Heather Clarke

Chickpea molecular breeding: New tools and concepts

SummaryChickpea is a cool season grain legume of exceptionally high nutritive value and most versatile food use. It is mostly grown under rain fed conditions in arid and semi-arid areas around the world. Despite growing demand and high yield potential, chickpea yield is unstable and productivity is stagnant at unacceptably low levels. Major yield increases could be achieved by development and use of cultivars that resist/tolerate abiotic and biotic stresses. In recent years the wide use of early maturing cultivars that escape drought stress led to significant increases in chickpea productivity. In the Mediterranean region, yield could be increased by shifting the sowing date from spring to winter. However, this is hampered by the sensitivity of the crop to low temperatures and the fungal pathogen Ascochyta rabiei. Drought, pod borer (Helicoverpa spp.) and the fungus Fusarium oxysporum additionally reduce harvests there and in other parts of the world. Tolerance to rising salinity will be a future advantage in many regions. Therefore, chickpea breeding focuses on increasing yield by pyramiding genes for resistance/tolerance to the fungi, to pod borer, salinity, cold and drought into elite germplasm. Progress in breeding necessitates a better understanding of the genetics underlying these traits. Marker-assisted selection (MAS) would allow a better targeting of the desired genes. Genetic mapping in chickpea, for a long time hampered by the little variability in chickpea’s genome, is today facilitated by highly polymorphic, co-dominant microsatellite-based markers. Their application for the genetic mapping of traits led to inter-laboratory comparable maps. This paper reviews the current situation of chickpea genome mapping, tagging of genes for ascochyta blight, fusarium wilt resistance and other traits, and requirements for MAS. Conventional breeding strategies to tolerate/avoid drought and chilling effects at flowering time, essential for changing from spring to winter sowing, are described. Recent approaches and future prospects for functional genomics of chickpea are discussed.

Low-Temperature Stress: Implications for Chickpea (Cicer arietinum L.) Improvement

Chickpea is the third major cool season grain legume crop in the world after dry bean and field pea. Chilling and freezing range temperatures in many of its production regions adversely affect chickpea production. This review provides a comprehensive account of the current information regarding the tolerance of chickpea to freezing and chilling range temperatures. The effect of freezing and chilling at the major phenological stages of chickpea growth are discussed, and its ability for acclimation and winter hardiness is reviewed. Response mechanisms to chilling and freezing are considered at the molecular, cellular, whole plant, and canopy levels. The genetics of tolerance to freezing in chickpea are outlined. Sources of resistance to both freezing and chilling from within the cultivated and wild Cicer genepools are compared and novel breeding technologies for the improvement of tolerance in chickpea are suggested. We also suggest future research be directed toward understanding the mechanisms involved in cold tolerance of chickpea at the physiological, biochemical, and molecular level. Further screening of both the cultivated and wild Cicer species is required in order to identify superior sources of tolerance, especially to chilling at the reproductive stages.

Geographical patterns of genetic variation in the world collections of wild annual Cicer characterized by amplified fragment length polymorphisms

Cicer reticulatum, C. echinospermum, C. bijugum, C. judaicum, C. pinnatifidum, C. cuneatum and C. yamashitae are wild annual Cicer species and potential donors of valuable traits to improve chickpea (C. arietinum). As part of a large project to characterize and evaluate wild annual Cicer collections held in the world gene banks, AFLP markers were used to study genetic variation in these species. The main aim of this study was to characterize geographical patterns of genetic variation in wild annual Cicer germplasm. Phylogenetic analysis of 146 wild annual Cicer accessions (including two accessions in the perennial C. anatolicum and six cultivars of chickpea) revealed four distinct groups corresponding well to primary, secondary and tertiary gene pools of chickpea. Some possible misidentified or mislabelled accessions were identified, and ILWC 242 is proposed as a hybrid between C. reticulatum and C. echinospermum. The extent of genetic diversity varied considerably and was unbalanced between species with greatest genetic diversity found in C. judaicum. For the first time geographic patterns of genetic variation in C. reticulatum, C. echinospermum, C. bijugum, C. judaicum and C. pinnatifidum were established using AFLP markers. Based on the current collections the maximum genetic diversity of C. reticulatum, C. echinospermum, C. bijugum and C. pinnatifidum was found in southeastern Turkey, while Palestine was the centre of maximum genetic variation for C. judaicum. This information provides a solid basis for the design of future collections and in situ conservation programs for wild annual Cicer.

Pollen selection for chilling tolerance at hybridisation leads to improved chickpea cultivars

The potential of pollen selection as part of the breeding efforts to increase chilling tolerance in chickpea was investigated. This alternative approach to apply selection pressure at the gametophytic stage in the life cycle has been proposed widely, but there are no reports of the technique being implemented in a crop improvement program. In this paper, we describe how we developed a practical pollen selection technique useful for chickpea improvement.Pollen selection improved chilling tolerance in crossbreds compared with the parental chickpea genotypes and compared with progeny derived without pollen selection. This is backed up by controlled environment assessments in growth rooms and by field studies. We also clearly demonstrate that chilling tolerant pollen ‘wins the race’ to fertilise the ovule at low temperature, using flower color as a morphological marker. Overall, pollen selection results in a lower threshold temperature for podding, which leads to pod setting two to four weeks earlier in the short season Mediterranean-type environments of Western Australia. Field testing at multiple sites across Australia, as part of the national crop variety testing program, shows that these breeding lines have a significant advantage in cool dryland environments.The major factors which affected the success of pollen selection are discussed in the paper, from generation of variability in the pollen to a means to recover hybrids and integration of our basic research with an applied breeding program. We conclude that chilling tolerance observed in the field over successive generations are the result of pollen selection during early generations.

Albinism in Plants: A Major Bottleneck in Wide Hybridization, Androgenesis and Doubled Haploid Culture

Albinism is a common problem encountered in interspecific crosses and tissue culture experiments including anther culture and generation of doubled haploids. It is characterized by partial or complete loss of chlorophyll pigments and incomplete differentiation of chloroplast membranes. This in turn impairs photosynthesis and the plants eventually die at a young stage without reaching maturity. Environmental conditions such as light, temperature, media composition and culture conditions play some role in determining the frequency of albino plant formation. Genetic factors are even more important, and are major determinants in albinism. Genetic studies in different crops show that it is a recessive trait governed by many loci. Both the nuclear and chloroplast genomes affect albinism and incompatibilities between the two are a probable cause of many pigment defects in hybrid progenies. Such incompatibility has been reported in a large number of angiosperms. The mechanisms behind these incompatibilities are poorly understood. Studies of plastid DNA inheritance together with observations using electron microscopy have established that the transmission of plastids can be maternal, paternal or biparental, even within the same genus, especially following wide crosses; contrary to the widespread belief that plastids are almost always transmitted from the maternal parent. Albinism has been overcome in some crop species through somatic hybridization and development of cybrids (cytoplasmic hybrids). However, the strict requirement of efficient protoplast regeneration is a major limitation of these techniques. This review focuses on albinism following interspecific crosses or development of doubled haploids facilitated by tissue culture experiments, underlying mechanisms, and the possibilities for dealing with this important biotechnological limitation.

Toward doubled haploid production in the Fabaceae: progress, constraints, and opportunities

The Fabaceae species have a major role to play in sustainable farming systems, but they have lagged behind other families in respect to the development of doubled haploid protocols for plant improvement. Currently, no plant improvement program uses doubled haploids on a routine basis for any member of the Fabaceae. There has recently been renewed interest in haploid research as the usefulness of doubled haploid material in molecular mapping has become clear. This review provides a comprehensive account of the current information regarding the development of haploid protocols in the Fabaceae. In the Fabaceae crop species there have been isolated reports of haploid plant induction in the phaseoloid clade; soybean, cowpea and pigeonpea, as well as promising progress towards haploidy in peanut and winged bean. As yet there have been no reports of haploid plant production in the galegoid clade, but early stage haploid embryogenesis has been achieved in chickpea, field pea, and lupin. Success in the production of haploid plants has also been reported within the pasture genera Lotus, Medicago, and Trifolium and the arboreal genera Cassia, Peltophorum, and Albizzia. A review of the literature has enabled us to identify some general similarities between the protocols developed for haploid plant induction across the various legumes. These are the culture of intact anthers; use of a cold pretreatment to induce sporophytic development; targeting of microspores at the uninucleate stage of development; and use of MS (Murashige and Skoog, 1962) based nutrient medium with plant growth regulators to encourage continued division following induction. These protocol commonalities will assist researchers to identify approaches suited to their target Fabaceae species. The paucity of research funding for haploid research in most Fabaceae species has highlighted the need for strong collaborative linkages between institutions and researchers. Referees: Professor Laima Kott, Crop Science Department, University of Guelph, Guelph ON N1G 2W1.

The potential of herbaceous native Australian legumes as grain crops: a review

Many agricultural systems around the world are challenged by declining soil resources, a dry climate and increases in input costs. The cultivation of plants that are better adapted than current crop species to nutrient poor soils, a dry climate and low-input agricultural systems would aid the continued profitability and environmental sustainability of agricultural systems. This paper examines herbaceous native Australian legumes for their capacity to be developed as grain crops adapted to dry environments. The 14 genera that contain herbaceous species are Canavalia, Crotalaria, Cullen, Desmodium, Glycine, Glycyrrhiza, Hardenbergia, Indigofera, Kennedia, Lotus, Rhynchosia, Swainsona, Trigonella and Vigna . A number of these genera (e.g., Glycine, Crotalaria, Trigonella and Vigna ) include already cultivated exotic grain legumes. Species were evaluated based on the extent to which their natural distribution corresponded to arid and semi-arid climatic regions, as well as the existing information on traits related to harvestability (uniformity of ripening, propensity to retain pod, pod shattering and growth habit), grain qualities (seed size, chemistry, color and the absence of toxins) and fecundity. Published data on seed yield were rare, and for many other traits information was limited. The Australian species of Vigna , Canavalia and Desmodium mainly have tropical distributions and were considered poorly suited for semi-arid temperate cropping systems. Of the remaining genera Glycyrrhiza and Crotalaria species showed many suitable traits, including an erect growth habit, a low propensity to shatter, flowers and fruits borne at the end of branches and moderate to large seeds (5 and 38 mg, respectively). The species for which sufficient information was available that were considered highest priority for further investigation were Glycine canescens, Cullen tenax, Swainsona canescens, Swainsona colutoides, Trigonella suavissima, Kennedia prorepens, Glycyrrhiza acanthocarpa, Crotalaria cunninghamii and Rhynchosia minima.

Growth, yield and seed composition of native Australian legumes with potential as grain crops

BACKGROUND Many Australian native legumes grow in arid and nutrient-poor environments. Yet few Australian herbaceous legumes have been investigated for domestication potential. This study compared growth and reproductive traits, grain yield and seed composition of 17 native Australian legumes with three commercial grain legumes. RESULTS Seed yields of seven native legumes were > 40% of Cicer arietnum, with highest seed yields and harvest indices in Glycine sp. (14.4 g per plant, 0.54 g g(-1) ) and Lotus cruentus (10.2 g per plant, 0.65 g g(-1) ). Five native species flowered earlier than field pea (Pisum sativa) (109 days), though many were slower to flower and set seed. Largest seeds were found in Glycine canescens (17 mg), with seed of other native species 14 times smaller than commercial cultivars. Seed composition of many native legumes was similar to commercial cultivars (200-330 g protein kg(-1) dry weight (DW), 130-430 g dietary fibre kg(-1) DW). Two Cullen species had high fat content (>110 g kg(-1) DW) and Trigonella sauvissima had the highest crude protein content (370 g kg(-1) DW). CONCLUSION The seed composition and reproductive traits of some wild native Australian legumes suggest they could offer potential as grain crops for soils and environments where the current grain legumes are uneconomic. Further evaluation of genetic diversity, especially for seed size, overall productivity, and reproductive development is needed.

Albinism does not correlate with biparental inheritance of plastid DNA in interspecific hybrids in Cicer species

Cultivated chickpea (Cicer arietinum) was crossed with its wild relatives from the genus Cicer to transfer favorable genes from the wider gene pool into the cultivar. Post-hybridization barriers led to yellowing and subsequent senescence from as early as 5 days after fertilization, however, the ovules of hybrid embryos could be rescued in vitro. Hybrids were classified as green, partially green or albino. The hybrid status of regenerated plantlets in vitro was confirmed by amplification of nuclear DNA markers. To check whether chloroplast development correlated with plastid DNA inheritance in these crosses, primers were designed using conserved plastid gene sequences from wild and cultivated species. All three possible plastid inheritance patterns were observed: paternal, maternal and biparental. This is the first report of biparental inheritance of plastid DNA in Cicer. No correlation was observed between parental origin of the plastid genome and degree of albinism, indicating that chloroplast development in hybrid genotypes was mostly influenced by nuclear factors.

Leaf type is not associated with ascochyta blight disease in chickpea (Cicer arietinum L.)

The three major leaf types in chickpea are normal compound leaf, simple leaf and multipinnate. Simple leaf types are less commonly cultivated worldwide and are often reputed to be susceptible to ascochyta blight disease, whereas other leaf types range from resistant to susceptible. This study determined the association between host plant resistance to ascochyta blight and different leaf types in segregating populations derived from crosses between disease resistant and susceptible chickpea genotypes. In addition, the inheritance of disease resistance and leaf type was investigated in intraspecific progeny derived from crosses between two resistant genotypes with normal leaf type (ICC 3996 and Almaz), one susceptible simple leaf type (Kimberley Large) and one susceptible multipinnate leaf type (24 B-Isoline). Our results showed that, in these segregating populations, susceptibility to ascochyta blight was not linked to multipinnate or simple leaf types; resistance to ascochyta blight depended more on genetic background than leaf shape; leaf type was controlled by two genes with a dihybrid supplementary gene action; normal leaf type was dominant over other leaf types; and inheritance of ascochyta blight resistance was controlled by two major genes, one dominant and one recessive. Since there was no linkage between ascochyta blight susceptibility and leaf type, breeding various leaf types with ascochyta blight resistance is a clear possibility. These results have significant implications for chickpea improvement, as most current extra large seeded kabuli varieties have a simple leaf type.