Establishment of an in vitro propagation and transformation system of Balanites aegyptiaca
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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.
KeywordsAgrobacterium tumefaciens Balanites aegyptiaca ERD10 Regeneration Rooting Salt stress
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.
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.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- 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 95:628–635CrossRefPubMedPubMedCentralGoogle 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 CrossRefPubMedGoogle 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 9:191–197Google 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 4:12–18Google 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 11:466–471Google 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) 8:186–192Google 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 12:570–579Google 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 5:58–60Google 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 701–704Google Scholar
- Hall JB, Walker DH (1991) B. aegyptiaca Del. A monograph. School of Agricultural and Forest Science, University of Wales, BangorGoogle Scholar
- Hwida MF, El-Kader EMA (2012) Slow growth conservation and molecular characterization of Balanites aegyptiaca L. Res J Agric Biol Sci 8:179–190Google 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
- 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 351Google Scholar
- Radwan UA, Springuel I, Biswas PK, Huluka G (2000) The effect of salinity on water use efficiency of Balanites aegyptiaca. Egypt J Biol 2:1–7Google Scholar
- Rathore JS, Rathore V, Shekhawat NS, Singh RP, Liler G, Phulwaria M, Dagla HR (2004) Micropropagation of woody plants. Plant Biotechnol Mol Markers 13:207Google 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 11:2529–2534Google Scholar
- Sokolowsky V, Kaldenhoff R, Ricci M, Russo VEA (1990) Fast and reliable mini-prep RNA extraction from Neurospora crassa. Fungal Genet Newsl 36:41–43Google 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 53:183–190Google 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 4:987–999Google Scholar