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Journal : ANNALES BOGORIENSES

Agrobacterium-mediated transformation of banana Musa acuminata AA cv "“Mas Lampung” with hpt gene using sterile corm as target tissue Witjaksono, Witjaksono; Nena, Ade; Estiati, Amy
ANNALES BOGORIENSES Vol 13, No 1 (2009): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (285.979 KB) | DOI: 10.1234/27

Abstract

The protocol for  Agrobacterium-mediated transformation of local banana plants cv  “Mas Lampung” (AA) has been established. A selectable marker gene (hpt) has been used to study the transformation using  in vitro corm slices as target tissues.  Banana  in vitro corm slices were co-cultivated with the EHA105 strain of Agrobacterium tumefaciens harbouring binary vector pCAMBIA 1301 containing hygromycin resistance gene (hpt) as a selectable marker and intron-containing β-Glucuronidase (gus-intron) gene as a reporter gene driven by  CaMV 35S promoter.   Polymerase Chain Reaction (PCR) were used to examine the existence of  hpt gene in plants resulted from the transformation.  Using primer pairs specific for hpt gene, our PCR analysis on leaves showed the presence of the  hpt   transgene in banana transgenic plants at first generation (T0) of transformation.  To prove the existence of hpt gene in the fruits of transgenic banana plants, PCR analysis were also carried out.  The data showed that the  hpt  gene could be amplified  from banana fruits of tested samples.   These result demonstrates that the Agrobacterium-mediated transformation method used in this experiment has been successful to transfer gene into banana plants. Thus, the transformation method reported here could be used as a standard protocol to  transfer another useful genes into local banana plants cv. “Mas Lampung”.  Furthermore, the presence of transgene in fruits of banana transgenic plants is important achievement  especially for transgene that is expected to be expressed in the fruit including to introduce vaccine genes into banana fruits for edible vaccine.Key words:  Agrobacterium,  hpt  gene, transgenic banana Mas Lampung, Musa acuminate corm slices, PCR
Tissue Culture Propagation of “Hausa potato” (Solenostemon rotundifolius (Poir) JK Morton) Leksonowati, Aryani; Witjaksono, Witjaksono
ANNALES BOGORIENSES Vol 16, No 1 (2012): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.1234/38

Abstract

We have developed an effective plant propagation method of “hausa potato‟ through shoot proliferation  in vitro, acclimatization in the greenhouse and transplantation of the plantlet on the individual media container for planting material production. The shoot cultures were initiated from shoots that grew from tuber collected from the field, disinfested using standard method and transferred to MS medium with 1 mg/l  of  BA for shoot proliferation. The proliferating shoots were then subcultured to MS medium with no growth regulators to induce rooting and enlarge the shoot. The plantlets were acclimatized in a plastic box containing medium of a mixture of cocopeat, sands, rice husk charcoal and top soil covered with plastic film to maintain humidity. The plantlets survived at 85% and almost 100% grew into planting materials ready for transplanting in the soil.  The planting materials survived in the field and produced tuber normally. Keywords: Solanostemon rotundifolius, hausa potato, kentang hitam, in vitro propagation, acclimatization, planting materials.