failure to isolate F. eumartii from infected leaflets suggests the involvement of a toxin in the disease. The fungus was grown in liquid Richard's medium and thereafter a filtrate was obtained dialyzing (MW cutoff 12,000–14,000) and sterilizing the culture by filtration (0.22 m). Potato leaves treated with both the pathogen or the filtrate showed symptoms of bronze spots and significantly higher electrolyte leakage when compared to controls. Tomato leaves showed neither bronze spots nor electrolyte leakage after plant inoculation with the pathogen or with the filtrate treatment. Both, the absence of visible symptoms and the lack of electrolyte leakage in tomato could be associated to a certain degree of host specificity of the F. eumartii filtrate towards potato. The filtrate also induced symptoms similar to infections by F. eumartii in adult plants and in vitro plantlets of cultivars Huinkul MAG and Kennebec. Callus responses to the filtrate were related to responses of the cultivars to the pathogen in greenhouse. These results show the potential of the culture filtrate of F. eumartii for use in screening for wilting resistance.     

In vitro techniques for genomic alteration in rice plants

Because of the explosive increase in world population, a sufficient food supply must be achieved by varietal improvement in the major cereal crops including rice. It is expected that new in vitro techniques incombination with conventional breeding methods may effectively raise the yield potential. On the other hand, there are many environmental problems to be solved world-wide such as, global warming, environmental pollution, ecological destruction, reduction in water supplies and so on. Therefore, it is necessary to rapidly develop new varieties for the future combining of higher yield potential with excellent grain quality, and resistance to both biotic and abiotic stresses for the promotion of sustainable agriculture. Although many efforts have been made to introduce useful traits from wild species to cultivated rice via hybridization, it is still difficult to overcome breeding barriers such as cross incompatibility and hybrid sterility and inviability in practical breeding. Now in vitro techniques are going to make it possible to use genetic manipulation and cell culture and fusion techniques to speed up the breeding process. For sustainable agriculture, it is important to utilize the useful genes from alien species. For this purpose, asymmetric protoplast fusions have already been used successfully to transfer disease resistance in Brassica napus. In this experiment, a high level of resistance to the rice blast disease was transferred from wild species through asymmetric fusions. It is also noted that manipulation of cytoplasmic genomes is possible through asymmetric fusions as shown in the induction of new cytoplasmic male sterility (CMS). This revised version was published online in August 2006 with corrections to the Cover Date.     

Development of methods using phytoalexin (resveratrol) assessment as a selection criterion to screen grapevine in vitro cultures for resistance to grey mould (Botrytis cinerea)

Summary The purpose of this research was, firstly to determine the ability of grapevine in vitro cultures to synthesize resveratrol, a stilbene-type phytoalexin that is considered to be a good marker for resistance of grapevines to Botrytis cinerea, the causal organism for grey mould. Secondly, this study sought to establish the relationship between phytoalexin production potential and resistance to Botrytis cinerea in grapevines. In this aim, resveratrol production was assessed in 13 Vitis species or cultivars. A good correlation appeared between resveratrol production by grapevine in vitro cultures and grey mould resistance except for two Vitis spp. for which no correlation was observed, thus suggesting that resistance of grapevines may sometimes be associated with factors other than stilbene phytoalexins. In view of the results obtained, the potential use of resveratrol induction and of in vitro methods as a tool for screening grapevines for resistance to B. cinerea was discussed.     

Disease resistance in protected crops and mushrooms

Summary Cultivars of tomatoes, cucumbers, lettuce and peppers have been bred for resistance to one or more pathogens. Some tomato and cucumber cultivars have resistance to a wide range of diseases. Resistance has been transient in many cases and a succession of cultivars with new genes or new combinations of resistance genes has been necessary to maintain control. There has been a number of notable exceptions and these have included durable resistance to such pathogens asFulvia fulva and tomato mosaic virus. With lettuce the resistance situation is complicated by the occurrence of fungicide resistant pathotypes. There are no strains ofAgaricus bisporus purposely bred for disease resistance.In protected flower crops only resistance to Fusarium wilt in carnations has been purposely bred but differences in disease resistance are apparent in cultivars of many ornamental crops. This is particularly so in chrysanthemums where there are cultivars with resistance to many of the major pathogens. Similar situations occur with other flower crops and pot plants. Cultivars of some species have not been systematically investigated for resistance.The need for genetic resistance will increase with the further reduction, in the limits on pesticide use and an increasing public awareness and importance of pesticide pollution.ADAS is an executive agency of the Ministry of Agiculture, Fisheries and Food and the Welsh Office.     

Breeding alfalfa cultivars resistant to the alfalfa weevil

A ten-year study consisting of nine field experiments was conducted to evaluate alfalfa (Medicago sativa L.) for resistance to larval feeding by the alfalfa weevil (Hypera postica Gyllenhal). The results were consistent over years and experiments. Only one source of alfalfa germplasm, designated Starnes, showed resistance to larval feeding in the field under natural infestations of the weevil. The cultivars Team, Arc, and Liberty have been developed from this germplasm source. The only effective method of selection for field resistance to the weevil was field selection under natural weevil infestations. Selection was based on low larval feeding on spaced plants. After six generations of such selection the Starnes germplasm showed only one-third as much defoliation as did the check cultivars. The heritability of the weevil resistance was moderately high. Field resistance was transferred to other strains through hybridization; hybrids were intermediate to the resistant and susceptible parents. Resistance was stable.Laboratory and greenhouse efforts to breed alfalfa weevil resistant alfalfa-using such techniques as adult cotyledon-nonpreference tests, adult leaf-feeding tests, larval-growth and survival tests, oviposition-stimulus tests, and oviposition-nonpreference tests-have failed to produce field resistance.Paper No. 5308 of the Journal series of North Carolina Agricultural Experiment Station, Raleigh, N. C., in cooperation wich ARS, USDA. Received 15 June 1977.     

Inheritance of trichomes and resistance to pod borer (Helicoverpa armigera) and their association in interspecific crosses between cultivated pigeonpea (Cajanus cajan) and its wild relative C. scarabaeoides

The legume pod borer, Helicoverpa armigera, is one of the most devastating pests of pigeonpea. High levels of resistance to pod borer have been reported in the wild relative of pigeonpea, Cajanus scarabaeoides. Trichomes (their type, orientation, density and length) and their exudates on pod wall surface play an important role in the ovipositional behavior and host selection process of insect herbivores. They have been widely exploited as an insect defense mechanism in number of crops. In the present investigation, inheritance of resistance to pod borer and different types of trichomes (A, B, C and D) on the pod wall surface in the parents (C. cajan and C. scarabaeoides) and their F₁, F₂, BC₁ (C. cajan × F₁), and F₃ generations has been studied. Trichomes of the wild parents (high density of the non-glandular trichomes C and D, and glandular trichome B and low density of glandular trichome A) were dominant over the trichome features of C. cajan. A single dominant gene as indicated by the segregation patterns individually will govern each trait in the F₂ and backcross generation. Segregation ratio of 3 (resistant): 1 (susceptible) for resistance to pod borer in the F₂ generation under field conditions was corroborated with a ratio of 1:1 in the backcross generation, and the ratio of 1 non-segregating (resistant): 2 segregating (3 resistant: 1 susceptible): 1 non-segregating (susceptible) in F₃ generation. Similar results were obtained for pod borer resistance under no-choice conditions. Resistance to pod borer and trichomes associated with it (low density of type A trichome and high density of type C) are governed individually by a dominant allele of a single gene in C. scarabaeoides. Following backcrossing, these traits can be transferred from C. scarabaeoides into the cultivated background.     

A novel source of resistance to the two-spotted spider mite in Lycopersicon pimpinellifolium (Jusl.) Mill.: its genetics as affected by interplot interference

We have previously found an accession of Lycopersicon pimpinellifolium (Jusl.) Mill. (`TO-937') that appeared to resist attack by the two-spotted spider mite (Tetranychus urticae Koch). L. pimpinellifolium is a very close relative of the cultivated tomato (Lycopersicon esculentum Mill.) and thereby a potential source of desirable traits that could be introgressed to the crop species. The objective of this study was to investigate the genetics of the resistance present in `TO-937'. Resistance to infestation by the spider mite was quantified in 24-plant plots of L. pimpinellifolium accessions `TO-937' and `PE-10', L. pennellii accession `PE-45', L. esculentum cultivars `Moneymaker', `Roma' and `Kalohi' (reported to be partially resistant: Stoner & Stringfellow, 1967), and the interspecific F₁ cross, L. esculentum `Moneymaker' × L. pimpinellifolium `TO-937'. Only `TO-937', the F<sub>1</sub>, and`PE-45' were found to be resistant. Resistance of `TO-937' was complete when evaluated in two small greenhouses completely planted with `TO-937' so as to simulate the genotypic homogeneity usual in commercial crops. Generations (P₁, P₂, F₁, F₂, BC₁P₁, and BC₁P₂) of a P₁ (susceptible) × P₂ (resistant) cross (`Moneymaker' × `TO-937') were studied for resistance in a single-plant per plot design. Resistance of `TO-937' was inherited with complete dominance and appeared to be controlled by either two or four genes according to whether segregation in the F₂ or the BC₁P₁, respectively, were considered. However, calculation of the number of genes involved in the resistance was complicated by negative interplot interference due to the high frequency of resistant genotypes within most of the generations. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic analysis of resistance to soybean cyst nematode in PI 438489B

Soybean (Glycine max L. Merr.) plant introduction PI 438489B is a unique source that has resistance to all known populations of soybean cyst nematode (Heterodera glycines Ichinohe, SCN). This PI line also has many desirable agronomic characteristics, which makes it an attractive source of SCN resistance for use in a soybean breeding program. However, characterization of SCN resistance genes in this PI line have not been fully researched. In this study, we investigated the inheritance of resistance to populations of SCN races 1, 2, 3, 5, and 14 in PI 438489B. PI 438489B was crossed to the susceptible cultivar ‘Hamilton’ to generate F₁ hybrids. The random F₂ plants and F₃ lines were evaluated in the greenhouse for reaction to these five populations of SCN races. Resistance to SCN races 1, 3, and 5 were mostly conditioned by three genes (Rhg Rhg rhg). Resistance to race 2 was controlled by four genes (Rhg rhg rgh rgh). Three recessive genes were most likely involved in giving resistance to race 14. We further concluded that resistance to different populations of SCN races may share some common genes or pleiotropic effects may be involved. This revised version was published online in July 2006 with corrections to the Cover Date.     

Partial resistance to a Fusarium root disease in Egyptian white lupin landraces

A Fusarium sp. root pathogen of lupin is the causal agent of the most important disease that affects the cultivation of white lupin (Lupinus albus L.) in Egypt. The aim of our research was to investigate whether host resistance to Fusariumroot disease was available in Egyptian landraces ofLupinus albus. Five Fusarium isolates collected from white lupin samples in Egypt were tested with an Egyptian landrace and a French cultivar. The most aggressive isolate was used to screen an additional 15 Egyptian landraces, two cultivars released in Egypt after selection among landraces, one Polish cultivar, and two French cultivars. The assessment of host response to Fusarium was performed in a field, and under controlled conditions in a greenhouse pot experiment. Most landraces and the two Egyptian cultivars showed better resistance with good accordance between field and pot experiment. This experiment showed that Egyptian genetic resources of white lupin possess partial resistance to Fusarium root rot. Egyptian germplasm may be an alternative genetic source for incorporating partial resistance to Fusarium root rot into the breeding pools. This revised version was published online in July 2006 with corrections to the Cover Date.     

Resistance to Septoria nodorum blotch in several Triticum species

Summary Resistance to septoria nodorum blotch in Triticum monococcum, T. tauschii, T. timopheevii, T. dicoccum and T. durum was evaluated on plants at the three-leaf stage in greenhouse tests. A high frequency of resistant genotypes was found in T. monococcum, T. tauschii and T. timopheevii, but not in T. dicoccum and T. durum. The resistance of F₁ plants of crosses of resistant T. monococcum (PI 289599) and T. timopheevii (PI 290518) accessions with susceptible common wheat cv. Park and durum wheat cv. Wakooma, respectively, was evaluated on the basis of percentage leaf necrosis, lesion number, lesion size and incubation period. No dominance was found for long incubation period, but various dominance relationships occurred for low percentage leaf necrosis, low lesion number and small lesion size, depending on the cross. Multiple regression analysis showed that lesion number contributed more to percentage leaf necrosis than lesion size or incubation period. Resistance to septoria nodorum blotch was transferred successfully from T. timopheevii to cultivated durum wheat. Resistant BC₁F₇ lines, recovered from the T. timopheevii (PI 290518) × Wakooma cross, showed normal chromosome behaviour at meiosis (14 bivalents) and were self-fertile. However, an effective level of resistance was not recovered in lines derived from the other interspecific crosses.     

In vitro techniques for selecting wheat (Triticum aestivum L.) for Fusarium-resistance. I. Double-layer culture technique

Summary Calluses of spring and winter wheats (Triticum aestivum L.) were selected for Fusarium resistance in vitro, using the double-layer culture technique. Potato-dextrose agar medium in vials was inoculated with mycelia of Fusarium graminearum and F. culmorum. After one week, fungal cells were killed by autoclaving and the agar medium containing the thermostable toxic metabolites was overlayered with MS callus-growing medium. Later, wheat calluses were placed on the upper medium for 4–5 weeks, and from the surviving calluses plants were regenerated. R₂ seedling populations from self-fertilized R₁ plants of 4 varieties were tested for Fusarium resistance by artificial infections in the greenhouse, and 3% of the regenerated R₂ plants have been found to be more resistant than the original cultivars.