SpringerLink
Log in

Comparative effects of plant growth regulators on leaf and stem explants of Labisia pumila var. alata

  • Published:
Journal of Zhejiang University SCIENCE B Aims and scope Submit manuscript

Abstract

Objective

Labisia pumila var. alata, commonly known as ‘Kacip Fatimah’ or ‘Selusuh Fatimah’ in Southeast Asia, is traditionally used by members of the Malay community because of its post-partum medicinal properties. Its various pharmaceutical applications cause an excessive harvesting and lead to serious shortage in natural habitat. Thus, this in vitro propagation study investigated the effects of different plant growth regulators (PGRs) on in vitro leaf and stem explants of L. pumila.

Methods

The capabilities of callus, shoot, and root formation were evaluated by culturing both explants on Murashige and Skoog (MS) medium supplemented with various PGRs at the concentrations of 0, 1, 3, 5, and 7 mg/L.

Results

Medium supplemented with 3 mg/L indole-3-butyric acid (IBA) showed the optimal callogenesis from both leaf and stem explants with (72.34±19.55)% and (70.40±14.14)% efficacy, respectively. IBA was also found to be the most efficient PGR for root induction. A total of (50.00±7.07)% and (77.78±16.47)% of root formation were obtained from the in vitro stem and leaf explants after being cultured for (26.5±5.0) and (30.0±8.5) d in the medium supplemented with 1 and 3 mg/L of IBA, respectively. Shoot formation was only observed in stem explant, with the maximum percentage of formation ((100.00±0.00)%) that was obtained in 1 mg/L zeatin after (11.0±2.8) d of culture.

Conclusions

Callus, roots, and shoots can be induced from in vitro leaf and stem explants of L. pumila through the manipulation of types and concentrations of PGRs.

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

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price includes VAT (Thailand)

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Abdul Kadir, A., Nik Hussain, N.H., Wan Bebakar, W.M., Mohd, D.M., Wan Mohammad, W.M.Z., Hassan, I.I., Shukor, N., Kamaruddin, N.A., Wan Mohd, W.N., 2012. The effect of Labisia pumila var. alata on postmenopausal women: a pilot study. Evid. Based Complement. Alternat. Med., 2012:216525. [doi:10.1155/2012/216525]

    Article  PubMed  Google Scholar 

  • Aggarwal, D., Kumar, A., Reddy, M.S., 2010. Shoot organogenesis in elite clones of Eucalyptus tereticornis. Plant Cell Tiss. Org. Cult., 102(1):45–52. [doi:10.1007/s11240-010-9703-y]

    Article  Google Scholar 

  • Auer, C.A., Motyka, V., Březinová, A., Kamínek, M., 1999. Endogenous cytokinin accumulation and cytokinin oxidase activity during shoot organogenesis of Petunia hybrida. Physiol. Plant., 105(1):141–147. [doi:10.1034/j.1399-3054.1999.105121.x]

    Article  CAS  Google Scholar 

  • Christensen, B., Sriskandarajah, S., 2008. In vitro culture of Hibiscus rosa-sinensis L.: influence of iron, calcium and BAP on establishment and multiplication. Plant Cell Tiss. Org. Cult., 93(2):151–161. [doi:10.1007/s11240-008-9354-4]

    Article  CAS  Google Scholar 

  • Conde, P., Sousa, A., Costa, A., Santos, C., 2008. A protocol for Ulmus minor Mill. micropropagation and acclimatization. Plant Cell Tiss. Org. Cult., 92(1):113–119. [doi:10. 1007/s11240-007-9310-8]

    Article  Google Scholar 

  • Cozza, R., Turco, D., Bati, C.B., Bitonti, M.B., 1997. Influence of growth medium on mineral composition and leaf histology in micropropagated plantlets of Olea europaea. Plant Cell Tiss. Org. Cult., 51(3):215–223. [doi:10.1023/A:1005966404642]

    Article  CAS  Google Scholar 

  • Cunha, A.C.G.D., Ferreira, M.F., 1996. Somatic embryogenesis, organogenesis and callus growth kinetics of flax (Linum usitatissium L.). Plant Cell Tiss. Org. Cult., 47(1):1–8. [doi:10.1007/BF02318959]

    Article  Google Scholar 

  • Debnath, S.C., 2008. Zeatin-induced one-step in vitro cloning affects the vegetative growth of cranberry (Vaccinium macrocarpon Ait.) micropropagules over stem cuttings. Plant Cell Tiss. Org. Cult., 93(2):231–240. [doi:10. 1007/s11240-008-9366-0]

    Article  Google Scholar 

  • Debnath, S.C., McRae, K.B., 2005. A one-step in vitro cloning procedure for cranberry (Vaccinium macrocarpon Ait.): the influence of cytokinins on shoot proliferation and rooting. Small Fruits Rev., 4(3):57–75. [doi:10.1300/J301v04n03_05]

    Article  CAS  Google Scholar 

  • Dieleman, J.A., Verstappen, F.W.A., Nicander, B., Kuiper, D., Tillberg, E., Tromp, J., 1997. Cytokinins in Rosa hybrida in relation to bud break. Physiol. Plant., 99(3):456–464. [doi:10.1111/j.1399-3054.1997.tb00560.x]

    Article  CAS  Google Scholar 

  • Eccher, T., Noe, N., 1989. Comparison between 2iP and zeatin in the micropropagation of highbush blueberry (Vaccinium corymbosum). Acta Hort., 241:185–190.

    Google Scholar 

  • Evans, D.E., Coleman, J.O.D., Kearns, A., 2003. Callus Cultures. In: Basics Plant Tissue Culture. BIOS Scientific Publishers, New York, p.63–67.

    Google Scholar 

  • Farouk, A.E., Nawi, M.N., Hassan, S., 2008. Antibacterial peptides from Euycoma longifolia (Tongkat Ali) and Labisia pumila (Kacip Fatimah) leaves in Malaysia. Sci. Brun., 9:55–63.

    Google Scholar 

  • Husen, A., Pal, M., 2007. Metabolic changes during adventitious root primordium development in Tectona grandis Linn. f. (teak) cuttings as affected by age of donor plants and auxins (IBA and NAA) treatment. New Forests, 33(3):309–323. [doi:10.1007/s11056-006-9030-7]

    Article  Google Scholar 

  • Imin, N., Nizamidin, M., Daniher, D., Nolan, K.E., Rose, R.J., Rolfe, B.G., 2005. Proteomic analysis of somatic embryogenesis in Medicago truncatula explant cultures grown under 6-benzylaminopurine and 1-naphthaleneacetic acid treatments. Plant Physiol., 137(4):1250–1260. [doi:10. 1104/pp.104.055277]

    Article  PubMed  CAS  Google Scholar 

  • Jamia, A.J., Houghton, P.J., Milligan, S.R., Ibrahim, J., 2003. The oestrogenic and cytotoxic effects of the extracts of Labisia pumila and Labisia pumila var. pumila in vitro. J. Sains Kesihatan Malaysia, 1:53–60.

    Google Scholar 

  • JIRCAS-SFD Joint Research Project, 2007. Agroforestry Approach to the Rehabilitation of Tropical Lands by Using Nurse Trees. JIRCAS-Sabah Forestry Department (SFD), Malaysia.

    Google Scholar 

  • Khalafalla, M.M., Gaali, E.E.I., Abbas, F.M., Ali, H.A., 2007. Neem (Azadirachta indica A. Juss) callus induction and its larvaecidal activity against Anopheles mosquito. Int. J. Biotechnol. Biochem., 3(1):85–94.

    Google Scholar 

  • Khalafalla, M.M., Daffalla, H.M., El-Shemy, H.A., Abdellatef, E., 2009. Establishment of in vitro fast-growing normal root culture of Vernonia amygdalina-a potent African medicinal plant. Afr. J. Biotechnol., 8(2):5952–5957.

    CAS  Google Scholar 

  • Kuroha, T., Satoh, S., 2007. Involvement of cytokinins in adventitious and lateral root formation. Plant Root, 1:27–33. [doi:10.3117/plantroot.1.27]

    Article  CAS  Google Scholar 

  • Leonardi, C., Ruggen, A., Malfa, S.I., 2001. Hormone effects on in vitro proliferation and rooting of Grevillea explants. Sci. Hort., 90(3–4):335–341. [doi:10.1016/S0304-4238 (01)00228-X]

    Article  CAS  Google Scholar 

  • Liu, Z.R., Sanford, J.C., 1988. Plant regeneration by organogenesis from strawberry leaf and runner tissue. HortScience, 23:1056–1059.

    Google Scholar 

  • Ludwig-Müller, J., Vertocnik, A., Town, C.D., 2005. Analysis of indole-3-butyric acid-induced adventitious root formation on Arabidopsis stem segments. J. Exp. Bot., 56(418):2095–2105. [doi:10.1093/jxb/eri208]

    Article  PubMed  Google Scholar 

  • Marks, T.R., Simpson, S.E., 2000. Rhizogenesis in Forsythia intermedia and Syringa vulgaris: application of a simple internode experimental system. Plant Cell Rep., 19(12): 1171–1176. [doi:10.1007/s002990000264]

    Article  CAS  Google Scholar 

  • Md Ariff, F.F., Hashim, S.S., Haja, M., Osman, M., 2013. An assessment of genetic relationship among superior accessions of Labisia pumila analyzed by amplified fragment length polymorphism (AFLP) markers. Open Sci. Reposit. Agric., e70081945. [doi:10.7392/Agriculture. 70081945]

    Google Scholar 

  • Minocha, S.C., 1987. Plant Growth Regulators and Morphogenesis in Cell and Tissue Culture of Forest Trees. In: Cell and Tissue Culture in Forestry. Martinus Nijhoff Publisher, Dordrecht, p.50–66. [doi:10.1007/978-94-017-0994-1_4]

    Chapter  Google Scholar 

  • Molina, R.V., Castello, S., Luis, G.A., Guardiola, J.L., 2007. Light cytokinin interaction in shoot formation in epicotyl cuttings of Troyer citrangr cultured in vitro. Plant Cell Tiss. Org. Cult., 89(2–3):131–140. [doi:10.1007/s11240-007-9221-8]

    Article  CAS  Google Scholar 

  • Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant., 15(3):473–497. [doi:10.1111/j.1399-3054.1962.tb08052.x]

    Article  CAS  Google Scholar 

  • Nandagopal, S., Kumari, R.B.D., 2007. Effectiveness of auxin induced in vitro root culture in chicory. J. Cent. Eur. Agric., 8(1):73–80.

    Google Scholar 

  • Nandi, S.K., de Klerk, G.J.M., Parker, C.W., Palni Nandi, L.M.S., 1990. Endogenous cytokinin levels and metabolism of zeatin riboside in genetic tumour tissue and non-tumorous tissues of tobacco. Physiol. Plant., 78(2):197–204. [doi:10.1111/j.1399-3054.1990.tb02081.x]

    Article  CAS  Google Scholar 

  • Nolan, K.E., Irwanto, R.R., Rose, R.J., 2003. Auxin up-regulates MtSERK1 expression in both Medicago truncatula root-forming and embryogenesis cultures. Plant Physiol., 133(1):218–230. [doi:10.1104/pp.103. 020917]

    Article  PubMed  CAS  Google Scholar 

  • Pérez-Francés, J.F., Valdés, F., Msrtín, R., 1995. Callus induction and culture from explants of Erysimum scoparium in a growth regulator-free medium. Plant Cell Tiss. Org. Cult., 43(3):223–228. [doi:10.1007/BF00039948]

    Google Scholar 

  • Pierik, R.L.M., Steegmans, H.H.M., 1975. Analysis of adventitious root formation in insolated stem explants of Rhododendron. Sci. Hort., 3(1):1–20. [doi:10.1016/0304-4238(75)90031-X]

    Article  Google Scholar 

  • Reed, B.M., Esquivel, A.A., 1991. The used of zeatin to initiate in vitro cultures of Vaccinium species and cultivars. HortScience, 26:1320–1322.

    Google Scholar 

  • Roberts, K., 2007. Auxin. In: Handbook of Plant Science. John Wiley & Sons Ltd., USA, p.352–360.

    Google Scholar 

  • Rose, R.J., Wang, X.D., Nolan, K.E., Rolfe, B.G., 2006. Root meristems in Medicago truncatula tissue culture arise from vascular-derived procambial-like cells in a process regulated by ethylene. J. Exp. Bot., 57(10):2227–2235. [doi:10.1093/jxb/erj187]

    Article  PubMed  CAS  Google Scholar 

  • Roy, A.T., De, D.N., 1990. Tissue culture and plant regeneration from immature embryo explants of Calotropis gigantean (L.) R. Br. Plant Cell Tiss. Org. Cult., 20:229–233. [doi:10.1007/BF00041886]

    CAS  Google Scholar 

  • Sahoo, Y., Chand, P.K., 1998. In vitro multiplication of a medicinal herb, Tridax procumbens L. (Mexican daisy, Coatbuttons): influence of explanting season, growth regulator synergy, culture passage and planting substrate. Phytomorphology, 48(2):195–205.

    Google Scholar 

  • Sreedhar, R.V., Venkatachalam, L., Thimmaraju, R., Bhagyalakshmi, N., Narayan, M.S., Ravishankar, G.A., 2008. Direct organogenesis from leaf explants of Stevia rebaudiana and cultivation in bioreactor. Biol. Plant., 52(2):355–360. [doi:10.1007/s10535-008-0073-9]

    Article  CAS  Google Scholar 

  • Srivastava, L.M., 2001. Auxins. In: Plant Growth and Development: Hormones and Environment. Academic Press, USA, p.155–171.

    Google Scholar 

  • Su, Y.H., Liu, Y.B., Zhang, X.S., 2011. Auxin-cytokinin interaction regulates meristem development. Mol. Plant, 4(4):616–625. [doi:10.1093/mp/ssr007]

    Article  PubMed  CAS  Google Scholar 

  • Thomas, C., Meyer, D., Himber, C., Steinmetz, A., 2004. Spatial expression of sunflower SERK gene during induction of somatic embryogenesis and shoot organogenesis. Plant Physiol. Biochem., 42(1):35–42. [doi:10. 1016/j.plaphy.2003.10.008]

    Article  PubMed  CAS  Google Scholar 

  • Vesperinas, E.S., 1998. In vitro root induction in hypocotyl and plumule explants of Helianthus annuus. Env. Exp. Bot., 39(3):271–277. [doi:10.1016/S0098-8472(98)00019-7]

    Article  CAS  Google Scholar 

  • Wang, B., Peng, D.X., Liu, L.J., Sun, Z.H., Zhang, N., Gao, S.M., 2007. An efficient adventitious shoot regeneration system for ramie (Boehmeria nivea Gaud) using thidiazuron. Bot. Stud., 93:173–180.

    Google Scholar 

  • Watas, A.A., Ben, J.J., Tal, E., Solomon, H., 1992. In vitro propagation of Grevillea species. Acta Hort., 316:51–54.

    Google Scholar 

  • Xu, Z., Um, Y.C., Kim, C.H., Lu, G., Guo, D.P., Liu, H.L., Bah, A.A., Mao, A., 2008. Effect of plant growth regulators, temperature and sucrose on shoot proliferation from the stem disc of Chinese jiaotou (Allium chinense) and in vitro bulblet formation. Acta Physiol. Plant, 30(4):521–528. [doi:10.1007/s11738-008-0150-x]

    Article  CAS  Google Scholar 

  • Yamada, Y., Sekiya, J., Koshimizu, K., 1972. Cytokinininduced shoot formation. Phytochemistry, 11(3):1019–1021. [doi:10.1016/S0031-9422(00)88447-2]

    Article  CAS  Google Scholar 

  • Zaizuhana, S., Puteri, J., Noor, M.B., Noral’ashikin, Y., Muhammad, H., Rohana, A.B., Zakiah, I., 2006. The in vivo rodent micronucleus assay of Kacip Fatimah (Labisia pumila) extract. Trop. Biomed., 23(2):214–219.

    PubMed  Google Scholar 

  • Zolman, B.K., Martinez, N., Millius, A., Adham, A.R., Bartel, B., 2008. Identification and characterization of arabidopsis indole-3-butyric acid response mutants detective in novel peroxisomal enzymes. Genetics, 180(1):237–251. [doi:10.1534/genetics.108.090399]

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Division of Human Biology, International Medical University, 57000, Kuala Lumpur, Malaysia

    Anna Pick Kiong Ling

  2. Department of Science, Faculty of Engineering and Science, Universiti Tunku Abdul Rahman, 53300, Setapak, Kuala Lumpur, Malaysia

    Kinn Poay Tan

  3. Agrotechnology and Bioscience Division, Malaysian Nuclear Agency, Bangi, 43000, Kajang, Selangor, Malaysia

    Sobri Hussein

Authors
  1. Anna Pick Kiong Ling
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kinn Poay Tan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Sobri Hussein
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Anna Pick Kiong Ling.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ling, A.P.K., Tan, K.P. & Hussein, S. Comparative effects of plant growth regulators on leaf and stem explants of Labisia pumila var. alata . J. Zhejiang Univ. Sci. B 14, 621–631 (2013). https://doi.org/10.1631/jzus.B1200135

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1631/jzus.B1200135

Key words

CLC number

Search

Navigation