Pendugaan Gen Bph1, Bph2, Bph3, Dan Bph4 Pada Galur-galur Padi Terpilih Tahan Hama Wereng Batang Cokelat (Nilaparvata Lugens[Stål])

Pests are major constraints to increasingrice production and brown planthoppers/BPH (Nilaparvatalugens [Stål]) is one of the major pests of rice plant.Resistance cultivar is one of the strategies for BPHmanagement. The objective of this research was to analyzethe Bph1, bph2, Bph3, and bph4 gene existence on theselected rice lines using the molecular markers. Thephenotype of the rice lines were tested based on theirresponse to BPH population collected from West Java andCental Java. Molecular markers linked to Bph1, bph2, Bph3,and bph4 were used to characterize the genotypic profilebased on PCR analysis. The results showed that there are sixgenotypes resistant to one of the BPH populations fromWest Java or Central Java. The six rice varieties weredetected to have not only allele of Bph3 gene, but also otherdifferent allele genes. B12344-2D-PN-42-1 and Inpari 13 weredetected to have the alleles of Bph3 dan Bph1 genes.B12512-18-SI-3-3-MR-3-PN-1, B12512-18-SI-3-3-MR-3-PN-1,BMIP46-4-1, and PTB33 were detected to have the alleles ofBph3 and bph2 genes. Meanwhile, B11007E-MR-3-2-PN-2-1-MR-1-2 was detected to have alleles of three genes: Bph3,bph2, and bph4. Nevertheless, this last line had mediumresistance to both BPH populations invested. There is apossibility that the interaction between two genes, Bph3 andbph4, occured which may affect the resistance responses ofrice varieties tested to BPH

Transformasi Gen Antisens ACC Oksidase pada Pepaya dengan Teknik Penembakan Partikel

Papaya (Carica papaya L.) is a climacteric fruit that exhibit avery fast ripening rate. Ethylene controls the ripening eventin the papaya fruit. 1-aminocyclopropane-1-carbocxylic acid(ACC) oxidase gene encodes a specific enzyme for ethylenebiosynthesis. The gene had become a target for manipulationto make a gene construct of an antisense ACC oxidaseto regenerate transgenic papaya that has a characteristic ofdelayed ripening. The objective of the experiment is to engineer transgenic papaya that has a delayed ripening characteristic by transforming papaya with the antisense ACC oxidase gene through particle bombardment technique. The immature embryos of papaya variety Burungwere used for the explants. Antisense ACC oxidase and reporter (gus) genes were co-transformed to papaya calli. Four hundreds eighteen calli were bombarded by the antisense ACC oxidase gene. The transformation experiment resulted 25 putatives transgenic plants out of fifty plantsacclimatized in a greenhouse. Gus gene expression assay observed at 9 days after bombardment showed that the papaya explants bombarded twice at 9 cm shoot distance had 53.3% transformation rate of gus positive and 5.25 blue spots number in average. The results of PCR analysis showed that four out of 25 transgenic putative papaya plants (TR6, TR9, TR20, dan TR24), indicated a positive PCR of the antisense ACC oxidase gene with the amplified fragment DNA size of 800 base pair

Pendugaan Gen Bph1, bph2, Bph3, dan bph4 pada Galur-galur Padi Terpilih Tahan Hama Wereng Batang Cokelat (Nilaparvata lugens[Stål])

Pests are major constraints to increasingrice production and brown planthoppers/BPH (Nilaparvatalugens [Stål]) is one of the major pests of rice plant.Resistance cultivar is one of the strategies for BPHmanagement. The objective of this research was to analyzethe Bph1, bph2, Bph3, and bph4 gene existence on theselected rice lines using the molecular markers. Thephenotype of the rice lines were tested based on theirresponse to BPH population collected from West Java andCental Java. Molecular markers linked to Bph1, bph2, Bph3,and bph4 were used to characterize the genotypic profilebased on PCR analysis. The results showed that there are sixgenotypes resistant to one of the BPH populations fromWest Java or Central Java. The six rice varieties weredetected to have not only allele of Bph3 gene, but also otherdifferent allele genes. B12344-2D-PN-42-1 and Inpari 13 weredetected to have the alleles of Bph3 dan Bph1 genes.B12512-18-SI-3-3-MR-3-PN-1, B12512-18-SI-3-3-MR-3-PN-1,BMIP46-4-1, and PTB33 were detected to have the alleles ofBph3 and bph2 genes. Meanwhile, B11007E-MR-3-2-PN-2-1-MR-1-2 was detected to have alleles of three genes: Bph3,bph2, and bph4. Nevertheless, this last line had mediumresistance to both BPH populations invested. There is apossibility that the interaction between two genes, Bph3 andbph4, occured which may affect the resistance responses ofrice varieties tested to BPH

Transformasi Gen Antisens ACC Oksidase Pada Pepaya Dengan Teknik Penembakan Partikel

Papaya (Carica papaya L.) is a climacteric fruit that exhibit avery fast ripening rate. Ethylene controls the ripening eventin the papaya fruit. 1-aminocyclopropane-1-carbocxylic acid(ACC) oxidase gene encodes a specific enzyme for ethylenebiosynthesis. The gene had become a target for manipulationto make a gene construct of an antisense ACC oxidaseto regenerate transgenic papaya that has a characteristic ofdelayed ripening. The objective of the experiment is to engineer transgenic papaya that has a delayed ripening characteristic by transforming papaya with the antisense ACC oxidase gene through particle bombardment technique. The immature embryos of papaya variety Burungwere used for the explants. Antisense ACC oxidase and reporter (gus) genes were co-transformed to papaya calli. Four hundreds eighteen calli were bombarded by the antisense ACC oxidase gene. The transformation experiment resulted 25 putatives transgenic plants out of fifty plantsacclimatized in a greenhouse. Gus gene expression assay observed at 9 days after bombardment showed that the papaya explants bombarded twice at 9 cm shoot distance had 53.3% transformation rate of gus positive and 5.25 blue spots number in average. The results of PCR analysis showed that four out of 25 transgenic putative papaya plants (TR6, TR9, TR20, dan TR24), indicated a positive PCR of the antisense ACC oxidase gene with the amplified fragment DNA size of 800 base pair

Regenerasi Pepaya Melalui Kultur in Vitro

A study was conducted in the Indonesian Center forAgricultural Biotechnology and Genetic Resources Researchand Development to optimize papaya regeneration systemsthrough in vitro culture. Four steps were done, i.e., callusinduction, callus regeneration, root formation, and acclimatization.Explant materials used were immature embryos ofpapaya cv. Burung. Immature papaya embryos were culturedon different media. The best medium for embryogeniccallus development was ½ MS + 10 mg/l 2.4-D + 60% sucrose+ 143 mg/l adenine sulphate + 50 mg/l myo inositol +400 mg/l glutamine, while that for callus embryo regenerationwas MS + 0.5 mg/l GA3 + 0.1 mg/l kinetin + Morel andWetmore Vitamin. Using this medium, the average of shootformation was three shoots per explant of embriogeniccallus, and the percentage of regenerated callus was 80%.The color of shoot derived from this treatment was green.Eighty percent of plants formed a complete root developmentusing ½ MS + 0.5 mg/l paclobutrazol media. Media hullof rice and compost was the best medium for papaya plantacclimatization. The percentage of survival on that acclimatizationstep was 65%

EVALUATION OF THE POTENTIAL WEEDINESS AND BIOCHEMICAL PROPERTIES OF TRANSGENIC RICE WITH GLU-1DX5 GENE

A Glu-1Dx5 allele, encoding for a high molecular weight glutenin sub unit Dx5, has been experimentally proven to be a major determinant for dough elasticity and functionality of bread wheat. Considering its important role, this gene has been successfully transferred to rice cv Fatmawati using particle bombardment. Some promising lines derived from consecutive generations have been obtained, however early biosafety assessment for transgenic plant is required in order to scientifically evaluate the equality of transgenic rice lines to their counterpart.Two promising lines (T3-19 and T3-20) which are supposed to be homozygous were subjected to weediness and biochemical evaluation. The evaluation of invasiveness as a measure of weediness was carried out in biosafety containment of BB Biogen, Bogor by growing a mixture of transgenic and non­transgenic. Biochemical testing was evaluated for nitrogen, carbohydrate, protein, fat, vitamin, ash, minerals (Ca and Mg), and 17 amino acids. Weediness testing revealed that competitiveness of transgenic rice plants grown in screen house as well as biosafety containment was equal with those of non transgenic plants, suggesting that transgenic rice plants have not high potency to be invasive. Transgenic rice seed had equal nutritive value with those of counterpart. These results indicate that transgenic rice plants (seeds) are substantially equivalent to non transgenic original lines except for the presence of Glu-1Dx5 gene.Keywords: Glutenin, Invasiveness, Nutrient composition, Transgenic rice

Regenerasi Pepaya melalui Kultur In Vitro

A study was conducted in the Indonesian Center forAgricultural Biotechnology and Genetic Resources Researchand Development to optimize papaya regeneration systemsthrough in vitro culture. Four steps were done, i.e., callusinduction, callus regeneration, root formation, and acclimatization.Explant materials used were immature embryos ofpapaya cv. Burung. Immature papaya embryos were culturedon different media. The best medium for embryogeniccallus development was ½ MS + 10 mg/l 2.4-D + 60% sucrose+ 143 mg/l adenine sulphate + 50 mg/l myo inositol +400 mg/l glutamine, while that for callus embryo regenerationwas MS + 0.5 mg/l GA3 + 0.1 mg/l kinetin + Morel andWetmore Vitamin. Using this medium, the average of shootformation was three shoots per explant of embriogeniccallus, and the percentage of regenerated callus was 80%.The color of shoot derived from this treatment was green.Eighty percent of plants formed a complete root developmentusing ½ MS + 0.5 mg/l paclobutrazol media. Media hullof rice and compost was the best medium for papaya plantacclimatization. The percentage of survival on that acclimatizationstep was 65%

Virulence of Brown Planthopper and Development of Core Collection of the Pest

Brown planthopper is the most important rice pest in Indonesia. Its high adaptability to feed and reproduce on previously introduced resistant varieties to form more virulent population often causes BPH outbreak and hopperburn that lead to total crop yield loss. Rice breeding for resistant to BPH requires information on the current status of BPH virulences in the fields to anticipate the virulence adaptation on new varieties. The objectives of this study were to investigate the degree of virulence of BPH populations and to cluster the BPH virulence to form BPH core collection. Thirteen BPH populations collected from paddy fields in six provinces (Banten, West Java, Central Java, East Java, South Kalimantan, and South Sulawesi) in 2011 and 2013 were tested on 10 differential rice varieties and seven host varieties of BPH populations, using the standard seedbox screening technique. Based on resistance reaction of four differential varieties (TN1, Mudgo, ASD7, and Rathu Heenathi), most BPH populations were identified as more virulent than biotype 4 (T1, Banten, PG, West Java; BY, East Java; B2 and B3, South Kalimantan; X1 and X3, South Sulawesi), four populations were biotype 4 (JWDL, Central Java; SD, East Java; X2 and X4, South Sulawesi), and one population each was biotype 3 (T2, Banten) and biotype 2 (S1, West Java). Populations X1 and B3 showed broad virulences to all varieties, whereas T2 was the least virulent. BPH field\u27s population had evolved into more virulence than biotype 4. Genotype resistance screening should use the BPH of this virulence population. Five BPH clusters which were further divided into 10 subclusters representing differential virulence toward 10 differential varieties were present in the tested BPH. Each virulence cluster was characterized by its ability to overcome four to eight single or double resistant genes. This BPH virulence core collection can be used in the characterization studies of candidate for resistant varieties or to form near-isogenic lines, or to study the insect and rice plant interaction

Mapping of Resistance Genes to Brown Planthopper in Untup Rajab, an Indonesian Local Rice Variety

Brown planthopper (BPH) is a major rice pest in Indonesia. The most economical and effective approach to control the insect pest is by using resistant varieties. Exploring for resistance genes is, therefore, a prerequisite for effective breeding program for BPH resistance. This study aimed to map BPH resistance genes in Untup Rajab, an Indonesian local rice variety. Genetic map was constructed using an F2 population from a cross between TN-1 and Untup Rajab, and SNP markers from RiceLD SNP Chip. Phenotyping was performed using bulk seedling test on F2:3 seedlings against two BPH populations, i.e. X1 and S1. Four QTLswere identified on chromosomes 5, 6, 8, and 11 with PVE values of 7.63%, 9.40%, 17.66%, and 3.05%, respectively. Relatively normal distribution of resistance phenotype and the relatively low PVE values indicate that Untup Rajab has a quantitative resistance to BPH with two different resistance loci identified for each BPH test population. The QTL on chromosome 8 overlaps with OsHI-LOX gene, which is associated with resistance to BPH, and adjacent to another QTL for resistance to green leafhopper. The QTL on chromosome 6 was found near OsPLDα4 and OsPLDα5 genes which are related to BPH resistance. Meanwhile, the QTL intervals on chromosome 5 and 11 did not overlap with any known BPH QTLs or genes, which make them attractive candidates for novel BPH resistance gene discovery