Overexpression of vacuolar transporters OsVIT1 and OsVIT2 reduces cadmium accumulation in rice.

Excessive cadmium in rice grain in agricultural production is an important issue to be addressed in some southern regions of China. In this study, we constructed transgenic rice overexpressing OsVIT1 and OsVIT2 driven by 35S promoter in the cultivar ZH11. Compared with ZH11, OsVIT1 expression in leaves was significantly increased by 3-6.6 times and OsVIT2 expression in leaves was significantly increased by 2-2.5 times. Hydroponic experiments showed that overexpression of OsVIT1 and OsVIT2 increased the tolerance to Fe deficiency, significantly reduced Cd content in shoot and xylem sap, and had no effect on Cd tolerance in rice. Two years of field trials showed that the Fe content in the grain of OsVIT1 and OsVIT2 overexpressed materials was significantly reduced by 20-40% and the straw Fe content was significantly increased by 10-45%, and the grain Fe content distribution ratio was significantly decreased and the straw Fe distribution ratio was significantly increased compared with the wild type. The OsVIT1 and OsVIT2 overexpressed materials significantly reduced the Cd content of grain by 40-80% and the Cd content of straws by 37-77%, and the bioconcentration factor of Cd was significantly reduced in both grains and straw of OsVIT1 and OsVIT2 overexpressed materials. Overexpression of OsVIT1 and OsVIT2 did not affect the concentration of other metal ions in rice straw and grain. qRT-PCR analysis showed that the expression of the low affinity cation transporter OsLCT1 was significantly downregulated in the OsVIT1 and OsVIT2 overexpressed materials. In conclusion, overexpression of OsVIT1 and OsVIT2 reduced Cd accumulation in straw and grains, providing a strategy for Cd reduction in rice.

Genome and Transcriptome Identification of a Rice Germplasm with High Cadmium Uptake and Translocation.

The safe production of food on Cd-polluted land is an urgent problem to be solved in South China. Phytoremediation or cultivation of rice varieties with low Cd are the main strategies to solve this problem. Therefore, it is very important to clarify the regulatory mechanism of Cd accumulation in rice. Here, we identified a rice variety with an unknown genetic background, YSD, with high Cd accumulation in its roots and shoots. The Cd content in the grains and stalks were 4.1 and 2.8 times that of a commonly used japonica rice variety, ZH11, respectively. The Cd accumulation in the shoots and roots of YSD at the seedling stage was higher than that of ZH11, depending on sampling time, and the long-distance transport of Cd in the xylem sap was high. Subcellular component analysis showed that the shoots, the cell wall, organelles, and soluble fractions of YSD, showed higher Cd accumulation than ZH11, while in the roots, only the cell wall pectin showed higher Cd accumulation. Genome-wide resequencing revealed mutations in 22 genes involved in cell wall modification, synthesis, and metabolic pathways. Transcriptome analysis in Cd-treated plants showed that the expression of pectin methylesterase genes was up-regulated and the expression of pectin methylesterase inhibitor genes was down-regulated in YSD roots, but there were no significant changes in the genes related to Cd uptake, translocation, or vacuole sequestration. The yield and tiller number per plant did not differ significantly between YSD and ZH11, but the dry weight and plant height of YSD were significantly higher than that of ZH11. YSD provides an excellent germplasm for the exploration of Cd accumulation genes, and the cell wall modification genes with sequence- and expression-level variations provide potential targets for phytoremediation.

Cyclic GMP signaling and regulation of SERCA activity during cardiac myocyte contraction.

We tested the hypothesis that cGMP-induced reductions in cardiac myocyte function were related to activation of the sarcoplasmic reticulum Ca2+-ATPase (SERCA) and cGMP-dependent phosphorylation of phospholamban. Ventricular myocyte function was measured using a video edge detector (n = 11 rabbits). Thapsigargin (TG) or cyclopiazonic acid (CPA) were used to inhibit SERCA. 8-Bromo-cGMP was added at 10(-6), 10(-5) M followed by TG 10(-8) M or KT5823 (cGMP-protein kinase inhibitor, 10(-6) M) prior to TG or CPA. Cyclic GMP-dependent protein phosphorylation and immunoblotting with anti-phospholamban antibody were examined. TG 10(-8) M significantly increased percent shortening (from 6.6+/-0.7 to 9.1+/-1.3%). Cyclic GMP 10(-5) M significantly decreased cell shortening from 9.3+/-0.9 to 5.1+/-0.6%. This was partially reversed by KT5823 (5.1+/-0.6 to 8.2+/-1.4%) suggesting that negative functional effects of cGMP were partially through the cGMP-dependent protein kinase. Addition of TG after cGMP also reduced the negative effects of cGMP on myocyte shortening suggesting involvement of SERCA in cGMP signaling. TG after cGMP and KT5823 treatment did not alter myocyte contractility (8.2+/-1.4 to 7.2+/-1.3%). CPA had similar effects as those of TG. Protein phosphorylation and immunoblotting showed that phospholamban was a target of the cGMP protein kinase. These results indicated that the cyclic GMP-induced reductions in myocyte function were partially mediated through the action of SERCA. It further suggested that cGMP signaling affects myocyte function through phosphorylation of phospholamban which regulates SERCA activity.