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  • 1
    ISSN: 1432-203X
    Keywords: Key words Agrobacterium tumefaciens ; Benzylisoquinoline alkaloids ; California poppy ; Eschscholzia californica Cham. ; Genetic transformation ; Somatic embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  An efficient Agrobacterium-mediated protocol for the stable genetic transformation of Eschscholzia californica Cham. (California poppy) via somatic embryogenesis is reported. Excised cotyledons were co-cultivated with A. tumefaciens strain GV3101 carrying the pBI121 binary vector. Except for the co-cultivation medium, all formulations included 50 mg l−1 paromomycin as the selective agent and 200 mg l−1 timentin to eliminate the Agrobacterium. Four to five weeks after infection, paromomycin-resistant calli grew on 80% of explants in the presence of 2.0 mg l−1 1-naphthaleneacetic acid (NAA) and 0.1 mg l−1 6-benzylaminopurine (BAP). Calli were cultured on somatic embryogenesis induction medium containing 1.0 mg l−1 NAA and 0.5 mg l−1 BAP, and somatic embryos were visible on 30% of the paromomycin-resistant calli within 3–4 weeks. Three to four weeks after the somatic embryos were transferred to phytohormone-free plant regeneration medium, 32% converted to paromomycin-resistant plants. Detection of the neomycin phosphotransferase gene and high levels of β-glucuronidase (GUS) mRNA and enzyme activity, and the cytohistochemical localization of GUS activity in all plant tissues confirmed the integrative transformation of the regenerated plants. The normal alkaloid profile of California poppy was unaffected by the transformation process; thus, the reported protocol could serve as a valuable tool to investigate the molecular and metabolic regulation of the benzophenanthridine alkaloid pathway.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell reports 19 (2000), S. 421-426 
    ISSN: 1432-203X
    Keywords: Key words California poppy ; Eschscholzia californica ; Plant regeneration ; Somatic embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The development of a rapid protocol for high-efficiency somatic embryogenesis and plant regeneration from seed-derived embryogenic callus cultures of California poppy (Eschscholzia californica Cham.) is reported. The optimized procedure required less than 13 weeks from the initiation of seed cultures to the recovery of plantlets and involved the sequential transfer of cultures onto solid Murashige and Skoog basal medium containing three different combinations of growth regulators. All steps were performed at 25  °C. Friable primary callus was induced from seeds of E. californica cultured on medium supplemented with 1.0 mg l−1 2,4-dichlorophenoxyacetic acid. The primary callus was transferred to medium containing 1.0 mg l−1 1-naphthaleneacetic acid and 0.5 mg l−1 6-benzylaminopurine to establish embryogenic callus and promote somatic embryogenesis. Regenerated plantlets were recovered after the conversion of somatic embryos on medium containing 0.05 mg l−1 6-benzylaminopurine and showed normal development. Embryogenic callus was induced at a frequency of 85%, an average of 45 somatic embryos were produced per callus, 90% of the somatic embryos converted, and about 70% of the plantlets were recovered in soil. The growth rate of somatic embryo-derived shoots could be increased by gibberellic acid treatment, but the resulting plantlets were hyperhydritic.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1573-5036
    Keywords: field bean ; nodulation ; N2 fixation ; nitrate ; Phaseolus spp. ; Rhizobium phaseoli
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Common bean,Phaseolus vulgaris L., is known to be ‘inefficient’ in nodulation and N2 fixation although it responds to applied nitrogen. An experiment was conducted to identify and to characterize bean cultivars nodulating in the presence of a high level of nitrogen. Sixteen cultivars and a check for inefficient nodulation, OAC Seaforth, were inoculated and grown for 40 days in replicated pots supplied with zero, 3.5 and 10.5 mM combined nitrogen as NO 3 − and NH 4 + . Seven traits relating to nodulation and N2 fixation were all significantly affected by N level (N), cultivar (Cv) and N × Cv interactions (except for root dry weight), indicating that cultivars responded differently to the N treatments. Total dry weight (W) and shoot to root ratio (S/R) increased with the increased N levels. Nodule dry weight (Wn), visual nodulation score (Nv) and nodulation index (Nx) decreased as the N increased. Percent N and N content per plant increased with the increased N level. Plant weight (W) was positively correlated with Wn, Nv and N content and negatively correlated with %N. Nodulation score was positively associated with Wn and plant N content. Genotypes superior in nodulation and N2 fixation in the presence of N were identified. Cultivars Italian Barlotti, California Light Red Kidney, Kentucky Wonder A and Pueblo 152 were selected for further testing and use in improving the nitrate tolerant nodulating characteristic of bean.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1573-5036
    Keywords: dry bean ; N2 fixation ; nodulation mutants ; Rhizobium leguminosarum bv. phaseoli
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Seed of white bean (Phaseolus vulgaris L. cv. OAC Rico) was treated with 0.04 M ethyl-methane sulphonate. Screening of the M2 progeny from 175 M1 lines in the presence of 1 mM nitrate revealed two nodulation mutants. One line was essentially non-nodulating in several tests, but small white nodules were observed occasionally in other tests with 16 separate Rhizobium strains. The other line formed tiny, creamy white, non-functional (ineffective) and tumor-like pseudo-nodules. The M3 and M4 progenies were fertile and bred true.
    Type of Medium: Electronic Resource
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