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All Journal Jurnal Agronomi Indonesia (Indonesian Journal of Agronomy) Indonesian Journal of Agricultural Science Indonesian Journal of Biotechnology Jurnal Penelitian Pertanian Tanaman Pangan
Suharsono, Suharsono
Departemen Biologi, Fakultas Matematika Dan Ilmu Pengetahuan Alam, Institut Pertanian Bogor (Bogor Agricultural University), Kampus IPB Darmaga, Bogor 16680, Indonesia
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Economics, Econometrics & Finance Immunology & microbiology Materials Science & Nanotechnology

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Analisis Keragaman Genetik Galur Kedelai Transgenik Toleran Cekaman Aluminium dan Varietas Non-Trasngenik Berdasarkan Marka SSR Pardal, Saptowo J.; Rahayu, V. R.; Nugroho, K.; Suharsono, Suharsono
Jurnal Penelitian Pertanian Tanaman Pangan Vol 4, No 3 (2020): Desember 2020
Publisher : Pusat Penelitian dan Pengembangan Tanaman Pangan

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jpptp.v4n3.2020.p171-177

Abstract

High genetic diversities are important factors in the development of new crop varieties. In vitro technique and Genetic Engineering are applicable for development of crop variability that is not found in the gene pool. Genetic variation may be derived from genetic variations in cells or in chromosomes. Variations in the cells may be obtained from cell mutations or polysomic mutations of a certain cells during the in vitro culture (plant regeneration in vitro). Genetic variations in chromosome may be caused by gene insertion, changes of chromosom structures (crossings), as well as changes of genes and cytoplasms. Changes of genetic characters may be improved by inserting novel gene into the cells target. To improve the plant tolerances to abiotic factors, tolerance gene constructs can be inserted to the target cells. For example, by inserting the Aluminum tolerance gene construct such as MaMt2 gene its can induce genetic diversity in transgenic soybean lines resulted from transformation. Research results showed that genetic diversity in transgenic soybean lines was found based on microsatellite marker (SSR= Simple Sequence Repeat). The genetic diversity produced by using genetic manipulation can provide chances to develop new plant genotipes that contain Al tolerance character.
Obtaining of transgenic potato (Solanum tuberosum L.) cultivar IPB CP3 containing LYZ‐C gene resistant to bacterial wilt disease Pasmawati Pasmawati; Aris Tjahjoleksono; Suharsono Suharsono
Indonesian Journal of Biotechnology Vol 26, No 1 (2021)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/ijbiotech.61682

Abstract

Bacterial wilt caused by Ralstonia solanacearum is one of the most important bacterial diseases in potato production. This study aimed to obtain the transgenic potato (Solanum tuberosum L.) cultivar IPB CP3, containing LYZ‐C gene encoding for lysozyme type C, resistant to bacterial disease caused by R. solanacearum. Genetic transformation using Agrobacterium tumefaciens LBA4404 to 124 internode explants resulted in the transformation efficiency of about 47.58% with a regeneration efficiency of approximately 30.51%. Gene integration analysis showed that 16 clones were confirmed as transgenic clones containing the LYZ‐C gene. Analysis of resistance to R. solanacearum of three transgenic clones showed that all three transgenic clones were more resistant than a non‐transgenic one. This result showed that the LYZ‐C gene integrated in the genome of transgenic potato increased the resistance of potato plants to R. solanacearum. We obtained two transgenic clones considered resistant to bacterial wilt disease.
CONSTRUCTION, EXPRESSION AND PURIFICATION OF RECOMBINANT PRE-MATURE PEPTIDE OF PLANTARICIN F FROM Lactobacillus plantarum S34 IN Escherichia coli Kusdianawati Kusdianawati; Apon Zaenal Mustopa; Suharsono Suharsono; Bugi Ratno Budiarto; Fatimah Fatimah; Hasim Danuri
Indonesian Journal of Agricultural Science Vol 16, No 1 (2015): April 2015
Publisher : Indonesian Agency for Agricultural Research and Development

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/ijas.v16n1.2015.p31-38

Abstract

Plantaricin is one of bacteriocins that have the potential to be used as food preservative. Plantaricin is safe for human consumption because it can be easily degraded by proteolytic enzymes. The objective of this study was to express and purify recombinant pre-mature peptide of plantaricin F from Lactobacillus plantarum S34 in Escherichia coli. Plantaricin gene-specific primer was used to obtain pln F structural gene amplicon from L. plantarum S34. This amplicon was cloned in pET32a vector and expressed in E. coli BL21 (DE3) pLysS. Pre-mature plantaricin F peptide was expressed as Histagged-fusion protein and separated by Co2+-chelating affinity chromatography. L. plantarum S34-derived pre-mature plantaricin F peptide fused with thioredoxin-(His)6tag had successfully been expressed in E. coli BL21 (DE3) pLysS using pET32a as an expression vector. The fused recombinant pln F as pre-mature state expressed had a molecular mass of +24 kDa, meanwhile the fused recombinant that contained only the leader peptide of pln F appeared as +20 kDa based on SDS-PAGE separations. The optimal production of fused recombinant pln F as soluble fraction was obtained when culture condition was added with 0.5 mM of IPTG and incubated at 22°C for 5 hours (OD~1). Furthermore, the expression of fused recombinant pln F as its pre-mature peptide pointed out that the pln F’s leader peptide could be proteolytically cleaved by a system in heterologous cells. Overall, heterologous pln F production as pre-mature peptide fused with thioredoxin-(His)6tag had been well established. From this research, we expect plantaricin F can be expressed and purified in E. coli.
Introgresi Gen CsNitr1-L dari Transgenik Nipponbare ke Ciherang dan Analisis Pewarisannya pada Generasi BC3F4 ,, Nazarudin; ,, Suharsono; ,, Sustiprijatno
Jurnal Agronomi Indonesia (Indonesian Journal of Agronomy) Vol. 42 No. 3 (2014): 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 (693.095 KB) | DOI: 10.24831/jai.v42i3.9160

Abstract

ABSTRACTCsNitr1-L gene is a gene encoding nitrites transporter and is included in the group of proton oligopeptide transporter (POT) gene family. The absorption of nitrites by plants expressing this transporter becomes efficient. The gene encoding this protein (CsNitr1-L) under the control of 35S CaMV Promoter had been introduced into rice plants (Oryza sativa L.) subspecies japonica cv. Nipponbare to transfer this gene. The japonica transgenic rice had been crossed with Ciherang variety followed by back-cross and self polination until BC3F4 generation. The aim of this study was to analyse introgression of CsNitr1-L gene in the transgenic rice BC3F4 generation. The transgenic rice plants in BC3F4 generation were selected based on the resistance to hygromicin. More than 90% population of BC3F4 are putative introgression rice lines carriying the transgene. The introgression of the transgene were indirectly confirmed by PCR analaysis using primer corresponding to hpt gene. The yield of introgression line was higher than these of original Ciherang cultivar. Four introgression lines (G3, G7, G8 and G11) that had higher yield were analysed by PCR. Result of the  analysis showed that these four transgenic plants carried the introgression.Keywords: proton oligopeptide transporter, nitrites transporter, efficient nitrogen, transgenic rice
Hubungan Metilasi DNA dengan Ekspresi Gen MADS-box pada Buah Mantel Tanaman Kelapa Sawit (Elaeis guineensis Jacq.) Anischan, Maharani; ,, Suharsono; Mathius, Nurita Toruan-; Kusnandar, Andree Sunanjaya
Jurnal Agronomi Indonesia (Indonesian Journal of Agronomy) Vol. 42 No. 3 (2014): 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 (488.105 KB) | DOI: 10.24831/jai.v42i3.9175

Abstract

ABSTRACT The presence of mantled fruit on large-scale clonal production of oil palm had trully decreased the oil productivity. Mantled phenotype is likely to be that of an epigenetic change involving DNA methylation and the MADS-box transcription factor gene which encoded floral organ homeotic transformation. The objectives of this research were to quantify the degree of methylation which determined fruit abnormality through Ultra Performance Liquid Chromatography (UPLC) and to study its correlation with the MADS-box EgAGL6, EgAG2, and EgAGA genes expressed on mantled fruit derived from oil palm clonal plants which have been quantified using Quantitative Real-Time PCR (qPCR). This research was arranged in two replications for each gene and cDNA. The expression of the target genes were compared to EF1-α1 as the reference gene. Through the Least Significant Difference (LSD) Test at 95% confidence level of qPCR result, EgAGL6 expression was significantly lower in mantled fruit which decreased from 1.88 fold in Abn m to 0.46 fold in Abn. EgAG2 expression was increased non-significantly from 0.91 fold in Abn m to 1.13 fold in Abn, while EgAGA expression was higher in mantled which increased significantly from 1.48 fold in Abn m to 1.71 fold  in Abn. Nuclease S1 digestion and UPLC revealed the genome-wide increase in DNA methylation on mantled fruit (18.33-19.55%) compared to its normal counterparts (5.67%). This increased in global DNA methylation was showed by the significant decreased in EgAGL6 transcript level of mantled fruit. This gene assumed to be involved in the development of mantled fruit. Keywords: DNA-methylation, MADS-box genes, Mantled fruit, Quantitative Real-Time PCR
Co-Authors Andree Sunanjaya Kusnandar Apon Zaenal Mustopa Aris Tjahjoleksono Bugi Ratno Budiarto Fatimah Fatimah Hasim Danuri Kusdianawati Maharani Anischan Nazarudin , Nugroho, K. Nurita Toruan- Mathius Pardal, Saptowo J. Pasmawati Pasmawati Rahayu, V. R. Sustiprijatno ,