Development of a high-throughput virus-induced gene-silencing vector in phenotypic screening of orchids
Journal
Orchid biotechnology III
Pages
331-340
Date Issued
2017
Author(s)
Abstract
The elegant and enchanting flowers of orchids have always captured hearts. Indeed, it is hard not to be curious about how orchid flowers evolved such astonishing shapes and colors. However, without proper research tools, the secrets of the orchid have remained hidden. We developed a Cymbidium mosaic virus (CymMV)-induced gene-silencing (VIGS) vector, pCymMV-pro60, for orchids, which allows investigation of orchid genetic information by loss-of-function assay. To keep pace with the advances in modern sequencing technology, which can now resolve the total genetic codes for a species in a short time, we improved pCymMV-pro60 for highthroughput cloning and inoculation. Besides, we developed an efficient gene functional analysis strategy for phenotypic screening. We demonstrated that the application of VIGS is not limited to the study of genes involved in observable phenotypes, but can also be carried out to study unobservable but detectable phenotypes such as transcription regulation. © 2017 by World Scientific Publishing Co. Pte. Ltd. All rights reserved.
SDGs
Other Subjects
Clone cells; Genetic engineering; Transcription; Viruses; Gene silencing; Genetic information; High-throughput; Orchid flower; Phenotypic screening; Research tools; Transcription regulations; Virus-induced gene silencing; Biodiversity
Type
book part