Query (optional)   in Class  All Allele Author Colleague Collection Assembly Gene Gene Class Gene Product Germplasm Image Journal Keyword Library Locus Map Map Data Marker Pathology Polymorphism Probe Protein QTL Rearrangement Reference Sequence Species Trait Trait Study Two Point Data  

---

GrainGenes Reference Report: PLC-57-207

[Submit comment/correction]

Reference	PLC-57-207
Title	Optimisation of procedures for microprojectile bombardment of microspore-derived embryos in wheat
Journal	Plant Cell
Year	2000
Volume	57
Pages	207-210
Author	Ingram HM Power JB Lowe KC Davey MR
Abstract	Summary: Using the PDS-1000/He Biolistic Particle Delivery System, the microprojectile travel distance, rupture disk pressure and DNA/gold particle concentrations were assessed in order to optimise short and longer-term beta-glucuronidase reporter gene expression in microspore-derived embryos of wheat. The effects were also evaluated of using sterile filter paper to support explants and treatment with a high osmoticum medium (0.2 M mannitol/0.2 M sorbitol or 0.4 M maltose). In the optimised procedure, wheat microspore-derived embryos (MDEs), were placed on filter paper and incubated on medium containing 0.4 M maltose, for 4 h pre- and 45 h post-bombardment. Five microliters pAHC25 (0.75 mg ml(-1) in TE buffer) was precipitated onto 25 microliters gold particles (60 mg ml(-1) in sterile water), using 20 microliters spermidine (0.1 M) and 50 microliters CaCl2 (2.5 M). The particles were centrifuged and resuspended in 75 microliters absolute ethanol prior to the preparation of 6 macrocarriers. A microprojectile travel distance of 70 mm, a rupture pressure of 1300 p.s.i., and a vacuum of 29' Hg were employed. Maltose at 0.4 M in the support medium was the most important factor influencing GUS activity in bombarded tissues. GUS activity, 1 day post-bombardment, reached 52 +/- 17 GUS-positive foci/MDE (mean +/- s.e.m, n = 3), with 17 +/- 4 foci/MDE at 15 days, giving a 3.0-fold increase (p < 0.05) compared to expression in MDEs bombarded on medium without a high osmoticum treatment
Keyword	[ Hide all but 1 of 18 ] 75 beta glucuronidase biolistics culture media embryo explants gene gene expression gene transfer genetic transformation in vitro culture methodology microprojectile bombardment micropropagation system transgenic plant triticum aestivum water

GrainGenes is a product of the Agricultural Research Service of the US Department of Agriculture.