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Lysine overproducer mutants with an altered dihydrodipicolinate synthase from protoplast culture of Nicotiana sylvestris (Spegazzini and Comes)

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Summary

Two S-(2-aminoethyl)L-cysteine (AEC) resistant lines were isolated by screening mutagenized protoplasts from diploid N. sylvestris plants. Both lines accumulated free lysine at levels 10 to 20-fold higher than in controls. Lysine overproduction and AEC-resistance were also expressed in plants regenerated from the variant cultures. A feedback insensitive form of dihydrodipicolinate synthase (DHPS), the pathway specific control enzyme for lysine synthesis, was detected in callus cultures and leaf extracts from the resistant lines. Aspartate kinase (AK), the other key enzyme in the regulation of lysine biosynthesis, was unaltered in the mutants. Crosses with wild type plants indicated that the mutation conferring insensitivity to feedback in DHPS, with as result overproduction of lysine and resistance to AEC, was inherited as a single dominant nuclear gene.

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Abbreviations

AK:

aspartate kinase (EC 2.7.2.4)

DHPS:

dihydrodipicolinate synthase (EC 4.2.1.52)

AEC:

S-(2-aminoethyl)L-cysteine

References

  • Aarnes H (1974) Aspartate kinase from some higher plants and algae. Physiol Plant 32:400–402

    Google Scholar 

  • Aarnes H (1977) A lysine-sensitive aspartate kinase and two molecular forms of homoserine dehydrogenase from barley seedlings. Plant Sci Lett 9:137–145

    Google Scholar 

  • Aarnes H, Rognes SE (1974) Threonine sensitive aspartate kinase and homoserine dehydrogenase from Pisum sativum. Phytochemistry 13:2717–2724

    Google Scholar 

  • Behrend J, Mateles RI (1975) Nitrogen metabolism in plant cell suspension cultures I. Effect of amino acids on growth. Plant Physiol 56:584–589

    Google Scholar 

  • Bielisky RL, Turner NA (1966) Separation and estimation of amino acids in crude extracts by thin-layer electrophoresis and chromatography. Anal Biochem 17:278–293

    Google Scholar 

  • Bourgin JP, Chupeau Y, Missonier C (1979) Plant regeneration from mesophyll protoplasts of several Nicotiana species. Physiol Plant 45:288–292

    Google Scholar 

  • Bourgin JP, Chupeau MC, Missonier C (1980) Amino acid-resistant plants from tobacco cells selected in vitro. In: Regeneration of plants from cell and tissue culture and genetic variability. Proc CNRS-FNS Workshop, July 1980. Orsay, France, pp 28–40

    Google Scholar 

  • Bright SWJ, Fetherstone LC, Miflin BJ (1979) Lysine metabolism in a barley mutant resistant to S-aminoethylcysteine. Planta 146:629–633

    Google Scholar 

  • Bright SWJ, Miflin BJ, Rognes SE (1982) Threonine accumulation in in seed of a barley mutant with an altered aspartate kinase. Biochem Genet 20:229–243

    Google Scholar 

  • Brock RD, Friederich EA, Landgridge J (1973) The modification of amino acid composition of higher plants by mutation and selection. In: Nuclear techniques for seed protein improvement. Int Atomic Energy Agency, Vienna, pp 329–338

    Google Scholar 

  • Bryan JK (1976) Amino acid biosynthesis and its regulation. In: Bonner J, Verner JE (eds) Plant biochemistry. Academic Press, New York, New York pp 525–560

    Google Scholar 

  • Bryan PA, Cawley D, Brunner CE, Bryan JK (1970) Isolation and characterization of a lysine-sensitive aspartate kinase from a multicellular plant. Biochem Biophys Res Commun 41:1211–1218

    Google Scholar 

  • Cattoir A, Degryse E, Jacobs M, Negrutiu I (1980) Inhibition of barley and Arabidopsis callus growth by lysine analogs. Plant Sci Lett 17:327–332

    Google Scholar 

  • Cattoir-Reynaerts A, Degryse E, Jacobs M (1981a) Isolation of mutants resistant to analogs of amino acids of the aspartate pathway. Arabidopsis. Inf Serv 18:59–65

    Google Scholar 

  • Cattoir-Reynaerts A, Degryse E, Negrutiu I, Aerts M, Jacobs M (1981b) Effects of aspartate-derived amino acids on growth of Arabodopsis plants and callus. Z Pflanzenphysiol 101:67–74

    Google Scholar 

  • Cattoir-Reynaerts A, Degryse E, Verbruggen I, Jacobs M (1983) Isolation and characterization of a lysine + threonine resistant variant in Daucus carota. Biochem Physiol Pflanz 178:81–90

    Google Scholar 

  • Chaleff RS, Carlson PS (1974) In vitro selection for mutants of higher plants. In: Ledoux L (ed) Genetic manipulation with plant material. Plenum Press, New York London, pp 351–357

    Google Scholar 

  • Cheshire RH, Miflin BJ (1975) The control of lysine biosynthesis in maize. Phytochemistry 14:695–698

    Google Scholar 

  • Davies HM, Miflin BJ (1977) Aspartate kinase from carrot cell suspension culture. Plant Sci Lett 9:323–332

    Google Scholar 

  • Demain AL (1975) Overproduction of microbial metabolites and enzymes due to alterations of regulation. In: Ghose TK, Fischer A (eds) Adv Biochem Engin, vol I. Springer, Berlin Heidelberg New York, pp 113–142

    Google Scholar 

  • Dierks-Ventling C (1983) Lysine biosynthesis and utilization during seed development of normal and opaque-2 Zea mays. Planta 157:233–238

    Google Scholar 

  • Durand J (1979) High and reproducible plating efficiencies of protoplasts isolated from in vitro grown haploid Nicotiana sylvestris. Z Pflanzenphysiol 93:283–295

    Google Scholar 

  • Filner P (1966) Regulation of nitrate reductase in cultured tobacco cells. Biochim Biophys Acta 118:229–310

    Google Scholar 

  • Heimer YM, Filner P (1971) Regulation of nitrate assimilation pathway of cultured tobacco cells. Biochim Biophys Acta 215:152–165

    Google Scholar 

  • Hibberd KA, Green CE (1982) Inheritance and expression of lysine plus threonine resistance selected in maize tissue culture. Proc Natl Acad Sci USA 79:559–562

    Google Scholar 

  • Hibberd KA, Walter T, Green CE, Gengenbach BG (1980) Selection and characterization of feedback-insensitive tissue culture of maize. Planta 148:183–187

    Google Scholar 

  • Lowry OH, Rosenbrough NL, Farr AL, Randall RJ (1951) Protein measurements with the Folin Phenol reagent. J Biol Chem 193:265–275

    Google Scholar 

  • Magnien E, Dalschaert X, Romengous M, Devreux M (1980) Improvement of protoplast isolation and culture technique from axenic plantlets of wild Nicotiana species. Acta Genet Sin 7:231–240

    Google Scholar 

  • Matthews BF, Widholm, JM (1978) Regulation of lysine and threonine synthesis in carrot cell suspension cultures and whole carrot roots. Planta 141:315–321

    Google Scholar 

  • Matthews BF, Shye SCH, Widholm JM (1980) Mechanism of resistance of a selected carrot cell suspension culture to S-aminoethylcysteine. Z Pflanzenphysiol 96:453–463

    Google Scholar 

  • Miflin BJ (1969) The inhibitory effects of various amino acids on the growth of barley seedlings. J Exp Bot 20:810–819

    Google Scholar 

  • Nakayama K, Tanaka H, Hagino H, Kinoshita S (1966) Studies on lysine fermentation. Agric Biol Chem 30:611–616

    Google Scholar 

  • Negrutiu I, Mousseau J (1980) Protoplast culture from in vitro grown plants of Nicotiana sylvestris. Z Pflanzenphysiol 100:373–376

    Google Scholar 

  • Negrutiu I, Mousseau J (1981) Culture sur grande échelle de protoplastes de Nicotiana sylvestris à partir de plantes cultivées in vitro. Z Pflanzenphysiol 103:367–372

    Google Scholar 

  • Negrutiu I, Muller JF (1981) Culture conditions of protoplast-derived cells of Nicotiana sylvestris for mutant selection. Plant Cell Rep 1:14–17

    Google Scholar 

  • Negrutiu I, Jacobs M, Cattoir A (1978) Arabidopsis thaliana, espèce modèle en génétique cellulaire. Physiol Veg 16:365–379

    Google Scholar 

  • Sano K, Shiio I (1970) Microbial production of lysine. 3. Production by mutants resistant to S-aminoethylcysteine. J Gen Appl Microbiol 16:373–391

    Google Scholar 

  • Schumacker W (1967) Die Fernleitung der Stoffe in Pflanzenkörpern: In: Ruhland W (Hrsg) Handb Pflanzenphysiol, Bd XIII. S 61–177

  • Shaeffer GW, Sharpe FT (1981) Lysine in seed protein from S-aminoethylcysteine resistant anther-derived cultures of rice. In Vitro 17:345–351

    Google Scholar 

  • Sharma AK, Sharma A (1972) Chormosome techniques. Butterworth, University Park Press, pp 155–156

    Google Scholar 

  • Shewry PR, Miflin BJ (1977) Properties and regulation of aspartate kinase from barley seedlings. Plant Physiol 59:69–73

    Google Scholar 

  • Stern R, Mehler AH (1965) Lysyl-tRNA synthetase from E. coli. Biochem Z 342:400–409

    Google Scholar 

  • Umbarger HE (1971) Metabolite analogs as genetic and biochemical probes. Adv Genet 16:119–140

    Google Scholar 

  • Widholm JM (1976) Selection and characterization of cultured carrot and tobacco cells resistant to lysine, methionine, and proline analogs. Can J Bot 54:1523–1528

    Google Scholar 

  • Widholm JM (1978) Problems in selecting mutant cells. IAPTC Newslett 23:2–6

    Google Scholar 

  • Wong KF, Dennis DT (1973) Aspartokinase from wheat germ. Plant Physiol 51:322–326

    Google Scholar 

  • Yugari Y, Gilvarg C (1965) The condensation step in diaminopimelate synthesis. J Biol Chem 240:4710–4715

    Google Scholar 

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Authors and Affiliations

  1. Plantengenetica, Instituut voor Moleculaire Biologie, Vrije Universiteit Brussel, St. Genesius Rode, B-1640, Belgium

    I. Negrutiu, A. Cattoir-Reynearts, I. Verbruggen & M. Jacobs

Authors
  1. I. Negrutiu
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  2. A. Cattoir-Reynearts
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  3. I. Verbruggen
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  4. M. Jacobs
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Communicated by G. Wenzel

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Negrutiu, I., Cattoir-Reynearts, A., Verbruggen, I. et al. Lysine overproducer mutants with an altered dihydrodipicolinate synthase from protoplast culture of Nicotiana sylvestris (Spegazzini and Comes). Theoret. Appl. Genetics 68, 11–20 (1984). https://doi.org/10.1007/BF00252303

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  • DOI: https://doi.org/10.1007/BF00252303

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