Skip to main content

Advertisement

Log in

Enhanced multiplication and improved ex vitro acclimatization of Decalepis arayalpathra

  • Original papers
  • Published:
Biologia Plantarum

Abstract

The proposed work describes a protocol for high-frequency in vitro regeneration through nodal segments and shoot tips in Decalepsis arayalpathra, a critically endangered medicinal liana of the Western Ghats. Nodal segments were more responsive than shoot tips in terms of shoot proliferation. Murashige and Skoog’s (MS) basal medium supplemented with 5.0 μM 6-benzyladenine (BA) was optimum for shoot initiation through both the explants. Among different combinations of plant growth regulators and growth additive screened, MS medium added with 5.0 μM BA + 0.5 μM indole-3-acetic acid + 20.0 μM adenine sulphate effectuated the highest response: 11.8 shoots per nodal segment and 5.5 shoots per shoot tip with mean shoot length of 9.2 and 4.8 cm, respectively. Half-strength MS medium with 2.5 μM α-naphthalene acetic acid was optimum for in vitro root induction. The plantlets with the well developed shoot and root were acclimatized in Soilrite™ with 92 % survival rate in the field conditions. During acclimatization, chlorophyll content, net photosynthetic rate, stomatal conductance, and transpiration rate were gradually changed in dependence of formation of new leaves. Further, the changes in activities of antioxidant enzymes, i.e., superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase (GR) as well as activity of carbonic anhydrase were also observed: a continuous rise in SOD activity, but a rise and fall in the activities of CAT, APX, and GR were also noticed. Maximum fresh mass (3.1 g plant-1), dry mass (0.35 g plant-1) of roots and 2-hydroxy-4-methoxybenzaldehyde content of 9.22 μg cm-3(root extract) were recorded after 8 weeks of acclimatization.

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

Abbreviations

APX:

ascorbate peroxidase

ADS:

adenine sulphate

BA:

6-benzyladenine

CA:

carbonic anhydrase

CAT:

catalase

Chl:

chlorophyll

E:

transpiration rate

Glu:

glutamine

GR:

glutathione reductase

gs :

stomatal conductance

2H4MB:

2-hydroxy-4-methoxybenzaldehyde

IAA:

indole-3-acetic acid

IBA:

indole-3-butyric acid

2iP:

2-isopentyladenine

Kn:

kinetin

MS:

Murashige and Skoog

NAA:

α-naphthalene acetic acid

PG:

phloroglucinol

PGR:

plant growth regulator

PN :

net photosynthetic rate

SOD:

superoxide dismutase

References

  • Aebi, H.: Catalase In vitro Methods. — Enzymol. 105: 121–12, 1984.

    Article  CAS  Google Scholar 

  • Anitha, S., Pullaiah, T.: In vitro propagation of Decalepis hamiltonii. — J. trop. med. Plants 3: 227–232, 2002.

    Google Scholar 

  • Asada, K.: The water-water cycle in chloroplasts: scavenging of active oxygen and dissipation of excess photons. — Annu. Rev. Plant Physiol. Plant mol. Biol. 50: 601–639, 1999.

    Article  CAS  PubMed  Google Scholar 

  • Datta, M.M., Mukherjee, P., Ghosh, B., Jha, T.B.: In vitro clonal propagation of biodiesel plant (Jatropha curcas L.). — Curr. Sci. 93: 1438–1442, 2007.

    CAS  Google Scholar 

  • Dhindsa, R.S., Plumb-Dhindsa, P., Thorpe, T.A.: Leaf senescence: correlated with increased levels of membrane permeability and lipid peroxidation and decreased levels of superoxide dismutase and catalase. — J. exp. Bot. 32: 93–10, 1981.

    Article  CAS  Google Scholar 

  • Dwivedi, R.S., Randhawa, N.S.: Evaluation of rapid test for the hidden hunger of zinc in plants. — Plant Soil 40: 445–451, 1974.

    Article  CAS  Google Scholar 

  • Faisal, M., Anis, M.: Changes in photosynthetic activity, pigment composition, electrolyte leakage, lipid peroxidation and antioxidant enzymes during ex vitro establishment of micropropagated Rauvolfia tetraphylla plantlets. — Plant Cell Tissue Org. Cult. 99: 125–132, 2009.

    Article  CAS  Google Scholar 

  • Faisal, M., Anis, M.: Effect of light irradiations on photosynthetic machinery and antioxidative enzymes during ex-vitro acclimatization of Tylophora indica plantlets. — J. Plant Interact. 5: 21–27, 2010.

    Article  CAS  Google Scholar 

  • Foyer, C.H., Halliwell, B.: The presence of glutathione and glutathione reductase in chloroplasts: a proposed role in ascorbic acid metabolism. — Planta 133: 21–25, 1976.

    Article  CAS  PubMed  Google Scholar 

  • Foyer, C.H., Mullineaux, P.M.: The presence of dehydroascorbate and dehydroascorbate reductase in plant tissues. — FEBS Lett. 425: 528–529, 1998.

    Article  CAS  PubMed  Google Scholar 

  • Gangaprasad, A., Decruse, W.S., Seeni, S., Nair, M.G.: Micropropagation and ecorestoration of Decalepis arayalpathra (Joseph & Chandra.) Venter an endemic and endangered ethnomedicinal plant of Western Ghats. — Ind. J. Biot. 4: 265–270, 2005.

    CAS  Google Scholar 

  • Giridhar, P., Rajasekaran, Y., Ravishankar, G.A.: Production of root specific flavour compound, 2-hydroxy-4-methoxy benzaldehyde by normal root cultures of Decalepis hamiltonii Wight & Arn (Asclepiadaceae). — J. Sci. Food Agr. 85: 61–64, 2005.

    Article  CAS  Google Scholar 

  • Husain, M.K., Anis, M.: Rapid in vitro multiplication of Melia azedarach L. (a multipurpose woody tree). — Acta Physiol. Plant 31: 765–772, 2009.

    Article  CAS  Google Scholar 

  • Husain, M.K., Anis, M., Shahzad, A.: In vitro propagation of Indian kino (Pterocarpus marsupium Roxb.) using thidiazuron. — In Vitro cell. dev. Biol. Plant 43: 59–64, 2007.

    Article  CAS  Google Scholar 

  • Joseph, J., Chandrasekharan, V.: Jankia arayalpathra–a new genus and species of Periplocaceae from Kerala, South India. — J. Indian Bot. Soc. 57: 308–312, 1978.

    Google Scholar 

  • Kaur, K., Kanta, U.: Clonal propagation of Acacia catechu wild by shoot tip culture. — Plant Growth Reg. 31: 143–145, 2000.

    Article  CAS  Google Scholar 

  • Kayihan, C., Eyidogan, F., Afsar, N., Oktem, H.A., Yucel. M.: Cu/Zn superoxide dismutase activity and respective gene expression during cold acclimation and freezing stress in barley cultivars. — Biol. Plant. 56: 693–698, 2012.

    Article  CAS  Google Scholar 

  • Malik, S.K., Chaudhury, R., Kalia, R.K.: Rapid in vitro multiplication and conservation of Garcinia indica: a tropical medicinal tree species. — Sci. Hort. 106: 539–553, 2005.

    Article  Google Scholar 

  • Mitrović, A., Bogdanović, J.: Activities of antioxidative enzymes during Chenopodium rubrum L. ontogenesis in vitro. — Arch. Biol. Sci. 60: 223–231, 2008.

    Article  Google Scholar 

  • Murashige, T.: Plant propagation through tissue culture. — Annu. Rev. Plant Physiol. 25: 135–166, 1974.

    Article  CAS  Google Scholar 

  • Murashige, T., Skoog, F.: A revised medium for rapid growth and bioassay with tobacco tissue culture. — Physiol. Plant 15: 473–497, 1962.

    Article  CAS  Google Scholar 

  • Nakano, Y., Asada, K.: Hydrogen peroxide is scavenged by ascorbate specific peroxidise in spinach chloroplasts. — Plant Cell Physiol. 22: 867–880, 1981.

    CAS  Google Scholar 

  • Naz, R., Anis, M., El Atta, H.A.: Micropropagation of Cassia occidentalis L. and the effect of irradiance on photosynthetic pigments and antioxidative enzymes. — Biol. Plant. 59: 1–10, 2015.

    Article  CAS  Google Scholar 

  • Pospíšilová, J., Tichá, I., Kadleček, P., Haisel, D., Plzáková, S.: Acclimatization of micropropagated plants to ex vitro conditions. — Biol. Plant. 42: 481–497, 1999.

    Article  Google Scholar 

  • Sahai, A., Shahzad, A.: High frequency in vitro regeneration system for conservation of Coleus forsklohlii: a threatened medicinal herb. — Acta Physiol. Plant 35: 473–481, 2013.

    Article  CAS  Google Scholar 

  • Sahai, A., Shahzad, A., Sharma. S.: Histology of organogenesis and somatic embryogenesis in excised root cultures of an endangered species Tylophora indica (Asclepiadaceae). — Aust. J. Bot. 58: 198–205, 2010.

    Article  Google Scholar 

  • Scalet, M., Federice, R., Guido, M.C., Manes, F.: Peroxidase activity and polyamine changes in response to ozone and simulated acid rain in Aleppo pine needles. — Environ. exp. Bot. 35: 417–425, 1995.

    Article  CAS  Google Scholar 

  • Sharma, S., Shahzad, A., Ahmad, A., Anjum. L.: In vitro propagation and the acclimatization effect on the synthesis of 2-hydroxy-4-methoxy benzaldehyde in Decalepis hamiltonii Wight and Arn. — Acta Physiol. Plant 36: 2331–2344, 2014a.

    Article  CAS  Google Scholar 

  • Sharma, S., Shahzad, A., Kumar, J., Anis, M.: In vitro propagation and synseed production of scarlet salvia (Salvia splendens). — Rend. Fis. Acc. Lincei 25: 359–368, 2014b.

    Article  Google Scholar 

  • Shine, V.J., Shyamal, S., Latha, P.G., Rajasekharan, S.: Gastric antisecretory and antiulcer activities of Decalepis arayalpathra. — Pharmac. Biol. 45: 210–216, 2007.

    Article  Google Scholar 

  • Sudha, C.G., Sherina, T.V., Anu-Anand, V.P., Reji, J.V., Padmesh, P., Soniya, E.V.: Agrobacterium rhizogenes mediated transformation of the medicinal plant Decalepis arayalpathra and production of 2-hydroxy-4-methoxy benzaldehyde. — Plant Cell Tissue Organ Cult. 112: 217–226, 2013.

    Article  CAS  Google Scholar 

  • Sudha, C.G., Krishnan, P.N., Pushpangadan, P., Seeni, S.: In vitro propagation of Decalepis arayalpathra, a critically endangered ethnomedicinal plant. — In Vitro cell. dev. Biol. Plant 41: 648–654, 2005.

    Article  Google Scholar 

  • Sudha, C.G., Seeni, S.: Establishment and analysis of fastgrowing normal root culture of Decalepis arayalpathra, a rare endemic medicinal plant. — Curr. Sci. 81: 371–374, 2001.

    CAS  Google Scholar 

  • Thiyagarajan, M., Venkatachalam, P.: A reproducible and high frequency plant regeneration from mature axillary node explants of Gymnema sylvestre (Gurmur)-an important antidiabetic endangered medicinal plant. — Ind. Crop Prod. 50: 517–524, 2013.

    Article  CAS  Google Scholar 

  • Varshney, A., Anis, M.: Improvement of shoot morphogenesis in vitro and assessment of changes of the activity of antioxidant enzymes during acclimation of micropropagated plants of Desert Teak. — Acta Physiol. Plant. 34: 859–867, 2012.

    Article  CAS  Google Scholar 

  • Van Huylenbroeck, J.M., Piqueras, A., Debergh, P.C.: Photosynthesis and carbon metabolism in leaves formed prior and during ex vitro acclimatization of micropropagated plants. — Plant Sci. 134: 21–30, 1998.

    Article  Google Scholar 

  • Venter, H.J.T., Veroeven, R.L.: A tribal classification of the Periplocoideae (Asclepiadaceae). — Taxon 46: 705–720, 1997.

    Article  Google Scholar 

  • Verma, R.S., Mishra, P., Kumar, A., Chauhan, A., Padalia, R.C., Sundaresan, V.: Chemical composition of root aroma of Decalepis arayalpathra (J. Joseph and V. Chandras.) Venter, an endemic and endangered ethnomedicinal plant from Western Ghats, India. — Nat. Prod. Res. 28: 1202–1205, 2014.

    Article  CAS  PubMed  Google Scholar 

  • Wang, H., Li, Y., Gao, Y., Zu, Y.: CO2, H2O exchange and stomatal regulation of regenerated Camptotheca acuminata plantlets during ex vitro acclimatization. — J. Forest Res. 17: 273–276, 2006.

    Article  Google Scholar 

  • Wang, J., Liu, H., Zhao, J., Gao, H., Zhou, L., Liu, Z., Chen, Y., Sui, P.: Antimicrobial and antioxidant activities of the root bark essential oil of Periploca sepium and its main component 2-hydroxyl-4-methoxybenzaldehyde. — Molecules 24: 5807–5817, 2010.

    Article  Google Scholar 

  • Xu, F.J., Li, G., Jin, C.W., Liu, W.J., Zhang, S.S., Zhang, Y.S., Lin, X.Y.: Aluminium induced changes in reactive oxygen species accumulation, lipid peroxidation and antioxidant capacity in wheat root tips. — Biol. Plant. 56: 89–96, 2012.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to A. Shahzad.

Additional information

Acknowledgments: Financial assistance in the form of University Non-net Fellowship to Zishan Ahmad is gratefully acknowledged. Dr. Shiwali Sharma gratefully acknowledges the financial assistance provided by the Department of Science and Technology (DST) in the form of Young Scientist Project under Fast Track Scheme, SERB (Vide No. SB/FT/LS-364/2012).

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ahmad, Z., Shahzad, A. & Sharma, S. Enhanced multiplication and improved ex vitro acclimatization of Decalepis arayalpathra. Biol Plant 62, 1–10 (2018). https://doi.org/10.1007/s10535-017-0746-3

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10535-017-0746-3

Additional key words

Navigation