André Toussaint

Role of myo-inositol phosphate synthase and sucrose synthase genes in plant seed development

The aim of this review is to highlight the role of myo-inositol phosphate synthase (MIPS), which catalyses the first step in inositol biosynthesis and of sucrose synthase (Sus), an enzyme involved in UDP-glucose formation, the principal nucleoside diphosphate in the sucrose cleavage reaction and in trehalose biosynthesis. These two enzymes are involved in various physiological processes including seed growth and resistance to biotic and abiotic stresses. The study of mutated MIPS and Sus genes in some crops, such as soybean and cotton, has shown that these two proteins are directly involved in embryogenesis. They exhibit several isoforms that are essential for normal seed development. The possible role of both genes in seed development is discussed in this review.

Identification and characterization of drought stress responsive genes in faba bean (Vicia faba L.) by suppression subtractive hybridization

Faba bean (Vicia faba L.) ranks fourth in food legume crop production in the world. However, drought is a potential major constraint to faba bean production and improved faba bean cultivars and development of drought-resistant varieties play a key role in enhancing faba bean crop production. In this study, suppression subtractive hybridization (SSH) technique was used to study differential expression in response to water stress and to identify genes involved in molecular mechanism of drought tolerance. A forward subtractive cDNA library induced by water deficit conditions was constructed used Hara faba bean cultivar grown in pots and treated with either well-watered (WW) or water-stressed (WS). A total of 28 clones were identified as drought stress induced. After sequencing, ten unique expressed sequence tags (ESTs) were obtained by clustering and blast analysis which showed homology to known drought responsive genes including heat shock protein (HSP), late embryogenic abundant (LEA), zinc finger protein transcription factors (ZFP), lipid transfer protein (LTP), chlorophyll a/b-binding protein (ChlBP), thioredoxin h (Trx h), and ATP synthase as well as some functionally unknown transcripts. Their expression was characterized in Leaf, root, flower, cotyledon, and stem tissue. Quantitative RT-PCR analysis revealed that eight genes were consistently up-regulated in Hara compared to Giza3 cultivar, known as drought-tolerant and sensitive respectively under water deficit treatment. The expression of six genes was differentially expressed in different stages of water stress faba bean plant. Drought responsive genes showed changed expression patterns, indicating that they may play important roles in faba bean water stress response. Furthermore, these results indicate that drought-induced genes are related to metabolic pathways and genetic regulation of stress and development and can serve as a foundation for future studies to elucidate drought stress mechanisms of faba bean.

In silico identification and characterization of putative differentially expressed genes involved in common bean (Phaseolus vulgaris L.) seed development

Two genotypes of common bean (Phaseolus vulgaris L.) were studied to determine the structural cause of seed abortion in this species. In the non-abortive control (wild-type, cultivar BAT93), the histological analysis revealed a classical pattern of seed development and showed coordinated differentiation of the embryo proper, suspensor, endosperm tissue and seed coat. In contrast, the ethyl methanesulfonate (EMS) mutant (cultivar BAT93) showed disruption in the normal seed development leading to embryo abortion. Aborted embryos from these degenerate seeds showed abnormalities in suspensor and cotyledons at the globular, heart, torpedo and cotyledon stages. Exploring the feasibility of incorporating the available online bioinformatics databases, we identified 22 genes revealing high homology with genes involved in Arabidopsisthaliana embryo development and expressed in common bean immature seeds. The expression patterns of these genes were confirmed by RT–PCR. All genes were highly expressed in seed tissues. To study the expression profiles of isolated genes during Phaseolus embryogenesis, six selected genes were examined by quantitative RT–PCR analysis on the developing embryos of wild-type and EMS mutant plants. All selected genes were expressed differentially at different stages of embryo development. These results could help to improve understanding of the mechanism of common bean embryogenesis.

Phaseolus Immature Embryo Rescue Technology

Predominant among the production constraints of the common bean Phaseolus vulgaris are infestation of Ascochyta blight, Bean Golden Mosaic virus (BGMV), and Bean Fly. Interbreeding with Phaseolus -coccineus L. and/or Phaseolus polyanthus Greenm has been shown to provide P. vulgaris with greater resistance to these diseases. For interspecific crosses to be successful, it is important to use P. coccineus and P. polyanthus as female parents; this prevents rapid reversal to the recurrent parent P. vulgaris. Although incompatibility barriers are post-zygotic, early hybrid embryo abortion limits the success of F1 crosses. While rescue techniques for globular and early heart-shaped embryos have improved in recent years, -success in hybridization remains very low. In this study, we describe six steps that allowed us to rescue 2-day-old P. vulgaris embryos using a pod culture technique. Our methods consisted of (i) pod culture, (ii) extraction and culture of immature embryos, (iii) dehydration of embryos, (iv) germination of embryos, (v) rooting of developed shoots, and (vi) hardening of plantlets.

Alien chromosome transmission and somatic elimination in monosomic addition lines of Gossypium australe F. Muell in G. hirsutum L.

The efficiency of using monosomic alien addition lines (MAALs) to introgress agronomical traits of interest carried by wild diploid Gossypium species into the main cultivated cotton species G. hirsutum depends on the opportunities of confronting the alien chromosome with the recipient background genome at each generation and on the occurrence of translocations and homoeologous recombinations. The selfed-progeny of five MAALs of G. australe in G. hirsutum was screened with SSR markers to determine the transmission frequency of the alien chromosome and monitor its integrity. Three MAALs revealed a transmission frequency significantly lower than the expected ratio and one MAAL presented an exclusive transmission of the additional chromosome. In these four MAAL the alien chromosome was transmitted almost unaltered. With the fifth MAAL the alien chromosome was normally transmitted but was altered in half of the plants carrying it. In one MAAL, normally carrying brown fiber, the emergence of some plants carrying white and brown fiber revealed the somatic elimination of the additional chromosome. The loss of this chromosome seems to be triggered by its deletion.