Abstract
Key message
The study describes an efficient regeneration system and encapsulation of nodal segments in F. carica accompanied with frequent root formation in microshoots under the influence of salicylic acid.
Abstract
The present study describes an improved protocol for clonal propagation and a short-term conservation strategy using an encapsulation technology for Ficus carica. Nodal segments were more responsive than shoot tips for shoot multiplication. Among the cytokinins tested, Murashige and Skoog’s (MS) medium augmented with 7.5 µM 6-benzyladenine (BA) was the best, but the leaves exhibited a rudimentary structure. Cytokinin–auxin combinations enhanced the multiplication rate; but leaf growth did not improve. However, the supplementation of 50 µM adenine sulphate (Ads) to MS + 7.5 µM BA + 0.5 µM α-naphthalene acetic acid (NAA) significantly enhanced mean shoot number per explant and overall growth of shoots and induced a maximum of 26.8 shoots per nodal segment and 15 shoots per shoot tip with mean shoot length of 8 and 4.3 cm, respectively, after 4 weeks of culture. Microshoots rooted on half-strength MS medium containing 2.5 µM salicylic acid (SA) either in semi-solid (mean of 4.6 roots per shoot) or liquid medium (mean of 5 roots per shoot). A gelling matrix of 4 % sodium alginate (Na-alginate) and 100 mM calcium chloride (CaCl2·2H2O) was used for the encapsulation of nodal segments. Maximum shoot growth (95.6 %) was recorded on MS medium supplemented with 7.5 µM BA + 0.5 µM NAA + 50 µM Ads. Rooting was induced upon transfer of sprouted microshoots to semi-solid half-strength MS medium supplemented with 2.5 µM SA. To examine the retention of encapsulated and non-encapsulated nodal segments’ viability, a low-temperature storage (4 °C) experiment was carried out. Encapsulated nodal segments stored at 4 °C for 1–6 weeks sprouted at variable frequencies in successive weeks of transfer. Plantlets were acclimatized and established in field where they grew without any detectable variations after 6 months of transfer.
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Abbreviations
- Ads:
-
Adenine sulphate
- BA:
-
6-Benzyladenine
- IAA:
-
Indole-3-acetic acid
- IBA:
-
Indole-3-butyric acid
- 2ip:
-
2-Isopentyl adenine
- Kn:
-
Kinetin
- MS:
-
Murashige and Skoog (1962)
- NAA:
-
α-Naphthalene acetic acid
- PGR:
-
Plant growth regulator
- SA:
-
Salicylic acid
References
Anonymous (2002) The wealth of India. A dictionary of Indian raw materials and industrial products, vol 4. NISCAIR and CSIR, New Delhi, p 26–32
Aref HL, Salah KB, Chaumont JP, Fekih A, Aouni M, Said K (2010) In vitro antimicrobial activity of four Ficus carica latex fraction against resistant human pathogens. Pak J Pharm Sci 23:53–58
Bapat VA, Rao PS (1962) Plantlet regeneration from encapsulated and non-encapsulated desiccated somatic embryos of forest tree Santalum album. J Plant Biochem Biotechnol 1:109–113
Campos VP (1997) Nematόides na cultura da figueira. Inf Agrop 18:33–38
Danso KE, Ford-Lloyd BV (2003) Encapsulation of nodal cuttings and shoot tips for storage and exchange of cassava germplasm. Plant Cell Rep 21:718–725
Fráguas CB, Pasqual M, Dutra LF, Cazetta JO (2004) Micropropagation of fig (Ficus carica L.) ‘Roxo de Valinhos’ plants. In Vitro Cell Dev Biol-Plant 40:471–474
Hepaksoy S, Aksoy U (2006) Propagation of Ficus carica L. clones by in vitro culture. Biol Plant 50:433–436
Husain MK, Anis M, Sahazad A (2008) In vitro propagation of a multipurpose leguminous tree (Pterocarpus marsupium Roxb.) using nodal explants. Acta Physiol Plant 30:353–359
İsfendiyaroğlu M, Özeker E (2008) Rooting of Olea europaea ‘Domat’ cuttings by auxin and salicylic acid treatments. Pak J Bot 40:1135–1141
Kim KM, Kim MY, Yun PY, Chandrasekhar T, Lee H-Y, Song P-S (2007) Production of multiple shoots and plant regeneration from leaf segments of fig tree (Ficus carica L.). J Plant Biol 50:440–446
Kumar V, Radha A, Kumar CS (1998) In vitro plant regeneration of Ficus carica L. cv. Gular using apical buds from mature trees. Plant Cell Rep 17:717–720
Lakshmanan P, Lee CL, Goh CJ (1997) An efficient in vitro method for mass propagation of a woody ornamental Ixora coccinea L. Plant Cell Rep 16:572–577
Murashige T (1974) Plant propagation through tissue culture. Annu Rev Plant Physiol 25:135–166
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue culture. Physiol Plant 15:473–497
Nieves N, Larenzo JC, Blanco MA, Gonzalez J, Perralta H, Hermandez M, Santos R, Concepcion O, Barroto CG, Taipa R, Martinez MF, Fundara GA (1998) Artificial endosperm of Cleopatra tangerine zygotic embryos, a mode for somatic embryo encapsulation. Plant Cell Tiss Org Cult 54:77–83
Qrunfleh IM, Shatnawi MM, Al-Ajlouni ZI (2013) Effect of different concentrations of carbon source, salinity and gelling agent on in vitro growth of fig (Ficus carica L.). African J Biotechnol 12:936–940
Rahman MM, Amin MN, Hossain MF (2004) In vitro propagation of Ficus benghalensis L. a multipurpose and keystone species of Bangladesh. Plant Tiss Cult 14:135–142
Redenbaugh K, Slade D, Vissa P, Fujii J (1987) Encapsulation of somatic embryos in synthetic seed coats. Hort Sci 22:803–809
Ribeiro IJA, Doenc ADF (1999) In: Corrêa LSD, Boliaani AC (eds) Cultura da figueira-do plantioá comercializacão. FAPESP, Ilha Solteira, pp 151–164
Rubnov S, Kashman Y, Rabinowitz R, Schlesinger M, Mechoulam R (2001) Suppressors of cancer cell proliferation from fig (Ficus carica) resin: isolation and structure elucidation. J Nat Prod 64:993–996
Sakhanokho HF, Kelley RY (2009) Influence of salicylic acid on in vitro propagation and salt tolerance in Hibiscus acetosella and Hibiscus moscheutos (cv ‘Luna Red’). African J Biotech 8:1474–1481
Sharma S, Shahzad A (2012) Encapsulation technology for short-term storage and conservation of a woody climber, Decalepis hamiltonii Wight and Arn. Plant Cell Tiss Org Cult 111:191–198
Sharma S, Shahzad A, Jan N, Sahai A (2009) In vitro studies on shoot regeneration through various explants and alginate-encapsulated nodal segments of Spilanthes mauritiana DC. an endangered medicinal herb. Int J Plant Dev Biol 3:56–61
Sharma S, Shahzad A, Teixeira da Silva JA (2013) Synseed technology-a complete synthesis. Biotechnol Adv 31:186–207
Sharma S, Shahzad A, Ahmad A, Anjum L (2014a) 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
Sharma S, Shahzad A, Kumar J, Anis M (2014b) In vitro propagation and synseed production of scarlet salvia (Salvia splendens). Rend Fis Acc Lincei 25:359–368
Shirin F, Rana PK, Mandal AK (2005) In vitro clonal propagation of mature Tectona grandis through axillary bud proliferation. J Forest Res 10:465–469
Vieitez AM, Carmen SM, Vieitez E (1985) In vitro plantlet regeneration from juvenile and mature Quercus robur L. J Hort Sci 60:99–106
Yakushiji H, Mase N, Sato Y (2003) Adventitious bud formation and plantlet regeneration from leaves of fig (Ficus carica L.). J Hort Sci Biotechnol 78:874–878
Author contribution statement
This work was carried out in collaboration among four authors. Author AS suggested the study and extended overall guidance during the experimentation. Author SS and SM equally performed the experimental work and wrote the first draft of the manuscript. Author TS analyzed the data. All the authors read and approved the final manuscript.
Acknowledgments
Dr. Shiwali Sharma is thankful to DST, for the award of Young Scientist under Fast Track Scheme, SERB (vide no. SB/FT/LS-364/2012) for providing research assistance. Dr. Anwar Shahzad gratefully acknowledges the financial support provided by UGC and UP-CST in the form of research projects (vide no. 39-369/2010 SR and vide no. CST/D3836). Taiba Saeed acknowledges the financial support provided by UGC under the scheme of Maulana Azad National Fellowship (file no. MANF-MUS-UTT-2624).
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The authors have no conflict of interest.
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Communicated by K. Klimaszewska.
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Sharma, S., Shahzad, A., Mahmood, S. et al. High-frequency clonal propagation, encapsulation of nodal segments for short-term storage and germplasm exchange of Ficus carica L.. Trees 29, 345–353 (2015). https://doi.org/10.1007/s00468-014-1114-y
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DOI: https://doi.org/10.1007/s00468-014-1114-y