Zulkarnain, Zulkarnain (2017) THE APPLICATION OF PLANT TISSUE CULTURE IN THE PROPAGATION OF Guichenotia macrantha Turcz. S2 thesis, University of Melbourne, Australia.

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Guichenotia macrantha Turcz. is an important woody ornamental species of Western Australia. This species is commonly grown as a cut flower, container or rockery plant and, is preferred in cultivation to other members of the genus. Clonal propagation of G. macrantha is conventionally by semi-hardwood cuttings that frequently takes many weeks to produce roots. Plant tissue culture technique offers a number of advantages and has been widely applied to a wide range of plant species. It is not yet reported as having been applied to this ornamental plant. Therefore, the feasibility of G. macrantha propagation by tissue culture is investigated in this study. Plant tissue culture is a technique to isolate plant tissue, organs or embryos, followed by growing them in an aseptic condition in order to regenerate complete plants. With this technique, the development of a specific nutrient formulation is often necessary for particular plant species. One of the components determining the success of proliferation and differentiation of tissue cultured materials are plant growth regulators, particularly auxins and cytokinins. Amongst the auxin group, indole-3-acetic acid (IAA) is commonly used in practice as well as benzylamino purine (BAP) of cytokinin group. Both regulators have been proven to be effective for in vitro culture of a wide range of woody species. This investigation is aimed at determining the level of IAA and/or BAP that results in satisfactory growth of G. macrantha nodal cuttings cultured in vitro, and to develop a practical procedure for mass propagation of this plant. It is expected that this investigation would provide a valuable contribution to agricultural biotechnology and to the conservation of this Australian native species. This study was conducted at the Plant Science Laboratory, Victorian College of Agriculture and Horticulture, Burnley Campus. The duration of this study was over ten months, from January to October 1994. Two factors were investigated: IAA levels (0.00, 0.57, 2.85, 5.71µM) and BAP levels (0.00, 2.22, 4.44, 8.88µM) in a completely randomised design. Analysis of variance was employed in data analysis, followed by multiple-comparison test using the studentisized range method. A full-strength Murashige and Skoog (MS) composition supplemented with 3% sucrose and solidified with 0.8% Bacto agar at pH 5.8 ± 0.02 was used as a basal medium. At the rooting stage, however, the medium used was half-strength MS supplemented with 1mg/l activated charcoal with no growth regulators. Data on culture contamination, shoot growth, in vitro rooting, plantlet regeneration, acclimatisation and callus formation were recorded from the first week of culture initiation until the end of experiment period. The results indicated that weekly treatments using 0.5mg/l Benlate on greenhouse-grown stock plants followed by the application of 0.05% Na-hypochlorite in surface sterilisation resulted in 98% explants cultured at the initiation stage which are contamination free. BAP at 4.44µM or higher was found to cause vitrification and necrotic symptoms on leaves and shoot tips, and reduced shoot elongation. BAP also showed significant effect on the percentage of transferable explants, average microcutting production and percentage of explant forming callus. However, neither IAA alone nor its interaction with BAP resulted in significant effect on these three parameters. Root formation was only observed for 10% of explants treated with 2.85µM IAA in the second subculture. At the rooting stage, root formation varied from 15.79 to 46.15% on either IAA and/or BAP treatments. The percentage of explants which survived acclimatisation varied from 80 to 100%. It was concluded that: mass propagation of G. macrantha via tissue culture is made possible by using single node explants; BAP significantly enhances proliferation and differentiation of cultured tissues; the best combination for a satisfactory growth of explants is 0.57µM IAA + 2.22µM BAP; and the use of 1mg/l activated charcoal is not effective to induce in vitro root formation on microcuttings. Further investigation was recommended for the improvement of the procedure developed in this study.

Type: Thesis (S2)
Subjects: S Agriculture > SB Plant culture
Depositing User: Zulkarnain
Date Deposited: 27 Dec 2017 00:50
Last Modified: 06 Jan 2022 06:29

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