HOMOLOGI FUNGSI GEN KNAT1 ( Knotted 1– like Arabidopsis thaliana) PADA ANGGREK BULAN Phalaenopsis amabilis (L.) Bl. DENGAN MEDIATOR Agrobacterium tumefaciens

Sulastri Isminingsih, Endang Semiarti, Aziz Purwantoro



To understand the function of KNAT1 gene Arabidopsis in the forming and developing of Indonesian origin orchid shoot Phalaenopsis amabilis through transform p35S::KNAT1 and pGreen to protocorm like bodies (plb) of the orchid mediated by Agrobacterium tumafaciens LBA4404. The plb transformants were grown on New Phalaenopsis selection medium containing 5 mg/l BAP, 0.15 mg/l NAA, 15 mg.l Kanamycin and addition of 300 mg/l Cefotaxim to eliminate the overgrowth of Agrobacterium. The analysis of positive transformant use Polimerase Chain Reaction (PCR) with the specific oligonucleotide primers for KNAT1 gene: KNAT1F1R1 and universal primers for pGreen: 35SO and Tnos. The result shows that 3 shoots of 1850 transformants positively carry out the 35S::KNAT1 construct (frequency of transformation was 0.16 %) while 5 shoot of 1850 transformants also positively carry out the pGreen vector, with the frequency of transformation was 0.27 %. The phenotype analysis of 35S::KNAT1 transformants show multiplication on forming of the leaf from a plb to + 10 shoots and forming of the leaf shape which has terompet like shapes and rectangular shape.

Key words: Arabidopsis, KNAT1 gene, Phalaenopsis amabilis, shoot


Arabidopsis, KNAT1 gene, Phalaenopsis amabilis, shoot

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Belarmino,M.M.,Mii,M.2000. Agrobacterium-Mediated Genetic Transformation of Phalaenopsis Orchid. Plant Cell Reports 19: 435-442.

Chuck, G., Lincoln, C., dan Hake, S. 1996. KNAT1 Induced Lobed Leaves with Ectopic Meristem when Overexspressed in Arabidopsis. Plant Cell 8: 1277-1289

Djaafarer, R. 2006. Trend dan Pasar Anggrek Phalaenopsis di Indonesia. Prosiding Seminar Nasional Anggrek (Kiat Pengembangan Anggrek Indonesia di era Globalisasi) p.10-12

Draper, J., R.Scott, P. Armitage and R. Walden. 1988. Plant Gene Transformation and Gene Expression : A Laboratory Manual. Blackwell Scientific publication. Oxford. 355p.

Lincoln, C.., Long,J., Yamaguchi, J., Serikawa, K., dan Hake, S. 1994. A Knotted1-like Homeobox Gene ini Arabidopsis is Expressed in the Vegetative Meristem and Dramatically Alters Leave Morphology when Overexpressed in Transgenic Plants. Plant Cell Reports 6: 1859-1876.

Nadeau, J. A., Zhang, X. S., Li, J., O’Neill, S.D. 1996. Ovule Development: Identification of Stage-Spesific and Tissue Spesific cDNAs. Plant cell 8: 213-239

Sambrook, J., E.F. Fritsch dan T. Maniatis. 1989. Molecular Cloning, A Laboratory Manual. Second Edition. Cold Spring Harbor Laboratory Press, New York.

Semiarti, E., 2006. Perkembangan Bioteknologi Anggrek. Prosiding Seminar Nasional Aanggrek (Kiat Pengembangan Anggrek Indonesia di era Globalisasi) p38-42

Semiarti, E., Y. Ueno, H. Tsukaya, C. Machida, & Y. Machida. 2001. The Asymmetric Leaves2 gene of Arabidopsis thaliana Regulates Formation of Asymmetric Lamina, Establishment of Venation and Eepression of Meristem-Related Homeobox Genes in Leaves. Plant Development 128. p. 1771-1783

Valvekens D., M.V. Montagu and M.V. Lusebettens 1988. Agrobacterium tumefaciens- Mediated Transformation of Arabidopsis thaliana Root Explants by using Kanamycin Selection. Proc. Natl. Acad. Sci. USA 5536-5540.

Venkatachalam, P., Geetha, N., Jayabalan, N., Saravanababu, and Sita, L. 1998. Agrobacterium-Mediated Genetic Transformation of Groundnut (Arachis hypogaea L): An Assessment of Factors Affecting Regeneration of Transgenic Plants., J. Plant Research. 111: 565-572

Watson, J.D., M. Gilman, J. Witkowski, M. Zoller. 1992. W.H. Freeman and Company. New York

DOI: http://dx.doi.org/10.33512/j.agrtek.v1i1.558


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