当前位置： SCI文献检索 > Plant Methods期刊下所有文献 > Methods for transient assay of gene function in floral tissues.

Methods for transient assay of gene function in floral tissues.

Abstract：

BACKGROUND:There is considerable interest in rapid assays or screening systems for assigning gene function. However, analysis of gene function in the flowers of some species is restricted due to the difficulty of producing stably transformed transgenic plants. As a result, experimental approaches based on transient gene expression assays are frequently used. Biolistics has long been used for transient over-expression of genes of interest, but has not been exploited for gene silencing studies. Agrobacterium-infiltration has also been used, but the focus primarily has been on the transient transformation of leaf tissue. RESULTS:Two constructs, one expressing an inverted repeat of the Antirrhinum majus (Antirrhinum) chalcone synthase gene (CHS) and the other an inverted repeat of the Antirrhinum transcription factor gene Rosea1, were shown to effectively induce CHS and Rosea1 gene silencing, respectively, when introduced biolistically into petal tissue of Antirrhinum flowers developing in vitro. A high-throughput vector expressing the Antirrhinum CHS gene attached to an inverted repeat of the nos terminator was also shown to be effective. Silencing spread systemically to create large zones of petal tissue lacking pigmentation, with transmission of the silenced state spreading both laterally within the affected epidermal cell layer and into lower cell layers, including the epidermis of the other petal surface. Transient Agrobacterium-mediated transformation of petal tissue of tobacco and petunia flowers in situ or detached was also achieved, using expression of the reporter genes GUS and GFP to visualise transgene expression. CONCLUSION:We demonstrate the feasibility of using biolistics-based transient RNAi, and transient transformation of petal tissue via Agrobacterium infiltration to study gene function in petals. We have also produced a vector for high throughput gene silencing studies, incorporating the option of using T-A cloning to insert the gene sequence of interest. These techniques should allow analysis of gene function in a much broader range of flower species.

journal_name

Plant Methods

journal_title

Plant methods

authors

Shang Y,Schwinn KE,Bennett MJ,Hunter DA,Waugh TL,Pathirana NN,Brummell DA,Jameson PE,Davies KM

doi

10.1186/1746-4811-3-1

subject

Has Abstract

pub_date

2007-01-08 00:00:00

pages

issn

1746-4811

pii

1746-4811-3-1

journal_volume

pub_type

杂志文章

在线工具

Methods for transient assay of gene function in floral tissues.