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Pembentukan Buah dan Perkecambahan Tanaman Jarak Pagar pada Transformasi Langsung Melalui Jalur Tabung Polen Zainudin, Agus; Purwoko, Bambang Sapta; Santoso, Tri Joko; Ardie, Sintho Wahyuning; Trikoesoemaningtyas, dan
Jurnal Agronomi Indonesia (Indonesian Journal of Agronomy) Vol. 46 No. 1 (2018): Jurnal Agronomi Indonesia
Publisher : Indonesia Society of Agronomy (PERAGI) and Department of Agronomy and Horticulture, Faculty of Agriculture, IPB University, Bogor, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (432.449 KB) | DOI: 10.24831/jai.v46i1.12897

Abstract

The genetic transformation via pollen-tube pathway in jatropha is the first alternative method that was applied in this plant. The objective of the research was to study fruit set and germination of three genotypes of jatropha following direct transformations via pollen-tube pathways. The research was conducted during April 2014 until January 2015 at jatropa’s experimental field, Pasuruan, and at green house of ICABIOGRAD, Bogor. Three genotypes of jatropha i.e., IP3A, IP3P and JcUMM18 were used. In the first experiment, split plot design was used where 3 levels of DNA plasmid concentration as a main plot and 5 levels of stigma-drip time of DNA plasmid as a subplot. In the second experiment randomized block design was used with single factor consisting of 15 combinations of concentration and stigma-drip time of DNA plasmid as treatments and control. The results demonstrated that interaction between concentration and application time of DNA plasmid did not significantly affect fruit and seeds formation of three Jatropha genotypes. Combination of DNA plasmid concentration with time of stigma-drip had significant effect on seed germination rate of IP3A genotype, but not significant on the other variables. The concentration of 0.05-0.5 µg µL-1 and application time of DNA plasmid at 1-10 hours after pollination could be applied on jatropha genetic transformation via pollen-tube pathways.Keywords: DNA plasmid, Jatropha curcas, pCAMBIA1301, pollen-tube, stigma-drip
Efficiency of Genetic Transformation via Pollen-Tube Pathway of Jatropha (Jatropha curcas L.) Based on Histochemical and Molecular Analysis Zainudin, Agus; Purwoko, Bambang Sapta; Santoso, Tri Joko; Ardie, Sintho Wahyuning; Trikoesoemaningtyas, and
Jurnal Agronomi Indonesia (Indonesian Journal of Agronomy) Vol. 45 No. 3 (2017): Jurnal Agronomi Indonesia
Publisher : Indonesia Society of Agronomy (PERAGI) and Department of Agronomy and Horticulture, Faculty of Agriculture, IPB University, Bogor, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (630.424 KB) | DOI: 10.24831/jai.v45i3.12925

Abstract

The genetic transformation via pollen-tube pathway is an alternative method to overcome the constraints imposed by genotype specificity in transformation and regeneration in jatropha (Jatropha curcas L.) tissue culture. Therefore, it is necessary to establish important parameters for efficient genetic transformation of jatropha via pollen-tube pathway. The objective of the research was to study the efficiency of direct transformation of jatropha via pollen-tube pathway based on histochemical and molecular analysis. Solution of purified pCAMBIA1301 DNA plasmid carrying a hptII marker gene and a gus reporter gene with concentration level of 0.05, 0.25, 0.50 µg µl-1 were applied to stigma of flowers at 1, 2, 4, 7, 10 h after pollination. Seedling of IP3A, IP3P and JcUMM18 jatropha’s genotypes derived from 15 combination treatments of plasmid DNA concentration and application time, also wild type was subjected to histochemical and molecular analyses. Based on those analyses, the efficiency of transformation via pollen-tube pathway of three jatropha genotypes ranged from 1.5-16.7%. PCR analysis showed that a number of positive plants were identified by using specific primers hptII and gus, i.e. 1-3 and 3-7 plants of the 15 combined treatments, respectively. It indicated that the transformation efficiency via the pollen-tube pathway varied in each jatropha genotype.Keywords: Jatropha curcas L., pCAMBIA1301, plasmid DNA, stigma-drip