Screening techniques and sources of resistance to parasitic angiosperms

Parasitic angiosperms cause great losses in many important crops under different climatic conditions and soil types. The most widespread and important parasitic angiosperms belong to the genera Orobanche, Striga, and Cuscuta. The most important economical hosts belong to the Poaceae, Asteraceae, Solanaceae, Cucurbitaceae, and Fabaceae. Although some resistant cultivars have been identified in several crops, great gaps exist in our knowledge of the parasites and the genetic basis of the resistance, as well as the availability of in vitro screening techniques. Screening techniques are based on reactions of the host root or foliage. In vitro or greenhouse screening methods based on the reaction of root and/or foliar tissues are usually superior to field screenings and can be used with many species. To utilize them in plant breeding, it is necessary to demonstrate a strong correlation between in vitro and field data. The correlation should be calculated for every environment in which selection is practiced. Using biochemical analysis as a screening technique has had limited success. The reason seems to be the complex host-parasite interactions which lead to germination, rhizotropism, infection, and growth of the parasite. Germination results from chemicals produced by the host. Resistance is only available in a small group of crops. Resistance has been found in cultivated, primitive and wild forms, depending on the specific host-parasite system. An additional problem is the existence of pathotypes in the parasites. Inheritance of host resistance is usually polygenic and its transfer is slow and tedious. Molecular techniques have yet to be used to locate resistance to parasitic angiosperms. While intensifying the search for genes that control resistance to specific parasitic angiosperms, the best strategy to screen for resistance is to improve the already existing in vitro or greenhouse screening 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     

The bread wheat races of bunt represented in the Australian bunt collection

Summary The Australian Bunt Collection, obtained from infected crops between 1962–1977, has been classified into races, based on the differential reactions to ten known bunt resistant genes. Eight physiologic races of Tilletia laevis and three of T. tritici were identified. No race had virulence against the genes Bt3, Bt5, Bt8, or Bt10, of wheat. Consequently four major genes are available for breeding Australian wheat cultivars with resistance to common bunt.     

Variation in some agronomically important characters in a germplasm collection of beet (Beta vulgaris L.)

Summary A geographically representative selection of germplasm of Beta vulgaris, section Beta has been assessed for characteristics important in sugarbeet breeding, including downy mildew resistance, resistance to aphid colonisation and infection by the beet virus yellow complex. The occurrence of maintainer lines for cytoplasmic male-steriles was also investigated. Desirable qualities were found in some accessions, including nothern European wild vulgaris ssp. maritima and some old multigerm cultivars of fodder beets.     

Molecular mapping of powdery mildew resistance genes in wheat: A review

Powdery mildew, caused by Blumeria graminis f. sp. tritici (syn. Erysiphe graminis f. sp. tritici), is one of the most important diseases of common wheat (Triticum aestivum L.) worldwide. Molecular mapping and cloning of genes for resistance to powdery mildew in hexaploid wheat will facilitate the study of molecular mechanisms underlying resistance to powdery mildew diseases and help understand the structure and function of powdery mildew resistance genes, and permit marker-assisted selection in breeding programs. So far, 48 genes/alleles for resistance to powdery mildew at 32 loci have been identified and located on 16 different chromosomes, of which 21 resistance genes/alleles have been tagged by restriction fragment length polymorphisms (RFLPs), random-amplified polymorphic DNAs (RAPDs), amplified fragment length polymorphisms (AFLPs), sequence characterized amplified regions (SCARs), sequence-tagged sites (STS) or simple sequence repeats (SSRs). Several quantitative trait loci (QTLs) for adult plant resistance (APR) to powdery mildew have been associated with molecular markers. The detailed information on chromosomal location and molecular mapping of these genes has been reviewed. Isolation of powdery mildew resistance genes and development of valid molecular markers for pyramiding resistance genes in breeding programs is also discussed.     

Screening techniques and sources of resistance to foliar diseases caused by major necrotrophic fungi in grain legumes

Summary Necrotrophic pathogens of the cool season food legumes (pea, lentil, chickpea, faba bean and lupin) cause wide spread disease and severe crop losses throughout the world. Environmental conditions play an important role in the development and spread of these diseases. Form of inoculum, inoculum concentration and physiological plant growth stage all affect the degree of infection and the amount of crop loss. Measures to control these diseases have relied on identification of resistant germplasm and development of resistant varieties through screening in the field and in controlled environments. Procedures for screening and scoring germplasm and breeding lines for resistance have lacked uniformity among the various programs worldwide. However, this review highlights the most consistent screening and scoring procedures that are simple to use and provide reliable results. Sources of resistance to the major necrotrophic fungi are summarized for each of the cool season food legumes. Marker-assisted selection is underway for Ascochyta blight of pea, lentil and chickpea, and Phomopsis blight of lupin. Other measures such as fungicidal control and cultural control are also reviewed. The emerging genomic information on the model legume, Medicago truncatula, which has various degrees of genetic synteny with the cool season food legumes, has promise for identification of closely linked markers for resistance genes and possibly for eventual map-based cloning of resistance genes. Durable resistance to the necrotrophic pathogens is a common goal of cool season food legume breeders.     

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.     

Screening for Fusarium resistance in seedling populations of Asiatic hybrid lily

A test to select Fusarium resistant seedlings of the Asiatic hybrid lily is described. Young seedlings of 28 populations, obtained from an incomplete diallel between eight parents with different levels of Fusarium resistance, were tested for resistance. Significant differences in Fusarium resistance between and within populations were detected. The average percentage of selected seedlings ranged from 34% in resistant × resistant crosses to 2% in susceptible × susceptible crosses. Although resistant descendants were obtained in susceptible × susceptible crosses, using at least one resistant parent produced higher percentages of resistant seedlings. The resistance level of the parents correlated highly with the general combining ability for Fusarium resistance based on the seedling test. For eight populations, seedlings selected for Fusarium resistance and non-tested (control) seedlings of the same cross were compared, after propagation, in a clonal test. Variation between and within populations, found at seedling level, was confirmed at clonal level. A positive selection response was found for all eight populations. In the seedling test, approximately 18% of the seedlings were selected as resistant of which 15% (2.7% of seedlings tested) appeared to be susceptible escapes. Comparison between selection at seedling level and at clonal level indicated that approximately 25% of the seedlings tested were missed (rejected resistant plants) in the seedling test. The practical use of a seedling test for Fusarium resistance in lily breeding programs is discussed.     

Current and future strategies in breeding lentil for resistance to biotic and abiotic stresses

Lentil production is limited by lack of moisture and unfavorable temperatures throughout its distribution. Waterlogging and salinity are only locally important. Progress has been made in breeding for tolerance to drought through selection for an appropriate phenology and increased water use efficiency and in breeding for winter hardiness through selection for cold tolerance.The diseases rust, vascular wilt, and Ascochyta blight, caused by Uromyces viciae-fabae, Fusarium oxysporum f. sp. lentis, and Ascochyta fabae f. sp. lentis, respectively, are the key fungal pathogens of lentil. Cultivars with resistance to rust and Ascochyta blight have been released in several countries and resistant sources to vascular wilt are being exploited. Sources of resistance to several other fungal and viral diseases of regional importance are known. In contrast, although the pea leaf weevil (Sitona spp.) and the parasitic weed broomrape (Orobanche spp.), and to a lesser extent the cyst nematode (Heterodera ciceri), are significant yield reducers of lentil, no sources of resistance to these biotic stresses have been found. Directions for future research in lentil on both biotic and abiotic stresses are discussed.     

Analysis of durable resistance to stem rust in barley

Summary Since the mid-1940's, barley cultivars grown in the northern Great Plains of the USA and Canada have been resistant to stem rust caused byPuccinia graminis f. sp.tritici. This durable resistance is largely conferred by a single gene,Rpg1, derived from a single plant selection of the cultivar Wisconsin 37 and an unimproved Swiss cultivar. At the seedling stage, barley genotypes withRpg1 generally exhibit low mesothetic reactions at 16–20° C and slightly higher mesothetic reactions at 24–28° C to many stem rust pathotypes. This resistance is manifested by a low level of rust infection and mostly incompatible type uredia on adult plants.Rpg1 reacts in a pathotype-specific manner since some genotypes ofP. g. f. sp.tritici are virulent on cultivars carrying this gene in the field. Several factors may have contributed to the longevity of stem rust resistance in barley, a) since barley is planted early and matures early, it can sometimes escape damage from stem rust inoculum carried from the south; b) one or more minor genes may augment the level of resistance already provided byRpg1; c) the cultivation of resistant wheat cultivars and eradication of barberry have reduced the effective population size and number of potential new pathotypes ofP. g. f. sp.tritici, respectively; and d) virulent pathotypes ofP. g. f. sp.tritici andP. g. f. sp.secalis have not become established. This situation changed in 1989 when a virulent pathotype (Pgt-QCC) ofP. g. f. sp.tritici became widely distributed over the Great Plains. However,Rpg1 may still confer some degree of resistance to pathotype QCC because stem rust severities have been low to moderate and yield losses light on barley cultivars carrying the gene during the last four seasons (1989–1992). Several sources of incomplete resistance to pathotype QCC have been identified in barley. To facilitate the transfer of resistance genes from these sources into advanced breeding lines, molecular marker assisted selection is being employed.     

SCAR and CAPS markers flanking the Brassica oleracea L. Pp523 downy mildew resistance locus demarcate a genomic region syntenic to the top arm end of Arabidopsis thaliana L. chromosome 1

We recently mapped the Pp523 locus that includes a single, dominant gene conferring resistance to downy mildew expressed in adult plants to a 75.1 cm long linkage group on a genetic linkage map of Brassica oleracea L. More recently, we identified a new AFLP marker 2.8 cm downstream from the resistance gene. The five DNA markers within an 8.5 cm region encompassing the Pp523 gene were cloned and sequenced. Three of these markers were transformed into SCARs (sequence characterised amplified regions), however, two among them were monomorphic and were analysed as CAPS (cleaved amplified polymorphic sequence) markers among the mapping population. Searched against genomic databases, the five B. oleracea DNA-marker sequences matched Arabidopsis thaliana L. gene sequences that delimit a conserved syntenic region in the top arm end of chromosome 1 of this last species. Considering the close genetic relatedness between both species, the information on this specific genomic region in A. thaliana is particularly useful for the construction of a fine-scale map of the corresponding genomic region in B. oleracea. The identified SCAR and CAPS markers can be used for marker assisted selection (MAS) in breeding programs aimed at the introgression of the Pp523 resistance locus, allowing the reliable indirect identification of plants harbouring the resistance gene with a margin of error of approximately six in ten-thousand selected plants.     

Identification of pathogenic races ofPhytophthora fragariae Hickman in Germany

Summary The fungusPhytophthora fragariae Hickman is the causal agent of red stele disease in strawberry (Fragaria × ananassa), this being a major disease in many areas with cool and moist conditions. Success of resistance breeding can be nullified by the appearence of specific races of the fungus capable of overcoming the introduced resistance. In some countries (USA, UK, Japan and Canada) races were identified by using a differential set of strawberry cultivars. The absence of an international standard differential set and the use of different test methods make a comparison of the identified races difficult or even lead to contradictory results for one variety/race combination. The aim of this study was to obtain information about the spectrum of pathogenic races in Germany as a basic contribution to research on resistance breeding against the fungus. The susceptibility of different strawberry cultivars to German isolates ofP. fragariae was evaluated. The inoculation was done by modifying a method described by Milholland et al. (1989). Rootedin vitro plants, four weeks after transferring them to the soil, were used for the investigation. Inoculation was done with a zoospore suspension of defined concentration. Up to now three German isolates, G-1, G-2, and G-3, can be separated by their ability to infect and produce oospores in the roots of the strawberry cultivars ‘Senga Sengana’, either ‘Saladin’, ‘Redgauntlet’ or ‘Climax’ andFragaria chiloensis clone ‘YaquinaB’. These results confirm the existence of pathogenic races ofP. fragariae in Germany and should be taken into consideration for resistance breeding against the fungus.     

Common bean breeding for resistance against biotic and abiotic stresses: From classical to MAS breeding

Summary Breeding for resistance to biotic and abiotic stresses of global importance in common bean is reviewed with emphasis on development and application of marker-assisted selection (MAS). The implementation and adoption of MAS in breeding for disease resistance is advanced compared to the implementation of MAS for insect and abiotic stress resistance. Highlighted examples of breeding in common bean using molecular markers reveal the role and success of MAS in gene pyramiding, rapidly deploying resistance genes via marker-assisted backcrossing, enabling simpler detection and selection of resistance genes in absence of the pathogen, and contributing to simplified breeding of complex traits by detection and indirect selection of quantitative trait loci (QTL) with major effects. The current status of MAS in breeding for resistance to angular leaf spot, anthracnose, Bean common mosaic and Bean common mosaic necrosis viruses, Beet curly top virus, Bean golden yellow mosaic virus, common bacterial blight, halo bacterial blight, rust, root rots, and white mold is reviewed in detail. Cumulative mapping of disease resistance traits has revealed new resistance gene clusters while adding to others, and reinforces the co-location of QTL conditioning resistance with specific resistance genes and defense-related genes. Breeding for resistance to insect pests is updated for bean pod weevil (Apion), bruchid seed weevils, leafhopper, thrips, bean fly, and whitefly, including the use of arcelin proteins as selectable markers for resistance to bruchid seed weevils. Breeding for resistance to abiotic stresses concentrates on drought, low soil phosphorus, and improved symbiotic nitrogen fixation. The combination of root growth and morphology traits, phosphorus uptake mechanisms, root acid exudation, and other traits in alleviating phosphorus deficiency, and identification of numerous QTL of relatively minor effect associated with each trait, reveals the complexity to be addressed in breeding for abiotic stress resistance in common bean.     

CO<Subscript>2</Subscript>-assimilation and chlorophyll fluorescence as indirect selection criteria for host tolerance against <Emphasis Type="Italic">Striga</Emphasis>

Striga hermonthica (Del.) Benth. is a parasitic weed on tropical cereals causing serious yield losses in Africa. The use of host crop varieties with improved resistance and tolerance against this parasite is a key component of an integrated control strategy. Breeding for tolerance is however seriously hampered by the absence of reliable and yet practical selection measures. The observation that the photosynthetic rate of tolerant genotypes is less sensitive to Striga infection was used as a starting point to search for suitable selection measures. In a greenhouse pot experiment the effect of Striga infection on the photosynthesis of four sorghum (Sorghum bicolor [L.] Moench) genotypes, differing in Striga tolerance level, was measured at three moments in time (26, 48 and 75 days after sowing). Genotypes were CK60-B, E36-1, Framida and Tiémarifing. Measurements involved CO₂-assimilation (A) and three chlorophyll fluorescence characteristics (electron transport rate through photosystem II [ETR], photochemical [Pq] and non-photochemical quenching [NPq]). Striga infection negatively affected A, ETR and Pq. Based on A and Pq, genotypes with superior levels of tolerance (Tiémarifing) could be discriminated from genotypes with superior level of resistance (Framida). Both A and Pq showed high heritabilities and consequently clear and predictable differences between genotypes. Using discriminative ability, heritability and cost effectiveness as main criteria, photochemical quenching (Pq) was concluded to possess the highest potential to serve as indirect selection measure for host plant tolerance to Striga. Screening should preferably be conducted at relatively high Striga infestation levels, between Striga emergence and host plant flowering.     

Breeding strategy in foxtail millet,Setaria italica (L.P. Beauv.), following interspecific hybridization

Summary An interspecific cross between foxtail millet Setaria italica and its wild relative Setaria viridis was undertaken in order to introduce from the latter triazine resistance in the cultivated plant. Four backcross generations obtained with S. italica as recurrent parent were studied. Results show that only two backcross generations associated with selection are enough to eliminate weedy characters and to return to the cultivated type. S. viridis could thus be an interesting source for improvement of foxtail millet without a complex and time consuming breeding strategy.     

Screening techniques and sources of resistance to rusts and mildews in grain legumes

Summary In this paper we review the existence of sources of resistance and the various available screening methods for resistance in grain legumes against the airborne pathogens powdery mildews, downy mildews and rusts. Available resistance against these pathogens is not abundant and is particularly in risk of erosion owing to the constant generation and introduction of new races of the pathogen. A continuous search for more resistance sources is therefore a priority in legume breeding and special emphasis should be paid to selection of resistance mechanisms that are likely to be durable and to implementation of strategies to prolong the durability of existing resistance.     

Resistance to Helminthosporium leaf blight and agronomic performance of spring wheat genotypes of diverse origins

Helminthosporium leaf blight (HLB), caused by a complex of Cochliobolus sativus (Ito & Kurib.) Drechsler ex Dastur and Pyrenophora tritici-repentis Died, is a serious disease of wheat (Triticum aestivum L.) in the warm lowlands of South Asia. Wheat cultivars grown in the area are either susceptible to HLB or possess low levels of resistance to it. A replicated field study was conducted in 1999 and 2000 at two sites in Nepal to determine the level of HLB resistance and other desirable traits in 60 wheat genotypes of diverse origin. The test genotypes were planted in main strips divided into two strips one of which was sprayed four times with Tilt (a.i. propiconazole) @ 125 g of a.i. ha^–1. Four readings of HLB were recorded to calculate the area under the disease progress curve (AUDPC). Other traits under investigation included biomass yield (BY), grain yield (GY), 1000-kernel weight (TKW), harvest index (HI), days to heading (DH) and maturity (DM), plant height (PHT), and effective tiller number (ETN). Wheat genotypes differed significantly for all traits. Mean AUDPC values ranged from 45 to 1268. A few exotic genotypes were highly resistant to HLB. Losses in GY due to HLB ranged from 2 to 26%, and TKW was reduced by up to 33%. A few genotypes showed HLB tolerance, i.e., relatively smaller GY and TKW reductions despite high levels of HLB. In general, medium to late maturity and higher levels of HLB resistance and low to high GY and TKW characterized genotypes exotic to South Asia. Biplot analysis identified several genotypes that were HLB-resistant and agronomically superior. Results suggest it is possible to improve HLB resistance of local wheat cultivars based on selective breeding using this pool of germplasm.     

Variation in forage grasses as hosts of the root-knot nematode Meloidogyne naasi (Franklin) and selection for resistance in ryegrass

Summary Reactions of 13 grasses to Meloidogyne naasi varied with species; ryegrasses, fescues and their hybrids were generally susceptible and cocksfoot and timothy resistant. Marked variation in host resistance levels occurred between genotypes within cultivars.Selection of single plants, followed by tests on replicate tillers, identified resistant and susceptible genotypes in both Italian and perennial ryegrass cultivars. Resistant plants had few nematode-induced galls and fewer females and eggs than susceptibles. There was more or less continuous variation, with many genotypes intermediate between extremes of resistance and susceptibility. Selected resistant and susceptible genotypes are of use in assessing variation in nematode populations and as controls for breeding and selection programmes.     

Inheritance of resistance to the bipartite Tomato chlorotic mottle begomovirus derived from Lycopersicon esculentum cv. ‘Tyking’

Severe outbreaks of bipartite begomoviruses (family Geminiviridae) have been observed on tomatoes after the introduction of the whitefly Bemisia tabaci (biotype B) in Brazil. The Lycopersicon esculentum line ‘TX 468-RG’ was identified as one of the best sources of broad-spectrum resistance to species comprising the tomato-infecting Begomovirus complex in Brazil. The genetic basis of resistance to one Begomovirus isolate was investigated using populations from the cross between ‘TX 468-RG’ (P₁) and the susceptible line ‘Ohio 8245’ (P₂). Parental lines, F₁, backcross (BC) to P₁ and BC to P₂ and F₂ generations were inoculated at the two true-leaf stage using 20 viruliferous whiteflies per plant. Assessment was done two weeks after inoculation based upon visual analysis of symptom expression. The ratio of resistant to susceptible plants closely fit to a single recessive gene (locus) model. The sequence analysis indicated that the Begomovirus isolate used in this assay was closely related to the bipartite Tomato chlorotic mottle virus. Therefore, this gene/locus, was tentatively named tcm-1 (tomato chlorotic mottle virus resistance-1). This locus has been transferred to distinct tomato cultivars and levels of resistance similar to that of ‘TX 468-RG’ were observed in advanced (F₈ and F₉) generations. In addition, breeding lines carrying the tcm-1 locus were also resistant to other Brazilian bipartite tomato-infecting Begomovirus species.     

Breeding for tolerance to beet-cyst eelworm Heterodera schachtii Schm. in sugarbeet

Summary Successive mass selections were made in sugarbeet varieties (Beta vulgaris L.) and in B. vulgaris × B. maritima L. hybrids for tolerance to wilting caused by beet-cyst eelworm attack. The selected lines showed improved tolerance to wilting, but no evidence of resistance to eelworm infection was demonstrated. By repeated back-crossing of the selected lines with the commercial varieties and concurrent selection root yield could be improved without substantially decreasing the tolerance to eelworm attack. These results indicate that tolerance is partly conditioned by complete and incomplete dominant genes.