Bioaccumulation,subcellular distribution and chemical forms of cadmium in Aster subulatus Michx.

【Objective】Through analyzing the bioaccumulation capacity,subcellular distribution and chemical forms of cadmium(Cd)in Aster subulatus Michx.,this study was to provide reference for revealing the Cd tolerance mechanism of A.subulatus Michx.【Method】After cultured for 24 d under the action of Hoagland nutrient solution and gradient Cd concentrations(0,30,60 and 90 mg/L),A.subulatus Michx.were harvested,and its leaf,stem and root were treated by differential centrifugation,chemical reagent extraction,and digested with graphite digester,respectively,then the Cd content in the root,stem and leaf were determined by atomic absorption spectroscopy.【Result】The experimental results indicated that the bioaccumulation capacity of Cd in A.subulatus Michx.was root>stem>leaf,and the maximum Cd concentration in the root,stem and leaf of A.subulatus Michx.were 130.74,78.69 and 56.62 mg/kg(fresh matter),respectively.Most of Cd stored in the cell wall and the soluble fractions of the root and leaf of A.subulatus Michx.,with only a smaller portion Cd in organelle fraction.Analysis result of subcellular Cd content showed that 52.27%-58.61%of Cd for root was mainly stored in the soluble fraction,but 42.10%-63.28%of Cd for leaf was mainly stored in the cell wall fraction.The concentration of pectates and protein integrated-Cd was higher in the root and leaf compared to other chemical forms Cd.Pectates and protein integrated-Cd was the main chemical forms Cd in the root and leaf of A.subulatus Michx.,and their percentages were 68.91%-74.80%and 57.38%-83.80%,respectively.Cd treatment could significantly increase the proportion of water-soluble organic acid Cd from 13.64%to 22.72%in root and undissolved phosphate Cd from 10.02%to 32.78%in leaf with increasing Cd concentration in the culture medium.【Conclusion】The root,stem and leaf of A.subulatus Michx.has strong bioaccumulation capacity to Cd,Cd is primarily stored in the soluble fractions of the root and cell wall fractions of the leaf,and less toxic pectates and protein integrated-Cd is the main chemical forms Cd in the root and leaf of A.subulatus Michx.,this might be the main mechanism of Cd tolerance in A.subulatus Michx.

Essential proteins identification method based on four-order distances and subcellular localization information

Essential proteins are inseparable in cell growth and survival. The study of essential proteins is important for understanding cellular functions and biological mechanisms. Therefore, various computable methods have been proposed to identify essential proteins. Unfortunately, most methods based on network topology only consider the interactions between a protein and its neighboring proteins, and not the interactions with its higher-order distance proteins. In this paper, we propose the DSEP algorithm in which we integrated network topology properties and subcellular localization information in protein–protein interaction(PPI) networks based on four-order distances, and then used random walks to identify the essential proteins. We also propose a method to calculate the finite-order distance of the network, which can greatly reduce the time complexity of our algorithm. We conducted a comprehensive comparison of the DSEP algorithm with 11 existing classical algorithms to identify essential proteins with multiple evaluation methods. The results show that DSEP is superior to these 11 methods.

Subcellular distribution of prohibitin 1 in rat liver during liver regeneration and its cellular implication

BACKGROUND The function of prohibitin 1(Phb1)during liver regeneration(LR)remains relatively unexplored.Our previous research identified downregulation of Phb1 in rat liver mitochondria 24 h after 70%partial hepatectomy(PHx),as determined by subcellular proteomic analysis.AIM To investigate the potential role of Phb1 during LR.METHODS We examined changes in Phb1 mRNA and protein levels,subcellular distribution,and abundance in rat liver during LR following 70%PHx.We also evaluated mitochondrial changes and apoptosis using electron microscopy and flow cytometry.RNA-interference-mediated knockdown of Phb1(PHBi)was performed in BRL-3A cells.RESULTS Compared with sham-operation control groups,Phb1 mRNA and protein levels in 70%PHx test groups were downregulated at 24 h,then upregulated at 72 and 168 h.Phb1 was mainly located in mitochondria,showed a reduced abundance at 24 h,significantly increased at 72 h,and almost recovered to normal at 168 h.Phb1 was also present in nuclei,with continuous increase in abundance observed 72 and 168 h after 70%PHx.The altered ultrastructure and reduced mass of mitochondria during LR had almost completely recovered to normal at 168 h.PHBi in BRL-3A cells resulted in increased S-phase entry,a higher number of apoptotic cells,and disruption of mitochondrial membrane potential.CONCLUSION Phb1 may contribute to maintaining mitochondrial stability and could play a role in regulating cell proliferation and apoptosis of rat liver cells during LR.

An efficient transient gene expression system for protein subcellular localization assay and genome editing in citrus protoplasts

Protoplast has been widely used in biotechnologies to circumvent the breeding obstacles in citrus, including long juvenility, polyembryony, and male/female sterility. The protoplast-based transient gene expression system is a powerful tool for gene functional characterization and CRISPR/Cas9 genome editing in higher plants, but it has not been widely used in citrus. In this study, the polyethylene glycol(PEG)-mediated method was optimized for citrus callus protoplast transfection, with an improved transfection efficiency of 68.4%. Consequently, the efficiency of protein subcellular localization assay was increased to 65.8%, through transient expression of the target gene in protoplasts that stably express the fluorescent organelle marker protein. The gene editing frequencies in citrus callus protoplasts reached 14.2% after transient expression of CRISPR/Cas9 constructs. We demonstrated that the intronic polycistronic tRNAgRNA(inPTG) genome editing construct was functional in both the protoplast transient expression system and epicotyl stable transformation system in citrus. With this optimized protoplast transient expression system, we improved the efficiency of protein subcellular localization assay and developed the genome editing system in callus protoplasts, which provides an approach for prompt test of CRISPR vectors.

Subcellular Distribution and Chemical Forms of Cadmium in the Medicine Food Homology Plant Platycodon grandiflorum(Jacq.)A.DC.

Although Platycodon grandiflorum(Jacq.)A.DC.is a renowned medicine food homology plant,reports of excessive cadmium(Cd)levels are common,which affects its safety for clinical use and food consumption.To enable its Cd levels to be regulated or reduced,it is necessary to first elucidate the mechanism of Cd uptake and accumulation in the plant,in addition to its detoxification mechanisms.This present study used inductively couple plasma-mass-spectrometry to analyze the subcellular distribution and chemical forms of Cd in different tissues of P.grandiflorum.The experimental results showed that Cd was mainly accumulated in the roots[predominantly in the cell wall(50.96%-61.42%)],and it was found primarily in hypomobile and hypotoxic forms.The proportion of Cd in the soluble fraction increased after Cd exposure,and the proportion of insoluble phosphate Cd and oxalate Cd increased in roots and leaves,with a higher increase in oxalate Cd.Therefore,it is likely that root retention mechanisms,cell wall deposition,vacuole sequestration,and the formation of low mobility and low toxicity forms are tolerance strategies for Cd detoxification used by P.grandiflorum.The results of this study provide a theoretical grounding for the study of Cd accumulation and detoxification mechanisms in P.grandiflorum,and they can be used as a reference for developing Cd limits and standards for other medicine food homology plants.

Analysis of Subcellular Localization and Pathogenicity of Plum Bark Necrosis Stem-Pitting Associated Virus Protein P6

Infection of plum bark necrosis stem pitting associated virus(PBNSPaV)has been reported in many Prunus species in several countries,causing significant economic losses.The very small proteins encoded by plant viruses are often overlooked due to their short sequences and uncertain significance.However,numerous studies have indicated that they might play important roles in the pathogenesis of virus infection.The role of small hydrophobic protein P6,encoded by the open reading frame 2 of PBNSPaV,has not been well explored.In this study,we amplified the P6 fragment from a PBNSPaV isolate by RT-PCR using specific primers and found that it is 174 bp long and encodes a protein of approximately 6.3 kD with a transmembrane domain.Subcellular localization analysis of P6 proteins in tobacco leaves showed that P6 localizes to the cytomembrane and nuclear membrane.To further clarify the pathogenicity of P6 proteins,we constructed a PVX-P6 expression vector by inserting the p6 fragment into a potato virus X(PVX)-based vector and transformed it into Agrobacterium tumefaciens GV3101.Infiltration of Nicotiana benthamiana(N.benthamiana)with the PVX vector-transformed A.tumefaciens led to slight mosaic symptoms at 14 days of post-inoculation.Meanwhile,infiltration with the PVX-P6 vector-transformed A.tumefaciens resulted in no significant symptoms.These results demonstrated that heterologous expression of P6 in N.benthamiana could not enhance the pathogenicity of PVX.Our study indicates that P6 may not be a potential pathogenic factor associate with the causing of symptoms,and the mode of action of PBNSPaV-P6 protein remains to be further studied.

Subcellular Distribution of Cadmium in Mining Ecotype Sedum alfredii

The mining ecotype Sedum alfredii Hance could tolerate and grow normally in a nutritive solution containing cadmium (Cd) as high as 400 mumol/L. Under such a high Cd concentration, the subcellular accumulation of Cd in root, stem and leaf of this plant was found to be the highest in the cell wall, less in the soluble fraction and lowest in the cell organs. The mode of subcellular distribution of Cd in the mining ecotype S. alfredii was similar to other hyper accumulators of heavy metals, in which Cd was distributed more in the aerial part of plant. The results suggest that the mining ecotype S. alfredii is a new species of Cd hyperaccumulator.

Prediction of Subcellular Localization of Eukaryotic Proteins Using Position-Specific Profiles and Neural Network with Weighted Inputs

Subcellular location is one of the key biological characteristics of proteins. Position-specific profiles （PSP） have been introduced as important characteristics of proteins in this article. In this study, to obtain position-specific profiles, the Position Specific lterative-Basic Local Alignment Search Tool （PSI-BLAST） has been used to search for protein sequences in a database. Position-specific scoring matrices are extracted from the profiles as one class of characteristics. Four-part amino acid compositions and lst-7th order dipeptide compositions have also been calculated as the other two classes of characteristics. Therefore, twelve characteristic vectors are extracted from each of the protein sequences. Next, the characteristic vectors are weighed by a simple weighing function and inputted into a BP neural network predictor named PSP-Weighted Neural Network （PSP-WNN）. The Levenberg-Marquardt algorithm is employed to adjust the weight matrices and thresholds during the network training instead of the error back propagation algorithm. With a jackknife test on the RH2427 dataset, PSP-WNN has achieved a higher overall prediction accuracy of 88.4% rather than the prediction results by the general BP neural network, Markov model, and fuzzy k-nearest neighbors algorithm on this dataset. In addition, the prediction performance of PSP-WNN has been evaluated with a five-fold cross validation test on the PK7579 dataset and the prediction results have been consistently better than those of the previous method on the basis of several support vector machines, using compositions of both amino acids and amino acid pairs. These results indicate that PSP-WNN is a powerful tool for subcellular localization prediction. At the end of the article, influences on prediction accuracy using different weighting proportions among three characteristic vector categories have been discussed. An appropriate proportion is considered by increasing the prediction accuracy.

Cd Distribution and Subcellular Localization in Leaf and Its Effects on Growth of Soybean(Glycine max) Seedlings

In order to investigate Cd accumulation, subcellular distribution, and local-ization in soybean seedlings leaves, soybean seedlings were cultivated in solution containing different concentrations of Cd. The results showed that most Cd associ-ated with the cellwal s and soluble fractions, and a minor part of Cd presented in mitochondria fractions, nuclear and chloroplast fractions, especial y exposure to high Cd concentrations. Under 20.00 mg/L Cd stress, Cd subcellular distribution fol owed a sequence as： soluble fractions （55.00%）＆gt;cellwal s （30.0%）＆gt;mitochondria fractions （8.21%）＆gt;nuclear and chloroplast fractions （6.79%）. Deposited Cd black particles were observed in cellwal s, chloroplasts, nuclei, and vacuoles through electrical microscope slice. This fact indicated that the cellwal s of soybean leaves were the first protecting organel es from Cd toxicity, and the cellwal s and soluble fractions were the main place for Cd storage. Due to Cd accumulated in the organel es, the intercellular space was enlarged and the subcellular structure was damaged, especial y for the chloroplasts.

Activities, Quantitative Changes and Subcellular Localization of α-Amylase During Development of Apple Fruit

Starch degradation in cells is closely associated with cereal seed germination, photosynthesis in leaves, carbohydrate storage in tuberous roots, and fleshy fruit development. α_Amylase is considered as one of the key enzymes catalyzing starch breakdown, but up to date its role in starch breakdown in living cells remains unclear because the enzyme was often shown extrachloroplastic in living cells. The present experiment showed that α_amylase activity was progressively increasing concomitantly with the decreasing starch concentrations during the development of apple ( Malus domestica Borkh cv. Starkrimson) fruit. The apparent amount of α_amylase assessed by Western blotting also increased during the fruit development, which is consistent with the seasonal changes in the enzyme activity. The enzyme subcellular_localization studies via immunogold electron_ microscopy technique showed that α_amylase visualized by gold particles was predominantly located in plastids, but the gold particles were scarcely found in other subcellular compartments. A high density of the enzyme was observed at the periphery of starch granules during the middle and late developmental stages. These data proved that the enzyme is compartmented in its functional sites in the living cells of the fruit. The predominantly plastid_distributed pattern of α_amylase in cells was shown unchanged throughout the fruit development. The density of gold particles (α_amylase) in plastids was increasing during the fruit development, which is consistent with the results of Western blotting. So it is considered that α_amylase is involved in starch hydrolysis in plastids of the fruit cells.

Transformation of Arabidopsis by Rice OsWRKY78::GFP Fusion Gene and Subcellular Localization of OsWRKY78 Protein

[Objective] The study was to understand the subcellular localization of OsWRKY78 protein in plants. [Method] Primers specific for OsWRKY78 gene were designed according to the OsWRKY78 full length sequence in Genbank. The gene was cloned by RT-PCR method. The gene was then recombined into a plasmid expression vector carrying green fluorescent protein (GFP) gene, pBinGFP. The recombinant was confirmed by PCR and enzyme digestion. The recombinant plasmid pBinGFP-OsWRKY was transformed into Arabidopsis through Agrobacterium tumefaciens strain GV3101 and transgenic plants were obtained. [Result] Measured by fluorescence microscopy, the expression of OsWRKY78 and GFP fusion protein in root tip cells was localized in the nucleus. [Conclusion] This study laid the foundation for further investigating the function of OsWRKY78 gene and its role in related signal transduction and provided theoretical basis for exploring the relation between OsWRKY78 gene and brown planthoppers.

Expression of Recombinant Tryptophan Decarboxylase in Different Subcellular Compartments in Tobacco Plant

The gene encoded for tryptophan decarboxylase (TDC), which is the key enzyme in terpenoil indole alkaloids pathway, was targeted to different subcellular compartments and stably expressed in transgenic tobacco (Nicotiana tabacum L.) plants at the levels detected by Western blot and tryptamine accumulation analysis. It was shown that the TDC was located in subcellular compartments, the chloroplasts and cytosol. The recombinant TDC targeted to chloroplasts and cytosol in tobacco plants was effectively expressed as soluble protein by Western blot analysis and enzymatic assay. The level of tryptamine accumulation in chloroplast was higher than that in cytosol and very low in vacuole and endoplasmic reticulum (ER) to be hardly detected by Western blot analysis. It was indicated that the highest amount of tryptamine was in chloroplasts, lower in endoplasmic reticula and the lowest in vacuoles as compared to those in wild type plants. The TDC targeted to different subcellular compartments of tobacco plants and its expression level were studied by different nucleotide sequences coding signal peptides at 5'-end of tdc gene in order to know the effects of the TDC in compartmentation on its functionality.

Subcellular Localization of Aquaporins OsPIP2-6 in Rice

[Objective] This study was to explore the subcellular localization of aquaporins OsPIP2-6 in rice. [Method] A key rice aquaporins gene OsPIP2-6 was cloned and used for construction of a transient expression vector,which was then transformed into onion epidermis via particle bombardment for confocal microscopy analysis using YFP gene as a reporter gene. [Result] The results showed that rice aquaporins OsPIP2-6 was mainly located in the plasma membrane. [Conclusion] Our results provided theoretical basis for further understanding plant aquaporins.

Expression of Strictosidine Synthase in Selected Subcellular Compartments of Tobacco Plant

Strictosidine synthase (STR) is a key enzyme involved in the biosynthesis of terpenoid indole alkaloids (TIA) by condensing tryptamine and secologanin into strictosidine. The transgenic tobacco plants targeting STR to subcellular compartments were established to express STR in chloroplast, vacuole and endoplasmic reticulum (ER) by the tobacco stable transformation. It was shown that STR was effectively expressed in the above subcellular compartments by Western blot analysis and STR enzymatic assay. In vitro , STR enzymatic assay was measured indirectly by fluorimetrically detecting depletion of tryptamine feeding on secologanin in the reaction mixture. The tryptamine were completely depleted by STR in the crude extract of leaves of transgenic tobacco plants targeting and expressing STR in the chloroplast, vacuole and ER, which ascertained the STR functionally targeted to the three subcellular compartments. To confirm STR correct targeting and expressing in chloroplast, the chloroplasts were isolated and the fractions of purified chloroplasts were analyzed by Western blot. The hypothesis of STR correct targeting to the chloroplast was tested. The results have implications on our understanding of the complex intracellular trafficking in metabolic intermediates of TIA biosynthesis.

Protective Actions of Blumea Flavanones on Primary Cultured Hepatocytes and Liver Subcellular Organelle against Lipid Peroxidation

To search for the protective actions of blumea flavanones (BFs) on hepatocytes and hepatic subcellular organelle against lipid peroxidation, monkey′s hepatocytes were isolated and cultured with or without blumea flavanones, then damaged by FeSO 4 cysteine or CCl 4. The lipid peroxidation (malondialdehyde production) and alteration in hepatocyte membrane (leakage of GPT) were estimated. Hepatic subcellular organelles were also isolated and incubated with or without blumea flavanones, then injured by FeSO 4 ascorbate. The generation of malondialdehyde(MDA) was measured. It was found that BFs 10 and 100 μmol·L 1 inhibited the MDA generation and GPT (glutamic pyruvic transaminase) leakage out of hepatocytes that were induced by CCl 4 or FeSO 4 cysteine. BFs could prevent lipid peroxidation initiated by FeSO 4 ascorbate in subcellular organelle suspension. Among BFs, BF 2 possessed the strongest activity. Conclusion: Blumea flavanones possess antioxidation activities that protect monkey′s hepatocytes and hepatic subcellular organelle against injuries induced by FeSO4 or CCl 4.

Study on Effects of Calcium on Photosynthetic Characteristics and Its Subcellular Distribution in Rice at Seeding Stage

Using the cultivars with high and low calcium in brown rice as tested ma- terials, the effects of calcium on photosynthetic characteristics and its subcellular dis- tribution in different rice cultivars at seedling stage were investigated with hydrocul- ture method. The results showed that the photosynthetic rate in leaves of high cal- cium cultivar was higher than that low calcium cultivar. The calcium content in leaves of rice was higher than that in roots. Among various tissues and organs, the calcium contents ranked as cell wall〉ribosome and intracellular soluble components〉 plastid （or chloroplasts）〉mitochondria〉nucleus. Cell wall is the most important distri- bution place for calcium, while calcium is rarely distributed in nuclei or mitochon- drias. In all the organelles, the calcium concentrations were all increased with the increased treatment concentration of calcium, especially in high calcium cultivar. The high calcium treatment showed more significant effects on calcium contents in im- portant organelles of high calcium cultivar, so it is speculated that the high calcium cultivar had higher calcium accumulation and tolerance capacities.

Subcellular localization of copper in tolerant and non-tolerant plant

The ability of Elsholtzia splendens Naki(E. splendens) to accumulate copper appears to be governed by its high degree of copper tolerance. However, the tolerance mechanism on the physiological basis is unknown. Using transmission electron microscope(TEM) and energy dispersive analysis of X-rays(EDX), the likely location of copper within the cells of the tolerant and non-tolerant was determined. Here the role of vacuolar and cell wall compartmentalization in this copper tolerant plant were investigated. A direct comparison of copper locations of E. splendens and the non-tolerant Astragalus sinicus L.(A. sinicus) showed that the majority of copper in the tolerant was localized primarily in the vacuolar, cell wall, on the plasmamembrane, beside lipid grains induced by copper pollution, in the chloroplasts and amyloids; but in the non-tolerant, copper precipitates only be observed on the plasmamembrane, in the chloroplasts and cytoplasm under copper exposure conditions that were toxic to both species. This revealed that the tolerant accumulates more copper in the vacuole and cell wall than the non-tolerant, where was regarded as the storage compartment of tolerant plant or hyperaccumulator for heavy metals.

Uptake, Subcellular Distribution, and Chemical Forms of Cadmium in Wild-Type and Mutant Rice

Wild-type （Zhonghua 11） and mutant rice （Oryza sativa L.） plants were used to investigate the effect of cadmium （Cd） application on biomass production, to characterize the influx of Cd from roots to shoots, and to determine the form, content, and subcellular distribution of Cd in the roots, leaf sheaths, and leaves of the rice plants. Seedlings were cultivated in a nutrient solution and were treated with 0.5 mmol L^-1 of Cd^2＋ for 14 d. The sensitivity of rice plants to Cd toxicity was tested by studying the changes in biomass production and by observing the onset of toxicity symptoms in the plants. Both the wild-type and mutant rice plants developed symptoms of Cd stress. In addition, Cd application significantly （P ≤ 0.01） decreased dry matter production of roots, leaf sheaths, and leaves of both types, especially the mutant. The Cd content in roots of the mutant was significantly （P ≤0.05） higher than that of the wild-type rice. However, there was no significant difference in the Cd content of roots, leaf sheaths, and leaves between the wild-type and mutant rice. Most of the Cd was bound to the cell wall of the roots, leaf sheaths, and leaves, and the mutant had greater Cd content in cell organelles than the wild type. The uneven subcellular distribution could be responsible for the Cd sensitivity of the mutant rice. Furthermore, different chemical forms of Cd were found to occur in the roots, leaf sheaths, and leaves of both types of rice plants. Ethanol-, water-, and NaCl-extractable Cd had greater toxicity than the other forms of Cd and induced stunted growth and chlorosis in the plants. The high Cd content of the toxic forms of Cd in the cell organelles could seriously damage the cells and the metabolic processes in mutant rice plants.

Zn Accumulation and Subcellular Distribution in the Zn Hyperaccumulator Sedurn alfredii Hance

Zn accumulation and subcellular distribution in leaves of the hyperaccumulating ecotype （HE） and non-hyperaccumulating ecotype （NHE） of Sedum alfredii Hance were studied using radiotracer and gradient centrifugation techniques. Leaf Zn accumulation in the HE of S. alfredii was 18.5-26.7 times greater than that in the NHE when the plants were grown at 1-500μmol Zn L-1. Leaf section uptake of 65Zn was highly dependent on external Zn levels. Greater 65Zn uptake in HE was noted only at external Zn levels 〉 100μmol L-1. Zinc subcellular distribution in the leaves of the two ecotypes of S. alfredii was： cell wall 〉 soluble fraction 〉 cell organelle. However, more Zn was distributed to the leaf cell wall and soluble fractions for HE than for NHE. In the leaf of HE, 91%-94% of the Zn was found in the cell walls and the soluble fraction and only 6%-9% Zn was distributed in the cell organelle fraction. For NHE, about 20%-26% Zn was recovered in the cell organelle fraction. In stems, Zn distribution to the ceil wail fraction was approximately two fold greater in the HE than that in the NHE. For the hyperaccumulating ecotype of S. alfredii, the cell wall and the vacuole played a very important role in Zn tolerance and hyperaccumulation.

Analysis of Specific Binding and Subcellular Localization of Wheat ERF Transcription Factor W17

The study aims to detect the subcellular localization of ERF （ethylene-responsive element binding factor） transcription factor W17 protein, the interaction between W 17 and cis-acting regulatory elements GCC-box and DRE in vitro, the binding and transactivating ability in vivo, and the role of W17 in higher plant stress-signal pathway. Recombinant plasmid W17/163hGFP was introduced into onion epidermal cells by the particle bombardment method with a PDS 1000/He. Transformed cells were incubated for 24 h at 22℃ in the dark and green fluorescence was monitored under a confocal microscope. The gene W17 was fused N-terminus of GST （glutathione-S-transferase） in prokaryotic expression vector pGEX-4T-1 and then transformed into E. coli strain BL21 （DE3）. IPTG （0.5 mmol L-1） was added to induce the expression of recombinant GST/W17 for 3 h. The fused proteins were purified by GST purification columns, and then subjected to gel retardation assay with a 32p-labeled GCC or DRE sequence. The different reporter and effector plasmids were introduced into tobacco leaves through agroinfiltration, then transformed leaves stained by X-Gluc, faded with 75% alcohol and monitored under a Stereozooming microscope. The GFP fused with W17 protein was localized in the nuclei; SDS-PAGE assay demonstrated that the fused protein GST/W17 could be induced and purified with molecular weight at around 42.2 kD under the induction of IPTG. Purified fused protein was able to specifically bind to both the wild-type GCC-box and DRE element, but had no interaction with either the mutant DRE or GCC-box; W17 protein can bind to GCC-box and transactive downstream GUS gene in vivo. W17 can localize into the nuclei, and it may be involved not only in biotic stresses controlled by GCC-box, but also in abiotic stresses （e.g., salt-） induced signaling pathway.