Plant Regeneration from Mesophyll Protoplasts of Alfalfa

Kao, K. N.; Michayluk, M. R.

doi:10.1016/s0044-328x(80)80051-1

Cited by 195 publications

(60 citation statements)

References 4 publications

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“…First, exogenously supplied ammonium ion has both a qualitative and quantitative effect on the pattern of alfalfa morphogenesis in vitro. Second, our observations are consistent with those of others [4,7,9] wlfich suggest that alfalfa regeneration in vitro involves the formation of somatic embryos.…”

Section: Introductionsupporting

confidence: 92%

Morphogenesis in callus tissue ofMedicago sativa: The role of ammonium ion in somatic embryogenesis

Walker

Sato

1981

Plant Cell Tiss Organ Cult

111

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Abstract. Exogenously supplied ammonium ion is critical to alfalfa morphogenesis in vitro. In alfalfa, the ability to induce the formation of either roots or somatic embryos provided an opportunity to examine the effects of ammonium ion on each pattern of morphogenesis. Somatic embryo formation required a minimum of 12.5 mM NH~ in regeneration medium for optimal expression. Root formation was inhibited by NH~ levels of 50 mM and above, and occurred in the absence of exogenous NH~. At high levels of NH~ somatic embryos were formed from ceils exposed to the root-inducing combination of hormones. This observation suggests that the growth regulators and exogenously supplied ammonium ion comprise an interactive system controlling the in vitro pattern of alfalfa morphogenesis.

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Section: Introductionsupporting

confidence: 92%

Morphogenesis in callus tissue ofMedicago sativa: The role of ammonium ion in somatic embryogenesis

Walker

Sato

1981

Plant Cell Tiss Organ Cult

111

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“…The activities of ODC and ADC in relation to pyrrolidine alkaloid formation were also investigated. (8), and 1% purified BBL agar (BBL Microbiology Systems, Becton Dickinson and Co., Cockeysville, MD) in sterilized Petri dishes (100 x 20 mm) at 25°C in the dark. The agar was purified as described elsewhere (23).…”

mentioning

confidence: 99%

Correlation between Polyamines and Pyrrolidine Alkaloids in Developing Tobacco Callus

Tiburcio¹,

Kaur‐Sawhney²,

Ingersoll³

et al. 1985

Plant Physiol.

109

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Since the diamine putrescine can be metabolized into the pyrrolidine ring of tobacco alkaloids as well as into the higher polyamines, we have investigated the quantitative relationship between putrescine and these metabolites in tobacco callus cultured in vitro. We measured levels of free and conjupted putrescine and spermidine, and pyrrolidine alkaloids, as well as activities of the putrescine-biosynthetic enzymes arginine and ornithine decarboxylase. In callus grown on high (11.5 micromolar) anaphthalene acetic acid, suboptimal for alkaloid biosynthesis, putrescine and spermidine conjugates were the main putrescine derivatives, while in callus grown on low (1.5 micromolar) a-naphthalene acetic acid, optimal for alkaloid formation, nornicotine and nicotine were the main putrescine derivatives. During callus development, a significant negative correlation was found between levels of perchloric acid-soluble putrescine conjugates and pyrrolidine alkaloids. The results suggest that bound putrescine can act as a pool for pyrrolidine alkaloid formation in systems where alkaloid biosynthesis is active. In addition, changes in arginine decarboxylase activity corresponding to increased alkaloid levels suggest a role for this enzyme in the overall biosynthesis of pyrrolidine alkaloids.Polyamines and their biosynthetic enzymes ADC3 (EC 4.1.1.19) and ODC (EC 4.1.1.17) may play an important role in many aspects of plant development including growth, differentiation, senescence, and response to stress (7). However, relatively little is known about their role in secondary plant metabolism. Putrescine is a precursor not only of PAs (7), but also of parts of important alkaloids such as the pyrrolidine rings of tobacco alkaloids (nicotine and nomicotine), tropane alkaloids (hyoscyamine, hyoscine, and meteloidine), pyrrolizidine alkaloids (retronecine), and possibly phenanthroindolizidines (tylophorine) (10). The putrescine-derived PAs (spermidine, homospermidine, and spermine) may be conjugated to cinnamic acids or to fatty acids, giving rise to other more complex alkaloids (18).PAs are present in the plant cell in both free and conjugated forms (19,20 been correlated with the onset of reproduction (4) and virus resistance (12), but not with alkaloid biosynthesis.Several publications support the view that auxin levels are involved in the regulation of nicotine production by tobacco callus cultures (22) and root cultures (21) as well as in the intact tobacco plant (25). Recently, we found (23) that the alkaloid level increased dramatically when tobacco callus grown on 11.5 ,gM NAA (suboptimal for alkaloid biosynthesis) was transferred to a medium containing 1.5 ;tM NAA (optimal for alkaloid biosynthesis). In the present study, we have used this experimental approach to investigate the relation between PAs (free and conjugated) and pyrrolidine alkaloids in tissues where alkaloid biosynthesis is active. The activities of ODC and ADC in relation to pyrrolidine alkaloid formation were also investigated.MATERIALS AND METHODS P...

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“…As previously suggested, high influence of the genotype on the in vitro response of alfalfa, especially for somatic embryogenesis was observed (see Bingham et al, 1975, Kao & Michayluk, 1980, Chen et al, 1987Atanassov & Brown, 1984;Du et al, 1994).…”

Section: In Vitro Proceduresmentioning

confidence: 53%

Plant regeneration from proroplasts of alfalfa (Medicago sativa) via somatic embryogenesis

Monteiro

Appezzato‐da‐Glória

Valarini

et al. 2003

Sci. agric. (Piracicaba, Braz.)

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Alfalfa is one of the most frequently studied species from the production of tissue culturederived embryos point of view. In this study, five alfalfa cultivars were analyzed with reference to their ability to regenerate plants from protoplast cultures via somatic embryogenesis. Plant regeneration from leafderived protoplasts isolated from the cultivar Rangelander was achieved using a protocol defined for alfalfa cell suspension-derived embryogenesis. Because of its high efficiency, this procedure is recommended for protoplast electroporation-mediated genetic transformation of alfalfa.

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Plant Regeneration from Mesophyll Protoplasts of Alfalfa

Cited by 195 publications

References 4 publications

Morphogenesis in callus tissue ofMedicago sativa: The role of ammonium ion in somatic embryogenesis

Morphogenesis in callus tissue ofMedicago sativa: The role of ammonium ion in somatic embryogenesis

Correlation between Polyamines and Pyrrolidine Alkaloids in Developing Tobacco Callus

Plant regeneration from proroplasts of alfalfa (Medicago sativa) via somatic embryogenesis

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