Skip to main content
SpringerLink
Log in

Establishment of an in vitro propagation and transformation system of Balanites aegyptiaca

  • Original Article
  • Published:
Plant Cell, Tissue and Organ Culture (PCTOC) Aims and scope Submit manuscript

Abstract

Balanites aegyptiaca (Balanitaceae) is a drought-tolerant but salt-sensitive tree species distributed in the tropical and arid lands in Africa and Asia. The tree contains many secondary metabolites and a high percentage of oil in the kernels that can be used for biodiesel production. This study aimed to establish an in vitro propagation system of two B. aegyptiaca provenances (El-Kharga and Wadi El-Alaqi) from nodal and cotyledon explants. The explants were placed on Murashige and Skoog medium supplemented with different concentrations of 6-benzyladenine (BA) and thidiazuron (TDZ) for shoot induction. BA was significantly more effective in shoot induction from nodal explants and treatment with BA also resulted in higher regeneration rates of about 40–60 % of adventitious shoots on cotyledon explants, whereas on TDZ-containing medium slightly higher shoot numbers per explant but a negative effect on shoot length were recorded. Rooting was achieved in 40–60 % of the shoots on medium containing between 1.2 and 4.8 µM indole-3-butyric acid. Three different Agrobacterium tumefaciens strains (EHA105, GV3101, and LBA4404) harboring the plasmid pCAMBIA2301 containing the nptII marker and gus reporter genes were used to establish a transformation system in B. aegyptiaca. Strain GV3101 resulted in the highest survival rates and highest number of explants positive in the GUS assay. This selected A. tumefaciens strain was used to introduce pBinAR containing the sequence encoding ERD10 (early responsive to dehydration 10) to produce salt-tolerant B. aegyptiaca plants. The presence of the ERD10 and the nptII gene were detected by PCR in transformed B. aegyptiaca plants.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Ahmad N, Anis M (2007) Rapid clonal multiplication of a woody tree, Vitex negundo L. through axillary shoots proliferation. Agrofor Syst 71:195–200. doi:10.1007/s10457-007-9078-1

    Article  Google Scholar 

  • Anis M, Varshney A, Siddique A (2010) In vitro clonal propagation of Balanites aegyptiaca (L.) Del. Agrofor Syst 78:151–158. doi:10.1007/s10457-009-9238-6

    Article  Google Scholar 

  • Bakhsh A, Anayol E, Ozcan SF (2013) Comparison of transformation efficiency of five Agrobacterium tumefaciens strains in Nicotiana tabacum L. Emir J Food Agric 26:259–264. doi:10.9755/ejfa.v26i3.16437

    Article  Google Scholar 

  • Bates S, Preece JE, Navarrette NE, Van Sambeek JW, Gaffney GR (1992) Thidiazuron stimulates shoot organogenesis and somatic embryogenesis in white ash (Fraxinus americana L.). Plant Cell Tissue Organ Cult 31:21–30

    Article  CAS  Google Scholar 

  • Brugnoli E, Lauteri M (1991) Effects of salinity on stomatal conductance, photosynthetic capacity, and carbon isotope discrimination of salt-tolerant (Gossypium hirsutum L.) and salt-sensitive (Phaseolus vulgaris L.) C (3) non-halophytes. Plant Physiol 95:628–635

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Caruso A, Morabito D, Delmotte F, Kahlem G, Carpin S (2002) Dehydrin induction during drought and osmotic stress in Populus. Plant Physiol Biochem 40:1033–1042. doi:10.1016/S0981-9428(02)01468-7

    Article  CAS  Google Scholar 

  • Chapagain BP, Wiesman Z (2008) Metabolite profiling of saponins in Balanites aegyptiaca plant tissues using LC (RI)-ESI/MS and MALDI-TOF/MS. Metabolomics 4:357–366

    Article  CAS  Google Scholar 

  • Chapagain BP, Yehoshua Y, Wiesman Z (2009) Desert date (Balanites aegyptiaca) as an arid lands sustainable bioresource for biodiesel. Bioresour Technol 100:1221–1226

    Article  CAS  PubMed  Google Scholar 

  • Chetty VJ, Ceballos N, Garcia D, Narváez-Vásquez J, Lopez W, Orozco-Cárdenas ML (2013) Evaluation of four Agrobacterium tumefaciens strains for the genetic transformation of tomato (Solanum lycopersicum L.) cultivar Micro-Tom. Plant Cell Rep 32:239–247. doi:10.1007/s00299-012-1358-1

    Article  CAS  PubMed  Google Scholar 

  • Cohen SN, Chang ACY, Hsu L (1972) Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci USA 69:2110–2114. doi:10.1073/pnas.69.8.2110

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Dawah AK, Ali MAM, El-Mekawey MA, El-Deeb MD, Hassan HMS (2013) Effect of sucrose concentrations and casein hydrolysate on multiplication of desert date (Balanites aegyptiaca, L.) plants. Res J Agric Biol Sci 9:191–197

    Google Scholar 

  • Deng Z, Pang Y, Kong W, Chen Z, Wang X, Liu X, Tang K (2005) A novel ABA-dependent dehydrin ERD10 gene from Brassica napus. DNA Seq 16:28–35. doi:10.1080/10425170500040180

    Article  CAS  PubMed  Google Scholar 

  • Distabanjong K, Geneve RR (1997) Multiple shoot formation from cotyledonary node segments of Eastern redbud. Plant Cell Tissue Organ Cult 47:247–254

    Article  Google Scholar 

  • Dubey PK, Yogi M, Bharadwaj A, Soni ML, Singh A, Sachan AK (2011) Balanites aegyptiaca Del. a semi arid forest tree: a review. Acad J Plant Sci 4:12–18

    Google Scholar 

  • Elfeel AA, Sherif ZH, Abohassan RA (2013) Stomatal conductance, mineral concentration and condensed tannin in three Balanites aegyptiaca (L.) Del. intra-specific sources affected by salinity stress. J Food Agric Environ 11:466–471

    CAS  Google Scholar 

  • El-Mekawy MAM, Ali MAA, Dawah AK, Hassan HMS (2012) Effect of some additives on micropropagation of Balanites aegyptiaca L. explants. World J Agric Sci (WJAS) 8:186–192

    CAS  Google Scholar 

  • El-Tahir A, Ibrahim AM, Satti GMH, Theander TG, Kharazmi A, Khalid SA (1998) Potential anti-leishmanial activity of some Sudanese medicinal plants. Phytother Res 12:570–579

    Google Scholar 

  • Gour VS, Kant T (2011) Efficacy of low cost gelling agents and carbon source alternatives during in vitro rooting of Balanites aegyptiaca and Phyllanthus emblica microshoots. Tree For Sci Biotechnol 5:58–60

    Google Scholar 

  • Gour VS, Emmanuel CJSK, Kant T (2005) Direct in vitro shoot morphogenesis in desert date Balanites aegyptiaca (L.) Del. from root segments. In: Multipurpose trees in the tropics: management and improvement strategies, pp 701–704

  • Gour VS, Sharma SK, Emmanuel CJSK, Kant T (2007) A rapid in vitro morphogenesis and acclimatization protocol for Balanites aegyptiaca (L.) Del.—a medicinally important xerophytic tree. J Plant Biochem Biotechnol 16:151–153

    Article  CAS  Google Scholar 

  • Gupta AK, Harish Rai MK, Phulwaria M, Agarwal T, Shekhawat NS (2014) In vitro propagation, encapsulation, and genetic fidelity analysis of Terminalia arjuna: a cardioprotective medicinal tree. Appl Biochem Biotechnol 173:1481–1494. doi:10.1007/s12010-014-0920-4

    Article  CAS  PubMed  Google Scholar 

  • Hall JB, Walker DH (1991) B. aegyptiaca Del. A monograph. School of Agricultural and Forest Science, University of Wales, Bangor

  • Hanin M, Brini F, Ebel C, Toda Y, Takeda S, Masmoudi K (2011) Plant dehydrins and stress tolerance: versatile proteins for complex mechanisms. Plant Signal Behav 6:1503–1509. doi:10.4161/psb.6.10.17088

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Höfgen R, Willmitzer L (1992) Transgenic potato plants depleted for the major tuber protein patatin via expression of antisense RNA. Plant Sci 87:45–54

    Article  Google Scholar 

  • Huetteman CA, Preece JE (1993) Thidiazuron: a potent cytokinin for woody plant tissue culture. Plant Cell Tissue Organ Cult 33:105–119. doi:10.1007/BF01983223

    Article  CAS  Google Scholar 

  • Husain MK, Anis M (2009) Rapid in vitro multiplication of Melia azedarach L. (a multipurpose woody tree). Acta Physiol Plant 31:765–772. doi:10.1007/s11738-009-0290-7

    Article  CAS  Google Scholar 

  • Hwida MF, El-Kader EMA (2012) Slow growth conservation and molecular characterization of Balanites aegyptiaca L. Res J Agric Biol Sci 8:179–190

    Google Scholar 

  • Jefferson RA (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5:387–405

    Article  CAS  Google Scholar 

  • Khan MI, Ahmad N, Anis M (2011) The role of cytokinins on in vitro shoot production in Salix tetrasperma Roxb.: a tree of ecological importance. Trees 25:577–584. doi:10.1007/s00468-010-0534-6

    Article  Google Scholar 

  • Kim SY, Nam KH (2010) Physiological roles of ERD10 in abiotic stresses and seed germination of Arabidopsis. Plant Cell Rep 29:203–209. doi:10.1007/s00299-009-0813-0

    Article  CAS  PubMed  Google Scholar 

  • Kim M, Schumann CM, Klopfenstein NB, Nebraska EC (1997) Effects of thidiazuron and benzyladenine on axillary shoot proliferation of three green ash (Fraxinus pennsylvanica Marsh.) clones. Plant Cell Tissue Organ Cult 48:45–52

    Article  CAS  Google Scholar 

  • Kovacs D, Kalmar E, Torok Z, Tompa P (2008) Chaperone activity of ERD10 and ERD14, two disordered stress-related plant proteins. Plant Physiol 147:381–390. doi:10.1104/pp.108.118208

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mante S, Scorza R, Cordts J (1988) Plant regeneration from mature plum (Prunus domestica) cotyledons. In Vitro Cell Dev Biol 24:39A (Abstr)

    Google Scholar 

  • Merkle SA, Dean JF (2000) Forest tree biotechnology. Curr Opin Biotechnol 11:298–302. doi:10.1016/S0958-1669(00)00099-9

    Article  CAS  PubMed  Google Scholar 

  • Mohamed AM, Wolf W, Spiess WEL (2002) Physical, morphological and chemical characteristics, oil recovery and fatty acid composition of Balanites aegyptiaca Del. kernels. Plant Foods Hum Nutr 57:179–189

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Ndoye M, Diallo I, Gassama-Dia YK (2003) In vitro multiplication of the semi-arid forest tree, Balanites aegyptiaca (L) Del. Afr J Biotechnol 2:421–424

    Article  CAS  Google Scholar 

  • NRC (2008). Lost Crops of Africa, volume 3: fruits: development, security and cooperation policy and global affairs. National academics press, Washington DC., ISBN-13:978-0309105965, pp 351

  • Peña L, Séguin A (2001) Recent advances in the genetic transformation of trees. Trends Biotechnol 19:500–506

    Article  PubMed  Google Scholar 

  • Phulwaria M, Rai MK, Patel AK, Kataria V, Shekhawat NS (2012) A genetically stable rooting protocol for propagating a threatened medicinal plant-Celastrus paniculatus. AoB Plants 5:plso54. doi:10.1093/aobpla/pls054

    Google Scholar 

  • Radwan UA, Springuel I, Biswas PK, Huluka G (2000) The effect of salinity on water use efficiency of Balanites aegyptiaca. Egypt J Biol 2:1–7

    Google Scholar 

  • Rathore JS, Rathore V, Shekhawat NS, Singh RP, Liler G, Phulwaria M, Dagla HR (2004) Micropropagation of woody plants. Plant Biotechnol Mol Markers 13:207

    Google Scholar 

  • Saharan V, Yadav RC, Yadav NR, Wiesman Z (2011) Somatic embryogenesis and plant regeneration of Balanites aegyptiaca Del (L.): an industrial important arid tree. J Cell Tissue Res 11:2529–2534

    CAS  Google Scholar 

  • Shirly RA, Sadhana PH (2002) In vitro propagation of cashew from young trees. In Vitro Cell Dev Biol Plant 38:152–156. doi:10.1079/IVP2001263

    Article  Google Scholar 

  • Siddique I, Anis M (2008) Direct plant regeneration from nodal explants of Balanites aegyptiaca L. (Del.): a valuable medicinal tree. New For 37:53–62. doi:10.1007/s11056-008-9110-y

    Article  Google Scholar 

  • Singh D, Buhmann AK, Flowers TJ, Seal CE, Papenbrock J (2014) Salicornia as a crop plant in temperate regions: selection of genetically characterized ecotypes and optimization of their cultivating conditions. AoB Plants 6:pluo71. doi:10.1093/aobpla/plu071

    Google Scholar 

  • Sivanesan I, Song JY, Hwang SJ, Jeong BR (2010) Micropropagation of Cotoneaster wilsonii Nakai a rare endemic ornamental plant. Plant Cell Tissue Organ Cult 105:55–63. doi:10.1007/s11240-010-9841-2

    Article  Google Scholar 

  • Sokolowsky V, Kaldenhoff R, Ricci M, Russo VEA (1990) Fast and reliable mini-prep RNA extraction from Neurospora crassa. Fungal Genet Newsl 36:41–43

    Google Scholar 

  • Torregrosa L, Iocco P, Thomas MR (2002) Influence of Agrobacterium strain, culture medium, and cultivar on the transformation efficiency of Vitis vinifera L. Am J Enol Vitic 53:183–190

    CAS  Google Scholar 

  • Varshney A, Anis M (2013a) Direct plantlet regeneration from segments of root of Balanites aegyptiaca Del. (L.)—a biofuel arid tree. Int J Pharm Biol Sci 4:987–999

    CAS  Google Scholar 

  • Varshney A, Anis M (2013b) Evaluation of clonal integrity in desert date tree (Balanites aegyptiaca Del.) by inter-simple sequence repeat marker assay. Acta Physiol Plant 35:2559–2565

    Article  CAS  Google Scholar 

  • Varshney A, Anis M (2014) Synseed conception for short-term storage, germplasm exchange and potentialities of regeneration genetically stable plantlets of desert date tree (Balanites aegyptiaca Del.). Agrofor Syst 88:321–329

    Article  Google Scholar 

Download references

Acknowledgments

We would like to thank Prof. Usama Radwan (Environmental Studies and Development Unit, Faculty of Sciences, Aswan University, Egypt) for the kind help to collect fruits of two genotypes (El-Kharga, Wadi El-Alaqi). We would like to thank the German Academic Exchange Service (DAAD) and the Ministry of Higher Education (MoHE) of the Arab Republic of Egypt cooperation agreement for the support and providing the PhD scholarship to Galal Khamis.

Author contributions

Galal Khamis has conducted all experiments. Jutta Papenbrock and Traud Winkelmann have planned, organized and supervised all experiments. Frank Schaarschmidt supported planning the experiments and evaluated the data statistically. The manuscript was written by all authors.

Author information

Authors and Affiliations

  1. Institute of Botany, Leibniz Universität Hannover, Herrenhäuser Str. 2, 30419, Hannover, Germany

    Galal Khamis & Jutta Papenbrock

  2. Department of Laser Applications in Metrology, Photochemistry and Agriculture (LAMPA), National Institute of Laser Enhanced Sciences, Cairo University, Giza, Egypt

    Galal Khamis

  3. Institute of Horticultural Production Systems, Leibniz Universität Hannover, Herrenhäuser Str. 2, 30419, Hannover, Germany

    Traud Winkelmann

  4. Institute of Biostatistics, Leibniz Universität Hannover, Herrenhäuser Str. 2, 30419 Hannover, Germany

    Frank Schaarschmidt

Authors
  1. Galal Khamis
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Traud Winkelmann
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Frank Schaarschmidt
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jutta Papenbrock
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jutta Papenbrock.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Khamis, G., Winkelmann, T., Schaarschmidt, F. et al. Establishment of an in vitro propagation and transformation system of Balanites aegyptiaca . Plant Cell Tiss Organ Cult 125, 457–470 (2016). https://doi.org/10.1007/s11240-016-0961-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11240-016-0961-1

Keywords

Search

Navigation