Zhenyan He

Arabidopsis NIP3;1 Plays an Important Role in Arsenic Uptake and Root-to-Shoot Translocation under Arsenite Stress Conditions

In Arabidopsis, the nodulin 26-like intrinsic protein (NIP) subfamily of aquaporin proteins consists of nine members, five of which (NIP1;1, NIP1;2, NIP5;1, NIP6;1, and NIP7;1) were previously identified to be permeable to arsenite. However, the roles of NIPs in the root-to-shoot translocation of arsenite in plants remain poorly understood. In this study, using reverse genetic strategies, Arabidopsis NIP3;1 was identified to play an important role in both the arsenic uptake and root-to-shoot distribution under arsenite stress conditions. The nip3;1 loss-of-function mutants displayed obvious improvements in arsenite tolerance for aboveground growth and accumulated less arsenic in shoots than those of the wild-type plants, whereas the nip3;1 nip1;1 double mutant showed strong arsenite tolerance and improved growth of both roots and shoots under arsenite stress conditions. A promoter-β-glucuronidase analysis revealed that NIP3;1 was expressed almost exclusively in roots (with the exception of the root tips), and heterologous expression in the yeast Saccharomyces cerevisiae demonstrated that NIP3;1 was able to mediate arsenite transport. Taken together, our results suggest that NIP3;1 is involved in arsenite uptake and root-to-shoot translocation in Arabidopsis, probably as a passive and bidirectional arsenite transporter.

The ER luminal binding protein (BiP) alleviates Cd2+-induced programmed cell death through endoplasmic reticulum stress–cell death signaling pathway in tobacco cells

Cadmium (Cd) is very toxic to plant cells and Cd(2+) stress induces programmed cell death (PCD) in Nicotiana tabacum L. cv. bright yellow-2 (BY-2) cells. In plants, PCD can be regulated through the endoplasmic reticulum (ER) stress-cell death signaling pathway. However, the mechanism of Cd(2+)-induced PCD remains unclear. In this study, we found that Cd(2+) treatment induced ER stress in tobacco BY-2 cells. The expression of two ER stress markers NtBLP4 and NtPDI and an unfolded protein response related transcription factor NtbZIP60 were upregulated with Cd(2+) stress. Meanwhile, the PCD triggered by prolonged Cd(2+) stress could be relieved by two ER chemical chaperones, 4-phenylbutyric acid and tauroursodeoxycholic acid. These results demonstrate that the ER stress-cell death signaling pathway participates in the mediation of Cd(2+)-induced PCD. Furthermore, the ER chaperone AtBiP2 protein alleviated Cd(2+)-induced ER stress and PCD in BY-2 cells based on the fact that heterologous expression of AtBiP2 in tobacco BY-2 cells reduced the expression of NtBLP4 and a PCD-related gene NtHsr203J under Cd(2+) stress conditions. In summary, these results suggest that the ER stress-cell death signaling pathway regulates Cd(2+)-induced PCD in tobacco BY-2 cells, and that the AtBiP2 protein act as a negative regulator in this process.

Arsenate reduces copper phytotoxicity in gametophytes of Pteris vittata

The fern Pteris vittata is an arsenic (As) hyperaccumulator and can take up very high concentrations of arsenic from the soil. However, little is known about its response to co-contamination with arsenic and copper (Cu). In this study, we used an in vitro model system of P. vittata gametophytes to investigate the impact of changes in As and Cu status on growth, chlorophyll (chl) concentration, metal accumulation, and subcellular localization. A remarkable inhibition of growth occurred when gametophytes were exposed to concentrations >or=1.0mM Na(3)AsO(4) or >or=0.5mM CuSO(4). chl concentration decreased significantly when gametophytes were exposed to >0.25mM of CuSO(4), but increased steadily with concentration to <or=2mM Na(3)AsO(4). Interestingly, the inhibitory effect caused by Cu was reduced in the presence of 0.25mM Na(3)AsO(4). However, the inhibition caused by exposure to 1.0mM Na(3)AsO(4) was not alleviated by 0.25mM CuSO(4). Further studies showed that 0.25mM Na(3)AsO(4) increased cell viability (CV) and chl concentration, while decreasing cell membrane permeability (CMP) of gametophytes with 1.0mM CuSO(4) stress. In contrast, 0.25mM CuSO(4) decreased CV and chl concentration, while increasing CMP when gametophytes were treated with 1.0mM Na(3)AsO(4). In addition, the subcellular distribution of As and Cu in P. vittata gametophytes differed. As was found primarily in the cytoplasm, while Cu was mainly localized in the cell wall. These results suggest that As can reduce Cu phytotoxicity in the As hyperaccumulator P. vittata, and that this may serve as a biological mechanism for the fern to adapt to soils co-contaminated with As and Cu.

A SAL1 loss-of-function Arabidopsis mutant exhibits enhanced cadmium tolerance in association with alleviation of endoplasmic reticulum stress

SAL1, as a negative regulator of stress response signaling, has been studied extensively for its role in plant response to environmental stresses. However, the role of SAL1 in cadmium (Cd) stress response and the underlying mechanism is still unclear. Using an Arabidopsis thaliana loss-of-function mutant of SAL1, we assessed Cd resistance and further explored the Cd toxicity mechanism through analysis of the endoplasmic reticulum (ER) stress response. The loss of SAL1 function greatly improved Cd tolerance and significantly attenuated ER stress in Arabidopsis. Exposure to Cd induced an ER stress response in Arabidopsis as evidenced by unconventional splicing of AtbZIP60 and up-regulation of ER stress-responsive genes. Damage caused by Cd was markedly reduced in the ER stress response double mutant bzip28 bzip60 or by application of the ER stress-alleviating chemical agents, tauroursodeoxycholic acid (TUDCA) and 4-phenyl butyric acid (4-PBA), in wild-type plants. The Cd-induced ER stress in Arabidopsis was also alleviated by loss of function of SAL1. These results identified SAL1 as a new component mediating Cd toxicity and established the role of the ER stress response in Cd toxicity. Additionally, the attenuated ER stress in the sal1 mutant might also shed new light on the mechanism of diverse abiotic stress resistance in the SAL1 loss-of-function mutants.

An Arabidopsis mutant of inositol pentakisphosphate 2‐kinase AtIPK1 displays reduced arsenate tolerance

Arsenate [As(V)] toxicity is considered to be derived from similarities in the chemical properties of As(V) and phosphate (Pi). An Arabidopsis thaliana mutant of inositol pentakisphosphate 2-kinase (AtIPK1), atipk1-1, has previously exhibited lower level of phytate and higher level of Pi, relative to wild-type (WT). Here, atipk1-1 displayed hypersensitivity to As(V) stress and less As(V) uptake when compared to WT. Overexpression of AtIPK1 controlled by the CaMV 35S promoter partially rescued the As(V)-sensitive phenotype of atipk1-1. When compared to control Pi status, addition of Pi enhanced As(V) tolerance of both WT and atipk1-1 plants, while the arsenic concentration was less reduced in the latter genotype. Despite the higher Pi level in atipk1-1 than did WT plants, the mutant suffered more severe Pi starvation under Pi limitation stress, indicating that Pi homeostasis was altered in the mutant. Gene expression analysis of WT and atipk1-1 plants showed the diverse effect of As(V) stress on Pi starvation-dependent regulation of Pi-responsive genes. Our study suggested that a particular mechanism of As(V) toxicity existed in atipk1-1 mutant, and may offer new insights into the interactions between Pi homeostasis and As(V) detoxification in plants.

Assessment of heavy metals pollution of soybean grains in North Anhui of China

Heavy metals may cause deleterious effects on human health through consumption of contaminated food crops. To assess the safety of soybean consumption, concentrations of heavy metals (Cu, Zn, Cr, Ni, Cd, As, Pb and Hg) in soybean grains sampled from the farmland in North Anhui (suitable for high-protein soybeans planting) were analyzed. The results showed that the concentrations of Ni, Cr, Cu and Pb in soybean gains exceeded the Chinese safety limits, with over-standard rate of 92.59%, 74.07%, 37.04% and 9.88% respectively. Spatial distribution of soybean pollution analysis showed that soybeans were severe polluted by Ni and concentrated in the west of North Anhui. SR-μXRF elemental maps of soybean gains indicated Cu, Cr, Ni, Pb and Hg specifically enriched at the embryo. In relation to health risks, there were no obvious non-carcinogenic risk and carcinogenic risk (CR) through ingesting soybeans from North Anhui. But it can induce a threat to peoples health if the consumption of soybean exceeds the routine dose.