AGRE-0051 Development of Chickpea Germplasm Resistant to Ascochyta Blight and Fusarium Wilt as A Winter Planting Alternative to Southern European Cereal Grains

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ECLAIR Cluster VII - Proteins : Fibre : Integrated Crop Protection & Biological Control : Plant Genetics : Protein/Amino Acid : Starch

In vitro regeneration of chickpea

CURRENT STATE OF THE PROJECT

1. Genetic improvement of chickpea yield through conventional and mutation breeding

The genetic improvement programme has completed the first year as was programmed. Breeding in Spain has been conducted by CIDA, up to F3 and thereafter by Koipesol. Selection has been done in two nurseries by each participant. One, in a cold location for winter hardiness selection and the other one as main nursery for yield and disease resistance selection. Breeding in Italy is based on a single seed descendant, with two generations per year combining nurseries in Chile and Italy. This method has proved to be viable. The mutagenesis breeding has begun by irradiating 2 million chickpea seeds with the goal of inducing resistance to the 6th pathotype of M. rabiei. Yield trials have been completed both in Italy and Spain with the most advanced lines of both programmes. Several hundred crosses have been made combining different lines.

2. Development of In vitro plant breeding techniques for chickpea

The in vitro regeneration of chickpea has been attempted by first inducing callus formation and later regenerating them in a different media. Stems have been obtained from calluses at low frequency. Presently new genotypes and media are under test.

3. Industrial uses of chickpea

Chickpea starches have been isolated and characterized. They were also examined for food application potential. The results show that chickpea starch has an excellent bland neutral taste. It gives similar viscosity instability on storage after exposure to heat, shear or low pH environments. It develops a low viscosity on cooking, but it sets back on overnight storage to give a rigid, opaque gel with syneresis of water. In terms of overall behaviour, chickpea starch can be considered a good alternative to commercial smooth pea starch.

Chickpea samples are classified as whole grain, by a commercial firm in Spain, in regard to their preference for direct human consumption. About l0 traits have been recognized for the evaluation of the preferred market qualities of chickpea. Chemical analysis is now undergoing to correlate these traits with the composition of the grain.

4. Identification of pathotypes of M. rabiei and F. oxysporum f. sp. ciceri and development of chickpea germplasm with resistance to them

One hundred and three chickpea isolates of arium oxysporum have been collected from a range of geographical origins. Twenty four isolates were not pathogenic of the 79 pathogenic isolates, 35 induce the yellowing syndrome and 44 induce the wilting syndrome. Single spore crops of F. o. ciceris and 170 isolates of M. rabiei collected from a range of locations in Spain have been stored for further uses.

Fifty nine F. o. ciceris isolates have been transferred to the U.K. team for APPCR studies including 10 isolates of race 0, 2 of race 5, 1 of each races l, 2, 3 and 4, 26 for which race identification was with held and 21 for which race has not been identified yet.

In blind testing by King's College group, race identification by DNA finger printing has been coincident with biological pathotyping for all isolates reported so far.

The mtDNA from races 07 of FOC has been characterized by RFLP analysis. The mtDNA of FOC is a circular molecule of 40.5 kibbases. The restriction fragment patterns of the mtDNA showed no polymorphism between the seven races.

Thirteen FLIP and four ILC lines from ICARDA have been identified with resistance to races 5 and 0 of FOC. Seven were resistant to the two races. Some 500ILC lines are currently been screened for resistance to race 5 under natural conditions.

The I Ha wiltsick plot is being established at the University farm near Cordoba.

5. Pathotype characterization of FOC and Mr through Randomly Amplified Polymorphic DNA (RAPD)) or Arbitrarily Primed PCR (APPCR) Techniques

The Randomly Amplified Polymorphic DNA (RAPD) or Arbitrarily PrimedPCR (AP PCR) techniques have facilitated the identification of polymorphism within and between different pathotypes of FOC.

TwentyThree primers have been assessed as to their suitability for generation of polymorphism between pathotypes. Primers 2 (from the Intergenic Spacer Region of Penicillium hordei) and KS (a pBluescript sequencing primer) both gave rise to specific polymorphism which were consistent between races. The specific bands found have enabled the correct identification of eight previously identified isolates as well as that of seven 'blind trial' isolates supplied by the plant pathology group of Cordoba. Experiments are in progress to analyze a further 26 isolates.