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GrainGenes Reference Report: PRT-23-9

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Reference	PRT-23-9
Title	A highly efficient plant regeneration system through multiple shoot differentiation from commercial cultivars of barley (Hordeum vulgare L.) using meristematic shoot segments excised from germinated mature embryos
Journal	Plant Cell Reports
Year	2004
Volume	23
Pages	9-16
Author	Sharma VK Hansch R Mendel RR Schulze J
Abstract	A fast and highly efficient short-term in vitro regeneration system was developed for barley (Hordeum vulgare L.) based on readily available explants. Clumps of multiple shoots and buds suitable for transformation were obtained 9-10 weeks after culture initiation from model and current commercial cultivars. Meristematic shoot segments (MSSs) excised from mature embryo-derived seedlings and subsequently cultured on MS-based medium containing 2 mg/l Picloram and 3 mg/l thidiazuron (TDZ) differentiated up to ten multiple shoots after 3-4 weeks with no or very little callus formation. Sectors of the already multiplied shoot clumps were further multiplied on proliferation-maintenance medium containing 2 mg/l Picloram and 2.5 mg/l TDZ. Biweekly subcultures resulted in a continuous process of multiplication of these highly differentiating green sectors without any loss of morphogenic potential. The differentiated small shoots and shoot buds gave rise to normal shoots on medium with 0.1 mg/l Picloram and 1 mg/l TDZ. After rooting on basal medium with 0.5 mg/l or 1 mg/l IBA the plants were transferred to soil and showed normal growth and fertility compared to the seed-grown plants. All of the genotypes tested formed multiple shoots. The percentage of relative MSS multiplication was 63-83%, and the average number of multiplied shoots per MSS ranged from 16 to 34 among the genotypes after 9-11 weeks
External Databases	Pubmed: 15221277
Keyword	[ Hide all but 1 of 24 ] barley callus differentiation efficient fertile barley genetic-transformation genotype growth hordeum vulgare immature inflorescences meristematic shoot segment microprojectile bombardment model multiple shoots particle bombardment plant regeneration regeneration sativa l. seedling shoot meristem soil somatic embryogenesis tissue-cultures transformation

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