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High-frequency shoot regeneration from leaf explants through organogenesis in bitter melon (Momordica charantia L.)

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Abstract

An efficient protocol for in vitro organogenesis was achieved from callus-derived immature and mature leaf explants of Momordica charantia, a very important vegetable and medicinal plant. Calluses were induced from immature leaf explants excised from in vitro (15-day-old seedlings) mature leaf explants of vivo plants (45 days old). The explants were grown on Murashige and Skoog (MS) medium with Gamborg (B5) vitamins containing 30 g l−1 sucrose, 2.2 g l−1 Gelrite, and 7.7 μM naphthalene acetic acid (NAA) with 2.2 μM thidiazuron (TDZ). Regeneration of adventitious shoots from callus (30–40 shoots per explant) was achieved on MS medium containing 5.5 μM TDZ, 2.2 μM NAA, and 3.3 μM silver nitrate (AgNO3). The shoots (1.0 cm length) were excised from callus and elongated in MS medium fortified with 3.5 μM gibberellic acid (GA3). The elongated shoots were rooted in MS medium supplemented with 4.0 μM indole 3-butyric acid (IBA). Rooted plants were acclimatized in the greenhouse and subsequently established in soil with a survival rate of 90%. This protocol yielded an average of 40 plants per leaf explant with a culture period of 98 days.

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Abbreviations

AgNO3 :

Silver nitrate

2,4-D:

2,4-Dichlorophenoxyacetic acid

BAP:

6-Benzylaminopurine

FAA:

Formaldehyde acetic acid alcohol

GA3 :

Gibberellic acid

IAA:

Indole-3-acetic acid

IBA:

Indole-3-butyric acid

MS:

Murashige and Skoog medium

NAA:

α-Naphthaleneacetic acid

TDZ:

Thiadiazuron

References

  • Agarwal M, Kamal R (2004) In vitro clonal propagation of Momordica charantia L. Indian J Biotechnol 3:426–430

    CAS  Google Scholar 

  • Ananthakrishnan G, Xia X, Elman C, Singer S, Paris HS, Gal-On A, Gaba V (2003) Shoot production in squash (Cucurbita pepo) by in vitro organogenesis. Plant Cell Rep 21:739–746

    PubMed  CAS  Google Scholar 

  • Baskaran P, Velayutham P, Jayabalan N (2009) In vitro regeneration of Melothria maderaspatana via indirect organogenesis. In Vitro Cell Dev Biol Plant 45:407–413

    Article  Google Scholar 

  • Beloin N, Gbeassor M, Akpagana K, Hudson J, Soussa KD, Koumaglo K, Arnason JT (2005) Ethnomedicinal uses of Momordica charantia (Cucurbitaceae) in Togo and relation to its phytochemistry and biological activity. J Ethnopharmacol 96:49–55

    Article  PubMed  Google Scholar 

  • Berlyn BP, Miksche JP (1976) Botanical microtechnique and cytochemistry. Iowa State University Press, Ames

    Google Scholar 

  • Bourinbaiar AS, Lee-Huang S (1996) The activity of plant derived antiretroviral proteins MAP30 and GAP31 against herpes simplex virus in vitro. Biochem Biophys Res Commun 219:923–929

    Article  PubMed  CAS  Google Scholar 

  • Curuk S, Ananthakrishnan G, Singer S, Xia X, Elman C, Nestel D, Cetiner S, Gaba V (2003) Regeneration in vitro from the hypocotyls of Cucumis species produces almost exclusively diploid shoots, and does not require light. HortScience 38:105–109

    Google Scholar 

  • Devendra NK, Subhash B, Seetharam YN (2009) Callus growth and plant regeneration in Momordica dioica (Roxb.) Willd. Cucurbitaceae. Am Eurasian J Sustain Agric 3:743–748

    Google Scholar 

  • Dong JZ, Jia SR (1991) High efficiency plant regeneration from cotyledons of watermelon (Citrullus vulgaris Schrad.). Plant Cell Rep 9:559–562

    Article  CAS  Google Scholar 

  • Gamborg O, Miller R, Ojima K (1968) Nutrients requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–158

    Article  PubMed  CAS  Google Scholar 

  • Ganasan K, Huyop F (2010) In vitro regeneration of Citrullus lanatus cv. Round Dragon. J Biol Sci 10:131–137

    Article  CAS  Google Scholar 

  • Han JS, Oh DG, Mok IG, Park HG, Kim CK (2004) Efficient plant regeneration from cotyledon explants of bottle gourd (Lagenaria siceraria Standl.). Plant Cell Rep 23:291–296

    Article  PubMed  CAS  Google Scholar 

  • Handley LW, Chambliss OL (1979) In vitro propagation of Cucumis sativus L. Sci Hortic 14:22–23

    CAS  Google Scholar 

  • Hoque A, Islam R, Joarder OI (1995) In vitro plantlets differentiation in kakrol (Momordica dioica Roxb.). Plant Tissue Cult 5:119–124

    Google Scholar 

  • Huda AKMN, Sikdar B (2006) In vitro plant production through apical meristem culture of bitter gourd (Momordica charantia L.). Plant Tissue Cult Biotech 16:31–36

    Google Scholar 

  • Islam R, Sarkar PK, Naderuzzaman ATM, Joarder OI (1994) In vitro regeneration of plants from cotyledons of Momordica charantia L. Plant Tissue Cult 4:105–109

    Google Scholar 

  • Kathal R, Bhatnagar SP, Bhojwani SS (1988) Regeneration of plants from leaf explants of Cucumis melo cv. Pusa sharbati. Plant Cell Rep 7:449–451

    Google Scholar 

  • Krug MGZ, Stipp LCL, Rodriguez APM, Mendes BMJ (2005) In vitro organogenesis in watermelon cotyledons. Pesq Agropec Bras Brasilia 40:861–865

    Google Scholar 

  • Lee YK, Chung WI, Ezura H (2003) Efficient plant regeneration via organogenesis in winter squash (Cucurbita maxima Duch.). Plant Sci 164:413–418

    Article  CAS  Google Scholar 

  • Leshem B, Ronen R, Soudry E, Lurie S, Gepstein S (1995) Cytokinin and white light coact to enhance polypeptide metabolism and shoot regeneration in cultured melon cotyledons. J Plant Physiol 145:291–295

    CAS  Google Scholar 

  • Malik S, Zia M, Rehman R, Chaudhary F (2007) In vitro plant regeneration from direct and indirect organogenesis of Momordica charantia. Pak J Biol Sci 10:4118–4122

    Article  PubMed  CAS  Google Scholar 

  • Mohiuddin AKM, Chowdhury MKU, Abdullah Zaliha C, Napis S (1997) Influence of silver nitrate (ethylene inhibitor) on cucumber in vitro shoot regeneration. Plant Cell Tissue Org Cult 51:75–78

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Nabi SA, Rashid MM, Al-Amin M, Rasul MG (2002) Organogenesis in teasle gourd (M. dioica Roxb.). Plant Tissue Cult 12:173–180

    Google Scholar 

  • Nishibayashi S, Kaneko H, Hayakawa T (1996) Transformation of cucumber (Cucumis sativus L.) plants using Agrobacterium tumefaciens and regeneration from hypocotyls explants. Plant Cell Rep 15:809–814

    Article  CAS  Google Scholar 

  • Ntui VO, Thirukkumaran G, Lioka S, Mii M (2009) Efficient plant regeneration via organogenesis in “Egusi” melon (Colocynthis citrullus L.). Sci Hortic 119:397–402

    Article  CAS  Google Scholar 

  • Ouma JP, Young MM, Reichert NA (2004) Optimization of in vitro regeneration of multiple shoots from hypocotyl sections of cotton (Gossypium hirsutum L.). Afr J Biotech 3:169–173

    CAS  Google Scholar 

  • Pal SP, Alam I, Anisuzzaman M, Sarker KK, Sharmin SA, Alam MF (2007) Indirect organogenesis in summer squash (Cucurbita pepo L.). Turk J Agric Forest 31:63–70

    CAS  Google Scholar 

  • Paul A, Mitter K, Raychaudhuri SS (2009) Effect of polyamines on in vitro somatic embryogenesis in Momordica charantia L. Plant Cell Tissue Org Cult 97:303–311

    Article  CAS  Google Scholar 

  • Punja ZK, Abbas N, Sarmento GG, Tang FA (1990) Regeneration of Cucumis sativus vars, sativus and hardwickii, C. melo and C. metuliferus from explants through somatic embryogenesis and organogenesis. Influence of explant source, growth regulator regime and genotype. Plant Cell Tissue Org Cult 21:93–102

    Article  CAS  Google Scholar 

  • Selvaraj N, Ganapathi A, Vasudevan A, Vengadesan G, Kasthuri Rengan S (2006a) In vitro morphogenesis of shoots from leaf explants of cucumber (Cucumis sativus L.). Acta Hortic 725:155–160

    CAS  Google Scholar 

  • Selvaraj N, Vasudevan A, Manickavasagam M, Ganapathi A (2006b) In vitro organogenesis and plant formation in cucumber. Biol Plant 50:123–126

    Article  CAS  Google Scholar 

  • Seo SH, Bai DG, Park HY (2000) High frequency shoot regeneration from leaf explants of cucumber. J Plant Biotechnol 2:51–54

    Google Scholar 

  • Shyamali S, Hattori K (2007) Effect of polyamines and silver nitrate on the high frequency regeneration from cotyledon explants of Bottle gourd (Lagenaria siceraria sp. Asiatica). Pak J Biol Sci 10:1288–1293

    Article  PubMed  CAS  Google Scholar 

  • Sikdar B, Shafiullah M, Chowdhury AR, Sharmin N, Nahar S, Joarder OI (2005) Agrobacterium-mediated GUS expression in bitter gourd (M. charantia L.). Biotechnology 4:149–152

    Google Scholar 

  • Singh A, Singh SP, Bamezai R (1998) Momordica charantia (Bitter gourd) peel, pulp, seed and whole fruit extract inhibits mouse skin papillomagenesis. Toxicol Lett 94:37–46

    Article  PubMed  CAS  Google Scholar 

  • Srivastava DR, Rianov VM, Piruzian ES (1989) Tissue culture and plant regeneration of watermelon (Citrullus vulgaris) Schrad. Cv. Melitopolski. Plant Cell Rep 8:300–302

    Article  Google Scholar 

  • Sultana RS, Bari Miah MA (2003) In vitro propagation of karalla (Momordica charantia L). J Biol Sci 3:1134–1139

    Article  Google Scholar 

  • Thiruvengadam M, Jayabalan N (2001) In vitro shoot multiplication and field establishment of kakrol (Momordica dioica Roxb.). J Ind Bot Soc 80:31–33

    Google Scholar 

  • Thiruvengadam M, Yang CH (2009) Ectopic expression of two MADS box genes from orchid (Oncidium Gower Ramsey) and lily (Lilium longiflorum) alters flower transition and formation in Eustoma grandiflorum. Plant Cell Rep 28:1463–1473

    Article  PubMed  CAS  Google Scholar 

  • Thiruvengadam M, Rekha KT, Jayabalan N (2006a) An efficient in vitro propagation of Momordica dioica Roxb. Ex. Willd. Philip Agric Sci 89:165–171

    Google Scholar 

  • Thiruvengadam M, Varisai Mohamed S, Yang CH, Jayabalan N (2006b) Development of an embryogenic suspension culture of bitter melon (Momordica charantia L.). Sci Hortic 109:123–129

    Article  CAS  Google Scholar 

  • Thiruvengadam M, Rekha KT, Yang CH (2007) Somatic embryogenesis and plant regeneration from petiole-derived callus of spine gourd (Momordica dioica Roxb. ex Willd). Funct Plant Sci Biotechnol 1:200–206

    Google Scholar 

  • Wang S, Tang L, Chen F (2001) In vitro flowering of bitter melon. Plant Cell Rep 20:393–397

    Article  CAS  Google Scholar 

  • Yadav RC, Saleh MT, Grumet R (1996) High frequency shoot regeneration from leaf explants of muskmelon. Plant Cell Tissue Org Cult 45:207–214

    Article  CAS  Google Scholar 

  • Zhang M, Cui H (2001) Stimulatory effects of different cytokinins on direct plant regeneration from cotyledon explants in Cucumis sativus L. Cucurbit Genet Cooper Rep 24:1–4

    Google Scholar 

  • Ziv M, Gadasi G (1986) Enhanced embryogenesis and plant regeneration from cucumber (Cucumis sativus L.) callus by activated charcoal in solid/liquid double layer cultures. Plant Sci 47:115–122

    Article  CAS  Google Scholar 

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Author notes

  1. Muthu Thiruvengadam

    Present address: PG and Research Department of Biotechnology, Marudupandiyar College, Thanjavur, 613 403, Tamil Nadu, India

Authors and Affiliations

  1. Graduate Institute of Biotechnology, National Chung-Hsing University, 250 Kuo-Kuang Road, Taichung, 402, Taiwan, ROC

    Muthu Thiruvengadam, K. T. Rekha & Chang-Hsien Yang

  2. Department of Plant Biotechnology, Bharathidasan University, Tiruchirappalli, 620 024, Tamil Nadu, India

    Narayanasamypillai Jayabalan

  3. Department of Applied Life Science, Konkuk University, Seoul, 143-701, South Korea

    Ill-Min Chung

Authors
  1. Muthu Thiruvengadam
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  2. K. T. Rekha
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  3. Chang-Hsien Yang
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  4. Narayanasamypillai Jayabalan
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  5. Ill-Min Chung
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Correspondence to Muthu Thiruvengadam.

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Thiruvengadam, M., Rekha, K.T., Yang, CH. et al. High-frequency shoot regeneration from leaf explants through organogenesis in bitter melon (Momordica charantia L.). Plant Biotechnol Rep 4, 321–328 (2010). https://doi.org/10.1007/s11816-010-0151-2

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  • DOI: https://doi.org/10.1007/s11816-010-0151-2

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