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INDONESIA
Jurnal AgroBiogen
Published by Kementerian Pertanian
ISSN : 19071094     EISSN : 25491547     DOI : -
Core Subject : Agriculture,
Agriculture, Biological Sciences & Forestry
Jurnal AgroBiogen memuat artikel primer dan sekunder hasil penelitian bioteknologi dan sumberdaya genetik tanaman, serangga, dan mikroba pertanian. Jurnal ini diterbitkan tiga kali setahun pada bulan April, Agustus dan Oktober oleh Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumberdaya Genetik Pertanian
Arjuna Subject : -
Articles 252 Documents
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Molecular and Phenotypic Analyses of Inpari HDB/K15 F2 Lines Containing sd1 Mutant Gene Resulted from Genome Editing Method Clara S. A. Fitriyanti; Suharsono Suharsono Suharsono; Tri Joko Santoso
Jurnal AgroBiogen Vol 16, No 1 (2020): June
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jbio.v16n1.2020.p35-44

Abstract

Inpari HDB variety is resistant to bacterial leaf blight (BLB) disease, but due to its tall stature, the variety is susceptible to lodging. Inpari HDB with semidwarf stature, therefore, is of high interest for lodging resistant performance. sd1 gene encoding GA20ox-2 enzyme is one of the genes responsible for imparting semidwarf stature of rice. In previous study, sd1 mutant rice cv. Kitaake (K15) was developed by using CRISPR/Cas9 technology. The objective of this study was to analyze molecularly and phenotypically F2 lines containing sd1 mutant gene resulted from a cross between Inpari HDB and K15 to develop semidwarfInpari HDB rice variety. Thirty F2 Inpari HDB/K15 lines were analyzed at molecular level using DNA sequencing method together with phenotypic assessment of the lines to verify the integration of sd1 mutant gene. DNA sequencing analysis showed that 9 out of the 30 F2 Inpari HDB/K15 lines were sd1 mutants. The remaining F2 lines contained 17 heterozygotes and 4 nonmutants. All the nine mutant lines demonstrated shorter plant stature and showed more tiller number per plant compared to the nonmutant lines. The sd1 mutant gene in the F2 lines showed pleiotropic effects on panicle number andshowed no effects on other traits, such as flowering time, panicle length, filled and unfilled grain percentages. This study showed the introduction of sd1 mutant gene generated semidwarf Inpari HDB lines. The semidwarf Inpari HDB lines obtained from this research should be further evaluated to confirm their lodging resistant performances.
Front Matter JA Vol 16 No 1 Agrobiogen, Jurnal
Jurnal AgroBiogen Vol 16, No 1 (2020): June
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jbio.v16n1.2020.p%p

Abstract

Back Matter JA Vol 16 No 1 Jurnal Agrobiogen
Jurnal AgroBiogen Vol 16, No 1 (2020): June
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jbio.v16n1.2020.p%p

Abstract

Genetic Diversity Analysis and Development of DNA Fingerprints of 20 Indonesian Local Chili Pepper Varieties Based on SSR Markers Rerenstradika Tizar Terryana; Nadia Della Savitri Ayu Ningrum; Kristianto Nugroho; Darmawan Saptadi; Helmi Kurniawan; Puji Lestari
Jurnal AgroBiogen Vol 16, No 2 (2020): December
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jbio.v16n2.2020.p45-58

Abstract

Chili pepper is one of the most valuable horticultural crops, widely cultivated in Indonesia. Analysis of its genetic diversity is needed to develop successful breeding programs of local varieties. Simple sequence repeat (SSR), a robust molecular marker used for genetic diversity analysis in plant species, offers potential, reliable DNA fingerprinting method to assess genetic variation and varietal identification of chili pepper. Fifteen SSR markers were used in this study to analyze the genetic diversity and develop profiling identification of DNA fingerprint of local chili pepper varieties. Twenty local and two improved varieties of three chili pepper species, consisting of 3, 1, and 18 varieties of Capsicum frutescens, C. chinense, and C. annuum, respectively, were assessed for their SSR polymorphism. A total of 87 alleles was obtained from the polymorphism analysis with high alleles variation (2–16 alleles) with average total allele of 5.8 and average polymorphism information content (PIC) of 0.59 (0.34–0.83). Clustering and Principle Coordinate Analyses (PCoA) classified the varieties into two groups with coefficient of similarity of 0.65 indicating their high genetic variability. Most local varieties belonged to the same cluster and separated from the two improved varieties. Based on PIC values and dendrogram with selected markers, five SSR markers, i.e. EPMS441, EPMS331, EPMS335, GPMS194, and CaSSRBio1.1, were identified as SSR marker set for DNA fingerprinting purposes. SSR marker set used in this study was successful in developing the varietal identity of local chili pepper varieties, as indicated by unique code of each variety.
Editing of PCNA Gene by CRISPR/Cas9 Technology to Improve the Red Chili Resistance to Yellow Leaf Curl Disease Kurniawati, Devi Ayu; Suharsono, nFN; Santoso, Tri Joko
Jurnal AgroBiogen Vol 16, No 2 (2020): December
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jbio.v16n2.2020.p79-88

Abstract

Yellow leaf curl disease caused by Pepper yellow leaf curl virus (PepYLCV), member of geminiviruses group, is responsible for heavy yield losses for chili pepper production. Resistant genes which can cause immunity to the disease have not been found in germplasm collection. The aim of the research was to edit proliferating cell nuclear antigen (PCNA) gene by using CRISPR/Cas9 technology for developing plant resistance against geminivirus in chili pepper. A CRISPR/Cas9 plasmid cassette construct harboring the guide RNA of PCNA gene was constructed by Golden Gate cloning strategy. The construct was then introduced into chili genome via in planta method using Agrobacterium tumefaciens EHA105. The transformed plants were bioassayed by virus inoculation and confirmed using PCR and DNA sequencing to identify a mutagenesis event in PCNA gene target. The results showed that CRISPR/Cas9 plasmid cassette harboring gRNA of PCNA gene was successfully constructed. In planta transformation using A. tumefaciens vector harboring CRISPR/Cas9-gRNA PCNA construct resulted in 307 and 193 transformed plants from chili var. Lingga and Ciko, respectively. Bioassay by using virus inoculation to the transformed plants obtained 6 and 14 lines of Lingga and Ciko, respectively, which were resistant to geminivirus (no symptom observed). The resistant lines of chili pepper var. Lingga and Ciko were mutated in PCNA gene with one base insertion or deletion mutation types. These results exhibit that the CRISPR/Cas9 genome editing can be used to induce mutant of PCNA gene in chili pepper. Further investigation is necessary to evaluate the selected chili lines resistant to PepYLCV infection.
Development of Transgenic Sugarcane Resistant to Stem Borer by Transforming cry1Ab-cry1Ac Fusion Gene through Agrobacterium tumefaciens Transformation Method Deden Sukmadjaja; Sri Koerniati; Auliya Lukman
Jurnal AgroBiogen Vol 17, No 1 (2021): JUNE
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jbio.v17n1.2021.p11-24

Abstract

Bulu Lawang (BL) is a sugarcane variety preferred by farmers in Indonesia due to its high yield, but this cultivar is susceptible to shoot and stem borer insect pests. Genetic engineering using cry1Ab and cry1Ac fusion gene is an effort to generate BL varieties resistant to the insect pests. This study aimed to 1) transform T-DNA containing cry1Ab-cry1Ac fusion gene into sugarcane genome by using Agrobacterium tumefaciens method, 2) obtain selection media composition of callus transformants, and 3) obtain transformation efficiency comparison of A. tumefaciens strains EHA105 and GV3101. The research was conducted at the Laboratory of Cell and Tissue Biology, the Laboratory of Molecular Biology, and the greenhouse of the Indonesian Center for Agricultural Biotechnology and Genetic Resources Research and Development, Bogor from March to August 2019. Research activities consisted of four parts namely 1) callus induction and Agrobacterium culture preparation containing pCambia-5300_OaRbcS-prom-cTP-cry1Ab-cry1Ac plasmid, 2) callus incubation in cocultivation and resting media, 3) selection and differentiation of shoots on regeneration media, and 4) molecular analysis using PCR method. Results showed that the composition of media, both for selection and regeneration processes of putative plant transformants, was the key to the success of this experiment. A. tumefaciens strain EHA105 resulted in higher transformation efficiency (11.1%) compared to that of strain GV3101 (9.0%). Molecular analysis showed that cryIAb-cryIAc fusion gene was successfully inserted into the sugarcane genome suggesting that the transgenic plant containing cry1Ab-cry1Ac fusion gene was obtained. The putative transgenic plants need further assay through bioassay tests to verify its resistance phenotype to the insect pests.
Improving Cassava Productivity by Soil Bioaugmentation with Phosphate-Solubilizing Actinomycetes and Fungi Alimuddin Ali; Muhammad Juanda; A. Farchan Sjahid; Herlina Rante
Jurnal AgroBiogen Vol 16, No 2 (2020): December
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jbio.v16n2.2020.p89-96

Abstract

Cassava is one of the most important food commodities, but its cultivation technique must be improved, especially the technology in increasing soil fertility. Lack of phosphate (P), one of the major biological nutrients in soil, can reduce agricultural production. Some P-solubilizing microbes can play an important role in increasing the availability of P for plants. The purpose of this study was to evaluate the ability of P-solubilizing microbes isolated from cassava rhizosphere in improving the growth of cassava after soil bioaugmentation with the formula of selected microbes. A total of 50 isolates were successfully isolated from cassava plant rhizosphere collected from several locations in South Sulawesi. In vitro screening on Pikovskaya agar media resulted in six Actinomycetes and six fungal isolates with the best P hydrolysis capacity. One Actinomycetes isolate (Streptomyces sp. A-SDR01) and one fungal isolate (Penicillium sp. F-SKG17) with nonantagonistic effect to each other based on in vitro test were able to improve the vegetative growth of cassava under in planta test. Combination of both isolates in a gum arabic formulation increased plant height and productivity compared to untreated plants when applied as soil bioaugmentation in limited field trials at four locations in South Sulawesi. Therefore, application of P-solubilizing microbes that possess soil bioaugmentation properties is recommended for increasing the growth of cassava plants and their use as microbial biofertilizers should be extended to wider areas.
Genetic Diversity Analysis of Indonesian Pigmented Rice Accessions Based on Functional Molecular Markers Andari Risliawati; Yusi N. Andarini; Rerenstradika T. Terryana; Kristianto Nugroho; Puji Lestari
Jurnal AgroBiogen Vol 17, No 1 (2021): JUNE
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jbio.v17n1.2021.p25-34

Abstract

Pigmented rice is functional staple food that becomes popular because of its anthocyanin content which is beneficial for health. Studies on the diversity of the local variety of Indonesian pigmented rice accessions have been carried out, but are still limited to one region of germplasm origin. This study aimed to analyze the genetic diversity of local varieties of pigmented rice collections of the IAARD-ICABIOGRAD Gene Bank. A total of 93 pigmented rice accessions from 16 provinces in Indonesia were analyzed using 15 functional molecular markers of SSR, STS, and indel. The total alleles detected were 115 with an average per locus of genetic diversity value of 0.71. There were five markers with PIC values >0.75, i.e. RM167, RM223, R8M33, R10M10, and GBSS1. The accessions were divided into two main groups based on their pericarp color. It is necessary to analyze the physicochemical content of the local rice accessions to complement the existing diversity information and identify potential pigmented rice accessions with high palatability.
Front Matter JA Vol 16 No 2 Jurnal Agrobiogen
Jurnal AgroBiogen Vol 16, No 2 (2020): December
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jbio.v16n2.2020.p%p

Abstract

Isolation and Homology Analysis of Alanine Aminotransferase Gene of Barley, Foxtail Millet, Cucumber, and Tomato Atmitri Sisharmini; Aniversari Apriana; Tri Joko Santoso; Bambang Sapta Purwoko; Nurul Khumaida; Kurniawan Rudi Trijatmiko
Jurnal AgroBiogen Vol 16, No 2 (2020): December
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jbio.v16n2.2020.p59-70

Abstract

Overexpression of alanine aminotransferase (AlaAT) gene can improve nitrogen use efficiency (NUE) in plants. The previous isolated AlaAT genes cannot be freely applied to generate NUE plants due to IPR restriction. Therefore, isolation of the gene from targeted monocot and dicot plants is necessary. The objectives of this study were to isolate AlaAT genes from barley, foxtail millet, cucumber, and tomato and analyze their homology to other isolated AlaAT genes in sequence databases (gene bank). Total RNA was isolated from roots of barley, foxtail millet, cucumber, and tomato, and then converted into cDNA using reverse transcription method. The cDNA was then cloned into pGEM®-T Easy plasmid and the verified clones were sequenced. The amino acid sequences were analyzed for their homologies using Clustal Omega software and phylogenetic tree was constructed. The results showed that the amino acids of AlaAT gene from barley was different from AlaAT genes of tomato and cucumber with homology level less than 80%. Phylogenetic tree predicted that AlaAT genes clustered into three groups with AlaAT genes of foxtail millet and barley clustered in one group together with other monocots in group I. AlaAT genes derived from dicots clustered into two groups in which AlaAT gene of tomato clustered in group II, while that of cucumber was in group III. The identity differences of AlaAT gene of tomato and that of cucumber as well as the estimates of the same enzymatic functions can open up enormous opportunities in genetic engineering research for the development of NUE rice.

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