Agrobacterium rhizogenes-mediated genetic transformation of Musa acuminata cv. Vaibalhla (AAA)
DOI:
https://doi.org/10.14719/pst.1612Keywords:
Agrobacterium rhizogenes, Genetic transformation, Musa acuminata, MizoramAbstract
Agrobacterium rhizogenes-mediated genetic transformation of Musa acuminata cv. Vaibalhla (AAA) was successfully carried out using A. rhizogenes strain A4 harboring the binary vector pCAMBIA2301with VrNHX1gene. In the study, male flowers of M. acuminata cv. Vaibalhla were used as explants to obtain white bud-like structures by culturing on MS basal medium supplemented with 2 mg/l 6-Benzylaminopurine (BAP) and 0.5 mg/l 1-Naphthaleneacetic acid (NAA). The subsequent shoot induction and plantlet regeneration were carried out in MS medium supplemented with Kinetin (2 mg/l) and NAA (0.5 mg/l). For the genetic transformation, in vitro raised plantlets were inoculated with A. rhizogenes strain A4 harboring pCAMBIA2301VrNHX1 for 30 min followed by 2 days of co-cultivation in the dark in a semi solid MS basal medium. The treated plants were then transferred and maintained in MS basal medium supplemented with ascorbic acid (75 mg/l), kanamycin (150 mg/l) and cefotaxime (400 mg/l). Initiation of hairy roots were observed within 2 days of transfer which were evaluated for GUS activity and the subsequent GUS+ roots were assayed for transient transformation by PCR using nptII and NHX1 gene specific primers. The transfer of the plasmid as well as the gene was confirmed with positive bands observed at 540 bp and 1.6 kb respectively.
Downloads
References
Broothaerts W, Mitchell HJ, Weir B, Kaines S, Smith LMA et al. Gene transfer to plants by diverse species of bacteria. Nature. 2005; 433: 629-33. https://doi.org/10.1038/nature03309
Venkatachalam L, Lokesh V, Bhagyalakshmi N. A rare event of Agrobacterium rhizogenes-assisted genetic transformation of ‘Silk’ banana (genotype-AAB). Journal of Microbial and Biochemical Technology. 2011; 3:1. https://doi.org/10.4172/1948-5948.1000043
Bosselut N, Ghelder CV, Claverie M, Voisin R, Onesto JP, Rosso MN, Esmenjaud D. Agrobacterium rhizogenes-mediated transformation of Prunus as an alternative for gene functional analysis in hairy-roots and composite plants. Plant Cell Reports. 2011; 30: 1313-26. https://doi.org/10.1007/s00299-011-1043-9
Chilton MD, Tepfer DA, Petit A, David C, Cassedelpart F, Tempe J. Agrobacterium rhizogenes inserts T DNA into the genomes of the host plants root cells. Nature. 1982; 295:432-34. https://doi.org/10.1038/295432a0
Moore L, Warren G, Strobel G. Involvement of a plasmid in hairy root disease of plants caused by Agrobacterium rhizogenes. Plasmid. 1979; 2: 617-26. https://doi.org/10.1016/0147-619X(79)90059-3
Kumar V, Satyanarayana KV, Sarala Itty S, Indu EP, Giridhar P et al. Stable transformation and direct regeneration of Coffea canephora P ex. Fr. by Agrobacterium rhizogenes mediated transformation without hairy-root phenotype. Plant Cell Reports. 2005;25:214-22. https://doi.org/10.1007/s00299-005-0045-x
Henzi MX, Christey MC, McNeil DL, Davis KM. Agrobacterium rhizogenes mediated transformation of broccoli (Brassica oleracea L. var. Italica) with an antisense 1- aminocyclopropane-1-carboxylic acid oxidase gene. Plant Science. 1999; 143:55-62. https://doi.org/10.1016/S0168-9452(99)00024-2
Yazaki K, Tanaka S, Matsuoka H, Sato F. Stable transformation of Lithospermumerythrorhizon by Agrobacterium rhizogenes and shikonin production of the transformants. Plant Cell Reports. 1998; 18: 214-19. https://doi.org/10.1007/s002990050559
Christey MC, Sinclair BK, Braun RH, Wyke L. Regeneration of transgenic vegetable brassicas (Brassica oleracea and B. campestris) via Ri mediated transformation. Plant Cell Reports. 1997;16:587-93. https://doi.org/10.1007/BF01275497
Liu S, Su L, Liu S, Zeng X, Zheng D, Hong L, Li L. Agrobacterium rhizogenes-mediated transformation of Arachis hypogaea: an efficient tool for functional study of genes. Biotechnology and Biotechnological Equipment. 30:869-78. https://doi.org/10.1080/13102818.2016.1191972
Khlifa HD, Klimek-Chodacka M, Baranski R, Combik M, Taha HS. Agrobacterium rhizogenes-mediated transformation of Hypericum sinaicum L. for the development of hairy roots containing hypericin. Brazilian Journal of Pharmaceutical Sciences. 2020; 56: e18327. http://dx.doi.org/10.1590/s2175-97902020000118327
Ishida JK, Yoshida S, Ito M, Namba S, Shirasu K. Agrobacterium rhizogenes-Mediated Transformation of the Parasitic Plant Phtheirospermum japonicum. PLoS ONE. 2011; 6: e25802. https://doi.org/10.1371/journal.pone.0025802
Leppyanen IV, Kirienko AN, Dolgikh EA. Agrobacterium rhizogenes-mediated transformation of Pisum sativum L. roots as a tool for studying the mycorrhizal and root nodule symbioses. PeerJ. 2019; 7: e6552. http://doi.org/10.7717/peerj.6552
Cheng Y, Wang X, Cao L, Ji J, Liu T, Duan K. Highly efficient Agrobacterium rhizogenes mediated hairy root transformation for gene functional and gene editing analysis in soybean. Plant Methods, 2021; 17:73. https://doi.org/10.1186/s13007-021-00778-7
Hrahsel L, Basu A, Sahoo L, Thangjam R. In vitro propagation and assessment of the genetic fidelity of Musa acuminata (AAA) cv. Vaibalhla derived from immature male flowers. Applied Biochemistry and Biotechnology. 2014; 172: 1530-39. https://doi.org/10.1007/s12010-013-0637-9
Mishra S, Alavilli H, Lee B, Panda SK, Sahoo L. Cloning and Functional Characterization of a Vacuolar Na+/H+ Antiporter Gene from Mungbean(VrNHX1) and Its Ectopic Expression Enhanced Salt Tolerance in Arabidopsis thaliana. PLOS ONE; 2014; 9: e106678. https://doi.org/10.1371/journal.pone.0106678
Höfgen R, Willmitzer L. Storage of competent cells for Agrobacterium transformation. Nucleic Acid Research. 1988; 16 (20): 9877. https://doi.org/10.1093/nar/16.20.9877
Jefferson RA. Assaying chimeric genes in plants: the GUS gene fusion system. Plant Molecular Biology. 1987;204:387-405. https://doi.org/10.1007/BF02667740
Thangjam R, Sahoo L. In vitro regeneration and Agrobacterium tumefaciens-mediated genetic transformation of Parkia timoriana (DC.) Merr.: a multipurpose tree legume. Acta Physiologiae Plantarum. 2012; 34: 1207-15. https://doi.org/10.1007/s11738-011-0917-3
May GD, Afza R, Mason HS, Wieko A, Novak, FJ, Arntzen CJ. Generation of transgenic banana (Musa acuminata) plants via Agrobacterium-mediated transformation. Nature Biotechnology. 1995; 13: 486-92. https://doi.org/10.1038/nbt0595-486
Becker BK, Dugdale B, Smith MK, Harding RM, Dale JL. Genetic transformation of Cavendish banana (Musa spp. AAA group) cv. ‘Grand Nain’ via micro projectile bombardment. Plant Cell Reports. 2000; 19: 229-34. https://doi.org/10.1007/s002990050004
Hrahsel L, Basu A, Sahoo L, Thangjam R. Genetic Transformation of Musa acuminata cv. Vaibalhla (AAA) using Agrobacterium tumefaciens. Science and Technology Journal. 2017;5:120-31. https://doi.org/10.22232/stj.2017.05.02.09
Downloads
Published
Versions
- 01-07-2022 (2)
- 18-06-2022 (1)
How to Cite
Issue
Section
License
Copyright (c) 2022 Lalremsiami Hrahsel, Robert Thangjam
This work is licensed under a Creative Commons Attribution 4.0 International License.
Copyright and Licence details of published articles
Authors who publish with this journal agree to the following terms:
- Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
Open Access Policy
Plant Science Today is an open access journal. There is no registration required to read any article. All published articles are distributed under the terms of the Creative Commons Attribution License (CC Attribution 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited (https://creativecommons.org/licenses/by/4.0/). Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).
