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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

Author-ID : 375592

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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Induction of Somatic Embryogenesis through Overexpression of ATRKD4 Genes in Phalaenopsis “Sogo Vivien” Exsyupransia Mursyanti; Aziz Purwantoro; Sukarti Moeljopawiro; Endang Semiarti
Indonesian Journal of Biotechnology Vol 20, No 1 (2015)
Publisher : Universitas Gadjah Mada

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

Selection of Phalaenopsis amabilis L. Blume Orchid Resistance to Hygromycin Ixora Sartika Mercuriani; Aziz Purwantoro; Sukarti Moeljopawiro; Seonghoe Jang; Endang Semiarti
Indonesian Journal of Biotechnology Vol 17, No 2 (2012)
Publisher : Universitas Gadjah Mada

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.

Early detection of the orchid flowering gene PaFT1 in tobacco cells using a GFP reporter Sri Wahyuningsih; Muhammad Dylan Lawrie; Budi Setiadi Daryono; Sukarti Moeljopawiro; Soenghoe Jang; Endang Semiarti
Indonesian Journal of Biotechnology Vol 21, No 1 (2016)
Publisher : Universitas Gadjah Mada

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.

Application of CRISPR/Cas9 genome editing system for molecular breeding of orchids Endang Semiarti; Sri Nopitasari; Yuli Setiawati; Muhammad Dylan Lawrie; Aziz Purwantoro; Jaka Widada; Yasushi Yoshioka; Shogo Matsumoto; Kana Ninomiya; Yuuki Asano
Indonesian Journal of Biotechnology Vol 25, No 1 (2020)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/ijbiotech.39485

Orchid is an important ornamental plant in Indonesia due to their natural beauty of flowers. In the tropical forest, orchids are being acquired for trading and commercial market. Thus, the effort is required to proliferate orchid in large quantities for conservation and improve the floral variation for plant breeding. The purpose of this study is to develop a firmed methodology of molecular breeding of orchids using CRISPR/Cas9 KO system. The plant material used was Phalaenopsis amabilis protocorms growth on NP medium+pepton (2 g/L). Protocorm were submerged in the culture of Agrobacterium tumefaciens that Ti‐plasmid had been filled with a T‐DNA construct of a pRGEB32 vector harboring sgRNA with PDS3 sequence. Detection for transformants was confirmed by PCR using HPT primers (545 bp), Cas9 primers (402 bp), PDS primers (280 bp) and trnL‐F (1200 bp) as an internal control. The results showed that 0.96% PDS transformants were obtained from PDS3T2 lines. Several transformant showed pale leaf color compared to non‐transformant plants. This study suggests that the target gene has successfully edited by CRISPR/Cas9 system and could be applied for that functional gene editing in orchids.

Cytoprotective activity of carrot and tomato callus extracts and the ex‐ pression of cytokines in UV‐B irradiated fibroblast cells Rumiyati Rumiyati; Sismindari Sismindari; Endang Semiarti; Sitarina Widyarani; Dewi Tika Sari; Brilliant Kharisma Apritadila; Anami Riastri
Indonesian Journal of Biotechnology Vol 24, No 2 (2019)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/ijbiotech.51734

Studies have suggested that both carrot (Daucus carota L.) and tomato (Solanum lycopersicum L.) callus extracts contain antoxidant compounds that might have the potental to protect cells from free radicals such as H2O2 that contribute to cell damage. The other sources of free radical exposure in human cells, such as UV‐B, should also be examined. UV‐B exposure can trigger increased expression of inﬂammatory cytokines such as cyclooxygenase‐2 (COX‐2) and tumor necrosis factor‐α (TNF‐α) and the antinﬂammatory cytokine interleukin‐10 (IL‐10), which causes photoaging. This study was conducted to investigate the cytoprotectve actvity of carrot and tomato callus aqueous extracts by observing cell viability using the MTT assay. Immunocytochemistry methods were used to examine the eﬀects of carrot and tomato callus aqueous extracts on the expression of COX‐2, TNF‐α, and IL‐10 in human dermal fibroblast adult (HDFa) cells exposed to UV‐B light. Carrot and tomato callus aqueous extracts were obtained by the maceration method using aqua bidistilled solvent. Results showed that both carrot and tomato callus aqueous extracts at 0.5 mg/mL exhibited the highest cytoprotective eﬀect in HDFa cells compared to that at other concentratons. Both carrot and tomato callus aqueous extracts could also decrease the expression of COX‐2 and TNF‐α, whereas carrot callus aqueous extract increased the expression of the anti‐inﬂammatory cytokine IL‐10 in HDFa cells.

Stability of T-DNA Integration in Phalaenopsis “Sogo Vivien” Transgenic Orchid Carrying 35S::Gal4::AtRKD4::GR Endang Semiarti; Exsyupransia Mursyanti; Ahmad Suyoko; Faiza Senja Widya Perdana; Catharina Tri Widyastuti; Aditya Nur Subchan
Biology, Medicine, & Natural Product Chemistry Vol 7, No 1 (2018)
Publisher : Sunan Kalijaga State Islamic University & Society for Indonesian Biodiversity

Orchid is an elegant ornamental plant and favoured by the society. Phalaenopsis "Sogo vivien" is a mini-sized orchid with an interesting white-striped purple petals. This study was aimed to analyze the stability of the integration of embryonic gene carrier T-DNA from Arabidobsis AtRKD4 into the P. "Sogo vivien" genome produced in 2016. The study was conducted in 3 stages: 1) Transgenic plant phenotype analysis (1 year old); 2) Examination of T-DNA integration in orchid genotypes using PCR. 3) Analysis of transgenic plant leaf explants’ ability to produce somatic embryo in vitro. In vitro cultures were performed on the base medium of New Phalaenopsis (NP), plus various concentrations of TDZ (0, 1, 2 mg.L-1) and IBA (0, 1, 2 mg.L-1) or without TDZ and IBA as controls. The transgenic Phalaenopsis ‘Sogo vivien’ were transferred to pot mediums via ex vitro with two treatments: the first leaves were cut as explants for in vitro culture, and the plants were transferred to the mixture of fern medium with shavings of bark. The integration of T-DNA in the genome was detected by DNA genome amplification from the second leaves using the AtRKD4 gene primers and the POH1 gene. The results showed that the highest number of somatic embryo (SE) propagules or protocorm like bodies (PLBs) amounted to 27 were derived from transgenic plant # 2 cultured on NP + 2 mg.L-1 TDZ +1 mg.L-1 IBA medium. The presence of AtRKD4 transgenes were detected with the amplification of 380 bp of the RKD4 gene from the genome of transgenic plant # 2 by using PCR. There were 2 out of 15 plants that positively carry the AtRKD4 gene and produce SE. Thus, the stability of the AtRKD4 carrier T-DNA integration in the genomes of transgenic plants was 13.3%.

KONSERVASI ANGGREK ALAM INDONESIA Vanda tricolor Lindl. varietas suavis MELALUI KULTUR EMBRIO SECARA IN-VITRO Rindang Dwiyani; Azis Purwantoro; Ari Indrianto; Endang Semiarti
Bumi Lestari Journal of Environment Vol 12 No 1 (2012)
Publisher : Environmental Research Center (PPLH) of Udayana University

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

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.

Karakterisasi Isolat Rhizoctonia sp. Patogenik dan Rhizoctonia Mikoriza Pada Tanaman Anggrek Tanah Spathoglottis plicata Soelistijono Soelistijono; Achmadi Priyatmojo; Endang Semiarti; Christanti Sumardiyono
Biota : Jurnal Ilmiah Ilmu-Ilmu Hayati Vol 16, No 2 (2011): June 2011
Publisher : Universitas Atma Jaya Yogyakarta

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24002/biota.v16i2.121

Rhizoctonia mikoriza merupakan jamur yang mampu berasosiasi dengan anggrek tanah. Selain sebagai mikoriza, terdapat isolat Rhizoctonia sp. patogen dan penyebab penyakit busuk akar pada Spathoglottis plicata. Penelitian ini bertujuan mengetahui perbedaan antara Rhizoctonia sp. patogen dan Rhizoctonia mikoriza secara morfologi dan molekular menggunakan teknik RAPD. Hasil penelitian secara morfologi menunjukkan bahwa warna koloni, panjang dan jumlah inti sel isolat Rhizoctonia sp. patogen dan Rhizoctonia mikoriza pada S. plicata tidak berbeda, tetapi berbeda pada ketebalan sel dan pengelompokan isolat berdasarkan uji anastomosis hifa. Teknik molekuler RAPD menunjukkan bahwa setiap isolat Rhizoctonia sp. patogen dan Rhizoctonia mikoriza memiliki perbedaan pada struktur DNA.

PENGARUH CAHAYA DAN TEMPERATUR TERHADAP PERTUMBUHAN TUNAS DAN PROFIL PROTEIN TANAMAN ANGGREK Phalaenopsis amabilis TRANSGENIK PEMBAWA GEN Ubipro::PaFT Rinaldi Rizal Putra; Ixora Sartika Mercuriani; Endang Semiarti
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

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.

Micropropagation of Mini Orchid Hybrid Phalaenopsis “Sogo Vivien” Exsyupransia Mursyanti; Aziz Purwantoro; Sukarti Moeljopawiro; Endang Semiarti
Journal of Tropical Biodiversity and Biotechnology Vol 1, No 1 (2016): June
Publisher : Universitas Gadjah Mada

Phalaenopsis “Sogo Vivien” is an orchid hybrid with mini size plant body, and exhibits numerous beautiful pink flowers, that is ideal as ornamental pot plant. Some plants of this orchid exhibit variegated leaves that improve the beauty of the plant, not only because of the flower but also as attracted leaves. This orchid has high economical value, but mass propagation of this orchid has not established yet. An effective method to propagate both the normal and variegated plants is worth to be generated. The objective of this research was to produce a large number of P. “Sogo Vivien” plants, including the variegated plants. The method used seeds from self pollinating variegated plant, and flower stalk nodes. The seeds were sown on three various medium: VW, NP and MS, and flower stalk nodes were planted on VW + BA 10 mg l-1 + active carbon. The results showed that the best medium for in vitro culture of P. “Sogo Vivien” was NP medium, in which all seeds could grew into plantlets. Most plantlets emerged from the seeds were non variegated, only one plantlet out of 1344 seeds was variegated (0.007%). Although all emerged plantlets from flower stalk exhibited variegated leaves. Particularly, the plantlets arised from the second and third basal nodes of flower stalk showed the highest growth rate than that from the other nodes. Histological analysis showed that at 11-13 days after shoot segment plantation on NP medium, the shape of apical cells in the nodes was changed, then followed by the change of cell shape in the basal part of the nodes, produced bipolar pattern, then gradually developed into shoot. These results suggest that mass propagation could be achieved using seed culture, but to get the variegated phenotypes, the second and third nodes of flower stalk from variegated plant were the best explants to be used.

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

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