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PHENOTYPIC AND MOLECULAR CHARACTERIZATION OF MULTISHOOTS DEVELOPMENT IN TRANSGENIC Phalaenopsis amabilis (L.) Blume HARBORING 35S::KNAT1 (KNOTTED-LIKE Arabidopsis thaliana 1)

BIOTROPIA · Vol. 25 No. 1 (2018) · hlm. 11–21

DOI
10.11598/btb.2018.25.1.615
ISSN
1907-770X, 0215-6334
Terbit
2018-04-26
Penerbit
SEAMEO BIOTROP
Bahasa
en
Lisensi
creativecommons.org/licenses/by-nc-nd/4.0

Abstrak

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.

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  • Universitas Gadjah Mada
  • Institut Teknologi Sepuluh Nopember
  • Universitas Airlangga
  • Universitas Pattimura
  • University of Warwick
  • Diponegoro University
  • University of Malaya
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Rekan penulisOrganisasiArtikel bersama
Sukarti MoeljopawiroUniversitas Gadjah Mada11
Ari IndriantoUniversitas Gadjah Mada7
Muhammad Dylan LawrieUniversitas Gadjah Mada6
Agus Budi SetiawanUniversitas Gadjah Mada5
Windi MoseUniversitas Pattimura4
Nintya SetiariDiponegoro University4
Exsyupransia MursyantiUniversitas Gadjah Mada4
Hery PurnobasukiUniversitas Airlangga3
Dini ErmavitaliniInstitut Teknologi Sepuluh Nopember3
Febri Yuda KurniawanUniversitas Gadjah Mada3
Jose Gutierrez‐MarcosUniversity of Warwick3
Jaka WidadaUniversitas Gadjah Mada3
Aries Bagus SasongkoUniversitas Gadjah Mada3
Dyah Weny RespatieUniversitas Gadjah Mada2
Chee How TeoUniversity of Malaya2
Budi Setiadi DaryonoUniversitas Gadjah Mada2
Aviesta LinggabuwanaUniversitas Gadjah Mada2
Saifa Usni PutriUniversitas Gadjah Mada2
Nuzlan RasjidUniversitas Gadjah Mada2
Fitriana PuspitasariUniversitas Gadjah Mada2
Naufal Ghozi Aditya PerdanaUniversitas Gadjah Mada2
Dika SundariUniversitas Gadjah Mada2
Ni'matuzahrohAfiliasi tidak diketahui1
Ratna Syifa’a RahmahanaInstitut Teknologi Sepuluh Nopember1
Bilqis Naura Safira Rizam RizamBandung Institute of Technology1
Belva Aria MahendraInstitut Teknologi Sepuluh Nopember1
Anisatus SolihahInstitut Teknologi Sepuluh Nopember1
Dianita IndahsariInstitut Teknologi Sepuluh Nopember1
Aulia Sunan FadilahInstitut Teknologi Sepuluh Nopember1
Wirdhatul MuslihatinInstitut Teknologi Sepuluh Nopember1
Ratchata ChokwiwatkulPrince of Songkla University1
Asadi AsadiAfiliasi tidak diketahui1
Sahiral YakubAfiliasi tidak diketahui1
Ari WibowoIndonesian Coffee and Cocoa Research Institute1
Tri Joko RaharjoState University of Semarang1
Stalis Norma EthicaUniversitas Muhammadiyah Semarang1
Oedjijono OedjijonoJenderal Soedirman University1

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Sumber metadata: OAI-PMH (metadata jurnal) http://journal.biotrop.org/index.php/biotropia/oai, pertama di-harvest 8 Nov 2026, terakhir terlihat 8 Nov 2026.
Metadata asli (oai_dc) 
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	<dc:title xml:lang="en">PHENOTYPIC AND MOLECULAR CHARACTERIZATION OF MULTISHOOTS DEVELOPMENT IN TRANSGENIC Phalaenopsis amabilis (L.) Blume HARBORING 35S::KNAT1 (KNOTTED-LIKE Arabidopsis thaliana 1)</dc:title>
	<dc:creator xml:lang="en">Saputro, Triono Bagus</dc:creator>
	<dc:creator xml:lang="en">Semiarti, Endang</dc:creator>
	<dc:creator xml:lang="en">Purwantoro, Aziz</dc:creator>
	<dc:subject xml:lang="en">35S::KNAT1 (Knotted-Like Arabidopsis thaliana1)</dc:subject>
	<dc:subject xml:lang="en">multishoots</dc:subject>
	<dc:subject xml:lang="en">Phalaenopsis amabilis</dc:subject>
	<dc:subject xml:lang="en">transgene</dc:subject>
	<dc:description xml:lang="en">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.</dc:description>
	<dc:publisher xml:lang="en">SEAMEO BIOTROP</dc:publisher>
	<dc:date>2018-04-26</dc:date>
	<dc:type>info:eu-repo/semantics/article</dc:type>
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	<dc:identifier>https://journal.biotrop.org/index.php/biotropia/article/view/615</dc:identifier>
	<dc:identifier>10.11598/btb.2018.25.1.615</dc:identifier>
	<dc:source xml:lang="en">BIOTROPIA; Vol. 25 No. 1 (2018): BIOTROPIA Vol. 25 No. 1 April 2018; 11 - 21</dc:source>
	<dc:source>1907-770X</dc:source>
	<dc:source>0215-6334</dc:source>
	<dc:source>10.11598/btb.2018.25.1.</dc:source>
	<dc:language>en</dc:language>
	<dc:relation>https://journal.biotrop.org/index.php/biotropia/article/view/615/380</dc:relation>
	<dc:rights xml:lang="en">Copyright (c) 2018 BIOTROPIA - The Southeast Asian Journal of Tropical Biology</dc:rights>
	<dc:rights xml:lang="en">http://creativecommons.org/licenses/by-nc-nd/4.0</dc:rights>
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