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.

Uptake,Depuration and Subcellular Distribution of Cadmium in Various Tissues of Perna viridis

Green-lipped mussels (Perna viridis) were collected from a local mariculture site and placed in pre-cleaned sea water tanks containing 0, 0.2, 0.5 and 1.3 μg/ml CdCl2. The level of Cd in the gill, viscera and gonad was measured. The concentration of Cd in the 3 tissues increased linearly over 4 days' of exposure. The rate of uptake depends on the concentration of Cd in the water. In the 3 tissues studied, the maximum rate of uptake (Vmax) was highest in the gill (500μg/g dry weight/day), followed by viscera (100μg/g dry weight/day) and gonad (56.8 μg/g dry weight/day). The majority of the Cd taken up was bound to a fraction of the heat-stable proteins similar to metallothioneins. The rate of Cd depuration from the tissues was poor. There was no change in Cd concentration over 8 days' depuration in clean water

Exogenous Nitric Oxide Involved in Subcellular Distribution and Chemical Forms of Cu^(2+) Under Copper Stress in Tomato Seedlings

Nitric oxide（NO）,a bioactive signaling molecule,serves as an antioxidant and anti-stress agent under abiotic stress.A hydroponics experiment was conducted to investigate the effects of sodium nitroprusside（SNP）,a NO donor,on tomato seedlings exposed to 50 μmol L-1CuCl 2.The results show that copper is primarily stored in the soluble cell sap fraction in the roots,especially after treatment with Cu＋SNP treatment,which accounted for 66.2% of the total copper content.The copper concentration gradually decreased from the roots to the leaves.In the leaves,exogenous NO induces the storage of excess copper in the cell walls.Copper stress decreases the proportion of copper integrated with pectates and proteins,but exogenous NO remarkably reverses this trend.The alleviating effect of NO is blocked by hemoglobin.Thus,exogenous NO is likely involved in the regulation of the subcellular copper concentrations and its chemical forms under copper stress.Although exogenous NO inhibited the absorption and transport of excess copper to some extent,the copper accumulation in tomato seedlings significantly increased under copper stress.The use of exogenous NO to enhance copper tolerance in some plants is a promising method for copper remediation.

Subcellular distribution and chemical form of Pb in hyperaccumulator Arenaria orbiculata and response of root exudates to Pb addition

Solution culture was conducted in order to understand accumulation characteristics and chemical forms of Pb in Arenaria orbiculata （A. orbiculata） and the response of root exudates to Pb addition. The results showed that： 1） Pb contents in the shoot and root of A. orbiculata increased with increasing in Pb concentrations in solution. 2） The contents of Pb chemical forms under Pb addition followed as： HAc extractable fraction＇（FriAC）〉 HC1 extractable fraction （FHcl）〉 NaCl extractable fraction （FNacl） 〉 ethanol-extractable fraction （FE） 〉 water extractable fraction （Fw）. 3） Increased Pb level in the medium caused increases in Pb contents in the four subcellular fractions of shoots and roots, with most accumulation in FIV （Fraction ＇IV, sbluble fraction） in shoots and FI （Fraction I, cell wall fraction） in roots. 4） Contents of soluble sugar and free amino acid of root exudates increased with increasing Pb concentration in solution. Significantly positive correlations between Pb and contents of soluble sugar and free amino acid were observed. 5） With Pb concentrations in solution, low molecular weight organic acids （LMWOAs） contents followed the tendency： tartaric acid 〉 acetic acid 〉 malic acid 〉 citric acid. Significantly positive correlation was observed between Pb and citric acid contents. The results indicate that soluble sugars, free amino acid and citric acid in root exudates of A.orbiculata facilitate the absorption and accumulation of Pb, which exist in NaCl-, HCI- and HAc- extractable Pb forms, FI and FIV fractions, resulting in tolerance of A.orbiculata to Pb.

Toxicity and subcellular distribution of cadmium in wheat as affected by dissolved organic acids

We aim to investigate the effects of humic acid （HA） and citric acid （CA） on the toxicity and subcellular distribution of Cd in wheat. Results show that the toxicity and uptake of Cd decreased with increasing HA. The EC50 values of Cd increased from 3.36 μmol/L to 4.96 and 7.33 μmol/L at 50 and 250 mg/L HA, respectively, but decreased to 1.39 μmol/L in the presence of CA based on free ion activity model （FIAM）. HA decreased the relative subcellular distribution of Cd in the heat-denatured proteins （decreased from 54% to 33%） but increased Cd in the heat-stable proteins in root （from 25% to 50%） at 7.61 μmol/L {Cd2＋} （free Cd activity）, which resulted in decreasing Cd toxicity. However, CA increased Cd toxicity due to the increased internalization of Cd although the relative subcellular distributions of Cd exhibited a decrease in the heat-denatured proteins and increase in the granule fraction compared to the control at high-level Cd. The FIAM could not predict the toxicity of Cd in the presence of organic acids. Alternatively, the internal Cd accumulation and subcellular Cd concentration were better to describe the toxicity of Cd to wheat.

Super-resolution imaging reveals the subcellular distribution of dextran at the nanoscale in living cells

Theranostic visualization of dextran at the nanoscale is beneficial for understanding the bioregulatory mechanisms of this molecule. In this study, we applied structured illumination microscopy(SIM) to capture the distribution of Cy5-Dextran at different incubation periods in living cells. The results showed that Cy5-Dextran could be absorbed by He La cells. In addition, we clarified that Cy5-Dextran exhibited differential organelle distribution(lysosomal or mitochondrial) in a time-dependent manner. Moreover,lysosomal Cy5-Dextran localization was found to be independent of the autophagy process, while Cy5-Dextran localized to the mitochondria triggered a pro-apoptotic event, upregulating the levels of reactive oxygen species(ROS) to accelerate mitochondrial fragmentation. This work uses a visualized strategy to reveal the anti-tumor bioactivity of dextran, which was achieved by regulating apoptosis and autophagy.

Subcellular Cd accumulation characteristic in root cell wall of rice cultivars with different sensitivities to Cd stress in soil

The variations of grain cadmiun（Cd） concentrations, translocation factors（TFs） of Cd from roots to shoots/grains of six rice cultivars, characterized with different Cd-sensitivities in polluted soil were studied, the selected rice cultivars were Xiangzao 17（R1）, Jiayu 211（R2）, Xiangzao 42（R3）, Zhuliangyou 312（R4）, Zhuliangyou 611（R5）, and Jinyou 463（R6）, respectively. The Cd subcellular distribution and Cd binding characteristics on subcellular fractions of rice root cell wall（CW） were further investigated. The results showed that the rice grain Cd contents varied significantly, with a maximum variation of 47.0% among the cultivars, the largest grain Cd content was observed with cultivar R1（Cd-sensitivity cultivar） and the smallest with R5（Cd-tolerance cultivar）. The translocation factors of Cd from roots to shoots（TF_（shoot）） and roots to grains（TF_（grain）） varied greatly among the cultivars. In general, the TFgrain of the cultivars followed the order of R1〉R2〉R3〉R4〉 R6-R5. The Cd concentration（mg kg^（–1） FW） in the fraction of root CW, the fraction of cell wall removing pectin（CW-P） and the fraction of cell wall removing pectin and hemicellulose（CW-P-HC） of the cultivars generally followed the order of CW-P〉CW〉CWP-HC; the ratios of Cd concentration（mg kg–1 FW） in the fraction of CW-P to that of CW were mostly more than 1.10, while the ratios of Cd concentration in the fraction of CW-P-HC to that of CW were mostly less than 0.60, indicating that Cd was mainly stored in the hemicellulose of the root CW. The ratios of Cd of CW-P-HC to CW generally followed the descending order of R1~R2〉R3〉R4〉R5~R6 for the cultivars, which implied that hemicellulose is probably the main subcellular pool for transferring Cd into rice grain, and it restrains the translocation of Cd from shoot to the grain, especially for the Cd-tolerance cultivars（R5 and R6）, the compartmentation of more Cd in hemicellulose in root CW is probably one of the main mechanisms for Cd tolerance of rice cultivars.

Calcium Forms, Subcelluar Distribution and Ultrastructure of Pulp Cells as Influenced by Calcium Deficiency in Apple (Malus pumila) Fruits

Calcium in Red Fuji and Starkrimson apples during storage were fractionated by sequentextracting. Localization and distribution of calcium and influence of calcium nutritionon cell ultrastructure were observed by transmission electron microscopy combined within situ precipitation of calcium with an improved method of potassium pyroantimonatetechnique. Results indicated that spraying calcium solution on surface of young fruitsincreased contents of calcium in all forms. During storage, contents of soluble calciumand pectic calcium declined and those in calcium phosphate, calcium oxalate and calciumsilicate increased. Calcium contents of Red Fuji in all forms were higher than those ofStarkrimson, indicating that calcium accumulating capability of Red Fuji fruits precededthat of Starkrimson. Under transmission electron microscopy, calcium antimonite precipitates(CaAP) was mainly distributed in cell wall, tonoplast, nuclear membrane and nucleoplasm,much more CaAP deposited in vacuole. Calcium deficiency during storage leads to decreaseof CaAP in locations mentioned above, disappearance of compartmentation, and entrance ofCaAP to cytoplasm. Transformation from soluble calcium and pectic calcium to calciumphosphate, oxalate and damages of biomembranes structuraly and functionally resultedfrom calcium deficiency during storage were the crucial causation of physiologicaldisorder.

Differential distribution of PINK1 and Parkin in the primate brain implies distinct roles

The vast majority of in vitro studies have demonstrated that PINK1 phosphorylates Parkin to work together in mitophagy to protect against neuronal degeneration.However,it remains largely unclear how PINK1 and Parkin are expressed in mammalian brains.This has been difficult to address because of the intrinsically low levels of PINK1 and undetectable levels of phosphorylated Parkin in small animals.Understanding this issue is critical for elucidating the in vivo roles of PINK1 and Parkin.Recently,we showed that the PINK1 kinase is selectively expressed as a truncated form(PINK1–55)in the primate brain.In the present study,we used multiple antibodies,including our recently developed monoclonal anti-PINK1,to validate the selective expression of PINK1 in the primate brain.We found that PINK1 was stably expressed in the monkey brain at postnatal and adulthood stages,which is consistent with the findings that depleting PINK1 can cause neuronal loss in developing and adult monkey brains.PINK1 was enriched in the membrane-bound fractionations,whereas Parkin was soluble with a distinguishable distribution.Immunofluorescent double staining experiments showed that PINK1 and Parkin did not colocalize under physiological conditions in cultured monkey astrocytes,though they did colocalize on mitochondria when the cells were exposed to mitochondrial stress.These findings suggest that PINK1 and Parkin may have distinct roles beyond their well-known function in mitophagy during mitochondrial damage.

Comparisons in subcellular and biochemical behaviors of cadmium between Iow-Cd and high-Cd accumulation cultivars of pakchoi （Brassica chinensis L.）

Subcellular distributions and chemical forms of cadmium （Cd） in the leaves, stems and roots were investigated in low-Cd accumulation cultivars and high-Cd accumulation cultivars ofpakchoi （Brassica chinensis L.）. Root cell wall played a key role in limiting soil Cd from entering the protoplast, especially in the low-Cd cultivars. The high-Cd cultivars had significantly higher leaf and stem Cd concentrations than the low-Cd cultivars in cell wall fraction, chloroplast/trophoplast fraction, organelle fraction and soluble fraction. In low-Cd cultivars, which were more sensitive and thus had greater physiological needs of Cd detoxification than high-Cd cultivars, leaf vacuole sequestrated higher proportions of Cd. Cd in the form of pectate/protein complexes （extracted by 1 tool. L~ NaC1） played a decisive role in Cd translocation from root to shoot, which might be one of the mechanisms that led to the differences in shoot Cd accumulation between the two types of cultivars. Furthermore, the formation of Cd- phosphate complexes （extracted by 2% HAc） was also involved in Cd detoxification within the roots of pakchoi under high Cd stress, suggesting that the mechanisms of Cd detoxification might be different between low- and high-Cd cultivars.