Biology Reference
In-Depth Information
Fig.
1
Schematic view on
137
Cs and
90
Sr uptake by soybean. Soybean shoot system (soybean plant without
roots, seed pods, and seeds) accumulated higher amounts of radioactivity than seeds during our experiments
in radioactive Chernobyl zone (Chistogalovka area)
agricultural areas remediated from radioactivity and provided
detailed characterization of fl ax seeds harvested from radioactive
and remediated Chernobyl areas [
1
,
2
,
6
].
Herein we describe the methodology that we use to analyze
seed proteome in radioactive Chernobyl area. Firstly, soils in
radioactive areas are often nutritiously poor and thus the fertiliza-
tion might be necessary. Secondly, selection of plant material
determines the level of radioactivity in the experiments. In our
experience, the uptake of radioactive elements differs between
plants and also between organs/tissues (Fig.
1
). For instance, soy-
bean shoot system (soybean without roots, seed pods, and seeds)
accumulated 3,600 ± 144 Bq/kg of
137
Cs and 54,000 ± 2,800 Bq/
kg of
90
Sr, during our experiments in radioactive Chernobyl zone
(Chistogalovka area). However, seeds accumulated only
2,130 ± 207 Bq/kg of
137
Cs and 11,840 ± 1,800 Bq/kg of
90
Sr
[
5
]. Furthermore, fl ax seeds accumulated even less radioactivity,
780 ± 39 Bq/kg of
137
Cs and 3,550 ± 360 Bq/kg of
90
Sr at the
same area.
2
Materials
1. Homogenization solution: 50 % (v/v) phenol, 0.45 M sucrose,
5 mM EDTA, 0.2 (v/v) 2-mercaptoethanol, 50 mM Tris-
HCl, pH 8.8 (
see
Note 1
).
2. 0.1 M ammonium acetate in 100 % methanol (store at −20 °C).
3. 80 % acetone (store at −20 °C).
4. 70 % ethanol (store at −20 °C).
2.1 Phenol-Based
Protein Extraction
