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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.
Isolasi dan Pengklonan Gen Penyandi H+-ATPase Membran Plasma dari Melastoma malabathricum L. Muzuni .; Didy Sopandie; Utut Widyastuti Suharsono; Suharsono .
Jurnal Agronomi Indonesia (Indonesian Journal of Agronomy) Vol. 42 No. 1 (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 (737.981 KB) | DOI: 10.24831/jai.v42i1.8159

Abstract

ABSTRACT Melastoma malabathricum L. is an Al-accumulating plant that grows well in acidic soils with high level of soluble aluminum in the tropics. One of the important proteins in the detoxifying Al stress is a plasma membrane H+-ATPase, a most abundant protein on the plasma membrane, encoded by PMA gene. The objective of this research was to isolate and characterize the gene encoding plasma membrane H+-ATPase from M. malabathricum L. Full length cDNA of MmPMA had been successfully isolated through a gradual isolation of the gene. The 5’ end and middle part of the MmPMA gene had been successfully isolated by PCR by using total cDNA as template and pma primers designed from some plants, while the 3’ end of Mmpma had been isolated by 3’ RACE. The parts of the gene had been successfully joined by PCR. The joining product was successfully inserted into pGEM-T Easy and the recombinant plasmid was successfully introduced into E. coli DH5α. Nucleotide sequence analysis showed that the length of MmPMA coding sequence was 2,871 bp encoding 956 amino acids with molecular weight of 105.29 kDa and a predicted pI value of 6.84. Local alignment analysis based on nucleotide of mRNA showed that MmPMA is 82% identical to pma Vitis vinifera; 81% to pma Juglans regia, pma Populus trichocarpa, pma Sesbania rostrata, and pma Prunus persica and 80% to pma Lycopersicon esculentum. Based on deduced amino acid sequence, MmPMA is 94% identical to PMA Vitis vinifera and PMA Juglans regia; 93% to PMA Populus trichocarpa; 92% to PMA Vicia faba, Lycopersicon esculentum, and Arabidopsis thaliana, AHA4. MmPMA has 10 transmembrane domains, 4 cytoplasm loops, 6 functional domains and 3 autoregulatory domains.Keywords: aluminum, cDNA, MmPMA, PCR, RACE
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.
Co-Authors Apon Zaenal Mustopa Aris Tjahjoleksono Bugi Ratno Budiarto Didy Sopandie Fatimah Fatimah Hasim Danuri Kusdianawati Muzuni, Muzuni Nugroho, K. Pardal, Saptowo J. Pasmawati Pasmawati Rahayu, V. R. Utut Widyastuti Suharsono