Identification of integrin-like in guard cells ofVicia faba

We addressed the existence and localization of integrin-like in guard cells ofVicia faba by using a probe of polyclonal antibody against the human integrin (α_vβ₃/β₅). Western blot results showed that three integrins-like of about 47.3, 43.7 and 41.1 ku were detected from the preparation of membrane fragments of purified guard cell protoplasts. Further research with immunofluorescent scanning microscopy indicated that those integrins-like were localized on plasma membrane of guard cells, most nearing the dorsal wall, which is consistent with the reception of signals from epidermal cells to guard cells. Thus our results indicate, for the first time, that integrins-like are present at guard cell plasma membrane ofVicia faba.     

50 ku keratin-like protein and β-microtublin coexist in higher plant cells

IF-like proteins have been obtained from suspension cells of Nicotiana tabacum by selective extraction. Western blot analysis shows that the major components of IF-like proteins are 6 keratin-like proteins of 64, 58, 55, 54, 50 and 45 ku. Specially the 50 ku ptotein also reacts with polyantibody against microtublin. Two-dimensional gel electrophoresis shows that the 50 ku protein is composed of two different proteins and their amino acid sequences have been determined. Part of the sequence of one protein is identical to that of β-microtublin and the other protein's sequence has no significant homologue, which should be a new sequence-unknown protein. These results suggest that 50 ku keratin-like protein and β-microtublin coexist in higher plant cells, and that may lead to the phenomenon of co-distribution of IF and microtuble in plant cells.     

50 ku keratin-like protein and -microtublin coexist in higher plant cells

IF-like proteins have been obtained from suspension cells of Nicotiana tabacum by selective extraction. Western blot analysis shows that the major components of IF-like proteins are 6 keratin-like proteins of 64, 58, 55, 54, 50 and 45 ku. Specially the 50 ku protein also reacts with polyantibody against microtublin. Two-dimensional gel electrophoresis shows that the 50 ku protein is composed of two different proteins and their amino acid sequences have been determined. Part of the sequence of one protein is identical to that of -microtublin and the other protein's sequence has no significant homologue, which should be a new sequence-unknown protein. These results suggest that 50 ku keratin-like protein and -microtublin coexist in higher plant cells, and that may lead to the phenomenon of co-distribution of IF and microtuble in plant cells.     

Immunochemical identification of gelsolin in maize pollen

Gelsolin is a representative of a type of actin-binding proteins (ABPs) universally found in eukaryotes. It plays role in nucleation, capping and severing of actin filamentsin vitro. In our experiment, gelsolin was purified from pig plasma and the polyclonal antibodies against it were prepared. The crude extracts of maize pollen were immunodetected by Western-blotting with polyclonal antibody and monoclonal antibody respectively. The immunodetection results show that gelsolin exists in maize pollen and its molecular weight is about 91 ku, similar to that of gelsolin found in animal tissues.     

甜菜碱处理种子对小麦和玉米幼苗抗盐性的效应

在不同盐度下用不同浓度的甜菜碱溶液处理小麦和玉米种子，结果表明，小麦幼苗的游离脯氨酸含量地上部分和根部都比对照高，玉米幼苗的游离脯氨酸含量地上部分高于对照，根部低于对照；小麦和玉米叶片的质膜透性都明显减小．初步证明，用甜菜碱处理小麦和玉米种子与其幼苗的抗盐性有一定关系     

结核杆菌19ku脂蛋白的表达与纯化

目的:通过基因工程技术获得重组结核分枝杆茵19 ku蛋白。方法:应用PCR技术扩增卡介苗的19 ku蛋白DNA序列;以质粒pET28a为表达栽体,构建19 ku重组质粒,然后转化大肠埃希菌BL21(DE3);在异丙基硫代-β-D-半乳糖苷(IPTG)诱导下,分别对不同诱导时间的表达产物进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE),凝胶经考马斯亮蓝染色检测蛋白。通过镍柱纯化后获得目的蛋白。结果:重组质粒pET28a-p19测序表明与报道的序列相同。它在大肠埃希菌BL21(DE3)细胞内以可溶性形式表达。不同IPTG诱导时间实验表明重组结核分枝杆茵l9 ku蛋白诱导4 h在大肠埃希菌中的表达量最高。结论:pET28a-p19大肠埃希菌工程株可高表达结核分枝杆菌重组19 ku蛋白。     

玉米大斑病菌HT-毒素特异性组分与原生质膜蛋白互作研究

以玉米叶片原生质为研究对象,以ANS(1-苯胺基-8-萘磺酸)作荧光探剂,用荧光标记法研究了毒素与质膜蛋白的互作.在被ANS标记后的原生质悬浮液中加入不同浓度HT-毒素后,ANS荧光强度明显增加,呈现出了剂量—效应关系,表明HT-毒素与质膜上ANS某些受体发生了互作.本实验采用2种试剂,从2个角度表明了ANS受体和质膜上存在的毒素受体具有蛋白特性.初步推测HT-毒素特异性组分的原初作用位点在质膜外侧.     

33 ku protein associated several polypeptides with nearly the same molecular weight but not the same isoelectric point

The 33 ku protein, prepared from NaCl-treated PS II particles, has shown an single band by SDS-PAGE. After being dialyzed against the low-osmotic medium at 4°C, it has been found that the 33 ku protein degraded into several small fragments. This result suggests that the preparations of 33 ku protein probably contain some latent proteinases. It has also been found, by the 2-D electrophoresis and IEF, that the preparations of 33 ku protein not dialyzed against the low-osmotic medium contain several polypeptides with nearly the same molecular weight but not the same isoelectric point as the 33 ku protein.     

低温胁迫下解放钟枇杷幼果细胞超微结构的变化

以解放钟枇杷幼果为试验材料,采用人工降温方法,研究低温胁迫下枇杷幼果超微结构的变化。结果表明：6℃、3℃低温胁迫时,解放钟果肉细胞质膜、液泡膜清晰,叶绿体、线粒体结构没有明显的变化;0℃低温处理条件下的原生质膜结构完整,少数叶绿体出现部分解体,双层膜有部分破裂现象,线粒体结构仍清晰可见;-3℃低温胁迫下,原生质膜、液胞膜均破裂,原生质体浓缩,叶绿体扭曲变形、相互融合,线粒体膜结构受损,脊消失。     

超滤技术在反渗透海水淡化预处理中的应用

该文介绍了超滤技术,分析了超滤技术应用于海水淡化预处理的技术优势,考察了超滤系统在反渗透海水淡化预处理工程中的运行情况.实验采用50 ku和10 ku聚砜中空纤维膜组件,讨论了超滤膜孔径、操作压力、过滤时间对预处理出水浊度、COD(Cr)和膜过滤通量的影响,分析了超滤系统出水污染指数(SDI)随时间的变化情况.     

Tn5转座子插入p95基因不影响AcMNPV的复制

从Tn5转座子介导的AcMNPV随机插入突变体库中,分离到一株复制正常的突变体AcApra41.突变定位发现Tn5转座子插入了病毒p95基因中.为了排除AcApra41中还有其他突变,利用同源重组法构建了p95基因定点插入突变的重组病毒AcGFP-P95in.PCR确认p95基因中插入了Tn5转座子;Western blot也证实AcApra41和AcGFP-P95in感染的细胞中,P95蛋白的分子量都因为插入突变而变小,由野生型的95 ku变为 55 ku.病毒复制动态曲线和荧光显微镜观察证实带有该插入突变的病毒能够在Sf9细胞中正常复制,并表达极晚期基因.这一结果表明完整的P95蛋白对病毒复制是非必须的.     

乙烯利对玉米生理效应的研究

经乙烯利处理后的玉米植株体内蛋氨酸和乙烯含量聚增,进而对玉米产生一系列生理效应:根系活力和气生根发育增强,根量在土壤深层的分布增多;地上部表现为先受抑后促进;伴随着植株矮化,玉米叶片细胞质膜透性增强和叶绿素降解;由于乙烯利对根系的促进作用,导致玉米生育后期绿叶数相对增多且功能期延长.本文拟将乙烯利对玉米的生理效应分成先后两个阶段——以抑制为主的初级效应和以促进为主的次级效应.     

Identification of a protein associated with circulative transmission of Barley yellow dwarf virus from cereal aphids, Schizaphis graminum and Sitobion avenae

Using 2-D electrophoresis and virus overlay assay,a 50-kDa protein(P50)exhibiting specific binding to purified virus particles of BYDV-GAV was found in the protein extracts from Schizaphis graminum and Sitobion avenae,two aphid species transmiting BYDV-GAV.P50 in the extracts of S.graminum was isolated by preparation electrophoresis and electro-eluted proteins from the gel slices for antiserum preparation.After feeding the antiserum through membrane,the transmission efficiencies of S.graminum and S.avenae for BYDV-GAV decreased significantly.It was suggested that P50 should be related with transmission process.Location of P50 was found at the plasma membrane surrounding the accessory salivary gland(ASG) in the head tissues of S.graminum by immunogold-labelling experiment.The ascertainment of the protein associated with virus transmission has a significance influence on further understanding the transmission mechanism and genetic engineering for resistant to vector transmission.     

Exposure of an antigen of chromaffin granules on cell surface during exocytosis

The synthesis rate of the membrane proteins of the catecholamine-storing vesicles (chromaffin granules) of the adrenal medulla is lower than that of the secretory proteins of the contents. Based on these results we proposed that after exocytosis the membranes of chromaffin granules are retrieved and are re-used for several secretion cycles (see also ref. 4). This concept of re-use of granule membranes has been further strengthened by the finding that exogenous markers which are taken up by secretory cells during stimulation can be traced to the Golgi region and to immature secretory organelles. However, one basic question remains: are the membranes of secretory organelles specifically and completely removed from the plasma membrane and if so, how fast is this process? By using an antiserum against a membrane glycoprotein of chromaffin granules we have now obtained quantitative data which demonstrate that during exocytosis this antigen becomes exposed on the cell surface and disappears again to a large degree within 30 min.     

哈维氏弧菌TS-628菌株抗原性研究

哈维氏弧菌(Vibrio harveyi)是海水鱼虾养殖中常见的致病菌,由该菌引发的病害给世界各地的养殖业带来重大经济损失,但对其有效抗原的筛选或相关疫苗的研究报道相当少,对其病害的防治也尚无有效措施.本文以患病青石斑鱼分离到的病原菌哈维氏弧菌TS-628菌株为研究对象,分别提取它的鞭毛蛋白、脂多糖(LPS)和外膜蛋白(OMP),并采用Western blot技术分析检测这3种成分的抗原性.结果显示,鞭毛蛋白主要的免疫印迹带约有4条,其中43、52 ku为主要免疫反应显色带;OMP主要的免疫印迹带约有5条,其中43 ku为最主要免疫反应显色带, 35、38、47和52 ku也具有较强的免疫显色反应.LPS没有检测到免疫印迹反应.这一研究结果将为灵敏检测哈维氏弧菌以及研制高效疫苗奠定基础.     

The tobacco aquaporin NtAQP1 is a membrane CO2 pore with physiological functions

Aquaporins, found in virtually all living organisms, are membrane-intrinsic proteins that form water-permeable complexes. The mammalian aquaporin AQP1 has also shown CO2 permeability when expressed heterologously in Xenopus oocytes, although whether this is a biochemical curiosity or of physiological significance is a matter of debate. Here we report that, in the same expression system, a CO2 permeability comparable to that of the human AQP1 is observed for the tobacco plasma membrane aquaporin NtAQP1. NtAQP1 facilitates CO2 membrane transport in the homologous plant system at the cellular level, and has a significant function in photosynthesis and in stomatal opening. NtAQP1 overexpression heightens membrane permeability for CO2 and water, and increases leaf growth. The results indicate that NtAQP1-related CO2 permeability is of physiological importance under conditions where the CO2 gradient across a membrane is small, as is the case between the atmosphere and the inside of a plant cell.     

Seed priming with chitosan improves maize germination and seedling growth in relation to physiological changes under low temperature stress

Low temperature stress during germination and early seedling growth is an important constraint of global production of maize. The effects of seed priming with 0.25%, 0.50%, and 0.75% (w/v) chitosan solutions at 15 °C on the growth and physiological changes were investigated using two maize (Zea mays L.) inbred lines, HuangC (chilling-tolerant) and Mo17 (chilling-sensitive). While seed priming with chitosan had no significant effect on germination percentage under low temperature stress, it enhanced germination index, reduced the mean germination time (MGT), and increased shoot height, root length, and shoot and root dry weights in both maize lines. The decline of malondialdehyde (MDA) content and relative permeability of the plasma membrane and the increase of the concentrations of soluble sugars and proline, peroxidase (POD) activity, and catalase (CAT) activity were detected both in the chilling-sensitive and chilling-tolerant maize seedlings after priming with the three concentrations of chitosan. HuangC was less sensitive to responding to different concentrations of chitosan. Priming with 0.50% chitosan for about 60~64 h seemed to have the best effects. Thus, it suggests that seed priming with chitosan may improve the speed of germination of maize seed and benefit for seedling growth under low temperature stress.