The expression analysis of AtRKD4 transgene in Dendrobium lineale Rolfe transgenic orchid carrying 35S::GR::AtRKD4 for micropropagation

Febryanti, Ni Luh Putu Kayika; Nurliana, Steffanie; Gutierrez-Marcos, José; Semiarti, Endang

doi:10.1063/5.0015876

Cited by 5 publications

(4 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Single treatment with 1 mg/L TDZ produced similar number of SEs (2.22 ± 1.39) but not all pseudobulb explants could produce SE (only 89%), the addition of 1 mg/L NAA pseudobulb was able to pro duce one SE in root explants (0.09 ± 0.37a). This is in accordance with the results of previous studies reported by Mose et al (2017), Febryanti et al (2020, Zulwanis et al (2020), andPuspitasari et al (2020), which showed that pseudobulb explants were the most responsive in SE formation. Although DMRT testing showed that in the for mation of SEs in pseudobulb explants from media with 1 mg/L TDZ have similar result with the media with a com bination of 1 mg/L TDZ + 1 mg/L NAA, and a significant difference in relation to other treatment media, but the per centage of SE formation the addition of a combination of 1 mg/L TDZ + 1 mg/L NAA was higher (100%).…”

Section: The Effect Of Naa and Tdz And Its Combinations On Induction ...supporting

confidence: 93%

“…Puspitasari et al (2020) found that NP + TDZ 2 mg/L + DEX 12 µM medium was the best treatment to pro duce SEs with 100% production in pseudobulb explants. In a study by Febryanti et al (2020) it was stated that the combination of PGRs increased the formation of SEs, in cluding DEX 15 µM + TDZ 1 mg/L which induced the formation of the best somatic embryos in stem explants of transformant Dendrobium lineale.…”

Section: Ilham Et Almentioning

confidence: 99%

See 1 more Smart Citation

The effectivity of thidiazuron and 1‐naphthaleneacetic acid on somatic embryo induction in transgenic Dendrobium phalaenopsis Fitzg. carrying 35S::GR::AtRKD4

Ilham

Puspitasari²,

Semiarti³

2022

Indones. J. Biotechnol.

Self Cite

View full text Add to dashboard Cite

Dendrobium phalaenopsis Fitzg. (also known as the Larat orchid) is an endemic orchid from Larat Island, Eastern Indonesia. Its beautiful flowers mean that many plants are taken for commercial purposes, leading to the rapid decline of populations in their natural habitats. The objectives of this study were to determine which organs of the transgenic Larat orchid carrying the 35S::GR::AtRKD4 construct, together with which concentrations of the plant growth regulators (PGRs) auxin and cytokinin, are suitable for the induction of somatic embryos (SEs). In this study, the AtRKD4 gene in Larat orchids was confirmed using PCR with specific primers for the AtRKD4 and HPT genes. Thidiazuron (TDZ) (1, 3 and 5 mg/L) in combination with 1‐naphthaleneacetic acid (NAA) (0.5 and 1 mg/L) were used on new phalaenopsis (NP) medium to induce SEs from leaves, pseudobulbs and roots. The AtRKD4 transgenes were detected as being stably integrated into the DNA genome of transformant plants using specific primers for AtRKD4 and HPT genes, and positive results were obtained using actin gene primers as internal controls for PCR. Pseudobulbs produced 19 to 20 SEs from 108 pseudobulb explants (89–100%), a higher number than produced in explants of the other organs studied. Among the PGR treatments, the best results were obtained in NP medium supplemented with a combination of 1 mg/L TDZ and 1 mg/L NAA, 100% of the explants of which produced SEs (2.11 ± 1.36). No significant difference was found between the morphology of the SEs produced from the non‐transformant Larat orchid pseudobulb explants and the 35S::AtRKD4 carrier transformant.

show abstract

Section: The Effect Of Naa and Tdz And Its Combinations On Induction ...supporting

confidence: 93%

Section: Ilham Et Almentioning

confidence: 99%

The effectivity of thidiazuron and 1‐naphthaleneacetic acid on somatic embryo induction in transgenic Dendrobium phalaenopsis Fitzg. carrying 35S::GR::AtRKD4

Ilham

Puspitasari²,

Semiarti³

2022

Indones. J. Biotechnol.

Self Cite

View full text Add to dashboard Cite

show abstract

“…Based on Zulwanis (2020), DNA bands that amplified from the explants were induced using DEX and TDZ during 1 to 9 th day, the highest transgene expression was found .in day 5 th of induction as the highest DNA band density. These results correspond to the results of gene expression analysed by Febryanti et al (2020) showing that AtRKD4 mRNA appeared in D.lineale transformant samples induced with medium NP +15 μM DEX or NP + 3 mgL-1 TDZ. This proves that the translation process goes well judging by the formation of AtRKD4 gene expression products in the form of proteins.…”

Section: Induction and Detection Of Atrkd4 Protein In Orchid Transformant Plants After Being Induced By Using Dex And Tdzsupporting

confidence: 89%

Detection of AtRKD4 Protein During Induction of Somatic Embryogenesis in Dendrobium lineale Rolfe Transgenic Orchids Carrying 35S::GR::AtRKD4

Wirajagat

Febryanti

Puspitasari

et al. 2021

J.Tropical Biodiversity Biotechnology

Self Cite

View full text Add to dashboard Cite

Dendrobium lineale is an Indonesian native orchid from the Spatulata section in Orchidaceae Family. This orchid is important because it is usually used as a parental plant in orchid breeding and is predicted to have a potential phytochemistry compound. In addition, in their natural habitat, this orchid is threatened due to forest exploitation and natural disaster. Therefore the precision mass propagation techniques for this orchid need to be conducted. Biotechnological approaches through inserting embryo gene such as AtRKD4 from Arabidopsis thaliana has already been successfully conducted. This study aims to check the integration stability of T-DNA harboring 35S::GR::AtRKD4 from ten selection transformants and to detect the existence of AtRKD4 protein after induction by Dexamethasone and/ Thidiazuron. The result showed that T-DNA were stably integrated into the genome of D. lineale transformants and the AtRKD4 protein with a molecular weight of 28.53 kDa was detected in D. lineale transformant plants after being induced by 15 µM DEX and 3 mgL-1 TDZ for 5 days.

show abstract

“…Because of the limitations and inefficiencies of the traditional approaches, hybridization and mutagenesis can not be used to get some desirable traits, like the spotted blooms and foliage of a single plant. Agrobacterium-mediated transformation and particle bombardment methods have been routinely used in transgenic molecular breeding, leading to significant progress in horticultural development (Liau et al, 2003;Men et al, 2003;Hsing et al, 2016;Khumkarjorn et al, 2017;Lin et al, 2018;Kayika Febryanti et al, 2020;Setiawati et al, 2020). Our understanding of orchid reproductive biology will undoubtedly change as a result of these efforts to enhance orchid genomeediting tools and the power of large-scale genome sequencing, which will enable us to better understand the inherent roles of orchid genes and changes to genes of interest for desired blooming and floral features (Molla and Yang 2019;Nopitasari et al, 2020;Tong et al, 2020;Guo et al, 2022).…”

Section: Introductionmentioning

confidence: 99%

In-depth analysis of genomes and functional genomics of orchid using cutting-edge high-throughput sequencing

et al. 2022

View full text Add to dashboard Cite

High-throughput sequencing technology has been facilitated the development of new methodologies and approaches for studying the origin and evolution of plant genomes and subgenomes, population domestication, and functional genomics. Orchids have tens of thousands of members in nature. Many of them have promising application potential in the extension and conservation of the ecological chain, the horticultural use of ornamental blossoms, and the utilization of botanical medicines. However, a large-scale gene knockout mutant library and a sophisticated genetic transformation system are still lacking in the improvement of orchid germplasm resources. New gene editing tools, such as the favored CRISPR-Cas9 or some base editors, have not yet been widely applied in orchids. In addition to a large variety of orchid cultivars, the high-precision, high-throughput genome sequencing technology is also required for the mining of trait-related functional genes. Nowadays, the focus of orchid genomics research has been directed to the origin and classification of species, genome evolution and deletion, gene duplication and chromosomal polyploidy, and flower morphogenesis-related regulation. Here, the progressing achieved in orchid molecular biology and genomics over the past few decades have been discussed, including the evolution of genome size and polyploidization. The frequent incorporation of LTR retrotransposons play important role in the expansion and structural variation of the orchid genome. The large-scale gene duplication event of the nuclear genome generated plenty of recently tandem duplicated genes, which drove the evolution and functional divergency of new genes. The evolution and loss of the plastid genome, which mostly affected genes related to photosynthesis and autotrophy, demonstrated that orchids have experienced more separate transitions to heterotrophy than any other terrestrial plant. Moreover, large-scale resequencing provide useful SNP markers for constructing genetic maps, which will facilitate the breeding of novel orchid varieties. The significance of high-throughput sequencing and gene editing technologies in the identification and molecular breeding of the trait-related genes in orchids provides us with a representative trait-improving gene as well as some mechanisms worthy of further investigation. In addition, gene editing has promise for the improvement of orchid genetic transformation and the investigation of gene function. This knowledge may provide a scientific reference and theoretical basis for orchid genome studies.

show abstract

The expression analysis of AtRKD4 transgene in Dendrobium lineale Rolfe transgenic orchid carrying 35S::GR::AtRKD4 for micropropagation

Cited by 5 publications

References 14 publications

The effectivity of thidiazuron and 1‐naphthaleneacetic acid on somatic embryo induction in transgenic Dendrobium phalaenopsis Fitzg. carrying 35S::GR::AtRKD4

The effectivity of thidiazuron and 1‐naphthaleneacetic acid on somatic embryo induction in transgenic Dendrobium phalaenopsis Fitzg. carrying 35S::GR::AtRKD4

Detection of <i>AtRKD4</i> Protein During Induction of Somatic Embryogenesis in <i>Dendrobium lineale</i> Rolfe Transgenic Orchids Carrying <i>35S::GR::AtRKD4</i>

In-depth analysis of genomes and functional genomics of orchid using cutting-edge high-throughput sequencing

Contact Info

Product

Resources

About