Monitoring by two‐dimensional electrophoresis somatic embryogenesis in leaf and petiole explants from Vitis

Abstract

A two-dimensional electrophoretic analysis of the total proteins was carried out in Vitis rupestris as model system in order to characterize the different developmental stages--from callus to plantlets--of somatic embryogenesis events in the grapevine. The patterns of callus, embryogenetic callus, somatic embryos and plantlets derived from leaf and petiole explants were compared. Each differentiation step was characterized by specific peptide spots.

Optimization of protein extraction and solubilization for mature grape berry clusters

“…Table 2 indicates the specificity of the excised spots (last column: spot specificity). For instance, 8,6, and 15 spots were specific to protocol 1, 2, and 4, respectively. All the 44 spots excised from 2-D pattern corresponding to protocol 3 were present on 2-D pattern from protocol 4 as well.…”

“…In 1992, Tesniere and Robin [7] adapted Hsu and Heatherbell's protocol to study proteins from mature red berries; they could visualize up to 52 spots on silver nitrate-stained 2-D gels. The same year, Gianazza et al [8] isolated proteins from callus regenerated from Vitis leaf and petiole and separated on silver nitrate-stained 2-DE gels, on which they could resolve up to 900 spots. The difference in number of spots between grapevine berry and callus may highlight the variation of protein extraction efficiency from one tissue to another within the same organism.…”

Somatic Embryogenesis in Grapevine

“…The knowledge of the biochemical changes occurring during morphogenesis would provide suitable markers for predicting the embryogenic potential of cultured explants and calli and for allowing an objective selection among cultures. The biochemical changes associated with morphogenesis, from explant to plantlet, were analyzed in Vitis rupestris S.: the analysis of the electrophoretic patterns of total proteins (Gianazza et al, 1992) and specific enzymes (acid phosphatase, alcohol dehydrogenase, esterase) (Martinelli et al, 1993 b) proved to be effective approaches to the characterization of the main steps of somatic embryogenesis.…”

Evaluation of Protein Extraction Methods for Vitis vinifera Leaf and Root Proteome Analysis by Two‐Dimensional Electrophoresis

“…Thus, Tesnières and Robin (1992) previously adapted Hsu and Heatherbell's protocol, based on phosphate buffer extraction, to study proteins from mature red berries and were able to visualize up to 52 spots on silver nitrate‐stained 2‐D gels. In the same year, Gianazza et al (1992) isolated proteins from callus regenerated from Vitis leaves and petioles and visualized on silver nitrate‐stained 2‐D gels, on which they could resolve up to 900 spots. In 2004, Sarry et al could detect up to 381 spots from the grape berry mesocarp using a trichloroacetic acid‐acetone (TCA‐acetone) based method.…”