Isolation and Characterization of Vanda Orchid Homeobox Gene from Vanda tricolor var. Suavis Lindl. form Merapi

DOI

10.2991/absr.k.220406.036How to use a DOI?

Keywords

Homeobox Gene; Homologous Gene; Vanda Orchid Homeobox; Vanda tricolor

Abstract

Vanda tricolor var. Suavis Lindl. form Merapi is one of the crucial Indonesian orchid species. Due to natural disasters or deforestation of their natural habitat, the population of this orchid continues to decline and is threatened to be extinct. Therefore, a strategy for mass propagation of this plant as a conservation effort is needed, both in-situ and ex-situ. Mass propagation using in vitro culture will greatly support ex situ conservation. It is well known that shoot growth in plants begins with activating of the homeobox gene in the Shoot Apical Meristem (SAM), which induces the activation of related genes to regulate the growth of plant organs. Information about the homeobox gene in V. tricolor is necessary to support the induction of optimal shoot growth in in vitro culture conditions. Previous studies have discovered the homeobox gene DOH1 in Dendrobium and POH1 in Phalaenopsis. We assumed that Vanda has a homologous gene to DOH1 and POH1. The objective of this study was to isolate and analyze Vanda homeobox gene with degenerate primers designed from DOH1 and POH1 sequences. Leaf from a mature V. tricolor was used as samples for DNA analysis. The PCR product amplified genomic DNA using primers from DOH1 and POH1 were analyzed by agarose gel electrophoresis. We found a similarity in the length of amplified product of Vanda homeobox gene using DOH1 primer within particular showed that the PCR fragment is aligned with the N-terminal region of DOH1. This confirmed the conserved area and promised a high similarity in structure and functions between both genes. Amplification with POH1 primer showed that high similarity in the length of PCR product as accumulated POH1 transcript found in P. amabilis from the previous study, which showed that it was the coding region. The subsequent sequence analysis on the candidate of DOH1 and POH1 homologous gene in V. tricolor showed that the gene has 77.27% similarity with DOH1 thus might be act as the key gene for shoot growth in V. tricolor orchids. The POH1 homologous gene in V. tricolor showed no significant similarity with any sequence in the database, suggesting it might be a new sequence that needs further study.

Copyright

© 2022 The Authors. Published by Atlantis Press International B.V.

Open Access

This is an open access article distributed under the CC BY-NC 4.0 license.

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TY  - CONF
AU  - Viantius Ruben
AU  - Muhammad Dylan Lawrie
AU  - Endang Semiarti
PY  - 2022
DA  - 2022/05/02
TI  - Isolation and Characterization of Vanda Orchid Homeobox Gene from Vanda tricolor var. Suavis Lindl. form Merapi
BT  - Proceedings of the 7th International Conference on Biological Science (ICBS 2021)
PB  - Atlantis Press
SP  - 255
EP  - 260
SN  - 2468-5747
UR  - https://doi.org/10.2991/absr.k.220406.036
DO  - 10.2991/absr.k.220406.036
ID  - Ruben2022
ER  -