Fig 1.
Amino acid sequence alignment and phylogenetic tree of Cu/Zn-SOD proteins from rice and other plant species.
(A) Highly homologous regions in the amino acid sequences of OsCu/Zn-SOD from different species. The black triangle stands for active site. (B) Phylogenetic tree of OsCu/Zn-SOD generated via NJ method.
Fig 2.
Analysis of OsCu/Zn-SOD gene expression.
(A) Expression patterns of OsCu/Zn-SOD gene in different rice tissues (i.e., root, leaf, tassel, shell, stem, flower, S5, and L5). (B) and (C) OsCu/Zn-SOD expression after treatment with 150 mM NaCl or 60 mM NaHCO3 for 0, 2, 4, 8, 16 and 24 h.
Fig 3.
Subcellular localization of the 35S::OsCu/Zn-SOD::GFP fusion protein.
(A) To interpret the color references in this figure legend, the reader should refer to the web version of this article. Fluorescence of OsCu/Zn-SOD-GFP fusion protein in onion; merged image of the subcellular localization of the chloroplast marker mcherry; superposition of green fluorescence and mCherry; DIC, bright field. (B) Targeting of GFP fusion proteins to Arabidopsis protoplast. Fluorescence of OsCu/Zn-SOD-GFP fusion protein in a protoplast derived from an Arabidopsis mesophyll cell; autofluorescence of chlorophyll; merged images from green and red channels; protoplast viewed under white light.
Fig 4.
Schematic of the expression vectors pCXSN::Cu/Zn-SOD and PCR detection of products.
(A). Schematic of the rice Cu/Zn-SOD gene two-plant expression vector. The vector starts with CaMV35S and is terminated by Tons. The PCXSN::OsCu/Zn-SOD expression vector contains the Hyg resistance gene. (B) PCR detection of the CaMV35S::OsCu/Zn-SOD vector in transgenic Longjing 11, generation T1. CK+- positive plasmid control; NT- nontransgenic Longjing 11. Lanes 1–5 contain PCR products from OsCu/Zn-SOD-overexpressing transgenic Longjing 11 T1 lines. (C) mRNA expression in OsCu/Zn-SOD-overexpressing lines, T3 generation (lanes 1–3) as detected by Northern blot.
Fig 5.
NaCl stress resistance of OsCu/Zn-SOD-overexpressing rice lines during germination.
(A) Phenotype. (B) Germination rate. (C) Plant fresh weight. (D) Plant height. (E) MDA content of NT and overexpression of OsCu/Zn-SOD Rice (T3-#1, #2, #3) after 9 days of germination and growth. Significant differences in parameters when a, b, c, d is p<0.05.
Fig 6.
Growth characteristics of OsCu/Zn-SOD-overexpressing lines treated with NaHCO3.
Phenotype A after 21 days of treatment with (A) 0 mM, (B) 5 mM, (C) 7.5 mM, (D) 10 mM NaHCO3. (E) Plant height. (F) Fresh weight. (G) SOD activity. (H) Chorophyll content.
Fig 7.
Analysis of active oxygen content in OsCu/Zn-SOD-overexpressing lines after 21 days of 0 mM, 5 mM, 7.5 mM, 10 mM stress.
NT- nontransgenic rice; 35S::SOD- OsCu/Zn-SOD-overexpressing rice.
Fig 8.
Growth conditions of OsCu/Zn-SOD-overexpressing and NT lines under simulated saline–sodic stress in the field.
(A) Phenotype. (B) Productive panicle. (C) Seed-setting rate. (D) Thousand-seed weight.