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MAJU, T.T. and SONIYA, E.V. (2012) In vitro regeneration system for multiplication and transformation in P i per nigrum L. International Journal of Medicinal and Aromatic Plants, 2 (1). pp. 178-184. ISSN 2249-4340

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Abstract

An efficient protocol for rapid multiplication of black pepper ( Piper nigrum L., Piperaceae) was developed. In vitro germ inated seedlings were used as the source of contamination free explants for the study. Direct induction of shoots was obtained from bulged portion of shoot tip and nodal segments cultured on Schenk and Hildebrand (SH) and Murashige and Skoog (MS) media con taining various combinations of cytokinins (0.5 - 4 mg l - 1 BA and 0.05 - 0.5 mg l - 1 TDZ) and auxins (0.2 - 1 mg l - 1 IAA, 0.05 – 0.5 mg l - 1 NAA, 0.5 - 6 mg l - 1 Kin, 0 . 2 - 0.6mg l - 1 2,4 - D). The best shoot prolif e- ration ( 18±1.7 shoots/explant) response was observed in SH medium with a combination of 2.5 mg l - 1 BA and 0.5 mg l - 1 IAA. The resulting shoots were subsequently rooted on half strength SH medium containing 0.5 mg l - 1 IBA within four weeks with 90% success. Genetic transformation system has been established for Piper nigrum L. var. Panniyur - 1 plants by infecting seedling derived explants with Agrobacterium tumefaciens strain EHA105 carrying binary plasmid pCA M- BIA1301, which contains scorable marker, β - l,3 - glucuronidase and selectable marker hygromycin phosphotransf e rase gene ( hpt ) under the control of CaMV 35S promoter. Infection was most effective when explants were infected with Agrobacterium for 15 min and co - cultivated for 2 days in the co - cultiva tion medium. Shoots were regenerated directly from the explants in the presence of hygromycin (20 mg m l - 1 ) and analysed. PCR and GUS histochemical analysis co n- firmed the transformation. This is the first report showing transformation is possible in this sp ice and this method will allow genetic improvement of this crop by the introduction of agronomically important genes.

Item Type: Article
Uncontrolled Keywords: Spice Black pepper; piperaceae; regeneration; seedlings; Agrobacterium tumefaciens .
Subjects: Plant Disease Biology & Biotechnology
Depositing User: Rgcb Library
Date Deposited: 23 Feb 2017 04:59
Last Modified: 04 Jul 2019 08:11
URI: http://rgcb.sciencecentral.in/id/eprint/260

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