Micropropagation of Mini Orchid Hybrid Phalaenopsis “Sogo Vivien”

Sidharta, Boy Rahardjo; Purwantoro, Aziz; Moeljopawiro, Sukarti; Semiarti, Endang

doi:10.22146/jtbb.12933

Cited by 6 publications

(6 citation statements)

References 26 publications

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“…Waki et al (2011) reported that in the Arabidopsis thaliana plant, the AtRKD4 gene produced the RKD4 protein triggers gene expression and pattern formation in early embryogenesis. This is also supported by research data on somatic embryogenesis induction on Phalaenopsis 'Sogo Vivien' from Mursyanti et al (2016) and research by Setiari et al (2018) in somatic embryogenesis of Dendrobium phalaenopsis. This is also in accordance with Mose et al (2020), who succeeded in inducing somatic embryogenesis by treating growth regulators thidiazuron and NAA from protocorms and P. amabilis leaf pieces as explants.…”

Section: Detection Of the Integration Of Atrkd4 Gene In The Genome Of P Amabilis Transformantsupporting

confidence: 59%

Stable Transformant of <i>Phalaenopsis amabilis</i> Somatic Embryo Carrying 35S::AtRKD4 Develops Into Normal Phenotype of Transgenic Plant

Perdana

Mose

Lawrie

et al. 2021

J.Tropical Biodiversity Biotechnology

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Phalaenopsis amabilis (L.) Blume orchid is an Indonesian national flower. The number of these orchids in their natural habitat is very limited, therefore plant propagation efforts are needed. One of the promising methods is plant propagation by inserting embryo gene AtRKD4 from a model plant Arabidopsis thaliana into the orchid genome to produce many somatic embryos. From previous research, we have obtained 28 plant P. amabilis transformants carrying the AtRKD4 gene, however, it was unknown whether these plants have normal phenotypes and growth similar to their parents. Therefore, descriptions on growth and morphology are needed. This research aimed to evaluate the phenotype of P. amabilis carrying 35S::AtRKD4 the transformants grown in greenhouse. To achieve it, AtRKD4 gene integration stability on transformants genome was analyzed. Morphology and cross-section anatomy structure on transformant and non-transformant plantswere described. The stability of AtRKD4 gene integration in the plant genome was confirmed by amplification of the AtRKD4 gene from genomic DNA with Polymerase Chain Reaction (PCR) using a specific primer for AtRKD4 and ACTIN genes as the internal control. The quantitative data from morphology and anatomy measurements were analyzed statistically using ANOVA. The results showed that AtRKD4 was stably integrated into the genome of P. amabilis transformants and all transformant plants showed similar morphology and anatomy characteristics as non-transformant plants. The AtRKD4 embryo gene was stably integrated into the orchid genome and the transformant plants grow normally without significant changes in phenotype.

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Section: Detection Of the Integration Of Atrkd4 Gene In The Genome Of P Amabilis Transformantsupporting

confidence: 59%

Stable Transformant of <i>Phalaenopsis amabilis</i> Somatic Embryo Carrying 35S::AtRKD4 Develops Into Normal Phenotype of Transgenic Plant

Perdana

Mose

Lawrie

et al. 2021

J.Tropical Biodiversity Biotechnology

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“…However, the percentage of mutation frequencies for the formation of this phenotype is only about 0.007% (Fig. 1, Mursyanti et al, 2016).…”

Section: Introductionmentioning

confidence: 97%

Stability of T-DNA Integration in Phalaenopsis “Sogo Vivien” Transgenic Orchid Carrying 35S::Gal4::AtRKD4::GR

et al. 2018

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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%.

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“…Variegation is a phenotype characterized by differences in color patterns. For example, Phalaenopsis 'Sogo Vivien' has variegated leaves and flowers (Mursyanti et al 2016). Several plant species have variegated patterns as well.…”

Section: Introductionmentioning

confidence: 99%

Isolation and Characterization of Chalcone Synthase (CHS) Gene in Variegated-Flower of Dendrobium 'Enobi' and Phalaenopsis Hybrid Orchids

Linggabuwana,

Putri,

Kurniawan

et al. 2023

HAYATI J Biosci

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Variegated flowers, characterized by the presence of different colors in flowers, have high economic and aesthetic values. The main pigment in the orchid's purple flowers is anthocyanin, while the chalcone synthase (CHS) gene is the key to anthocyanin biosynthesis. Analysis of the CHS gene can reveal some changes, including mutations, in the process of color patterning in flowers. This study aims to determine the structure of the CHS gene related to color patterning in Dendrobium 'Enobi' and Phalaenopsis hybrid with variegated flowers. The methods applied in this study are floral morphology observation, DNA isolation, CHS gene amplification, anthocyanin measurement, and bioinformatic analysis. Morphologically, the variegated pattern has appeared since the flowers were still in the bud on both orchids. Based on the anthocyanin content analysis, the difference in the genus is not directly related to the differences in the flower's anthocyanin content. In addition, the purple zone in the D. 'Enobi' and Phalaenopsis hybrid has a longer fragment of CHS than the white zone. Our analysis suggested several mutations in the white zone and differences in the type and location of several conserved domain proteins. Mutations at the CHS gene fragment might cause decreased anthocyanin pigment formation in the white region.

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Micropropagation of Mini Orchid Hybrid Phalaenopsis “Sogo Vivien”

Cited by 6 publications

References 26 publications

Stable Transformant of <i>Phalaenopsis amabilis</i> Somatic Embryo Carrying 35S::AtRKD4 Develops Into Normal Phenotype of Transgenic Plant

Stable Transformant of <i>Phalaenopsis amabilis</i> Somatic Embryo Carrying 35S::AtRKD4 Develops Into Normal Phenotype of Transgenic Plant

Stability of T-DNA Integration in Phalaenopsis “Sogo Vivien” Transgenic Orchid Carrying 35S::Gal4::AtRKD4::GR

Isolation and Characterization of Chalcone Synthase (CHS) Gene in Variegated-Flower of Dendrobium 'Enobi' and Phalaenopsis Hybrid Orchids

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