Screening techniques and sources of resistance to root rots and wilts in cool season food legumes

Soilborne, fungal pathogens of cool season food legumes, including seed and seedling blights, rot rots, and wilts are described. Seed and seedling diseases are caused primarily by Pythium and Rhizoctonia spp. The most important fungi causing root rots include Aphanomyces euteiches, Fusarium solani, Pythium spp., Sclerotium rolfsii, and Macrophomina phaseolina. Wilt is caused primarily by various host-specific forms of Fusarium oxysporum. This paper discusses these diseases and screening procedures that emphasize standardization of inoculum levels, maintenance of virulent pathogen cultures, inoculum growth media, environmental conditions, and host plant age. Sources of resistance to these diseases are discussed.     

Screening techniques and sources of resistance to nematodes in cool season food legumes

Identification of sources of resistance in cool season legumes to cyst (Heterodera spp.), root-knot (Meloidogyne spp.), and stem nematode (Ditylenchus dipsaci) is generally based on number of cysts on roots, root-knot nematode induced gall index, and stem nematode reproduction in shoot tissue, respectively. Various levels of resistance to cyst nematodes have been detected in chickpea and pea. Resistance has also been identified in chickpea, faba bean, and pea to the root-knot nematodes. Broad based durable sources of resistance to plant parasitic nematodes are required. Basic research is needed to develop transgenic plants with resistance based on hatch stimulants, inhibitors, toxins, or repellents found in antagonistic rhizosphere microorganisms. Selection of genotypes that favor development of beneficial rhizosphere microorganisms or root endophytes that increase the plant resistance to nematode infection deserves attention.     

Screening techniques and sources of resistance to foliar diseases caused by fungi and bacteria in cool season food legumes

Screening techniques are an important component of the overall strategy of breeding for resistance to diseases in cool season food legumes. Suitable screening methods have been developed for several major foliar diseases of chickpea, pea, faba bean, and lentil, and sources of resistance have been identified. International cooperation plays an important role in promoting research and keeping collections of cultivated species and their wild relatives. New biotechnological approaches are promising for enhancing the practical use of genes for resistance.     

Screening techniques and sources of tolerance to salinity and mineral nutrient imbalances in cool season food legumes

A large global land area is affected by saline, alkali (sodic), and acid soil conditions. Cool season food legumes are important crops in many countries with such adverse soils. Tolerant genotypes have been identified in many crops, including legumes. However, very little has been published on selection of tolerant cool season food legume crops. The inadequate knowledge and understanding of the responses of cool season food legume crops to these abiotic stresses, necessitates action by a collaborative network of interdisciplinary teams to make rapid progress in identifying tolerant germplasm and developing cultivars better adapted to unfavorable soil conditions.     

Screening techniques and improved biological nitrogen fixation in cool season food legumes

Dinitrogen fixation and legume productivity are greatly influenced through the interactions of legume host, Rhizobium, and the above- and below-ground environment. The benefits of improving legume N₂ fixation include reduced reliance on soil N, leading to more sustainable agricultural systems and reduced requirements for fertilizer N, enhanced residual benefits to subsequent crops, and increased legume crop yields. Most of the gains in N₂ fixation to date have been derived from management of legume cropping systems and through inoculation of legume seed with competitive and symbiotically effective rhizobia. Further gains are possible by developing plant cultivars with tolerance to soil abiotic factors, increased plant yield, and a broader and more effective matching of plant host and rhizobia. Techniques for screening material for superior N₂ fixation and examples of programs to increase fixed N, with attention to the major abiotic stresses, are discussed.     

Screening techniques and sources of resistance to abiotic stresses in cool-season food legumes

Summary The adaptability and productivity of cool-season food legumes (chickpea, faba bean, lentil, pea) are limited by major abiotic stresses including drought, heat, frost, chilling, waterlogging, salinity and mineral toxicities. The severity of these stresses is unpredictable in field experiments, so field trials are increasingly supplemented with controlled-environment testing and physiological screening. For drought testing, irrigation is used in dry fields and rain-out shelters in damp ones. Carbon isotope discrimination (Δ¹³C) is a well-established screen for drought tolerance in C3 cereal crops which is now being validated for use in grain legumes, but it is relatively expensive per sample and more economical methods include stomatal conductance and canopy temperature. Chickpea lines ICC4958 and FLIP87-59C and faba bean line ILB938 have demonstrated good drought tolerance parameters in different experiments. For frost tolerance, an efficient controlled-environment procedure involves exposing hardened pot-grown plants to sub-zero temperatures. Faba beans Cote d’Or and BPL4628 as well as lentil ILL5865 have demonstrated good freezing tolerance in such tests. Chilling-tolerance tests are more commonly conducted in the field and lentil line ILL1878 as well as derivatives of interspecific crosses between chickpea and its wild relatives have repeatedly shown good results. The timing of chilling is particularly important as temperatures which are not lethal to the plant can greatly disrupt fertilization of flowers. Salinity response can be determined using hydroponic methods with a sand or gravel substrate and rapid, efficient scoring is based on leaf symptoms. Many lines of chickpea, faba bean and lentil have shown good salinity tolerance in a single article but none has become a benchmark. Waterlogging tolerance can be evaluated using paired hydroponic systems, one oxygenated and the other de-oxygenated. The development of lysigenous cavities or aerenchyma in roots, common in warm-season legumes, is reported in pea and lentil but is not well established in chickpea or faba bean. Many stresses are associated with oxidative damage leading to changes in chlorophyll fluorescence, membrane stability and peroxidase levels. An additional factor relevant to the legumes is the response of the symbiotic nitrogen-fixing bacteria to the stress.     

Research achievements in plant resistance to insect pests of cool season food legumes

Plant resistance to at least 17 field and storage insect pests of cool season food legumes has been identified. For the most part, this resistance was located in the primary gene pools of grain legumes via conventional laboratory, greenhouse, and field screening methods. The use of analytical techniques (i.e., capillary gas chromatography) to characterize plant chemicals that mediate the host selection behavior of pest insects offers promise as a new, more rapid way to differentiate between insect-resistant and susceptible plant material. Examples of research achievements in mechanisms of resistance and host-plant resistance within the context of integrated control programs are discussed. Accelerating the development and subsequent releases of insect-resistant cultivars to pulse farmers requires more involvement from interdisciplinary teams of plant breeders, entomologists, plant pathologists, plant chemists, molecular biologists, and other scientists.     

Using host plant resistance to manage biotic stresses in cool season food legumes

Summary Kabuli chickpea (Cicer arietinum L.) is the common cultivated type of chickpea in arid and semi-arid environments in the Mediterranean region. Ascochyta blight, (Ascochyta rabiei (Pass.) Labr.), leaf miner (Liriomyza cicerina, Rond.) and cold, are the three most important stresses on chickpea grown under semi-arid conditions in this region. Phenotypic frequencies for responses to these stresses in the eight major chickpeagrowing regions of the world were estimated from 5,672 kabuli chickpea accessions assembled from these regions. In addition, the accessions were evaluated for 12 morpho-physiological and three phenological characters under semi-arid Mediterranean conditions at the International Center for Agricultural Research in the Dry Areas (ICARDA), Aleppo, Syria. Considerable regional differences in frequency distributions for response to the three stresses were observed. Average phenotypic diversity for responses to the three stresses was lower (H_o=0.474) than for morpho-physiological (H_o=0.754) and phenological (H_o=0.812) characters. The highest frequencies of accessions resistance to Ascochyta-blight and leaf-miner were found in South Asia and South Central Asia, respectively. A small number of chickpea breeding materials of ICARDA showed a moderate level of tolerance to cold. A group of four characters showing the strongest bivariate association with each of the three stresses was identified from the latter group. Then, a discrete multivariate log-linear analysis of the five-way frequency table was performed for each group. The simplest log-linear model for each group included both two- and three-factor association terms, but no independent factors. This suggested the potential for indirect selection for stress tolerance using one or more of these associated characters. The roles of these characters in ideotype breeding of kabuli chickpea for arid and semi-arid Mediterranean conditions deserves careful assessment.     

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.     

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.     

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.     

Breeding methods and selection indices for improved tolerance to biotic and abiotic stresses in cool season food legumes

The objective of breeding for stress tolerance is to improve productivity for a target level of stress. If tolerance is viewed as resistance to change in productivity with increasing stress, productivity under stress depends not only on stress tolerance, but also on maximum productivity. Index selection theory indicates that selection in non-stress environments will be more effective than direct selection for productivity under stress whenever the correlation between the two types of environments exceeds the heritability of productivity under stress. With high genetic correlation, selection should be conducted within a level of stress that maximizes heritability. In cases where heritability under non-stress is much higher than under stress, an index combining data from stress and non-stress environments is expected to be more efficient than selection based on evaluation only within stress environments.Secondary traits will be useful in breeding for productivity under stress whenever they have high heritability and high genetic correlation with productivity under stress. For some abiotic stresses and many biotic stresses, heritability will be highest in the presence of stress and indirect or index selection will be of limited value.     

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.     

Water use and water use efficiency of cool season grain legumes in low rainfall Mediterranean-type environments

The water use (E_t) and water use efficiency (WUE) of a range of cool season grain legume species (field pea [Pisum sativum L.], faba bean [Vicia faba L.], chickpea [Cicer arietinum L.], lentil [Lens culinaris Med.], albus lupin [Lupinus albus L.], dwarf chickling [Lathyrus cicera L.], ochrus chickling [Lathyrus ochrus L.], grass pea [Lathyrus sativus L.], narbon bean [Vicia narbonensis L.], common vetch [Vicia sativa L.], and purple vetch [Vicia benghalensis L.]) were examined on fine textured neutral to alkaline soils in the low to medium rainfall Mediterranean-type environments in south-western Australia at Merredin and Mullewa in two seasons. There was no difference in the total E_t between grain legumes at either site in either year. There was also no variation in soil water extraction between species on the shallow sandy loam soil at Merredin. However, C. arietinum, L. sativus and L. cicera had greater water extraction and P. sativum the least water extraction at Mullewa where soil conditions were less hostile and root penetration was not restricted. The pattern of water use varied markedly between the grain legume species examined. Grain yield was positively correlated with post-flowering water use (E_tpa) in both erect (r=0.59) and prostrate (r=0.54) grain legume species. Water use efficiencies for dry matter production (WUE_dm) of up to 30 kg ha⁻¹ mm⁻¹ for V. faba and V. narbonensis at Merredin, and water use efficiencies for grain yield (WUE_gr) of up to 16 kg ha⁻¹ mm⁻¹ for P. sativum and 13 kg ha⁻¹ mm⁻¹ for V. faba at Mullewa, were comparable to those reported for cereals and other grain legumes in previous studies in this and other environments. Potential transpiration efficiencies (TE) of 15 kg ha⁻¹ mm⁻¹ together with soil evaporation (E_s) values of 100–125 mm were estimated in this and associated studies, and can be used as benchmark values to assess the yield potential of cool season grain legume crops in low rainfall Mediterranean-type environments. The major traits of adaptation for grain legume species producing large yields in this short season environment are early flowering, and pod and seed set before the onset of terminal drought. Early phenology together with rapid ground cover and dry matter production allows greater water use in the post flowering period. This leads to greater partitioning of dry matter into seed, which is reflected in greater harvest index (HI) and WUE_gr, as was observed for V. faba and P. sativum. Improvement in the adaptation of other grain legume species to short season Mediterranean-type environments requires increased early growth for rapid ground cover and improved tolerance to low temperatures (especially for C. arietinum) during flowering and podding.     

Recurrent selection for quantitative resistance to soil-borne diseases in beans in the Mixteca region, Mexico

Most of the bean breeding for disease resistance has been done considering it as a qualitative trait. This work reports on bean breeding for resistance as a quantitative trait. Recurrent mass selection was used in order to simultaneously increase the polygenically inherited resistance to all locally important bean pathogens but with emphasis on bean common mosaic virus (BCMV) and bacterial blight (BB) in the Mixteca Region, Mexico. Qualitative resistances against soil-borne diseases, induced mainly by facultative pathogens, possessing little parasitic specialization, are unlikely to occur, and although breeding against these kinds of diseases was not originally planned, it turned out to be part of collateral field screening. The accumulation of quantitative resistance against soil-borne diseases in advanced lines of this program is described and includes two field experiments, a cycle for seed augmentation in large plots and two greenhouse experiments. The majority of lines of the advanced breeding cycles produced consistently higher yields and higher survival rates, when compared with the original parents and commercial varieties. However, the regional land races still have higher survival rates but they have lower yields. Soil-borne diseases were present during all breeding cycles carried out under field conditions in the Mixteca Region. This work indicates that breeding for quantitative resistance involves all locally important pathogens. It is then feasible to accumulate high levels of quantitative resistance, in a relatively short time, even to these intermittently severe, soil-borne diseases that are otherwise so difficult to manage. This revised version was published online in July 2006 with corrections to the Cover Date.     

Screening lentil for resistance to fusarium wilt: methodology and sources of resistance

Host-plant resistance is the best means to control the key disease of lentil-vascular wilt, caused by Fusarium oxysporum Schlecht. emend. Snyder & Hansen f.sp. lentis Vasudeva and Srinivasan. Systematic screening for resistance to lentil wilt was initiated in the field in 1993, in a wilt-sick plot in North Syria, with a core collection of 577 germplasm accessions from 33 countries. A subset (88 accessions) of mostly resistant accessions was re-screened in 1994. The most resistant accessions came from Chile, Egypt, India, Iran and Romania. Variation among accessions in the temporal pattern of wilting was analyzed. The limited wilting in resistant accessions followed a linear model through time, whereas the pattern for susceptible accessions was better described with an exponential model. This temporal variation emphasizes the need for repeated scoring during screening for resistance to lentil vascular wilt to identify ‘late-wilters’. To overcome spatial variation in inoculum density, the efficacy of using wilt scores from a systematically-repeated susceptible control in covariate analysis was tested. Covariance analysis significantly improved overall screening by 3% in 1993, but the improvement was non-significant in 1994. The results emphasize the relative uniformity of disease pressure in the wilt-sick plot and suggest that covariance analysis of a systematically arranged control will be of greater benefit in land which is less uniformly infected. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic improvement of legumes using somatic cell and molecular techniques

Summary The merits and limitations of somatic cell techniques involving Agrobacterium-mediated transformation, direct gene transfer and protoplast fusion, are discussed in relation to the genetic improvement of forage and grain legumes. Whilst progress with legumes is limited compared to that with plants of other families such as the Solanaceae, the fact that many legumes are readily amenable to tissue culture now permits somatic cell techniques to be targetted to these species. Future development of the subject will necessitate close collaboration between molecular biologists and plant breeders to enable novel plants generated by in vitro technologies to be incorporated into conventional breeding programmes.     

Integrated “omics” approaches to sustain global productivity of major grain legumes under heat stress

Grain legumes serve as key sources of dietary protein to the global human population. Consequence of high‐temperature (HT) stress is increasingly evident as drastically lost production of different crops including grain legumes worldwide, thus putting the global food security under great threat. In a changing climate scenario, cool season‐adapted grain legumes frequently encounter heat stress (HS) during their reproductive phase, thus witnessing serious yield losses. To combat the emerging challenges of HT stress, an integrated approach demanding collaborative efforts from various disciplines of plant science should be in place. This review summarizes major impacts of HT stress on grain legume, and captures the relevance of crop genetic resources to HS tolerance in these crops. Measurement of physiological traits assumes key place in view of ever‐increasing precision of next‐generation phenotyping assays. We also discuss the significance of genetic inheritance and QTL discovery and evolving “omics” science for developing HS tolerance grain legume crops.     

Heat tolerance in food legumes as evaluated by cell membrane thermostability and chlorophyll fluorescence techniques

Summary The genotypic variation for heat tolerance in chickpea, groundnut, pigeonpea, and soya bean was evaluated by testing membrane stability and photosystem (PSII) function in leaves at high temperatures. The legumes could be ranked from heat-tolerant to sensitive in the order: groundnut, soya bean, pigeonpea and chickpea. The damage to cell membranes (as reflected by an increased leakage of electrolytes) and PS II (as reflected by a decrease in the ratio of variable to maximum fluorescence) was less, and recovery from heat stress was faster in groundnut than in other crops. Prior exposure of plants to 35°C for 24h led to a reduced leakage of electrolytes at high temperatures in all crops but the differences among legumes were consistent. Substantial genotypic variation for heat tolerance was found in all legumes. Membrane injury was negatively associated with specific leaf weight in groundnut (r=–0.69^**) and soya bean (r=–0.56^**) but not in the pulses. Electrolyte leakage and fluorescence ratio were negatively correlated in all legumes. The potential use of electrolyte leakage and fluorescence tests as screening procedures for breeding heat-tolerant legumes is discussed.Abbreviations RI relative injury - Fo initial fluorescence - Fm maximum fluorescence - Fv variable fluorescence - PS II photosystem II - PAR photosynthetically active radiation