Sucrose- and H+-Dependent Charge Movements Associated with the Gating of Sucrose Transporter ZmSUT1

Background

In contrast to man the majority of higher plants use sucrose as mobile carbohydrate. Accordingly proton-driven sucrose transporters are crucial for cell-to-cell and long-distance distribution within the plant body. Generally very negative plant membrane potentials and the ability to accumulate sucrose quantities of more than 1 M document that plants must have evolved transporters with unique structural and functional features.

Methodology/Principal Findings

To unravel the functional properties of one specific high capacity plasma membrane sucrose transporter in detail, we expressed the sucrose/H⁺ co-transporter from maize ZmSUT1 in Xenopus oocytes. Application of sucrose in an acidic pH environment elicited inward proton currents. Interestingly the sucrose-dependent H⁺ transport was associated with a decrease in membrane capacitance (C_m). In addition to sucrose C_m was modulated by the membrane potential and external protons. In order to explore the molecular mechanism underlying these C_m changes, presteady-state currents (I_pre) of ZmSUT1 transport were analyzed. Decay of I_pre could be best fitted by double exponentials. When plotted against the voltage the charge Q, associated to I_pre, was dependent on sucrose and protons. The mathematical derivative of the charge Q versus voltage was well in line with the observed C_m changes. Based on these parameters a turnover rate of 500 molecules sucrose/s was calculated. In contrast to gating currents of voltage dependent-potassium channels the analysis of ZmSUT1-derived presteady-state currents in the absence of sucrose (I = Q/τ) was sufficient to predict ZmSUT1 transport-associated currents.

Conclusions

Taken together our results indicate that in the absence of sucrose, ‘trapped’ protons move back and forth between an outer and an inner site within the transmembrane domains of ZmSUT1. This movement of protons in the electric field of the membrane gives rise to the presteady-state currents and in turn to C_m changes. Upon application of external sucrose, protons can pass the membrane turning presteady-state into transport currents.     

商州产大马士革玫瑰精油研究

采用ISO9842:2003玫瑰油标准,对陕西商州引种的保加利亚主栽玫瑰品种“Kazanluk”(RosadamascenaMiller)精油进行分析。结果表明,该玫瑰精油香气具有天然玫瑰花香,香气细腻、柔和、飘逸。油为液体,淡黄色,相对密度(20/20℃)0.8599,折光指数(20℃)1.4592,旋光度(20℃)-4.3°,冻点20.6℃,酯值11.9,这些物理指标达到了标准要求。大量主香成分相对含量:乙醇0.5%,香茅醇45.5%,橙花醇4.5%,香叶醇7.6%,苯乙醇2.9%,十七烷烃1.13%,十九烷烃15.5%,二十一烷烃5.0%。这些成分含量与原产地保加利亚玫瑰油成分范围有一定差异,但与摩洛哥同类玫瑰油标准相符,可以作为商品油在国际市场销售。     

The organic cation transporters rOCT1 and hOCT2 are inhibited by cGMP

The electrogenic cation transporters OCT1 and OCT2 in the basolateral membrane of renal proximal tubules mediate the first step during secretion of organic cations. Previously we demonstrated stimulation and change of selectivity for rat OCT1 (rOCT1) by protein kinase C. Here we investigated the effect of cGMP on cation transport by rOCT1 or human OCT2 (hOCT2) after expression in human embryonic kidney cells (HEK293) or oocytes of Xenopus laevis. In HEK293 cells, uptake was measured by microfluorimetry using the fluorescent cation 4-(4-(dimethyl-amino)styryl)-N-methylpyridinium iodide (ASP + ) as substrate, whereas uptake into Xenopus laevis oocytes was measured with radioactively labelled cations. In addition, ASP +-induced depolarizations of membrane voltages (Vm) were measured in HEK293 cells using the slow whole-cell patch-clamp method. Incubation of rOCT1-expressing HEK293 cells for 10 min with 100 mM 8-Br-cGMP reduced initial ASP + uptake by maximally 78% with an IC50 value of 24 +/- 16 mM. This effect was not abolished by the specific PKG inhibitor KT5823, indicating that a cGMP-dependent kinase is not involved. An inhibition of ASP + uptake by rOCT1 in HEK293 cells was also obtained when the cells were incubated for 10 min with 100 mM cGMP, whereas no effect was obtained when cGMP was given together with ASP +. ASP + (100 mM)-induced depolarizations of Vm were reduced in the presence of 8-Br-cGMP (100 mM) by 44 +/- 11% (n = 6). Since it could be demonstrated that [3H]cGMP is taken up by an endogeneous cyanine863-inhibitable transporter, the effect of cGMP is probably mediated from inside the cell. Uptake measurements with [14C]tetraethylammonium and [3H]2-methyl-4-phenylpyridinium in Xenopus laevis oocytes expressing rOCT1 performed in the absence and presence of 8-Br-cGMP showed that cGMP does not interact directly with the transporter. The data suggest that the inhibition mediated by cGMP observed in HEK293 cells occurs most likely via a mammalian cGMP-binding protein that interacts with OCT1-2 transporters.     

豆科黄结属模式及其相关种名的考证

长期以来,Ｔermopsis lanceolata R.Br.被误认为是黄华属的模式。根据有关文献考证和国际植物命名法规,作者为认Ｔ.;lanceoltata是一个不合法名,黄华属的合法模式应为Ｔhermopsis Lupinoides(L.)Link.分布于东北亚,花具互生花的黄华长期被误定为Ｔ。lupinoides(L.)Link,应更正为Ｔhermopsis fabacea(Pall)DC.。     

Reaction of monoclonal antibodies with plasma membrane proteins after binding on nitrocellulose: renaturation of antigenic sites and reduction of nonspecific antibody binding

The immunochemical reaction of monoclonal antibodies directed against native membrane proteins was investigated after their separation in sodium dodecyl sulfate polyacrylamide gels and electrotransfer to nitrocellulose. Nonspecific binding of antibodies to membrane proteins, which was increased by beta-mercaptoethanol treatment or heat denaturation of the antibodies, could be significantly reduced if 1 M D-glucose plus 10% (v/v) glycerol was added during the incubation with the antibodies. It was found that specific antibody binding was drastically reduced by SDS treatment of the membrane proteins. During the electrotransfer to nitrocellulose and the simultaneous removal of SDS, some increase in antibody binding was observed. Considerable renaturation of antigenic sites in the blotted proteins could be induced if the nitrocellulose blots were incubated for 16 h at 37 degrees C in phosphate-buffered saline. With the introduction of both modifications, the renaturation step, and the addition of D-glucose and glycerol to reduce nonspecific antibody binding, the immunoblot technique may be successfully applied to detect conformational antibodies against membrane proteins.     

高通量扩增子测序分析生防细菌对丹参根际和根表土壤真菌群落多样性的影响

利用高量测序方法探究生防细菌对丹参植株根际和根表土壤真菌群落多样性的影响。

向丹参植株根部施入生防细菌DS-R5,培养45 d后采集根际和根表土壤样品提取总DNA,扩增样品基因组DNA的V4―V5区后进行双端测序,利用生物信息学解析生防细菌对丹参植株根际和根表土壤真菌群落多样性的影响。

菌株DS-R5处理后增加了根际和根表土壤真菌群落的多样性和丰度;根际土壤共有物种种类大于根表土壤,说明菌株DS-R5处理后根际土壤处理与对照物种种类更接近,而对根表土壤中的微生物物种影响较大。真菌群落结构组成分析结果表明,不同土壤样品在门水平上共有优势真菌主要有子囊菌门、接合菌门、担子菌门和未分类;相比根表土壤对照样品,根表土壤处理样品中子囊菌门丰度下降了13.0%,接合菌门丰度升高了69.2%;根际土壤处理样品相比根际土壤对照样品,子囊菌门和接合菌门丰度分别升高了5.9%和8.9%,但二者差异无统计学意义。在属水平上,根表土壤样品经菌株DS-R5处理后提高了有益菌属的丰度,同时降低了有害菌属的丰度。

丹参植株施入生防细菌后,改变了根际土壤和根表土壤中微生物群落结构和多样性,本研究结果可以为利用生防细菌防控丹参根腐病提供理论参考。

     

日光温室栽培对杏花及果实生长发育的影响

对日光温室内与露地栽培的金太阳杏的花期物候、花型及果实生长发育进行了系统观察与分析,结果表明,日光温室栽培比露地栽培的始花期提前33d,花期延长4d,不完全花比例上升33.25%;果枝上花的有效性顺序由露地时的中果枝>长果枝>短果枝>花束状果枝变为温室栽培的短果枝>花束状果枝>中果枝>长果枝;日光温室栽培使杏果实发育的第 、 阶段延长,第 阶段缩短,整个生育期延长15d。统计分析认为:日光温室栽培的第 阶段生长速率显著低于露地,第 阶段的累积生长量显著高于露地。较低的夜间温度是造成温室内杏果实第 阶段较长、生长较慢以及果个变大的原因。