Aspects of isolation underpinning mitotic behaviour in lupin protoplasts

Anupam Sinha; Peter D. S. Caligari

doi:10.1071/BT03153

RESEARCH ARTICLE

Previous Next Contents Vol 52(5)

Aspects of isolation underpinning mitotic behaviour in lupin protoplasts

Anupam Sinha ^A and Peter D. S. Caligari ^A ^B

+ Author Affiliations

- Author Affiliations

^A School of Plant Science, The University of Reading, Whiteknights, PO Box 221, Reading, Berkshire RG6 6AS, UK.

^B Corresponding author; present address: Instituto de Biología Vegetal y Biotecnología, Universidad de Talca, 2 Norte 685, Talca, Chile. Email: pcaligari@utalca.cl

Australian Journal of Botany 52(5) 669-676 https://doi.org/10.1071/BT03153
Submitted: 6 November 2003 Accepted: 24 June 2004 Published: 25 October 2004

Abstract

This study reports on the influence of critical isolation factors on the subsequent culture of protoplasts of Lupinus albus L. Protoplasts were isolated from in vitro seedling cotyledons of five early maturing accessions in which protoplast yields and division frequencies appeared to be correlated as a high protoplast yield corresponded with a high division frequency. The overall difference among the accessions for mitosis was non-significant, although the highest yield and division frequency were observed in accession LA132, with Alban giving a significantly lower level. Accession Lucrop produced the lowest number of protoplasts, all of which collapsed during culture. Of the enzyme types used for tissue maceration, Pectolyase Y23, was significantly inferior to Macerase in terms of giving way to mitosis. The extent of division in Macerase-isolated protoplast population was 266% higher than that in the Pectolyase Y23-isolated one. The physiological maturity level of the explant, expressed in terms of developmental age, was optimal when 14–18-day-old seedling cotyledons were used for protoplast production and culture, rather than more mature ones, despite higher protoplast yields in the latter. On K8p medium, the protoplast division frequency was 129% greater when 18-day-old seedling cotyledons were used, than that with any other treatment. This work on protoplast culture of the potentially important lupin species, which is a pulse rich in dietary protein, oil and fibre, allows a further understanding of the biology, with an aim to advance lupin biotechnology.

Acknowledgments

We thank Dr Andrew C. Wetten for his help with this work. Anupam Sinha received an ORS award, a Felix Fellowship (administered by University of Oxford, UK) and research grants from Northbrook Society, Sidney Perry Foundation and Leche Trust in partial support of this work.

References

van Ark HF, Hall RD, Creemers-Molenaar J, Krens FA (1992) High yields of cytoplasts from protoplasts of Lolium perenne and Beta vulgaris using gradient centrifugation. Plant Cell Tissue and Organ Culture 31, 223–232.

Atkins CA, Smith PMC, Gupta S, Jones MGK, Caligari PDS (1998) Genetics, cytology and biotechnology. In ‘Lupins as crop plants—biology, production and utilization’. (Eds JS Gladstones, CA Atkins, J Hamblin) pp. 67–92. (Centre for Agriculture and Biosciences International: Wallingford, UK)

Banks W, Muir DD (1980) Structure and chemistry of the starch granule. In ‘The biochemistry of plants—a comprehensive treatise, Vol. 3’. (Eds PK Stumpf, EE Conn) pp. 321–369. (Academic Press, Inc.: San Diego, CA)

Bateman DF, Basham HG (1976) Degradation of plant cell walls and membranes by microbial enzymes. In ‘Physiological plant pathology’. Encyclopaedia of plant physiology. (Eds R Heitefuss, PH Williams) pp. 316–355. (Springer-Verlag: Berlin)

Bergounioux C, Perennes C, Brown SC, Sarda C, Gadal P (1988) Relation between protoplast division, cell-cycle stage and nuclear chromatin structure. Protoplasma 142, 127–136.

Buirchell BJ (1994) Domestication and potential of the rough seeded lupins. In ‘Advances in lupin research’. (Eds JM Neves-Martins, ML Beirão da Costa) pp. 19–24. (Instituto Superior de Agronomia Press: Lisbon)

Burgess J (1983) Wall regeneration around isolated protoplasts. In ‘International review of cytology, supplement 16’. (Ed. KL Giles) pp. 55–77. (Academic Press, Inc.: New York)

Butenko RG (1979) Cultivation of isolated protoplasts and hybridisation of somatic plant cells. International Review of Cytology 59, 323–373.

Butt AD, Bestwick CS (1997) Generation of chemiluminescence during enzymatic isolation of protoplasts from leaves of Oryza sativa. Journal of Plant Physiology 150, 729–733.

Cailloux M, Do CB (1998) Osmoregulation and water regulation in protoplasts of the radish tuber (Raphanus sativus). Canadian Journal of Botany 76, 1835–1842.
| Crossref | GoogleScholarGoogle Scholar |

Caligari PDS, Rahim MA, Huyghe C, Hardy A, Römer P, Neves-Martins JM, Sawicka-Sienkiewicz E (1997) Adaptation of Lupinus mutabilis to European soil and climate conditions. Contract report number AIR3-CT93-0865. (Commission of European Communities: Brussels)

Cowling WA, Gladstones JS (2000) Lupin breeding in Australia. In ‘Linking research and marketing opportunities for pulses in the 21st century’. (Ed. R Knight) pp. 541–547. (Kluwer Academic Publishers: Dordrecht, The Netherlands)

Cowling WA, Huyghe C, Świeçicki W (1998) Lupin breeding. In ‘Lupins as crop plants—biology, production and utilization’. (Eds JS Gladstones, CA Atkins, J Hamblin) pp. 93–120. (Centre for Agriculture and Biosciences International: Wallingford, UK)

Cutler AJ, Saleem M, Coffey MA, Loewen MK (1989) Role of oxidative stress in cereal protoplast recalcitrance. Plant Cell, Tissue and Organ Culture 18, 113–127.

DeDonato M, Perucco E, Chiavazza P, Panei E (1989) Activation of nuclear DNA replication and mitosis in excised cotyledons of Capsicum annuum. Plant Science 63, 217–225.
| Crossref | GoogleScholarGoogle Scholar |

Delmer DP, Stone BA (1988) Biosynthesis of plant cell walls. In ‘The biochemistry of plants—a comprehensive treatise, Vol. 14’. (Eds PK Stumpf, EE Conn) pp. 373–420. (Academic Press, Inc.: San Diego, CA)

Dhir SK, Dhir S, Widholm J (1992) Regeneration of fertile plants from protoplasts of soybean Glycine max L.Merr. Genotypic difference in culture response. Plant Cell Reports 11, 285–289.
| Crossref |

Esau K (1979) ‘Anatomy of seed plants.’ (Wiley Eastern Ltd: New Delhi)

Evans PK, Cocking EC (1977) Isolated plant protoplasts. In ‘Plant tissue and cell culture’. (Ed. HE Street) pp. 103–135. (Blackwell Scientific Publications: Oxford, UK)

Firoozabady E (1986) The effects of cell cycle parameters on cell wall regeneration and cell division of cotton protoplasts (Gossypium hirsutum L.). Journal of Experimental Botany 37, 1211–1217.

Fish N, Karp A (1986) Improvement in regeneration from protoplast of potato and studies on chromosome stability. I. The effect of initial culture media. Theoretical and Applied Genetics 72, 405–412.
| Crossref |

Gilmour DM, Davey MR, Cocking EC, Pental D (1987) Culture of low numbers of forage legume protoplasts in membrane chambers. Journal of Plant Physiology 126, 457–465.

Gladstones JS, Atkins C, Hamblin J (1998) ‘Lupins as crop plants—biology, production and utilization.’ (Centre for Agriculture and Biosciences International: Wallingford, UK)

Gram T, Mattsson O, Joersbo M (1996) Division frequency of pea protoplasts in relation to starch accumulation. Plant Cell, Tissue and Organ Culture 45, 179–183.

Hahne G, Lörz H (1988) Release of phytotoxic factors from plant cell-walls during protoplast isolation. Journal of Plant Physiology 132, 345–350.

Hammatt N, Kim H, Davey MR, Nelson RS, Cocking EC (1987) Plant regeneration from cotyledon protoplasts of Glycine canescens and G. clandestina. Plant Science 48, 129–135.
| Crossref | GoogleScholarGoogle Scholar |

Ishii S (1988) Factors influencing protoplast viability of suspension-cultured rice cells during isolation process. Plant Physiology 88, 26–29.

Ishii S (1989) Enzymes for the isolation of protoplasts. In ‘Biotechnology in agriculture and forestry, vol. 8. Plant protoplasts and genetic engineering, part I’. (Ed. YPS Bajaj) pp. 23–33. (Springer-Verlag: Berlin)

Jung G, Wernicke W (1993) Cell-cycle in potentially dedifferentiating cereal mesophyll protoplast culture in vitro. I. Abnormalities in cycle kinetics. Journal of Plant Physiology 141, 444–449.

Kao KN (1977) Chromosomal behavior in somatic hybrids of soybean—Nicotiana glauca. Molecular and General Genetics 150, 225–230.
| Crossref |

Krautwig B, Lazzeri PA, Lörz H (1994) Influence of enzyme solution on protoplast culture and transient gene expression in maize (Zea mays L.). Plant Cell, Tissue and Organ Culture 39, 43–48.

Lee N, Wetzstein HY, Bornman CH (1989) Cortical microtubule organisation in Vitis protoplasts as affected by concentration of enzyme isolation medium and duration of incubation. Physiologia Plantarum 77, 27–32.

Li X, Xu Z, Wei Z (1995) Plant regeneration from protoplasts of immature Vigna sinensis cotyledons via somatic embryogenesis. Plant Cell Reports 15, 282–286.
| Crossref |

Lu DY, Cooper-Bland S, Pental D, Cocking EC, Davey MR (1983) Isolation and sustained division of protoplasts from cotyledons of seedlings and immature seeds of Glycine max L. Zeitschrift für Pflanzenphysiologie 111, 389–394.

Maheshwari SC, Gill R, Maheshwari N, Gharyal PK (1986) Isolation and regeneration of protoplasts from higher plants. In ‘Differentiation of protoplasts and of transformed plant cells’. (Eds J Reinert, H Binding) pp. 3–36. (Springer-Verlag: Berlin)

McDonald R, Fieuw S, Patrick JW (1996) Sugar uptake by the dermal transfer cells of developing cotyledons of Vicia faba L. Planta 198, 54–63.

Mende P, Wink M (1987) Uptake of the quinolizidine alkaloid lupanine by protoplasts and isolated vacuoles of suspension-cultured Lupinus polyphyllus cells. Diffusion or carrier-mediated transport? Journal of Plant Physiology 129, 229–242.

Millam S, Burns AT, Hocking TJ (1991) A comparative assessment of purification techniques for mesophyll protoplasts of Brassica napus L. Plant Cell, Tissue and Organ Culture 24, 43–48.

Muzquiz M (1992) ‘Lupinus mutabilis: its adaptation and production under European pedoclimatic conditions.’ (Commission of the European Communities: Luxembourg)

Nyman M, Wallin A (1988) Plant regeneration from strawberry (Fragaria × ananassa) mesophyll protoplasts. Journal of Plant Physiology 133, 375–377.

O’Brien IEW, Lindsay GC (1993) Protoplasts to plants of Gentianaceae—regeneration of lisianthus (Eustoma grandiflorum) is affected by calcium-ion, preconditioning, osmolality and pH of the culture media. Plant Cell, Tissue and Organ Culture 33, 31–37.

Ochatt SJ, Power JB (1992) Plant regeneration from cultured protoplasts of higher plants. In ‘Plant biotechnology—comprehensive biotechnology, 2nd supplement’. (Eds MW Fowler, GS Warren, M Moo-Young) pp. 99–127. (Pergamon Press Plc: Oxford, UK)

Parihar DS, Maheshwari SC, Khurana P (1995) High frequency somatic embryogenesis and plantlet regeneration from hypocotyl protoplast cultures of Brassica napus. Plant Cell, Tissue and Organ Culture 42, 113–115.

Perera SC, Ozias-Akins P (1991) Regeneration from sweet-potato protoplast and assessment of growth conditions for flow–sorting of fusion mixtures. Journal of the American Society for Horticultural Science 116, 917–922.

Piccirilli M, Pupilli F, Arcioni S (1988) Lotus tenuis Wald & Kit: In vitro conditions for plant regeneration from protoplasts and callus of various explants. Plant Science 55, 77–82.
| Crossref | GoogleScholarGoogle Scholar |

Pollock CJ, Kingston-Smith AH (1997) The vacuole and carbohydrate metabolism. Advances in Botanical Research 25, 195–215.

Preiss J, Levi C (1980) Starch biosynthesis and degradation. In ‘The biochemistry of plants—a comprehensive treatise, Vol. 3’. (Eds PK Stumpf, EE Conn) pp. 371–423. (Academic Press, Inc.: San Diego, CA)

Repka V (2001) Elicitor-stimulated induction of defense mechanisms and defense gene activation in grapevine cell suspension cultures. Biologia Plantarum 44, 555–565.
| Crossref | GoogleScholarGoogle Scholar |

Revilla MA, Tárrago JF (1988) A histological and histochemical study of starch granule degradation in the cotyledons of Lens culinaris. STARD 40, 294–296.

Roset S, Gilissen LJW (1993) Regeneration from protoplasts—a supplementary literature review. Acta Botanica Neerlandica 42, 1–23.

Schäfer-Menuhr A, Stürmer S (1987) Isolation und kultur von lupinenprotoplasten. II. Modifikation von nährmedien zur beschleunigten teilung von protoplasten aus blättern von Lupinus angustifolius sorte Kubesa. Landbauforschung Völkenrode 37, 231–234.

Shahin EA (1984) Isolation and culture of protoplasts—tomato. In ‘Cell culture and somatic cell genetics of plants, vol. 1’. (Ed. IK Vasil) pp. 370–380. (Academic Press, Inc.: Orlando, FL)

Shimizu JI (1985) Cell regeneration and division of grape mesophyll protoplasts. Journal of Plant Physiology 119, 419–424.

Sinha A, Wetten AC, Caligari PDS (2003a) Effect of biotic factors on the isolation of Lupinus albus protoplasts. Australian Journal of Botany 51, 103–109.
| Crossref | GoogleScholarGoogle Scholar |

Sinha A, Wetten AC, Caligari PDS (2003b) Optimisation of protoplast production in white lupin. Biologia Plantarum 47, 21–25.
| Crossref | GoogleScholarGoogle Scholar |

Simmonds DH (1991) Microtubules in cultured plant protoplast. Acta Botanica Neerlandica 40, 183–195.

Smith DL (1983) Cotyledon anatomy in the Leguminosae. Botanical Journal of the Linnean Society 86, 325–355.

Świeçicki WK, Olejinczak J (1999) Variability of white lupin induced by sodium azide (SA). In ‘Towards the 21st century’. (Ed. GD Hill) pp. 370–373. (International Lupin Association: Canterbury, New Zealand)

Tegeder M, Gebhardt D, Schieder O, Pickardt T (1995) Thidiazuron-induced plant regeneration from protoplasts of Vicia faba cv Mythos. Plant Cell Reports 15, 164–169.
| Crossref |

Vasil IK, Vasil V (1980) Isolation and culture of protoplasts. In ‘International review of cytology, supplement 11B’. (Ed. IK Vasil) pp. 1–19. (Academic Press, Inc.: New York)

Warren G (1991) Protoplast isolation and fusion. In ‘Plant cell and tissue culture’. (Eds A Stafford, G Warren) pp. 50–81. (Open University Press: Milton Keynes, UK)

Webb JK, Woodcock S, Chamberlain DA (1987) Plant regeneration from protoplasts of Trifolium repens and Lotus corniculatus. Plant Breeding 98, 111–118.

Wetten A, Sinha A, Caligari PDS (1999) Electrofusion of lupin protoplasts for the production of interspecific hybrids. In ‘Towards the 21st century’. (Ed. GD Hill) pp. 270–272. (International Lupin Association: Canterbury, New Zealand)

Wood RKS (1976) Killing of protoplasts. In ‘Biochemical aspects of plant–parasite relationship’. (Eds J Friend, DR Threlfall) pp. 105–116. (Academic Press: London)

Xu ZH, Davey MR, Cocking EC (1982) Organogenesis from root protoplasts of the forage legumes Medicago sativa, Trigonella foenum-graecum. Zeitschrift für Pflanzenphysiologie 107, 231–235.

Yang YP, Juang YS, Hsu BD (2002) A quick method for assessing chloroplastic starch granules by flow cytometry. Journal of Plant Physiology 159, 103–106.

Zhao K, Bittisnich DJ, Halloran GM, Whitecross MI (1995) Studies of cotyledon protoplast cultures from B. napus, B. campestris and B. oleracea. I. Cell wall regeneration and cell division. Plant Cell, Tissue and Organ Culture 40, 59–72.

Aspects of isolation underpinning mitotic behaviour in lupin protoplasts

Abstract

Acknowledgments

Subscriber Login