GENETIC TRANSFORMATION MEDIATED BY Agrobacterium AND TRANSGENIC SOYBEAN REGENERATION CONTAINING COAT PROTEIN GENE OF PEANUT STRIPE VIRUS

Gustian, Gustian GENETIC TRANSFORMATION MEDIATED BY Agrobacterium AND TRANSGENIC SOYBEAN REGENERATION CONTAINING COAT PROTEIN GENE OF PEANUT STRIPE VIRUS. In: Conference Programme Papers Abstracts, Andalas University and Bogor Agricultural University. (Unpublished)

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Abstract

The development of plant genetic enginering technique has contributed to a new prospect in overcoming low plant production. This technique has been proved to complement a non conventional one to introduce a superior gene from other organism that is isolated and contructed on a plasmid vector into plant tissue. Thus, this technique could be used to overcome the limited genetic resources in plant breeding programs. We proposed the study about development of genetic transformation and transgenic soybean regeneration. The objective of this research was to develop method which transform peanut stripe virus coat protein gene mediated agrobacterium tumefaciens into soybean. The reasech was conducted at plant molecular biology laboratory, and plant breeding study center laboratory, faculty of agriculture, bogor agricultural university. Four types of coat protein gene constructions of peanut stripe virus namely coat protein 1, coat protein 2, coat protein 3, and coat protein 4 were transformed into three Indonesian soybean varieties (Kerinci, Krakatau and Dempo) mediated by agrobacterium tumafaciens strain strain AGLI Carrying binary Ti-plasmid by using immmature cotyledonary explant is while two types of coat protein gene cntructions of peanut stripe virus namely coat protein 2 and coat protein 4 were transformed into MSC8606 line soybean using secondary somatic embryos explants. The observed variables for immature cotyledonary explant is (1) frequency of somatic embryo that appear, (2) frequency of somatic embryos sprouts which are kanamycin resistent, (3) frequency of plantlets which are kanamycin resistent. While the other experiment using secondary somatic embryos namely (1) transgene frequency, (2) clonal amount arising from the transgene, (3) frequency of mature somatic embryos that are kanamycin resistent, and (4) frequency of sprouts that are kanamycin resistent. The result of this reaserch received 5 plantlets which are kanamycin resistent. All the five plantlets consisted of two Kerinci varieties, one of which contains coat protein 1 and the other containing caot protein 4, still another one is Krakatau variety containing cp4, and the last two belong to Dempo variety, one of which contains cp2 and the other cp4. The transformation of the coat protein 2 and coat protein 4 by using secondary somatic embryo explants was more effective than immature cotyledonary. From 1965 explants which were transformed for coat protein 2 we received 113 transgene. From 1029 secondary somatic embryos that were culture on maturing media we received 665 mature somatic embryos. From 665 mature somatic embryos we received 291 sprouts that are Kanamycin resistent. Key words: genetic transformation, agrobacterium, transgenic, soybean, peanut stripe virus

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