European project 5 th PCRDT GARLIC AND EALTH

European project « 5 th PCRDT » GARLIC AND EALTH Second annual report Partner P 8 CIRAD-FLHOR

ADVANCEMENT OF THE Works

Scientific and administrative team • L. FEREOL scientist – Duties: somatic embryogenesis, histology and flow cytometry • S. CAUSSE technical assistant – Duties: somatic embryogenesis, maintenance of the material • M. Roux-Cuvelier – Duties: field trials • R. Kahane – Duties: management

Objectives • Somatic embryogenesis • Comparative characterisation of plantlets from somatic embryos

Work plan • Establishment of somatic embryogenesis procedure for four varieties – Rouge Reunion – Messidrome – Morasol – Printanor • Production and provision of somatic embryos via callus and via cell suspensions of these varieties

Work plan (continue) • Genetic evaluation of somatic embryos derived plants by : – – flow cytometry (by P 8) biochemistry (by P 5) molecular markers (by P 1) morphological and physiological analyses (by P 8, P 9)

Milestones of the first year ( = achieved; partly achieved) Provision of 200 plantlets from embryogenic calli « Rouge Reunion » Provision of 200 plantlets from embryogenic cell suspension « Rouge Reunion » Production of embryogenic callus « Messidrome » , « Morasol » Initiation of embryogenic cell suspension « Messidrome » , « Morasol » Production and germination of callus and cell suspension derived somatic embryos

Milestones of the second year Genetic characterisation of somatic embryos derived plantlets « Rouge Reunion » By flow ctytometry(P 8), finger printing(P 1), dry matter and sulphur content(P 5), morpho-physiology(P 9) Provision of 200 plantlets from embryogenic calli of « Messidrome » , « Morasol » Provision of 200 plantlets from embryogenic cell suspension of « Messidrome » , « Morasol » Genetic characterisation of these plants by flow cytometry

Milestones of the second year (continue) Production of embryogenic callus « Printanor » Initiation of embryogenic cell suspension « Printanor » Production and germination of callus and cell suspension derived somatic embryos « Printanor »

Deliverables DP 5 paper on: Evidence of a somaic embryogenesis process and plant regeneration and acclimatisation in garlic (Allium sativum L. ) L. FEREOL, V. CHOVELON, S. CAUSSE, N. MICHAUX FERRIERE and R. KAHANE DP 13 paper on: Establishment of garlic (Allium sativum L. ) cell suspensions and plant regeneration L. FEREOL, V. CHOVELON, S. CAUSSE, D. TRIAIRE, N. MICHAUX FERRIERE and R. KAHANE S U B M I Ted In P R O G R ESS

RESEARCH ACTIVITIES

Different morphological and histological aspect of garlic calluses Compact non embryogenic Semi-friable non embryogenic Friable non embryogenic Compact with embryogenic potentiel Semi-friable with embryogenic potentiel Friable with embryogenic potentiel

Established cell suspension Macroscopical aspect Microscopical aspect Histological aspect

Cell suspension before plating Macroscopical aspect Microscopical aspect Histological aspect

Plating after 16 days Macroscopical aspect Histological aspect

Plating after 30 days Macroscopical aspect Histological aspect

Plating after 45 days Macroscopical aspect Histological aspect

Media used for garlic somatic embryogenesis via callus and via cell suspension

Growth of cell suspension cultures

Effect of age of cell suspension culture on its multiplication rate

Effect of age of cell suspension culture on its regeneration

Effect of cell density at plating on number of embryos regenerated