Isolation and Characterization of Mutant Plant Cell Cultures

Widholm, Jack M.

doi:10.1007/978-1-4684-4379-0_9

Cited by 11 publications

(3 citation statements)

References 63 publications

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“…Recent achievements in the selection of variant cell lines have been covered by several recent reviews [4,5,7,9,10,18,20]. It appears that isolation of variant cell lines from leguminous crop is limited to a few reports, all of which used tissue and cell cultures of Glycine max.…”

Section: Introductionmentioning

confidence: 99%

Isolation of L-ethionine resistant cell lines in Vigna sinensis L. after mutagen treatment

Ahmed

Gupta

Ghosh

1986

Plant Cell Tiss Organ Cult

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Chemical mutagenesis was employed for the isolation of variant cell lines resistant to L-Ethionine (L-Eth) in Vigna sinensis L. We have measured cell survival after treatment of Vigna sinensis cell suspensions with different mutagens: Ethyl methanesulfonate (EMS), N-methyl-N-nitrosoguanidine (NTG) and Acridine Orange (AO). NTG was more toxic than EMS and AO.L-Eth resistant colonies were isolated by plating on selective medium after NTG treatment. The frequencies of appearance of resistant cells of MS-3 media supplemented with 20 t~g/ml L-Eth were 1.3 × 10 -6 to 1.8 × 10 -5. The highest number of L-Eth resistant caUi were recorded in cells treated with 10t~g/ml NTG. Few resistant colonies also appeared spontaneously from non-mutagenized cultures with a frequency of 2.9 × 10 -7 to 5.7 × 10 -7. However, a number of isolated colonies were discarded after successive retesting. The number of resistant calli dropped from 204 to 22 during successive retests. The significance of these observations has also been discussed.

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Section: Introductionmentioning

confidence: 99%

Isolation of L-ethionine resistant cell lines in Vigna sinensis L. after mutagen treatment

Ahmed

Gupta

Ghosh

1986

Plant Cell Tiss Organ Cult

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“…Free amino acids accumulate in the mutant tissues selected by amino acid analogs (Widholm, 1972) . Various kinds of mutants resistant to amino acid analogs have, in the past, been taken from cell cultures (Widholm, 1972(Widholm, ,1976(Widholm, ,1978(Widholm, ,1983Palmer & Widholm, 1975 ;Wakasa & Widholm, 1987) and these resistant cell lines have contained increased amounts of specific free amino acids . However, plant regeneration from these cell lines has been difficult because regeneration capacity is lost with subculturing over long periods (Lee & Kameya, 1989) .…”

Section: Introductionmentioning

confidence: 99%

Selection and characterization of a 5-methyltryptophan resistant mutant in Zea mays L.

Kang

Kameya

1993

Euphytica

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A maize plant resistant to 5-methyltrytophan (5MT) was selected from M 2 seeds (Zea mays L . Danggin, inbred line) originating from ears treated with ethylmethane sulfonate (0 .2%) at 6 hr after self-pollination .Genetic analysis of the progeny of plants selected from a medium containing 50 ppm 5MT showed that 5MT resistance was inherited as a single dominant nuclear gene . This resistance was also expressed in callus and seedling. Analysis of the free amino acids in kernels and calli showed that homozygous resistant plants (MR1) contained higher levels of total free amino acids than sensitive plants and calli . In particular, the their kernels the levels of tryptophan, threonine and serine were, respectively, 4 .5, 5 .9 and 6.3 times higher than those of the sensitive plants . From the results, it may be expected that mutants resistant to amino acid analogs will be useful not only for studying amino acid biosynthesis but also for improving the nutritional quality of maize .Abbreviations: EMS -Ethylmethane sulfonate, 5MT -5-methyltryptophan, 2,4-D -2-4-dichlorophenoxyacetic acid

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“…It is advantageous to use tissue culture systems for such selections because they supply, in uniform and pestfree environments, large numbers of cells that can be subjected to the selection pressure of toxic amino acid analogs and then regenerated into plants [5,11]. One means of surviving exposure to an amino acid analog is to overproduce the analog's corresponding amino acid [16,18]. Such amino acid overproduction potentially could alter the nutritional quality of a plant.…”

Section: Introductionmentioning

confidence: 99%

Selection of regenerable maize callus cultures resistant to 5-methyl-DL-tryptophan, S-2-aminoethyl-L-cysteine and high levels of L-lysine plus L-threonine

Miao

Duncan

Widholm

1988

Plant Cell Tiss Organ Cult

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Tissues resistant to lethal levels of equimolar L-lysine plus L-threonine (LT), 5-methyl-DL-tryptophan (5MT, a tryptophan analog), or S-2-aminoethyl-L-cysteine (AEC, a lysine analog) were selected from maize callus capable of plant regeneration (H99 and W77-R3019 genotypes).Resistance to LT resulted from resistant calli having a 19 times greater level of free threonine than wild type tissues. The resistance was expressed in roots of whole plants; threonine levels were two to nine times greater in leaves and kernels of resistant plants than in wild type plants. Slightly greater levels of isoleucine, lysine and methionine were also noted, particularly in the kernel. Genetic studies with individual resistant plants did not always produce inheritance ratios typical of simple Mendelian inheritance, but by the third generation after plant regeneration a trend towards homozygosity was apparent and the data suggests that LT resistance is inherited as a single dominant nuclear gene.Resistance to 5MT resulted from resistant calli having a 133 to 161 times greater level of free tryptophan than wild type tissues. Also, phenylalanine was 22 to 30 times as great and histidine, tyrosine and valine were about two times as great as in wild type tissues. Resistance was expressed in roots of whole plants, and tryptophan levels were at least 2000 times greater in resistant than in wild type plants. Phenylalanine was also 32 times greater. All regenerant plants resistant to 5MT were both male and female sterile.Resistance to AEC was caused by decreased AEC uptake by the callus tissue and was not due to increased levels of free lysine. Plants were not regenerated from this callus.

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Isolation and Characterization of Mutant Plant Cell Cultures

Cited by 11 publications

References 63 publications

Isolation of L-ethionine resistant cell lines in Vigna sinensis L. after mutagen treatment

Isolation of L-ethionine resistant cell lines in Vigna sinensis L. after mutagen treatment

Selection and characterization of a 5-methyltryptophan resistant mutant in Zea mays L.

Selection of regenerable maize callus cultures resistant to 5-methyl-DL-tryptophan, S-2-aminoethyl-L-cysteine and high levels of L-lysine plus L-threonine

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