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Effects of cytokinins, carbohydrates and amino acids on induction and maturation of somatic embryos in kodo millet (Paspalum scorbiculatum Linn.)

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Abstract

The effects of cytokinins, carbohydrates and amino acids were studied on maturation and regeneration of embryogenic callus (EC) in kodo millet (Paspalum scorbiculatum Linn.) using a two-step culture procedure. The highest percentage (87.6%) of EC induction was obtained in the induction medium containing Murashige and Skoog (MS) basal salts supplemented with 9.0 μM 2,4-dichlorophenoxy acetic acid and 2.25 μM kinetin. This EC was subcultured in the maturation medium for somatic embryo (SE) maturation and plantlet formation. Addition of cytokinins, carbohydrates and amino acids in the maturation medium promoted the SE maturation and plantlet formation. The maturation medium containing MS basal salts amended with 4.50 μM thidiazuron, 120 mM maltose and 200 μM l-proline gave the maximum number of SEs (39.4) and plantlets (31.3). Plantlets were successfully grown to maturity after hardening in the soil.

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Abbreviations

BA:

Benzyladenine

2,4-D:

2,4-Dichlorophenoxy acetic acid

EC:

Embryogenic callus

Kn:

Kinetin

MS:

Murashige and Skoog

PGR:

Plant growth regulator

SE:

Somatic embryo

2,4,5-T:

2,4,5-trichlorophenoxyacetic acid

TDZ:

Thidiazuron or N-phenyl-N′-1,2,3-thiadiazol-5-ylurea

Zn:

Zeatin

References

  • Armstrong CC, Green CE (1985) Establishment and maintenance of friable, embryogenic maize callus and the involvement of L-proline. Planta 164:207–214

    Article  CAS  Google Scholar 

  • Arockiasamy S, Prakash S, Ignacimuthu S (2001) High regenerative nature of Paspalum scrobiculatum L., an important millet crop. Curr Sci 80:496–498

    Google Scholar 

  • Asano Y, Ito Y, Ohara M, Sugiura K, Fujiie A (1994) Improved regeneration response of creeping bentgrass and japonica rice by maltose and lactose. Plant Cell Tiss Org Cult 39:101–103

    Article  CAS  Google Scholar 

  • Bela J, Shetty K (1999) Somatic embryogenesis in anise (Pimpinella anisum L.): the effect of proline on embryogenic callus formation and ABA on advanced embryo development. J Food Biochem 23:17–32

    Article  CAS  Google Scholar 

  • Bellettre C, Vasseur J (1999) Effects of glycerol on somatic embryogenesis in Cichorium leaves. Plant Cell Rep 19:26–31

    Article  CAS  Google Scholar 

  • Blanc G, Michaux-Ferri`ere N, Teisson C, Lardet L, Carron MP (1999) Effects of carbohydrate addition on the induction of somatic embryogenesis in Hevea brasiliensis. Plant Cell Tiss Organ Cult 59:103–112

    Article  CAS  Google Scholar 

  • Borlaug NE (2002) Feeding a world of 10 billion people: the miracle ahead. In Vitro Cell Dev Biol Plant 8:221–228

    Google Scholar 

  • Capelle S, Mok D, Kirchner S, Mok S (1983) Effects of thidiazuran on (Δ2-isopentyl) (8–14C) adenosine in callus tissues of Phaseolus linatus. L. Plant Physiol 73:796–802

    Article  CAS  PubMed  Google Scholar 

  • Ceasar SA, Ignacimuthu S (2008) Efficient somatic embryogenesis and plant regeneration from shoot apex explants of different Indian genotypes of finger millet (Eleusine coracana (L.) Gaertn.). In Vitro Cell Dev Biol Plant 44:427–435

    Article  CAS  Google Scholar 

  • Ceasar SA, Ignacimuthu S (2009) Genetic engineering of millets: current status and future prospects. Biotechnol Lett 31:779–788

    Article  CAS  PubMed  Google Scholar 

  • Chandra N, Kothari SL (1995) Advances in tissue culture and genetic transformation of cereals. J Indian Bot Soc 74:323–342

    Google Scholar 

  • Claparols I, Santos MA, Torne JM (1993) Influence of some exogenous amino acids on the production of maize embryogenic callus and on endogenous amino acid content. Plant Cell Tiss Org Cult 34:1–11

    Article  CAS  Google Scholar 

  • Gallego P, Hita O, Villalobos N, Dorado Martin L, Guerra H (2001) Somatic embryogenesis and plant regeneration in Medicago arborea L. In Vitro Cell Dev Biol Plant 37:199–203

    Article  CAS  Google Scholar 

  • George EF (1993) Plant propagation by tissue culture. Part 1. The technology, 2nd edn. Exegetics, Eddington

    Google Scholar 

  • Hita O, Gallego P, Villalobos N, Lanas I, Blazquez A, Martin JP, Fernandez J, Martin L, Guerra H (2003) Improvement of somatic embryogenesis in Medicago arborea. Plant Cell Tiss Org Cult 72:13–18

    Article  CAS  Google Scholar 

  • Jeannin G, Bronner R, Hahne G (1995) Somatic embryogenesis and organogenesis induced on the immature zygotic embryo of sunflower (Helianthus annuus L.) cultivated in vitro: role of the sugar. Plant Cell Rep 15:200–204

    Article  CAS  Google Scholar 

  • Karsai I, Bedo Z, Hayes PM (1994) Effect of induction medium pH and maltose concentration on in vitro androgenesis of hexaploid winter triticale and wheat. Plant Cell Tiss Org Cult 39:49–53

    Article  CAS  Google Scholar 

  • Kaur P, Kothari SL (2003) Embryogenic callus induction and efficient plant regeneration from root cultures of kodo millet. Phytomorphology 53:49–56

    Google Scholar 

  • Kaur P, Kothari SL (2004) In vitro culture of kodo millet: influence of 2, 4-D and picloram in combination with kinetin on callus initiation and regeneration. Plant Cell Tiss Org Cult 77:73–79

    Article  CAS  Google Scholar 

  • Kavi Kishor PB, Rao AM, Dhar AC, Naidu KR (1992) Plant regeneration in tissue cultures of some millets. Ind J Exp Biol 30:729–733

    Google Scholar 

  • Kothari SL, Satish K, Vishnoi RK, Kothari A, Watanabe KN (2005) Applications of biotechnology for improvement of millet crops: review of progress and future prospects. Plant Biotechnol 22:81–88

    CAS  Google Scholar 

  • Kothari-Chajer A, Sharma M, Kachhwaha S, Kothari SL (2008) Micronutrient optimization results into highly improved in vitro plant regeneration in kodo (Paspalum scrobiculatum L.) and finger (Eleusine coracana (L.) Gaertn.) millets. Plant Cell Tiss Org Cult 94:105–112

    Article  CAS  Google Scholar 

  • Latha MA, Dasvantha Reddy V, Madavi latha A, Venkateswara Rao K (2005) Production of transgenic plants resistant to leaf blast disease in finger millet (Eleusine coracana (L.) Gaertn.). Plant Sci 169:657–667

    Article  CAS  Google Scholar 

  • Latha MA, Rao KV, Reddy TP, Reddy VD (2006) Development of transgenic pearl millet (Pennisetum glaucum (L.) R. Br.) plants resistant to downy mildew. Plant Cell Rep 25:927–935

    Article  CAS  PubMed  Google Scholar 

  • Loiseau J, Marche C, Deunff YL (1995) Effects of auxins, cytokinins, carbohydrates and amino acids on somatic embryogenesis induction from shoot apices of pea. Plant Cell Tiss Org Cult 41:267–275

    Article  CAS  Google Scholar 

  • Marchant R, Davey MR, Lucas JA, Power JB (1996) Somatic embryogenesis and plant regeneration in Floribunda rose (Rosa hybrida L.) cvs. Trumpeter and Glad Tidings. Plant Sci 120:95–105

    Article  Google Scholar 

  • Mohamed SV, Wang CS, Thiruvengadam M, Jayabalan N (2004) In vitro plant regeneration via somatic embryogenesis through cell suspension cultures of horsegram [Macrotyloma uniflorum (Lam.) Verdc.]. In Vitro Cell Dev Biol Plant 40:284–289

    Article  CAS  Google Scholar 

  • Mohanty BD, Gupta SD, Ghosh PD (1985) Callus initiation and plant regeneration in ragi (Eleusine coracana (L). Gaertn). Plant Cell Tiss Org Cult 5:147–150

    Article  CAS  Google Scholar 

  • Mok MC, Mok DW, Amstrong DJ, Shudo K, Isogai Y, Okamanto T (2005) Cytokinin activity of N-phenyl-N’-1, 2, 3-thiadiazol-5- urea. (thidiazuron). Phytochemistry 21:1509–1511

    Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Mythili P, Madhavi A, Reddy VD, Seetharam N (2001) Efficient regeneration of pearl millet Pennisetum glacum (L.) from shoot tip cultures. Ind J Exp Biol 39:1274–1279

    CAS  Google Scholar 

  • Navarro-Alvarez W, Baenziger PS, Eskridge KM, Shelton DR, Gustafson VD, Hugo M (1994) Effect of sugars in wheat anther culture media. Plant Breed 112:53–62

    Article  CAS  Google Scholar 

  • Nayak P, Sen SK (1989) Plant regeneration through somatic embryogenesis from suspension cultures of a minor millet, Paspalum scrobiculatum L. Plant Cell Rep 8:296–299

    Article  Google Scholar 

  • Pareddy DR, Petolino JF (1990) Somatic embryogenesis and plant regeneration from immature inflorescences of several elite inbreds of maize. Plant Sci 67:211–219

    Article  Google Scholar 

  • Rengel Z, Jelaska S (1986) The effect of L-proline on somatic embryogenesis in long-term callus culture of Hordeum vulgar. Acta Bot Croat 45:71–75

    CAS  Google Scholar 

  • Santos M, Claparols I, Torne′ JM (1993) Effect of exogenous arginine, ornitine, methionine and -amino butyric acid on maize (Zea mays L.) embryogenesis and polyamine content. J Plant Physiol 142:74–80

    CAS  Google Scholar 

  • Scott P, Lyne RL (1994) The effect of different carbohydrate sources upon the initiation of embryogenesis from barley microspores. Plant Cell Tiss Org Cult 36:129–133

    Article  CAS  Google Scholar 

  • Seabrook JEA, Douglass LK (2001) Somatic embryogenesis on various potato tissues from a range of genotypes and ploidy levels. Plant Cell Rep 20:175–182

    Article  CAS  Google Scholar 

  • Shimizu K, Nagaike H, Yabuya T, Adachi T (1997) Plant regeneration from suspension culture of Iris germanica. Plant Cell Tiss Org Cult 50:27–31

    Article  Google Scholar 

  • Srivastav S, Kothari SL (2002) Embryonic callus induction and high frequency plant regeneration in pearl millet. Cer Res Commun 30:69–74

    Google Scholar 

  • Sticklen MB, Orabya HF (2005) Shoot apical meristem: a sustainable explant for genetic transformation of cereal crops. In Vitro Cell Dev Biol Plant 41:187–200

    Article  CAS  Google Scholar 

  • Suprasanna P, Rao KV, Reddy GM (1994) Embryogenic callus in maize: genotypic and amino acid effects. Cer Res Commun 22:79–82

    Google Scholar 

  • Thomas JC, Katterman FR (1986) Cytokinin activity induced by thidiazuran. Plant Physiol 81:681–683

    Article  CAS  PubMed  Google Scholar 

  • Thorpe TA (1995) In vitro embryogenesis in plants. Current plant science and biotechnology in agriculture. Kluwer, Dordrecht

    Google Scholar 

  • Tirajoh A, Kyung TS, Punja ZK (1998) Somatic embryogenesis and plantlet regeneration in American ginseng (Panax quinquefolium L.). In Vitro Cell Dev Biol Plant 34:203–211

    Google Scholar 

  • Vasil IK (2008) A short history of plant biotechnology. Phytochem Rev 7:387–394

    Article  CAS  Google Scholar 

  • Vikrant A, Rashid A (2001) Direct as well as indirect somatic embryogenesis from immature (unemerged) inflorescence of a minor millet Paspalum scrobiculatum L. Euphytica 120:167–172

    Article  CAS  Google Scholar 

  • Vikrant A, Rashid A (2002a) Somatic embryogenesis from immature and mature embryos of a minor millet Paspalum scrobiculatum L. Plant Cell Tiss Org Cult 69:71–77

    Article  CAS  Google Scholar 

  • Vikrant A, Rashid A (2002b) Induction of multiple shoots by thidiazuron from caryopsis cultures of minor millet (Paspalum scrobiculatum L.) and its effect on the regeneration of embryogenic callus cultures. Plant Cell Rep 21:9–13

    Article  CAS  Google Scholar 

  • Vikrant A, Rashid A (2003) Somatic embryogenesis from mesocotyl and leaf base segments of Paspalum scrobiculatum L., minor millet. In Vitro Cell Dev Biol Plant 39:485–489

    Article  Google Scholar 

  • Xie DY, Hong Y (2001) Regeneration of Acacia mangium through somatic embryogenesis. Plant Cell Rep 20:34–40

    Article  CAS  Google Scholar 

  • Xu Z, Wang D, Yang L, Wei Z (1984) Somatic embryogenesis and plant regeneration in callus cultured immature inflorescence of Seturia italica. Plant Cell Rep 3:144–150

    Article  Google Scholar 

  • Zhang CL, Chen DF, Kubalakova M, Zhang J, Scott NW, Elliott MC, Slater A (2008) Efficient somatic embryogenesis in sugar beet (Beta vulgaris L.) breeding lines. Plant Cell Tiss Org Cult 93:209–221

    Article  CAS  Google Scholar 

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Acknowledgments

The authors thank Dr. K. T. Krishne Gowda, Project Coordinator, Small Millets, University of Agricultural Sciences, Bangalore, India, for the generous supply of kodo millet seeds, and Dr. M. Gabriel Paulraj, Entomology Research Institute, Loyola College, Chennai for his help in photography.

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  1. Division of Plant Biotechnology, Entomology Research Institute, Loyola College, Chennai, Tamilnadu, 600034, India

    S. Antony Ceasar & S. Ignacimuthu

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  1. S. Antony Ceasar
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  2. S. Ignacimuthu
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Correspondence to S. Ignacimuthu.

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Ceasar, S.A., Ignacimuthu, S. Effects of cytokinins, carbohydrates and amino acids on induction and maturation of somatic embryos in kodo millet (Paspalum scorbiculatum Linn.). Plant Cell Tiss Organ Cult 102, 153–162 (2010). https://doi.org/10.1007/s11240-010-9716-6

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