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KARAKTERISTIK ISOLAT Rhizoctonia sp. PATOGENIK DAN Rhizoctonia MIKORIZA PADA TANAMAN ANGGREK TANAH (Spathoglottis plicata) Soelistijono, Soelistijono; Sumardiyono, Christanti; Priyatmojo, Achmadi; Semiarti, Endang
Agrineḉa Vol 12, No 1 (2012): jurnal AGRINEÇA
Publisher : Agrineḉa

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Abstract

Mycorrhizal Rhizoctonia is a fungus that capable to associate with terrestrial orchids. Apart from being mycorrhizal, there are isolates of Rhizoctonia sp. that are pathogenic and caused root rot disease on Spathoglottis plicata. This study aimed to know the differences between pathogenic Rhizoctonia sp. and mycorrhizal Rhizoctonia in morphology and molecular structure using RAPD technique. The results showed that colony colour,  cell lenght and nucleus number a several isolates of pathogenic Rhizoctonia sp. and of mycorrhizal Rhizoctonia on S. plicata had no differences, but had differences on cell thickness and isolate grouping based on hyphal anastomosis test. RAPD molecular technique showed that each isolate of pathogenic Rhizoctonia sp. and mycorrhizal Rhizoctonia had differences on DNA structure.
KONSERVASI ANGGREK ALAM INDONESIA Vanda tricolor Lindl. varietas suavis MELALUI KULTUR EMBRIO SECARA IN-VITRO Dwiyani, Rindang; Purwantoro, Azis; Indrianto, Ari; Semiarti, Endang
Bumi Lestari Journal of Environment Vol 12, No 1
Publisher : Udayana University

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Abstract

Vanda tricolor Lindl. var. suavis is an Indonesian wild orchid which is now extremely rare in nature due to its habitat destruction. Development of an appropriate method for propagation of this species through in vitro culture could be nessecary for conservation purposes. High phenolic content of plant tissue is a serious problem for V. tricolor research in the laboratory, which inhibits germination and growth of the embryo. To overcome this problem, seeds were sown in a medium with the addition of tomato extract as an antioxidant. The aim of this research is to find the most suitable concentration of tomato extract for germination and growth of the embryo of V. tricolor form Bali and Merapi in order to obtained healthy seedlings for conservation purposes. Orchid pods (7 months after polination) of V. tricolor Bali and Merapi were used as plant material. The treatment consisted of 5 concentrations of tomato extract which is 0, 50 100, 150, 200, 250 g L-1. Observation was done by counting the number of protocorms for each stage of growth at 4 weeks after seed sowing. The study concluded that V. tricolor Bali is more responsive to the tomato extract compared with Merapi. Concentration of 150 gL-1 tomato extract gave the highest percentage of protocorms for Bali form, whereas Merapi form did not give significant differences either with or without tomato extract added in the culture medium.
PHENOTYPIC AND MOLECULAR CHARACTERIZATION OF MULTISHOOTS DEVELOPMENT IN TRANSGENIC Phalaenopsis amabilis (L.) BLUME HARBORING 35S::KNAT1 (KNOTTED-LIKE Arabidopsis thaliana 1) Saputro, Triono Bagus; Semiarti, Endang; Purwantoro, Aziz
BIOTROPIA - The Southeast Asian Journal of Tropical Biology Vol 25, No 1 (2018)
Publisher : SEAMEO BIOTROP

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11598/btb.2018.25.1.615

Abstract

Phalaenopsis amabilis (L.) Blume is one of Indonesian natural orchid which has an aesthetic flower and possesses high economic value. The low multiplication rate and long periods of life cycle are the main obstacles to conventionally propagate this orchid. The aims of this research were to analyze the stability of transgenic plant P. amabilis harboring 35S::KNAT1 based on morpho-genomic characterization. KNAT1 gene is reported as a gene that involved in the shoot formation, and it  had been successfully introduced into Phalaenopsis amabilis (L.) Blume genome. After seven times regeneration, the confirmation of the transgene existence in the genom is needed to ensure whether the plant could consistently maintain the transgene in its genome and to characterize the shoot development. The experiment was carried out in 3 steps:  1) Co-integration analysis of 35S::KNAT1 into P. amabilis genom; 2) Phenotypic analysis on the multiplication rate, morphological variation and venation pattern; and 3) Protein profile analysis of transgenic plants. The results showed that the survival rate of putative transgenic was 58.7% on NP0 medium and 62.5% on NP SIM medium. PCR analysis confirmed that 82.5% transgenic growth on NP0 and 93.33% on NP SIM contained DNA fragment of KNAT1 gene, NPTII gene and trnL-F intergenic spacer, indicating that those plants are positive transgenic. The 35S::KNAT1 transgenes and phytohormone were independently involved in multishoots formation of P. amabilis transgenic plants. The phenotypic of plantlets were classified into six main criteria, i.e. normal shape, lobed leaves, rosette, elongated stem, cup shoot and widened leaves. The normal type was the most abundant type of variation (± 29%) in both medium. Protein profile showed that all transgenic plants produced 45,8 kDa protein and that was equivalent with molecular weight of KNAT1 protein. Taken together, all those data indicated that 35S::KNAT1 transgene were consistently integrated into the transgenic plant genome.
KARAKTERISTIK ISOLAT Rhizoctonia sp. PATOGENIK DAN Rhizoctonia MIKORIZA PADA TANAMAN ANGGREK TANAH (Spathoglottis plicata) Soelistijono, S.; Sumardiyono, Christanti; Priyatmojo, Achmadi; Semiarti, Endang
AGRINECA Vol 12, No 1 (2012): JURNAL AGRINECA
Publisher : AGRINECA

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Abstract

Mycorrhizal Rhizoctonia is a fungus that capable to associate with terrestrial orchids. Apart from being mycorrhizal, there are isolates of Rhizoctonia sp. that are pathogenic and caused root rot disease on Spathoglottis plicata. This study aimed to know the differences between pathogenic Rhizoctonia sp. and mycorrhizal Rhizoctonia in morphology and molecular structure using RAPD technique. The results showed that colony colour,  cell lenght and nucleus number a several isolates of pathogenic Rhizoctonia sp. and of mycorrhizal Rhizoctonia on S. plicata had no differences, but had differences on cell thickness and isolate grouping based on hyphal anastomosis test. RAPD molecular technique showed that each isolate of pathogenic Rhizoctonia sp. and mycorrhizal Rhizoctonia had differences on DNA structure.
PRODUKSI GALUR MURNI MELALUI INDUKSI EMBRIOGENIK MIKROSPORA CABAI MERAH BESAR DENGAN STRES Indrianto, Ari; Semiarti, Endang; Surifah, ,
Zuriat Vol 15, No 2 (2004)
Publisher : Zuriat

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Abstract

Induksi embriogenik mikrospora cabai merah besar dapat dilakukan dengan stres panas dan pelaparan (starvasi karbohidrat). Stres diperlukan untuk mengubah perkembangan gametofitik (mikrospora membelah asimetrik) kearah sporofitik (mikrospora membelah simetrik) untuk membentuk embrio. Mikrospora pada stadium uninukleat akhir dikulturkan secara aseptik didalam medium starvasi karbohidrat (medium B) masing-masing pada 25oC; 33oC dan 35oC selama 2; 4 dan 6 hari. Setelah praperlakuan stres, mikrospora disubkultur kedalam medium embriogenesis (medium A2) tanpa zat pengatur tumbuh dan diinkubasi pada suhu 25oC. Pengamatan sitologis terhadap tipe pembelahan inti dan mikrospora multiseluler (proembrio) dilakukan dengan pengecatan DAPI. Hasil penelitian menunjukkan, praperlakuan starvasi selama 4 hari dapat menginduksi 3.4% dari populasi mikrospora menjadi embriogenik, dimana 4% diantaranya tumbuh menjadi proembrio. Tambahan stres panas 33oC selama 4 hari meningkatkan persentase induksi embriogenik mikrospora menjadi 29%, dimana 30% diantaranya berkembang menjadi proembrio. Induksi embriogenik mikrospora cabai merah besar merupakan langkah awal untuk memproduksi tanaman haploid, penggandaan kromosom selanjutnya akan menghasilkan tanaman dobel haploid yang merupakan galur murni.
Selection of Phalaenopsis amabilis L. Blume Orchid Resistance to Hygromycin Mercuriani, Ixora Sartika; Purwantoro, Aziz; Moeljopawiro, Sukarti; Jang, Seonghoe; Semiarti, Endang
Indonesian Journal of Biotechnology Vol 17, No 2 (2012)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/ijbiotech.16000

Abstract

Examination of Phalaenopsis amabilis orchid resistance to hygromycin antibiotic is an important step to doprior to Agrobacterium-mediated genetic transformation in this orchids using Hygromycin phosphotransferase(HPT) gene as a selection marker in the T-DNA that harboring a desired gene to be transfered. We exposedthe plant on hygromycin containing medium. The experiment was conducted using 6 weeks old P. amabilisprotocorms. These protocorms were subcultured onto NP medium supplemented with various concentrationof Hygromycin (0, 5, 10, 20, 1nd 40 mg/l). The number of survival protocorms were examined every week for4 weeks after subcultured (WAS). The resistancy of hygromycin was calculated as ratio of death protocormsper total protocorms). The result showed that 10 mg/l hygromycin with 1 weeks of application caused deathclose to LD 50. This data indicate that P. amabilis resistance to hygromycin treatment on the appropriateconcentration 10 mg/l, and this concentration can be used for other purposes in orchid system.
GENOTYPIC AND PHENOTYPIC CHARACTERIZATION OF Alcaligenes javaensis JG3 POTENTIAL AS AN EFFECTIVE BIODEGRADER Ethica, Stalis Norma; Oedjijono, Oedjijono; Semiarti, Endang; Widada, Jaka; Raharjo, Tri Joko
BIOTROPIA - The Southeast Asian Journal of Tropical Biology Vol 25, No 1 (2018)
Publisher : SEAMEO BIOTROP

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11598/btb.2018.25.1.583

Abstract

Utilization of glycerol by lipase producing bacteria offers great benefits for fat and oil waste degradation and waterwaste treatment. Nevertheless, there have been lack of reports about the availability of non-pathogenic, lipase producing bacteria, which could naturally degrade glycerol produced from the lipolysis process by lipase. This study reported a newly identified species of rhizobacteria, Alcaligenes javaensis JG3, which is not only able to produce high level of lipase, but also able to degrade glycerol molecules. Identification of strain JG3 was carried out using SEM (Scanning Electron Microscope), BD Phoenix 100 Automated Microbiology System and 16S rRNA gene analysis to determine its taxonomy status. The ability of the strain to metabolize glycerol was investigated both genotypically and phenotypically using degenerate PCR and a glycerol minimal medium. Identification test results showed that strain JG3 belongs to genus Alcaligenes, with the closest relationship with A. faecalis and A. aquatilis (96% nucleotide similarity maximum). Degenerate PCR resulted in a 248-bp sequence showing 93% similarity with glpK of Candidatus Sodalis pierantonius SOPE, a key gene involved in glycerol metabolism. In vitro glycerol utilization test result showed that Alcaligenes sp. JG3 was able to grow on glycerol aerobically, but not anaerobically. It is concluded that Alcaligenes sp. JG3 possesses genes coding for glycerol metabolism and this trait is phenotypically expressed, thus making the strain potential to be used as an effective fat and oil biodegrader.
HOMOLOGI FUNGSI GEN KNAT1 ( Knotted 1– like Arabidopsis thaliana) PADA ANGGREK BULAN Phalaenopsis amabilis (L.) Bl. DENGAN MEDIATOR Agrobacterium tumefaciens Isminingsih, Sulastri; Semiarti, Endang; Purwantoro, Aziz
Jurnal Agroekoteknologi Vol 1, No 1 (2009)
Publisher : Jurusan Agroekoteknologi Fakultas Pertanian Untirta

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Abstract

ABSTRACTTo understand the function of KNAT1 gene Arabidopsis in the forming and developing of Indonesian origin orchid shoot Phalaenopsis amabilis through transform p35S::KNAT1 and pGreen to protocorm like bodies (plb) of the orchid mediated by Agrobacterium tumafaciens LBA4404. The plb transformants were grown on New Phalaenopsis selection medium containing 5 mg/l BAP, 0.15 mg/l NAA, 15 mg.l Kanamycin and addition of 300 mg/l Cefotaxim to eliminate the overgrowth of Agrobacterium. The analysis of positive transformant use Polimerase Chain Reaction (PCR) with the specific oligonucleotide primers for KNAT1 gene: KNAT1F1R1 and universal primers for pGreen: 35SO and Tnos. The result shows that 3 shoots of 1850 transformants positively carry out the 35S::KNAT1 construct (frequency of transformation was 0.16 %) while 5 shoot of 1850 transformants also positively carry out the pGreen vector, with the frequency of transformation was 0.27 %. The phenotype analysis of 35S::KNAT1 transformants show multiplication on forming of the leaf from a plb to + 10 shoots and forming of the leaf shape which has terompet like shapes and rectangular shape.Key words: Arabidopsis, KNAT1 gene, Phalaenopsis amabilis, shoot
Induction of Somatic Embryogenesis through Overexpression of ATRKD4 Genes in Phalaenopsis “Sogo Vivien” Mursyanti, Exsyupransia; Purwantoro, Aziz; Moeljopawiro, Sukarti; Semiarti, Endang
Indonesian Journal of Biotechnology Vol 20, No 1 (2015)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/ijbiotech.15276

Abstract

Phalaenopsis “Sogo Vivien “is a mini orchid hybrid with beautiful flowers and numerous inflorescences.Mass propagation of this orchid is needed to meet the market demand. Objective of this research was toinduce somatic embryogenesis of P.”Sogo Vivien” through insertion of AtRKD4 gene into orchid. T-DNAcontaining 35S::GAL4::AtRKD4::GR was inserted into 16-22 days after sowing orchid protocorms mediated byAgrobacterium tumefaciens EHA 105. Activation of the AtRKD4 gene was induced by glucocorticoid inductionsystem, using 15μM Dexamethasone (Dex). The results showed that 34 out of 2,648 orchid embryos developedinto protocorms on hygromycin selection medium, whereas only 4 out of 2,897 non-transformant protocormsdeveloped from embryos. A 500 bp of HPT genes was amplified from transformant candidates using specificprimers for HPT (HygF1 and HygR1) and 380 bp was amplified using specific primers for AtRKD4 (AtRKD4F1 and AtRKD4 R1), indicated that transgenes have been integrated into orchid genomes. Finally, 17 plantletswere positively carrying AtRKD4 and HPT genes, the efficiency of transformation was 0.63 %. Somatic embryoswere also emerged from leaf explants of transformant on hormone-free NP medium and became normalplantlets. It is probably due to the high activity of AtRKD4 genes in orchids
Early detection of the orchid flowering gene PaFT1 in tobacco cells using a GFP reporter Wahyuningsih, Sri; Lawrie, Muhammad Dylan; Daryono, Budi Setiadi; Moeljopawiro, Sukarti; Jang, Soenghoe; Semiarti, Endang
Indonesian Journal of Biotechnology Vol 21, No 1 (2016)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/ijbiotech.26781

Abstract

Here we describe a novel method of using green fluorescence protein (GFP) as a reporter gene for early detection of an integrated T­DNA containing the orchid flowering gene, PaFT1 (Phalaenopsis aphrodite Flowering locus T1) in the tobacco genome. Functional assays that report the presence of exogenous DNA early in development are especially useful in plants where the desired phenotype is only apparent after long periods of vegetative growth. The objective of this study is to establish a method for detecting an inserted Phalaenopsis orchid flowering gene and examining its function in tobacco. The p35S::PaFT1­ 35S::GFP construct was introduced into Agrobacterium tumefaciens strain EHA101. Transformed tobacco leaves were cultured on MS medium with addition of 1 mgL-1 NAA+3 mgL-1 BAP+50 mgL-1 Kanamycin+300 mgL-1 timentin for selection. Results showed bright green GFP fluorescent signals in 11 out of 15 (73%) tobacco leaf cells at a 2­month time point after transformation. GFP and PaFT1 fragments were amplified in genomic PCR using GFP and PaFT1 specific primers. The accumulated PaFT1 transcripts were observed in 3 month­old transgenic tobacco plants containing p35S::PaFT1­35S::GFP. Green florescence was observed only in the transgenic plants at the 5 month­old stage but not in the wild type controls.