Improvement in Lupinus luteus (Fabaceae) protoplast culture – stimulatory effect of agarose embedding and chemical nursing on protoplast divisions
Alina Wiszniewska A B and Anna Pindel AA Department of Botany and Plant Physiology, Faculty of Horticulture, University of Agriculture, Krakow, Poland.
B Corresponding author. Email: czuraa@ogr.ar.krakow.pl
Australian Journal of Botany 57(6) 502-511 https://doi.org/10.1071/BT09068
Submitted: 8 April 2009 Accepted: 2 September 2009 Published: 9 November 2009
Abstract
Comprehensive studies were undertaken to determine optimal conditions for yellow-lupin (Lupinus luteus L.) protoplast culture. The adopted isolation procedure resulted in a high yield of protoplasts obtained from hypocotyls, cotyledons and young leaves. The usefulness of liquid and agarose-solidified media was evaluated in relation to promoting survivability and morphogenetic responses of protoplasts. The influence of different plant-growth regulators and anti-oxidative agents was also examined. Cultivar ‘Parys’ was the most promising material for manipulations. The solidification of medium enhanced the development of cultures initiated from hypocotyls and cotyledons, significantly increasing the division rate. In both liquid and solid media, mitoses were arrested after the initial division and daughter cells failed to divide. An important breakthrough in the development of cultures appeared in media supplemented with 0.1% activated charcoal. Bypassing the mitosis suppression led to the formation of small aggregates from hypocotyl protoplasts. This is the first report on protocolony formation in protoplast cultures of L. luteus. Notable progress achieved here indicates that, although this species is considered as recalcitrant to in vitro manipulations, it is possible to obtain responsive cultures with higher morphogenetic potential and thus provide new material for breeding programs of lupins.
Aftab F, Iqbal J
(1999) Plant regeneration from protoplasts derived from cell suspension of adventive somatic embryos in sugarcane (Saccharum spp. hybrid cv. CoL-54 and cv. CP-43/33). Plant Cell, Tissue and Organ Culture 56, 155–162.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
Babaoglu M
(2000) Protoplast isolation in lupin (Lupinus mutabilis Sweet): determination of optimum explant sources and isolation conditions. Turkish Journal of Botany 24, 177–185.
Barthou H,
Petitprez M,
Briere C,
Souvre A, Alibert G
(1999) RGD-mediated membrane-matrix adhesion triggers agarose-induced embryoid formation in sunflower protoplasts. Protoplasma 206, 143–151.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
Binding H
(1974) Regeneration von haploiden und diploiden Pflanzen aus Protoplasten von Petunia hybrida. Zeitschrift für Pflanzenphysiologie 74, 327–356.
Ciesiołka D,
Gulewicz P,
Martinez-Villaluenga C,
Pilarski R,
Bednarczyk M, Gulewicz K
(2005) Products and biopreparations from alkaloid-rich lupin in animal nutrition and ecological agriculture. Folia Biologica 53, 59–66.
| Crossref | GoogleScholarGoogle Scholar |
Czura A, Pindel A
(2007) Limitation of callus browning during its development in in vitro cultures of yellow lupin. Advances of Agricultural Sciences – Problem Issues [in Polish] 522, 449–455.
Davey MR,
Anthony P,
Power JB, Lowe KC
(2005a) Plant protoplasts: status and biotechnological perspectives. Biotechnology Advances 23, 131–171.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Davey MR,
Anthony P,
Power JB, Lowe KC
(2005b) Plant protoplast technology: current status. Acta Physiologiae Plantarum 27, 117–130.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
Dhir SK,
Dhir S, Widholm JM
(1991) Plantlet regeneration from immature cotyledon protoplasts of soybean (Glycine max L.). Plant Cell Reports 10, 39–43.
Duranti M
(2006) Grain legume proteins and nutraceutical properties. Fitoterapia 77, 67–82.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Frearson EM,
Power JB, Cocking EC
(1973) The isolation, culture and regeneration of Petunia leaf protoplasts. Developmental Biology 33, 130–137.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Gaff DF, Okang’o-Ogola O
(1971) The use of non-permeating pigments for testing the survival of cells. Journal of Experimental Botany 22, 756–758.
| Crossref | GoogleScholarGoogle Scholar |
Gamborg OL,
Miller RA, Ojima K
(1968) Nutrient requirements of suspension cultures of soybean root cells. Experimental Cell Research 50, 151–158.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Guo Y,
Bai J, Zhang Z
(2007) Plant regeneration from embryogenic suspension-derived protoplasts of ginger (Zingiber officinale Rosc.). Plant Cell, Tissue and Organ Culture 89, 151–157.
| Crossref | GoogleScholarGoogle Scholar |
Humphries JM, Hughes SJ
(2006) Pharmaceutical, nutraceutical and industrial potential of temperate legumes. CRC Salinity Bulletin 2, 1–174.
Kao KN
(1977) Chromosomal behaviour in somatic hybrids of soybean–Nicotiana glauca. Molecular & General Genetics 150, 225–230.
| Crossref | GoogleScholarGoogle Scholar |
Khatun A,
Davey MR,
Cocking EC, Power JB
(2002) Organogenesis from protoplasts of tossa jute (Corchorus olitorius). Online Journal of Biological Sciences 2, 612–615.
Kunitake H,
Nakashima T,
Mori K,
Tanaka M, Mii M
(1995) Plant regeneration from mesophyll protoplasts of lisianthus (Eustoma grandiflorum) by adding activated charcoal into protoplast culture medium. Plant Cell, Tissue and Organ Culture 43, 59–65.
| Crossref | GoogleScholarGoogle Scholar |
Li X-B,
Xu Z-H, Wei Z-M
(1995) Plant regeneration from protoplasts of immature Vigna sinensis cotyledons via somatic embryogenesis. Plant Cell Reports 15, 282–286.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
Lin M-Y,
Lin C-Y, Chen Y-M
(1991) Biochemical studies on the enhancement of suspension cells and protoplasts proliferation in rice (Oryza sativa L.) by heat shock treatment. Botanical Bulletin of Academia Sinica 32, 205–214.
|
CAS |
Logemann E,
Wu SC,
Schroder J,
Schmelzer E,
Somssich IE, Hahlbrock K
(1995) Gene activation by UV light, fungal elicitor or fungal infection in Petroselinum crispum is correlated with repression of cell cycle related genes. The Plant Journal 8, 856–876.
Luo JP, Jia JF
(1998) Plant regeneration from callus protoplasts of the forage legume Astragalus adsurgens Pall. Plant Cell Reports 17, 313–317.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
Lynch TM, Linthilhac PM
(1997) Mechanical signals in plant development: a new method for single cell studies. Developmental Biology 181, 246–256.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Marchant R,
Davey MR, Power JB
(1997) Isolation and culture of mesophyll protoplasts from Rosa hybrida. Plant Cell, Tissue and Organ Culture 50, 131–134.
| Crossref | GoogleScholarGoogle Scholar |
Mizuhiro M,
Kenichi Y,
Ito K,
Kadowaki S,
Ohasi M, Mii M
(2001) Plant regeneration form cell suspension-derived protoplasts of Primula malacoides and Primula obconica. Plant Science 160, 1221–1228.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Mizukami Y,
Kato M,
Takamizo T,
Kanbe M,
Inami S, Hattori K
(2006) Interspecific hybrids between Medicago sativa L. and annual Medicago containing alfafa weevil resistance. Plant Cell, Tissue and Organ Culture 84, 80–89.
| Crossref | GoogleScholarGoogle Scholar |
Murashige T, Skoog F
(1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum 15, 473–497.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
Nagata T, Takebe I
(1970) Cell wall regeneration and cell division in isolated tobacco mesophyll protoplasts. Planta 92, 301–308.
| Crossref | GoogleScholarGoogle Scholar |
Pan ZG,
Liu CZ,
Zobayed SMA, Saxena PK
(2004) Plant regeneration from mesophyll protoplasts of Echinacea. Plant Cell, Tissue and Organ Culture 77, 251–255.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
Papadakis AK, Roubelakis-Angelakis KA
(2002) Oxidative stress could be responsible for the recalcitrance of plant protoplasts. Plant Physiology and Biochemistry 40, 549–559.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
Phoplonker MA, Caligari PDS
(1993) Cultural manipulations affecting callus formation from seedling explants of the pearl lupin (Lupinus mutabilis Sweet). Annals of Applied Biology 123, 419–432.
| Crossref | GoogleScholarGoogle Scholar |
Puonti-Kaerlas J, Eriksson T
(1988) Improved protoplast culture and regeneration of shoots in pea (Pisum sativum L.). Plant Cell Reports 7, 242–245.
| Crossref | GoogleScholarGoogle Scholar |
Radionenko MA,
Kuchuk NV,
Khvedynich OA, Gleba YY
(1994) Direct somatic embryogenesis and plant regeneration from protoplasts of red clover (Trifolium pratense L.). Plant Science 97, 75–81.
| Crossref | GoogleScholarGoogle Scholar |
Reichheld JP,
Vernoux T,
Lardon F,
van Montagu M, Inze D
(1999) Specific checkpoints regulate plant cell cycle progression in response to oxidative stress. The Plant Journal 17, 647–656.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
Rybczyński JJ
(1997) Plant regeneration from highly embryogenic callus, cell suspension and protoplasts of Trifolium fragiferum. Plant Cell, Tissue and Organ Culture 51, 159–170.
| Crossref | GoogleScholarGoogle Scholar |
Saxena PK, Gill R
(1986) Removal of browning and growth enhancement by polyvinylpolypyrrolidone in protoplast cultures of Cyamopsis tetragonoloba L. Biologia Plantarum 28, 313–315.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
Schäfer-Menuhr A
(1987) Isolation und Kultur von Lupinenprotoplasten. I. Protoplasten aus Blättern von Lupinus angustifolius Sorte Kubesa. Landbauforschung Völkenrode 37, 117–120.
Schäfer-Menuhr A
(1988) Isolation und Kultur von Lupinenprotoplasten. III. Protoplasten aus Zellsuspensionskulturen von Lupinus polyphyllus. Landbauforschung Völkenrode 38, 99–102.
Schäfer-Menuhr A
(1989) Isolation und Kultur von Lupinenprotoplasten. IV. Regeneration von Pflanzen aus Protoplasten. Landbauforschung Völkenrode 39, 133–136.
Scheres B, Benfey PN
(1999) Asymmetric cell division in plants. Annual Review of Plant Physiology and Plant Molecular Biology 50, 505–537.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Schuppler U,
He P-H,
John PC, Munns R
(1998) Effect of water stress on cell division and cell-division-cycle 2-like cell-cycle kinase activity in wheat leaves. Plant Physiology 117, 667–678.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Shibaya T, Sugawara Y
(2007) Involvement of arabinogalactan proteins in the regeneration process of cultured protoplasts of Marchantia polymorpha. Physiologia Plantarum 130, 271–279.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
Shillito RD,
Paskowski J, Potrykus I
(1983) Agarose plating and a bead type culture technique enable and stimulate development of protoplast-derived colonies in a number of plant species. Plant Cell Reports 2, 244–247.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
Sinha A, Caligari PDS
(2004) Aspects of isolation underpinning mitotic behaviour in lupin protoplasts. Australian Journal of Botany 52, 669–676.
| Crossref | GoogleScholarGoogle Scholar |
Sinha A, Caligari PDS
(2005) Enhanced protoplast division by encapsulation in droplets: an advance towards somatic hybridisation in recalcitrant white lupin. Annals of Applied Biology 146, 441–448.
| Crossref | GoogleScholarGoogle Scholar |
Sinha A,
Wetten AC, Caligari PDS
(2003) Effect of biotic factors on the isolation of Lupinus albus protoplasts. Australian Journal of Botany 51, 103–109.
| Crossref | GoogleScholarGoogle Scholar |
Sonntag K,
Ruge-Wehling B, Wehling P
(2009) Protoplast isolation and culture for somatic hybridisation of Lupinus angustifolius and L. subcarnosus. Plant Cell Tissue and Organ Culture 96, 297–305.
| Crossref | GoogleScholarGoogle Scholar |
Sun Y,
Zhang X,
Nie Y, Guo X
(2005) Production of fertile somatic hybrids of Gossypium hirsutum + G. bickii and G. hirsutum + G. stockii via protoplast fusion. Plant Cell, Tissue and Organ Culture 83, 303–310.
| Crossref | GoogleScholarGoogle Scholar |
Święcicki W,
Rybczyński J, Święcicki WK
(2000) Domestication and genetics of the yellow lupin (Lupinus luteus L.) and the biotechnological improvement of lupins. Journal of Applied Genetics 41, 11–34.
Thomas TD
(2008) The role of activated charcoal in plant tissue culture. Biotechnology Advances 26, 618–631.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Thompson JA,
Abdullah R,
Chen W-H, Gartland KMA
(1987) Enhanced protoplast divisions in rice (Oryza sativa L.) following heat shock treatment. Journal of Plant Physiology 127, 367–370.
Tian D,
Niu C, Rose RJ
(2002) DNA transfer by highly asymmetric somatic hybridisation in Medicago truncatula + Medicago rugosa and Medicago truncatula + Medicago scutellata. Theoretical and Applied Genetics 104, 9–16.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Vázquez S,
Carpena RO, Bernal MP
(2008) Contribution of heavy metals and As-loaded lupin root mineralisation to the availability of the pollutants in multi-contaminated soils. Environmental Pollution 152, 373–379.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Vessabutr S, Grant WF
(1995) Isolation, culture and regeneration of protoplasts from birdsfoot trefoil (Lotus corniculatus). Plant Cell, Tissue and Organ Culture 41, 9–15.
| Crossref | GoogleScholarGoogle Scholar |
White CL,
Staines VE, Staines MH
(2007) A review of the nutritional value of lupins for dairy cows. Australian Journal of Agricultural Research 58, 185–202.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
Wiszniewska A, Pindel A
(2009) Protoplast culture utilisation in studies on legume crops. Acta Agriculturae Scandinavica Section B. Soil and Plant Science in press ,
| Crossref | GoogleScholarGoogle Scholar |
Xiao W,
Huang X-L,
Huang X,
Chen Y-P,
Dai X-M, Zhao J-T
(2007) Plant regeneration from protoplasts of Musa acuminata cv. Mas (AA) via somatic embryogenesis. Plant Cell, Tissue and Organ Culture 90, 191–200.
| Crossref | GoogleScholarGoogle Scholar |
Zhou J,
Wang B, Zhu L
(2005) Conditioned culture for protoplasts isolated from chrysanthemum: an efficient approach. Colloids and Surfaces. B, Biointerfaces 45, 113–119.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
PubMed |
Zhu Y-M,
Hoshino Y,
Nakano M,
Takahashi E, Mii M
(1997) Highly efficient system of plant regeneration from protoplasts of grapevine (Vitis vinifera L.) through somatic embryogenesis by using embryogenic callus culture and activated charcoal. Plant Science 123, 151–157.
| Crossref | GoogleScholarGoogle Scholar |
CAS |
