Development of an efficient regeneration system for bombarded calli from immature embryos of Moroccan durum wheat varieties

ArticleOne of the biggest obstacles limiting genetic transformation of durum wheat is the lack of an efficient regeneration system for bombarded tissues. Our study aims to optimize culture conditions for regenerating bombarded calli from immature embryos of four durum wheat varieties ‘Amria’, ‘Chaoui’, ‘Isly’ and ‘Marouane’, through comparing the effects of phytohormones (IAA, zeatin and their interaction) and nitrogen amount and sources on callus induction and plant regeneration. Both tested induction media induced approximately the same rate of induced calli for all the tested varieties. However, the interaction of the induction and the regeneration media showed a highly significant effect on plantlet regeneration for all tested varieties. After bombardment, IM1/RM2 combination proved to be the favourable medium with up to 200% and 120% plantlets regenerated for ‘Chaoui’ and ‘Isly’ varieties respectively. Encouraging results obtained in this study will help to promote the research in genetic transformation and its improvement

Optimising Subsidiary Crop Applications in Rotations (OSCAR): A Perspective for the North Africa Region

Background: For sustainable improvement of wheat-based farming systems in the North Africa, there is a need to improve plant health, soil health and fertility. One approach to achieve this is by integrating subsidiary crops (SC) as living or dead mulches or cover crops with the main crops in rotations, which will increase plant species and microbial diversity and reducing water demand in dry climates. Methods: A collaborative research project funded by EU FP 7 (Project No. 289277) was initiated in April 2012, in partnership with European Union (public research organizations and private sector small and medium enterprises of Germany, Norway, Sweden, Denmark, the Netherland, UK, Poland, Switzerland and Italy), Brazil and Morocco (INRA-Morocco and ICARDA), in order to enhance understanding and use of SC systems, develop suitable farm technologies, increase the range of SC species, and enhance understanding of impact of SC on soil ecology, biology and microbial diversity and crop pests and diseases. Results: Based on two field experiments in Morocco (Sidi AI-Aidi and Sidi Allal-Tazi), the project will assess the economic and ecological impact including legume root health and soil health, fertility and microbial diversity and compare the results with other sites in Europe. Screening of new species and genotypes will result in identification of range of potentially useful plant species for SC for the North Africa and Europe. The identified SC species will be tested for their potential as forage and for extraction of useful biochemicals. Conclusion: The results of the project as a whole will be of use for and improve sustainability in low-input, organic, and conventional farming systems in the North Africa, Latin America and Europe

Genetic structure, diversity, and allelic richness in composite collection and reference set in chickpea (Cicer arietinum L.)

Background Plant genetic resources (PGR) are the basic raw materials for future genetic progress and an insurance against unforeseen threats to agricultural production. An extensive characterization of PGR provides an opportunity to dissect structure, mine allelic variations, and identify diverse accessions for crop improvement. The Generation Challenge Program http://www.generationcp.org conceptualized the development of "composite collections" and extraction of "reference sets" from these for more efficient tapping of global crop-related genetic resources. In this study, we report the genetic structure, diversity and allelic richness in a composite collection of chickpea using SSR markers, and formation of a reference set of 300 accessions. Results The 48 SSR markers detected 1683 alleles in 2915 accessions, of which, 935 were considered rare, 720 common and 28 most frequent. The alleles per locus ranged from 14 to 67, averaged 35, and the polymorphic information content was from 0.467 to 0.974, averaged 0.854. Marker polymorphism varied between groups of accessions in the composite collection and reference set. A number of group-specific alleles were detected: 104 in Kabuli, 297 in desi, and 69 in wild Cicer; 114 each in Mediterranean and West Asia (WA), 117 in South and South East Asia (SSEA), and 10 in African region accessions. Desi and kabuli shared 436 alleles, while wild Cicer shared 17 and 16 alleles with desi and kabuli, respectively. The accessions from SSEA and WA shared 74 alleles, while those from Mediterranean 38 and 33 alleles with WA and SSEA, respectively. Desi chickpea contained a higher proportion of rare alleles (53%) than kabuli (46%), while wild Cicer accessions were devoid of rare alleles. A genotype-based reference set captured 1315 (78%) of the 1683 composite collection alleles of which 463 were rare, 826 common, and 26 the most frequent alleles. The neighbour-joining tree diagram of this reference set represents diversity from all directions of the tree diagram of the composite collection. Conclusion The genotype-based reference set, reported here, is an ideal set of germplasm for allele mining, association genetics, mapping and cloning gene(s), and in applied breeding for the development of broad-based elite breeding lines/cultivars with superior yield and enhanced adaptation to diverse environments

Allelic richness and diversity in global composite collection and reference sets in chickpea (Cicer arietinum L.)

Chickpea is the fourth largest grain legume crop globally. A composite collection of 3000 accessions was formed and genotyped using 50 SSR markers. The accessions were also field evaluated for seven qualitative traits. Analysis of 48 SSR markers data on 2915 accessions detected 1683 alleles, of which 935 were rare and 748 common. Gene diversity varied from 0.533 to 0.975. Kabuli chickpea as a group were genetically more diverse than other seed types. Several group-specific unique alleles were detected: 104 in Kabuli, 297 in desi, and 69 in wild Cicer; 114 each in West Asia and Mediterranean, 117 in South and South East Asia, and 10 in African region accessions. A genotype-based reference set captured 1315 alleles compared to 1237 alleles in the reference set based on qualitative traits or 1354 alleles based on SSRs and qualitative traits data. The relative usefulness of these reference sets in chickpea breeding and genomics studies are being further investigated

Agrobacterium-mediated transformation of mature embryo tissues of bread wheat (Triticum aestivum L.) genotypes

Although significant progress has been made on Agrobacterium-mediated wheat transformation, current methodologies using immature embryos as recipient tissues are labor intensive, time consuming and expensive. The use of mature embryos as explants is increasingly being recognized as an optimal method for developing regenerable cell lines during wheat transformation. Therefore, we have developed an Agrobacterium-based transformation protocol using mature embryos while adjusting several factors that influence genetic transformation efficiency. In this study, we focussed on acetosyringone concentrations, genotypes and different types of mature embryos (intact or longitudinally halved-embryos or fragmented into four latitudinal pieces) used as a source of explants for the genetic transformation. A. tumefaciens strain EHA101 harboring the plasmid vector pTF101.1 carrying the barley HVA1 gene and bar-selectable marker gene were used. Mature intact-embryos and longitudinally halved-embryos yielded the highest number of putative transgenic plantlets on the selection medium. However, no plantlets were obtained from latitudinal fragmented mature embryos. ‘Amal’ and ‘Rajae’ genotypes regenerated the highest number of putative transgenic plants and 200 μM acetosyringone was found to be the optimal concentration for their transformation. A total of 47 transgenic plants were selected with 11 plantlets showing resistance to leaf painting. Molecular analysis revealed that 1% and 0.66% of T0 regenerated plantlets were successfully transformed and carried the HVA1 gene for the ‘Amal’ and ‘Rajae’ genotypes, respectively. Additional analysis shows the transgene is stably inherited in the T1 generation. Based on the results, we conclude that among the influencing factors tested, genotypes, mature embryo explant types and acetosyringone concentration contribute significantly to the success of bread wheat transformation

Towards the first linkage map of the Didymella rabiei genome.

A genetic map was developed for the ascomycete Didymella rabiei (Kovachevski) v. Arx (anamorph: Ascochyta rabiei Pass. Labr.), the causal agent of Ascochyta blight in chickpea (Cicer arietinum L.). The map was generated with 77 F1 progeny derived from crossing an isolate from the U.S.A. and an isolate from Syria. A total of 232 DAF (DNA AmplificationFingerprinting) primers and 37 STMS (Sequence-Tagged Microsatellite Site) primer pairs were tested for polymorphism between the parental isolates; 50 markers were mapped, 36 DAFs and 14 STMSs. These markers cover 261.4cM in ten linkage groups. Nineteen markers remained unlinked. Significant deviation from the expected 1:1 segregation ratios was observed for only two markers (Prob. of x2 <0.05). The implications of our results on ploidy level of the asexual spores are discussed

Improvement of haploid and doubled haploid plants using wheat anther culture

The production of haploids and double haploids, are widely used techniques in advanced breeding programs to speed-up the production of homozygote lines. Anther culture is one of the processes of induction and regeneration of haploids and double haploids from male gametic cells. Due to its high effectiveness and applicability in numerous plant species, it has outstanding potential for plant breeding and commercial exploitation of double haploids. Therefore, the objective of this revue is to discuss recent publications on improvement of wheat anther culture and compare it with older literature. Those publications will be of great utility in yield improvement of haploid and double haploid plants and will contribute for future research on anthers culture