Skip to main content
SpringerLink
Log in

In vitro Culture of Medicago arborea L. Anthers: Initial Response

  • Published:
Plant Growth Regulation Aims and scope Submit manuscript

Abstract

High production of viable somatic embryos was obtained from cultured anthers in the second phase of meiosis, using microscopic level observations of tetrads. The medium with the greatest embryogenic efficiency was H6, composed of Murashige and Skoog (MS) medium with 2 mg l−1 of 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 mg l−1 of kinetin. All (100%) of the somatic embryos obtained germinated and produced 63% green and 37% albino seedlings. In general, embryogenic calli had a higher ion concentration than non-embryogenic calli, with the exception of calcium whose concentration was higher in non-embryogenic calli. The calli induced in the different media differed in their sucrose and starch compositions. The most embryogenic medium H6-induced calli with the highest sucrose concentration and the lowest starch concentration, before visible embryos were observed. In the leaves of the albino seedlings, sucrose concentrations were very high while those of starch were very low. Ion concentrations were also lower in albino plants than in the leaves of green seedlings, with the exception of calcium, whose concentration was higher. Most of the albino individuals were homozygous, even when their progenitors were heterozygous, thereby confirming their haploid nature.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

2,4-D:

2,4-dichlorophenoxyacetic acid

dw:

dry weight

α,β-EST:

α,β-esterases

IBA:

indole-butyric acid

IDH:

isocitrate dehydrogenases

Kin:

kinetin

LAP:

leucin aminopeptidases

MDH:

malate dehydrogenases

MNR:

menadione reductases

NAA:

1-naphthalene acetic acid

PER:

peroxidases

References

  • R. Bacilieri T. Labbe A. Kremer (1994) ArticleTitleIntraspecific genetics structure in a mixed population of Quercus petraea (Matt.) Leibl and Q. robur L Heredity 73 130–141 Occurrence Handle10.1038/hdy.1994.112

    Article  Google Scholar 

  • U.S. Bishnoi R.K. Jain R. Gupta V.K. Chowdhury J.B. Chowdhury (2000) ArticleTitleHigh frequency androgenesis in indica × Basmati rice hybrids using liquid culture media Plant Cell Tiss. Org. Cult. 61 153–159 Occurrence Handle10.1023/A:1006407912576

    Article  Google Scholar 

  • L. Bouvier P.H. Guerif M. Djulbic C.H.E. Durel E. Chevreau Y. Lespinasse (2002) ArticleTitleChromosome doubling of pear haploid plants and homozygosity assessment using isozyme and microsatellite markers Euphytica 123 255–262 Occurrence Handle10.1023/A:1014998019674 Occurrence Handle1:CAS:528:DC%2BD38Xkt1aksLo%3D

    Article  CAS  Google Scholar 

  • C. Branca A. Trolley P. Firm M.M. Altamura M. Bassi (1994) ArticleTitleEarly phases in vitro culture of tomato cotyledons: starch accumulation and protein pattern in relation to the hormonal treatment Protoplasma 182 59–64 Occurrence Handle10.1007/BF01403689

    Article  Google Scholar 

  • C. Chu C. Wang C. Sun C. Hsu K. Yin C. Chu (1975) ArticleTitleEstablishment of an efficient medium for anther culture of rice through comparative experiments on the nitrogen sources Sci. Sin. 18 659–668

    Google Scholar 

  • C. Di-Vaio A. Petito M. Buccheri (2001) ArticleTitleEffect of girdling on gas exchanges and leaf mineral content in the ‘independence’ nectarine J. Plant Nutr. 24 1047–1060 Occurrence Handle10.1081/PLN-100103802 Occurrence Handle1:CAS:528:DC%2BD3MXltlyjsrw%3D

    Article  CAS  Google Scholar 

  • D.B. Duncan (1955) ArticleTitleMultiple range and multiple F tests Biometrics 11 1–42 Occurrence Handle10.2307/3001478

    Article  Google Scholar 

  • J.A. Elena-Roselló R. LLumaret E. Cabrera H. Michaud (1992) ArticleTitleEvidence for hybridation between sympatric holm-oak and cork-oak in Spain bases on diagnostic enzyme markers Vegetatio 99/100 115–118 Occurrence Handle10.1007/BF00118216

    Article  Google Scholar 

  • J.A. Elena-Roselló E. Cabrera (1996) ArticleTitleIsozyme variation in natural populations of cork-oak (Quercus suber L.). Population structurediversity, differentiation and gene flow Silvae Genetica 45 229–235

    Google Scholar 

  • L.A. Elkonin N.V. Pakhomova (1997) ArticleTitlePhosphate as an efficient stimulator of somatic embryogenesis in sorghum tissue culture Int. Sorghum Millets Newsl 38 101–102

    Google Scholar 

  • P. Gallego O. Hita N. Villalobos A. Dorado L. Martin H. Guerra (2001) ArticleTitleSomatic embryogenesis and plant regeneration with Medicago arborea L. plantlets In vitro Cell. Dev. Biol. 37 199–203 Occurrence Handle1:CAS:528:DC%2BD3MXkvVOmurk%3D Occurrence Handle10.1007/s11627-001-0034-x

    Article  CAS  Google Scholar 

  • O.L. Gamborg P.A. Miller K. Ojima (1968) ArticleTitleNutrient requirements of suspension cultures of soybean root cells Exp. Cell. Res. 50 151–158 Occurrence Handle5650857 Occurrence Handle10.1016/0014-4827(68)90403-5 Occurrence Handle1:CAS:528:DyaF1cXktVyqtLw%3D

    Article  PubMed  CAS  Google Scholar 

  • A.J. Gordon G.J.A. Ryle D.F. Mitchell K.H. Lowry C.E. Powell (1986) ArticleTitleThe effect of defoliation on carbohydrateprotein and Leg-Haemoglobin content of white clover nodules Ann. Bot. 58 141–154 Occurrence Handle1:CAS:528:DyaL28XlsV2rsrk%3D

    CAS  Google Scholar 

  • A.K. Jain A. Sarkar R.K. Datta (1996) ArticleTitleInduction of haploid callus and embryogenesis in vitro cultured anthers of mulberry (Morus indica) Plant Cell Tissue Org. Cult. 44 143–147 Occurrence Handle10.1007/BF00048192

    Article  Google Scholar 

  • G.B. Kiss G. Csanádi K. Kálmán P. Kaló L. Ökrész (1990) ArticleTitleConstruction of a basic genetic map for alfalfa using RFLP, RAPD, isozyme and morphological markers Mol. Genet. Genom. 238 129–137

    Google Scholar 

  • L.F. La Cour (1954) ArticleTitleSmear and squash techniques in plant cytology Lab. Pract. 3 326–330

    Google Scholar 

  • S.Y. Lee J.I. Cheong T.S. Kim (2003) ArticleTitleProduction of doubled haploids through anther culture of M1 rice plants derived from mutagenized fertilized egg cells Plant Cell Rep. 22 218–223 Occurrence Handle12845474 Occurrence Handle10.1007/s00299-003-0663-0 Occurrence Handle1:CAS:528:DC%2BD3sXnvFaltr4%3D

    Article  PubMed  CAS  Google Scholar 

  • K.A. Lesins I. Lesins (1979) Medicago arborea W. Junk (Eds) Genus Medicago (Leguminosae). A Taxogenetic Study Kluwer Boston 130–133

    Google Scholar 

  • R. Lichter (1981) ArticleTitleAnther culture of Brassica napus rape in a liquid culture medium Z. Pflanzenphysiol. 103 229–237 Occurrence Handle1:CAS:528:DyaL3MXlvVCjsL4%3D

    CAS  Google Scholar 

  • Y.B. Linhart (1988) Ecological and evolutionary studies of Ponderosa pine in the Rocky Mountains D.M. Baumgartner J.E. Lotan (Eds) Ponderosa Pine: The Species and its Management. Cooperative Extensión Washington State University Pullman, WA

    Google Scholar 

  • W. Liu M.Y. Zheng C.F. Konzak (2002) ArticleTitleImproving green plant production via isolated microspore culture in bread wheat (Triticum aestivum L.) Plant Cell Rep. 20 821–824 Occurrence Handle10.1007/s00299-001-0408-x Occurrence Handle1:CAS:528:DC%2BD38XhtVCgurw%3D

    Article  CAS  Google Scholar 

  • A.B. Martín Y. Cuadrado H. Guerra P. Gallego O. Hita L. Martín A. Dorado N. Villalobos (2000) ArticleTitleDifferences in the contents of total sugars, reducing sugars, starch and sucrose in embryogenic and non-embryogenic calli from Medicago arborea L Plant Sci. 154 143–151 Occurrence Handle10729613 Occurrence Handle10.1016/S0168-9452(99)00251-4

    Article  PubMed  Google Scholar 

  • M. Medina N. Villalobos P.J. la Cruz ParticleDe A. Dorado H. Guerra (1998) ArticleTitleEffect of culture medium and lighting conditions on the morphological characteristics and carbohydrate contents of Medicago strasseri calli Acta Physiol. Plant. 20 383–392 Occurrence Handle1:CAS:528:DyaK1MXhtlykt78%3D Occurrence Handle10.1007/s11738-998-0024-2

    Article  CAS  Google Scholar 

  • G. Muller-Starck J. Jorgensen (1991) ArticleTitleEnzyme gene markers as indicators of the initial ploidy in anther cultures of trees Can. J. For. Res. 21 1141–1144 Occurrence Handle10.1139/x91-159

    Article  Google Scholar 

  • T. Murashige F. Skoog (1962) ArticleTitleA revised medium for rapid growth and bioassays with tobacco tissue cultures Physiol. Plant. 15 473–497 Occurrence Handle10.1111/j.1399-3054.1962.tb08052.x Occurrence Handle1:CAS:528:DyaF3sXksFKm

    Article  CAS  Google Scholar 

  • S.T. Nguyen R. Paquin (1971) ArticleTitleMéthodes d’extraction et de purification des acides aminés libres et des protéines de tissus végétaux J. Chromatogr. 61 349–351 Occurrence Handle10.1016/S0021-9673(00)92432-4 Occurrence Handle1:CAS:528:DyaE3MXlsVygs7Y%3D

    Article  CAS  Google Scholar 

  • E.A. Nichols F.H. Ruddle (1973) ArticleTitleA review of enzyme polymorphismlinkage and electrophoretic conditions for mouse and somatic cell hybrids in starch gels J. Histochem. Cytochem. 21 1066–1081 Occurrence Handle4587225 Occurrence Handle1:CAS:528:DyaE2cXptlCgsg%3D%3D

    PubMed  CAS  Google Scholar 

  • J.P. Nitsch C. Nitsch (1969) ArticleTitleHaploid plants from pollen grains Science 163 85–87 Occurrence Handle17780179 Occurrence Handle10.1126/science.163.3862.85 Occurrence Handle1:STN:280:DC%2BC3cvgslKltA%3D%3D

    Article  PubMed  CAS  Google Scholar 

  • K.Y. Paek S. Canderlerdy A. Thorpe (1988) ArticleTitlePhysiological effects of Na2SO4NaCl on callus cultures of Brassica campestris (Chinese cabbage) Physiol. Plant 72 160–166 Occurrence Handle10.1111/j.1399-3054.1988.tb06638.x Occurrence Handle1:CAS:528:DyaL1cXht1yktbc%3D

    Article  CAS  Google Scholar 

  • M. Panaia T. Senaratna E. Bunn K.W. Dixon K. Sivasithamparam (2000) ArticleTitleMicropropagation of the critically endangered Western Australian species, Symonanthus bancroftii (F. Muell.) L. Haegi (Solanaceae) Plant Cell Tissue Organ Cult. 63 23–29 Occurrence Handle10.1023/A:1006457624282 Occurrence Handle1:CAS:528:DC%2BD3MXisV2rsbc%3D

    Article  CAS  Google Scholar 

  • Pasteur N., Pasteur G., Bonhomme F., Catalan J. and Britton-Davidian J. 1987. Manuel technique de genetique par electrophorese des proteines. Technique et Documentation (Lavoisier), Montpellier, 217 pp.

  • M. Peng D.J. Wolyn (1999) ArticleTitleImproved callus formation and plant regeneration for shed microspore culture in asparagus (Asparagus officinalis L.) Plant Cell Rep. 18 954–958 Occurrence Handle10.1007/s002990050690 Occurrence Handle1:CAS:528:DyaK1MXltlaju7w%3D

    Article  CAS  Google Scholar 

  • C. Pérez (2001) Técnicas Estadísticas con SPSS Prentice Hall Madrid 357–404

    Google Scholar 

  • S. Rasco-Gaunt A. Riley P. Barcelo P.A. Lazzeri (1999) ArticleTitleAnalysis of particle bombardment parameters to optimise DNA delivery into wheat tissues Plant Cell Rep. 19 118–127 Occurrence Handle10.1007/s002990050721 Occurrence Handle1:CAS:528:DC%2BD3cXjsVehs70%3D

    Article  CAS  Google Scholar 

  • M. Sadiq W.A. Abdurrehman (1999) ArticleTitleElemental composition of alfalfa leaves and its relation to soil composition and irrigation water quality in the Eastern Province of Saudi Arabia J. Plant Nutr. 22 1269–1278 Occurrence Handle1:CAS:528:DyaK1MXkslClur4%3D Occurrence Handle10.1080/01904169909365711

    Article  CAS  Google Scholar 

  • J.G. Scandalious (1969) ArticleTitleGenetic control of multiple molecular forms of enzymes in plants: a review Biochem. Genet. 3 37–79 Occurrence Handle10.1007/BF00485973

    Article  Google Scholar 

  • R.K. Selander M.M. Smith S.Y. Yang W.E. Johnson J.B. Gentry (1971) ArticleTitleBiochemical polimorphism in the genus Peromyscus. I. Variation of the old-field mouse Stud. Genet. Univ. Texas 7103 49–90

    Google Scholar 

  • P. Shanmughavel L. Sha Z. Zheng M. Cao (2001) ArticleTitleNutrient cycling in a tropical seasonal rain forest of ZishuangbannaSouthwest China. 1. Tree species: nutrient distribution and uptake Bioresour. Technol. 80 163–170 Occurrence Handle11601539 Occurrence Handle10.1016/S0960-8524(01)00095-5 Occurrence Handle1:CAS:528:DC%2BD3MXmvVGnsL0%3D

    Article  PubMed  CAS  Google Scholar 

  • C.R. Shaw R. Prasad (1970) ArticleTitleStarch gel electrophoresis of enzymes a compilation of recipes Biochem. Genet. 4 297–320 Occurrence Handle4193186 Occurrence Handle10.1007/BF00485780 Occurrence Handle1:CAS:528:DyaE3cXkt1Wms7c%3D

    Article  PubMed  CAS  Google Scholar 

  • L.D. Talbott E. Zeiger (1998) ArticleTitleThe role of sucrose in guard cell osmoregulation J. Exp. Bot. 49 329–337 Occurrence Handle10.1093/jexbot/49.suppl_1.329

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Departamento de Fisiología Vegetal, Facultad de Biología y Farmacia, Universidad de Salamanca, Plaza de los Doctores de la Reina s/n, E-37007, Salamanca, Spain

    Itziar Lanas, Piedad Gallego, Juana Elena-Rosello, Nieves Villalobos & Hilario Guerra

  2. Laboratorio de Fisiología Vegetal, Facultad de Biología y Escuela Técnica Superior de Ingenieros Agrónomos, Universidad Complutense y Politécnica de Madrid, E-28040, Madrid, Spain

    Luisa Martin

  3. Departamento de Botánica, Facultad de Biología, Universidad de Salamanca, Plaza de los Doctores de la Reina s/n, E-37007, Salamanca, Spain

    Javier Fernandez

  4. Departamento de Química Analítica, Facultad de Químicas, Universidad de Salamanca, Spain

    Angel Alonso

  5. Departamento de Estadística, Facultad de Economía y Empresa, Universidad de Salamanca, Spain

    Antonio Blazquez

Authors
  1. Itziar Lanas
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Piedad Gallego
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Luisa Martin
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Javier Fernandez
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Angel Alonso
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Juana Elena-Rosello
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Antonio Blazquez
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Nieves Villalobos
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Hilario Guerra
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Nieves Villalobos.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lanas, I., Gallego, P., Martin, L. et al. In vitro Culture of Medicago arborea L. Anthers: Initial Response. Plant Growth Regul 49, 49–60 (2006). https://doi.org/10.1007/s10725-006-0007-5

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10725-006-0007-5

Keywords

Search

Navigation