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All Journal HAYATI Journal of Biosciences Jurnal Hortikultura Indonesia (JHI) Journal of Tropical Life Science : International Journal of Theoretical, Experimental, and Applied Life Sciences Bumi Lestari Zuriat Jurnal Ilmiah Aplikasi Isotop Radiasi BIOTROPIA - The Southeast Asian Journal of Tropical Biology AL KAUNIYAH Biota: Jurnal Ilmiah Ilmu-Ilmu Hayati Pelita Perkebunan AGRIVITA, Journal of Agricultural Science Jurnal Agroekoteknologi Indonesian Journal of Biotechnology Biology, Medicine, & Natural Product Chemistry Bioeksperimen: Jurnal Penelitian Biologi Journal of Tropical Biodiversity and Biotechnology ANNALES BOGORIENSES Jurnal Pertanian Agros Biosaintifika: Journal of Biology & Biology Education Scripta Biologica Jurnal Riset Biologi dan Aplikasinya JURNAL ILMIAH AGRINECA Agrosaintifika: Jurnal Ilmu-Ilmu Pertanian Annales Bogorienses
Endang Semiarti
Fakultas Biologi, Universitas Gadjah Mada, Indonesia
Aerospace Engineering Agriculture, Biological Sciences & Forestry Humanities Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Decision Sciences, Operations Research & Management Earth & Planetary Sciences Education Energy Environmental Science Immunology & microbiology Materials Science & Nanotechnology Medicine & Pharmacology Public Health Veterinary

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Morphology and molecular characterization of Vanda tricolor × Vanda limbata orchid hybrid based on VOH1 gene characters Alydarafa, Hafshah; Melati, Chrisnanda Ayu; Semiarti, Endang
Indonesian Journal of Biotechnology Vol 29, No 4 (2024)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/ijbiotech.91456

Abstract

Vanda is a monopodial epiphyte orchid that spreads throughout Asia and Southeast Asia reaching 70 species. Indonesia itself has its own endemic Vanda orchid such as Vanda tricolor and Vanda limbata. A hybrid of V. tricolor and V. limbata is predicted to form a new specific character in the flower and leaf. The purpose of this study was to determine the morphological and molecular differences between V. tricolor, V. limbata, and Vanda hybrids resulting from crosses between that two orchids, by analysing the morphological characteristics of the roots, leaves, flowers and the structure of the Vanda Orchid Homeobox1 (VOH1) shoot‐forming gene isolated from V. tricolor, V. limbata, and their hybrids. The morphological analysis was conducted using RHS colour chart, size measurement of plants, and the transversal preparation of the leaf. Molecular analysis was performed by PCR using Dendrobium Orchid Homeobox 1 (DOH1) primers, followed by sequencing and bioinformatic analysis. Morphologically, the flower’s colour of the hybrid is most similar to V. limbata but the flower’s patterns are more similar to V. tricolor meanwhile the leaf colour of the hybrid is brighter than the both parents. The slides illustrate the sclerenchyma tissue is made up of strongly thickened walls containing lignin indicates the presence of homeobox DOH1 gene homolog, namely VOH1. The molecular result displayed by the phylogenetic tree of the VOH1 indicates that the hybrid has more similarities with V. limbata.
Establishment of an efficient primary callus induction for embryogenic potential of Coffea arabica Arimarsetiowati, Rina; Daryono, Budi Setiadi; Astuti, Yohana Theresia Maria; Semiarti, Endang
Pelita Perkebunan (a Coffee and Cocoa Research Journal) Vol. 38 No. 2 (2022)
Publisher : Indonesian Coffee and Cocoa Research Institute

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iccri.jur.pelitaperkebunan.v38i2.510

Abstract

Coffea arabica L. is a species of coffee that contribute for more than seventy percent of world coffee production. Various attempts have been made to obtain large quantities of planting material through propagation in vitro somatic embryogenesis technology. The objective of this experiment was to evaluate the effect of different plant growth regulators (PGRs) on callus induction (indirect somatic embryogenesis) in AS2K clone of Arabica coffee. Mother plants of Arabica coffee were established in coffee experimental field of Indonesian Coffee and Cocoa Research Institute at Andung Sari, Bondowoso, East Java, Indonesia (-7Ëš55'' ' S, 113Ëš41'' ' E) at an altitude of 1380,1 m dpl. Leaf explants were cultured on a half-strength Murashige and Skoog (MS) medium supplemented with various concentration (1.0, 2.0, 3.0 mg/L) of 2,4-D and (1.0, 2.0, 3.0 mg/L) thidiazuron in combination with 1.0 mg/L BAP. All the experiments were organized in completely random design (CDR) and repeated three times, each using minimum seven replicates (a total of 21 explants per treatment). The morphologycal and histological analysis of the different types of callus were observed. The percentage of callus formation was recorded every two weeks until eight weeks. The highest percentage of callus formation (almost 60%) was in medium containing 1 mg/L 2,4-D dan 1 mg/L BAP. Morphological and histological studies prove that the callus has a friable and embryogenic texture and begins to develop various stages of somatic embryo formation, starting with the globular, heart, torpedo and cotyledonary phases.
Phenotype and genotype characterization of Phalaenopsis amabilis (L.) Blume Orchid Transformant Harboring Construct UBI::Cas9::U3::PDS3 Andani Kesuma, Amarilis; Nopitasari, Sri; Yoshioka, Yasushi; Matsumoto, Shogo; Semiarti, Endang
Jurnal Hortikultura Indonesia (JHI) Vol. 11 No. 3 (2020): Jurnal Hortikultura Indonesia
Publisher : Indonesian Society for Horticulture / Department of Agronomy and Horticulture

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.29244/jhi.11.3.212-220

Abstract

Phalaenopsis amabilis (L.) Blume adalah tanaman hias “Puspa Pesona Indonesia” yang dapat ditingkatkan kualitasnya dengan teknik rekayasa genetika. Transformasi genetik dengan perantara Agrobacterium tumefaciens dan CRISPR/Cas9 digunakan dalam penelitian ini untuk pengeditan genom secara lebih spesifik dan presisi pada target sekuen gen PHYTOENE DESATURASE3 (PDS3) yaitu gen yang berperan penting pada biosintesis kloroplas. Dalam penelitian ini digunakan tanaman transforman umur 12 bulan yang ditumbuhkan dari protokorm yang telah diintegrasi dengan T-DNA pembawa konstruksi UBI::Cas9::U3::PDS3/plasmid pRGEB32. Pembuktian tanaman transforman tersebut masih mengandung konstruksi T-DNA tersebut perlu dilakukan, yaitu dengan karakterisasi secara genotipe dan fenotipe. Tujuan penelitian ini adalah untuk mengkarakterisasi P. amabilis transforman pembawa T-DNA dengan konstruksi UBI::Cas9::U3::PDS3 secara genotip dan fenotip dibandingkan dengan P. amabilis non-transforman. Karakterisasi genotipe dilakukan dengan mendeteksi integrasi T-DNA pembawa konstruksi UBI::Cas9::U3::PDS3 pada genom anggrek P. amabilis menggunakan beberapa primer yaitu HPT, Cas9, PDS3 dan trnL-F (primer kontrol internal). Analisis karakter fenotipe dilakukan dengan pengamatan morfologi dan analisis kadar klorofil menggunakan metode spektrofotometri. Hasil penelitian menunjukkan bahwa genom anggrek P. amabilis transforman pembawa konstruksi UBI::Cas9::U3::PDS3 umur 12 bulan dapat teramplifikasi oleh semua primer. Analisis fenotipe P. amabilis transforman menunjukkan adanya perubahan warna tanaman dari hijau menjadi albino dengan kadar klorofil lebih rendah jika dibandingkan dengan P. amabilis non-transforman. Hal ini menunjukkan bahwa teknologi CRISPR/Cas9 dapat digunakan untuk mengedit genom tanaman anggrek. Kata kunci: Anggrek, CRISPR/Cas9, klorofil, Phalaenopsis amabilis (L.) Blume, PHYTOENE DESATURASE 3 (PDS3), Transforman
PENGARUH CAHAYA DAN TEMPERATUR TERHADAP PERTUMBUHAN TUNAS DAN PROFIL PROTEIN TANAMAN ANGGREK Phalaenopsis amabilis TRANSGENIK PEMBAWA GEN Ubipro::PaFT Putra, Rinaldi Rizal; Mercuriani, Ixora Sartika; Semiarti, Endang
Bioeksperimen: Jurnal Penelitian Biologi Vol 2, No 2: September 2016
Publisher : Universitas Muhammadiyah Surakarta

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.23917/bioeksperimen.v2i2.2483

Abstract

Penelitian ini bertujuan untuk mencari kondisi yang tepat dalam percepatan pembungaan tanaman Phalaenopsis amabilis transgenik yang telah disisipi gen penentu waktu pembungaan Ubipro::PaFT. Metode penelitian ini menggunakan tanaman transgenik pembawa gen Ubipro::PaFT umur 18 bulan setelah penanaman. Tanaman ditumbuhkan pada inkubator dengan pencahayaan menggunakan lampu LED putih dan kombinasi LED putih biru, dengan fotoperiodisitas 8 jam terang 16 jam gelap, suhu 25ºC pada fase terang dan 20ºC pada fase gelap selama 20 minggu. Setelah 20 minggu pertumbuhan tanaman, dilakukan analisis profil protein dengan metode SDS-PAGE untuk mengetahui protein yang diproduksi pada setiap fase pertumbuhan yang diamati.Hasil penelitian menunjukkan kombinasi cahaya LED putih dan biru meningkatkan pembentukan daun sebesar 60%, panjang daun 70,58%, tetapi belum diperoleh kemunculan infloresen. Analisis profil protein menunjukkan terbentuknya protein dengan ukuran 108,57; 71,30; 56,16; 40,85; 26,79; 13,27; dan 13,12 kilodalton pada tanaman transgenik, tetapi tidak terdeteksi protein dengan ukuran 19,65 kDa yang sesuai dengan berat molekul protein PaFT, sementara protein dengan ukuran sekitar 56,16 kDa sesuai dengan berat molekul protein POH1(Phalaenopsis Orchid Homeobox1). Hal ini menunjukkan bahwa gen vegetatif POH1 mampu menghambat aktivasi gen PaFT pada tanaman P. amabilis transgenik umur 20 minggu, sehingga tanaman masih dalam fase juvenil dan belum mampu diinduksi untuk berbunga.
Improvement of Genetic Transformation Efficiency in Vanda tricolor Orchid Using Acetosyringone Dwiyani, Rindang; Purwantoro, Azis; Indrianto, Ari; Semiarti, Endang
Annales Bogorienses Vol. 14 No. 2 (2010): Annales Bogorienses
Publisher : BRIN

Show Abstract | Download Original | Original Source | Check in Google Scholar

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 improving Vanda orchid through genetic modification could be valuable for horticulture and, indirectly, also for conservation. In this research, a method of Agrobacterium-mediated transformation of two V. tricolor obtained from Salak Mount, West Java and Merapi Mount, Yogyakarta in Indonesia protocorms was improved using acetosyringone (AS). Concentrations of 0 and 25 ppm AS were used in transformation of pG35S binary vector containing kanamycin resistance geneinto V. tricolor protocorms. The result showed that 25 ppm AS was required on inoculation with Agrobacterium solution, without AS on cocultivation. Five weeks after treatment on the 300 ppm kanamicyn containing medium, green protocorms were obtained, that was 11.01% for V. tricolor from Salak Mount with pre-culture treatment prior to inoculation, 9.39% for V. tricolor from Merapi Mount with pre-culture treatment prior to inoculation, and 1.37% for V. tricolor from Merapi Mount without pre-culture treatment prior to inoculation. The best condition to set high efficiency of transformation is pre-culture protocorms prior inoculation, soaking protocorm on 25 ppm AS for 30 minutes, then co-cultivate its on AS-free callus induction medium.
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 Vol. 25 No. 1 (2018): BIOTROPIA Vol. 25 No. 1 April 2018
Publisher : SEAMEO BIOTROP

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (104.448 KB) | 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.
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 Vol. 25 No. 1 (2018): BIOTROPIA Vol. 25 No. 1 April 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 orchids which has an aesthetic flower and possesses high economic value. The low multiplication rate and long period 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 is involved in 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 genome 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 genome; 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 fragments 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 multishoot formation of P. amabilis transgenic plants. The phenotypes 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 media. 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 was consistently integrated into the transgenic plant genome.
Co-Authors , Surifah A.A. Ketut Agung Cahyawan W Achmadi Priyatmojo Aditya Nur Subchan Agus Slamet Ahmad Suyoko Alisa Julia Nurulita Alydarafa, Hafshah Aminatun Munawarti Aminatun Munawarti, Aminatun Amru Rizal Basri Anami Riastri Andani Kesuma, Amarilis Annisa, RR Rifka Ari Indrianto Ari Indrianto Ari Indrianto Ari Indrianto Ari Indrianto Ari Indrianto Ari Indrianto Aries Bagus Sasongko Arimarsetiowati, Rina Arum, Dyah Ayu Puspita Asri Fajar Milasari Azis Purwantoro Azis Purwantoro Aziz Purwantoro Bekti Sulistya Utami Binti Tsulsiyah Brilliant Kharisma Apritadila BUDI SETIADI DARYONO Cahya Lembayung Sutra Catharina Tri Widyastuti Christanti Sumardiyono Christanti Sumardiyono Della Rosiana Ningtias Devi Bunga Pagalla Dewi Tika Sari Dewi Yuliana Rizqi Dika Sundari Eka Fitriana Candra Ningrum El Hakim, Alim Erwin Prastowo Evilili Usmanti Exsyupransia Mursyanti Exsyupransia Mursyanti Exsyupransia Mursyanti Faiza Senja Widya Perdana Falah Nur Alifianto Febri Yuda Kurniawan Fhea Putri Cristy Fitriana Puspitasari Frisca Damayanti Gde Cahyadi Wirajagat Gildantia, Elke Hanifa, Yumna Rahmadias Ikhsanudin Nur Rosyidi Ixora Sartika Mercuriani JAKA WIDADA Jose Gutierrez-Marcos Jose Gutierrez-Marcos Kana Ninomiya Kinasih, Anggiresti Kurniawan, Febri Yuda Linggabuwana, Aviesta Maryani Maryani Matsumoto, Shogo Maura Indria Meidianing Meidianing, Maura Indria Melati, Chrisnanda Ayu Muhammad Dylan Lawrie Muhammad Dylan Lawrie Muhammad Dylan Lawrie Muhammad Dylan Lawrie Naufal Ghozi Aditya Perdana Ni Luh Putu Kayika Febryanti Nintya Setiari Nintya Setiari Nopitasari, Sri Oedjijono Oedjijono, Oedjijono Oktaviana Herawati Pauline Destinugrainy Kasi Prasojo, Ireneus Seno Pratiwi, Apriliana Putra, Rinaldi Rizal Putri, Saifa Usni R Soelistijono Rahayu Sulistianingsih Ramadhani, Aulia Noor Rasjid, Nuzlan Rinaldi Rizal Putra, Rinaldi Rizal Rindang Dwiyani Rizka Riliant Puspasari Rumiyati Rumiyati Rumiyati, Rumiyati Seonghoe Jang Setiaji, Arkan Shogo Matsumoto Sismindari . Sismindari Sismindari Sismindari Sismindari Sitarina Widyarani Soenghoe Jang Sri Nopitasari Sri Wahyuningsih Stalis Norma Ethica Sukarti Moeljopawiro Sukarti Moeljopawiro Sukarti Moeljopawiro Sukarti Moeljopawiro Sukarti Moeljopawiro Sukarti Moeljopawiro Sukarti Moeljopawiro Sulastri Isminingsih Taryono, Taryono Thoyibatul Farida Tri Joko Raharjo Triono Bagus Saputro, Triono Bagus Umi Kulsum Nur Qomariah Windi Mose Windi Mose Woerjono Mangoendidjojo Woro Anindito Sri Tunjung Yasushi Yoshioka Yohana Theresia Maria Astuti, Yohana Theresia Maria Yoshioka, Yasushi Yuli Setiawati Yuuki Asano Zulfa Layina