青稞B组醇溶蛋白基因与上游调控区的克隆及基因表达

高等植物种子胚乳贮藏蛋白是种子发芽时的主要氮源，也是人类和动物食用植物蛋白的主要来源。大麦种子胚乳贮藏蛋白主要是醇溶蛋白（hordeins），占大麦胚乳总蛋白的50–60%。根据大麦醇溶蛋白的大小和组成特点，大麦醇溶蛋白被划分为三种类型：富硫蛋白亚类(B，γ-hordeins)、贫硫蛋白亚类（C-hordeins)以及高分子量蛋白亚类(D-hordeins)。B组和C组醇溶蛋白是大麦胚乳的两类主要贮藏蛋白，它们分别占大麦总醇溶蛋白成分的70–80%和10–12%。遗传分析表明，大麦B、C、D和γ-组醇溶蛋白分别是由位于大麦第五染色体1H（5）上的Hor2、Hor1、Hor3和Hor5位点编码。Hor2位点编码大量分子量相同但组成不同的B组醇溶蛋白（B-hordein）。B-hordein的种类、数量和分布是影响大麦酿造、食用及饲养品质的重要因素之一。为深入了解B-hordein基因家族的结构和染色体组织，探明Hor2位点基因表达的发育调控机制，最终达到改良禾谷类作物籽粒品质的目的，本研究以青藏高原青稞为材料，采用同源克隆法，分别克隆B-hordein基因和启动子，通过原核生物表达验证B-hordein基因功能，并利用实时定量PCR探索B-hordein基因表达时空关系，取得如下研究结果： 1. 以具有特殊B组醇溶蛋白亚基组成的9份青藏高原青稞为材料，根据GenBank中三个B-hordein基因序列（GenBank No. X03103, X53690和X53691）设计一对引物，通过PCR扩增，获得23个B-hordein基因克隆并对其进行了序列分析。核苷酸序列分析表明，所有克隆均包含完整的开放阅读框。有11个克隆都存在一个框内终止密码子，推测这11个克隆可能是假基因。推测的氨基酸序列分析表明，所有大麦B-hordein具有相似的蛋白质基本结构，均包括一个高度保守的信号肽、中间重复区以及C-端结构域。不同大麦种重复区内重复基元的数目有较大差异。青稞材料Z07–2和Z26的B-hordeins仅具有12个重复基元结构，更接近于野生大麦。这些重复基元数目的差异导致了重复区序列长度和结构的变异。这种现象极可能是由于醇溶谷蛋白基因在进化过程中染色体的不平衡交换或复制滑动所造成的。对所克隆基因和禾本科代表性醇溶谷蛋白基因进行聚类分析，结果表明所有来自栽培大麦的B-hordeins聚类成一个亚家族，来自野生大麦的B-hordeins以及普通小麦的LMW-GS聚类成另外一个亚家族，表明这两个亚家族的成员存在显著差异。此外，我们发现B-hordein基因推测的C-末端序列具有一些有规律的特征：即具有相同C-末端序列的B-hordein基因在系统发生树中聚类为同一个亚组（除BXQ053，BZ09-1，BZ26-5分别单独聚为一类外）。这个特征将有助于我们对所有B组醇溶蛋白基因家族成员进行分类，避免了在SDS-PAGE电泳图谱上仅依靠大小分类的局限性。 2. 根据上述克隆的青稞B-hordein基因的5’端序列设计三条基因特异的反向引物，以青稞Z09和Z26的基因组DNA为模板，采用SON-PCR和TAIL-PCR技术分离克隆出8个B-hordein基因的上游调控序列（命名为Z09P和Z26P）。序列分析表明，推测的TATA box位于–80 bp，CAAT–like box位于–140 bp处。此外，Z09P和Z26P中有六个序列在–300 bp处均存在一个由高度保守的EM基序和类GCN4基序构成的胚乳盒(Endosperm Box，EB)，在约–560 bp处存在一个胚乳盒类似结构。而Z09P-2和Z26P-3不存在保守的胚乳盒或其类似结构，预示着这两个启动子所调控的基因表达可能受不同类型反式作用因子的调节，推测该启动子对基因的表达调控具有多样性。 3. 将B-hordein基因的开放阅读框定向克隆到表达载体pET-30a中，将其导入大肠杆菌表达菌株BL21中进行外源基因的诱导表达以验证所克隆基因的功能。结果表明仅含重组子pET-BZ07-2和pET-BZ26-5的BL21细菌有目的表达蛋白产生。在诱导3 h时的蛋白表达量最高；3 mM IPTG诱导的蛋白表达量要高于1 mM IPTG诱导的表达量。这为分离纯化B-hordein蛋白以及进一步研究其对大麦籽粒品质的影响奠定基础。 4. 根据从青稞Z09和Z26中分离克隆的B-hordein基因序列设计一对基因特异的引物，同时，选择大麦α-微管蛋白基因（GenBank no. U40042）为看家基因并设计特异引物，利用实时荧光定量PCR检测了青稞籽粒4个胚乳发育时间段的B-hordein基因表达，荧光定量结果显示：两份材料中B-hordein基因的表达量均随发育过程的进行而逐渐升高。Z09中B-hordein基因在开花后7天开始转录，而Z26开花4天后就有低水平B-hordein的表达，这表明Z26中B-hordein基因可能比Z09表达的较早或者Z09中B-hordein基因表达水平较低以致于不能被检测到。此外，在4个不同的胚乳发育时期中，Z26中B-hordein基因的表达量均高于Z09材料。在开花12天到18天的过程中，Z09和Z26中B-hordein基因的表达水平有一个急剧性的升高。这说明在不同胚乳发育时期，Hor2位点的B-hordein等位基因变异体存在mRNA的差异表达。 Seed endosperm storage proteins in higher plants are the main resources of nitrogen for germinating and plant proteins for human and animals. Barley prolamins (also called hordeins) are the major storage proteins in the endosperm and account for 50–60% of total proteins. Hordeins are classically divided into three groups: sulphur-rich (B, γ-hordeins), sulphur-poor (C-hordeins) and high molecular weight (HMW, D-hordeins) hordeins based on the size and composition. B-hordeins and C-hordeins are two major groups and each respectively account for about 70-80% and 10-12% of the total hordein fraction in barley endosperm. Genetic analysis showed that B-, C-, C-, γ-hordeins are encoded by Hor2, Hor1, Hor3 and Hor5 locus on the chromosome 1H (5). Hor2 locus is rich in alleles that encode numerous heterogeneous B-hordein polypeptides. It is reported that B-hordein species, quantity and distribution are significant factors affecting malting, food and feed quality of barley. To understand comprehensively the structure and organization of B-hordein gene family in hull-less barley and explore the developmental control mechanisms of Hor2 locus gene expression and eventually to better exploitation in crop grain quality improvement, we isolated and cloned B-hordein genes and promotors of hull-less barley from Qinghai-Tibet Plateau by PCR, and testified their expression founction in bacteria expression system and explore their spatial and temporal expression pattern by quantitative real time PCR. Our results are as followed, 1. Twenty-three copies of B-hordein gene were cloned from nine hull-less barley cultivars of Qinghai-Tibet Plateau with special B-hordein subunits and molecularly characterized by PCR, based on three B-hordein genes published previously (GenBank No. X03103, X53690 and X53691). DNA sequences analyses confirmed that the six clones all contained a full-length coding region of the barley B-hordein genes. Eleven clones all contain an in-frame stop codon and they are probably pseudogenes. The analysis of deduced amino acid sequences of the genes shows that they have similar structures including signal peptide domain, central repetitive domain, and C-terminal domain. The number of the repeats was largerly variable and resulted in polypeptides in different sizes or structures among the genes. Twelve such repeated motifs were found in Z07–2 and Z26, and they are close to those of the wild barleys, and it is most probably caused by unequal crossing-over and/or slippage during replication as suggested for the evolution of other prolamins. The relatedness of prolamin genes of barley and wheat was assessed in the phylogenetic tree based on their polypeptides comparison. Our phylogenetic analysis suggested that the predicted B-hordeins of cultivated barley formed a subfamily, while the B-hordeins of wild barleys and the two most similar sequences of LMW-GS of T. aestivum formed another subfamily. This result indicated that the members of the two subfamilys have a distinctive difference. In addition, we found the B-hordeins with identical C-terminal end sequences were clustered into a same subgroup (except BXQ053，BZ09-1 and BZ26-5 as a sole group, respectively), so we believe that B-hordein gene subfamilies possibly can be classified on the basis of the conserved C-terminal end sequences of predicted polypeptide and without the limit of SDS-PAGE protein banding patterns. 2. The specific primers were designed according to the published sequences of barley B-hordein genes from Z09 and Z26. Using total DNA isolated from them as the templates, eight clones (designated Z09Pand Z26P) of upstream sequences of the known B-hordein genes was obtained by TAIL-PCR and SON-PCR. Sequences analysis shows that the putative TATA box was present at position –80 bp and CAAT-like box at position –140 bp. Besides, a putative Endosperm Box including an Endosperm Motif (EM) and a GCN4-Like Motif was found at position –300 bp in six clones, and another Endosperm-like box was found at positon –560 bp. While the Endosperm Box or Endosperm-like box was not found in Z09P-2 and Z26P-3. This may indicate that gene expression drived by the two promtors was probably controlled by different trans-acting factors and the genetic control mechanism of corresponding gene expression may be diverse. 3. The B-hordein genic region coding for the mature peptide was cloned into expression vector pET-30a and transformed into bacterial strain BL21 for identifying gene expression fountion. Protein SDS–PAGE analysis showed that only the transformed lysate with the pET-BZ07-2 and pET-BZ26-5 constructs produced proteins related to B-group hordeins of barley, and the mounts of proteins induced by 3 mM IPTG and 3 h were higher than other conditions. This established a base for isolating and putifying B-hordein and further exploring their effects on barley grain quality. 4. The gene-specific primers of B-hordein genes from Z09 and Z26 were used for the quantification of B-hordein gene expression. The α-tubulin gene from Hordeum vulgare subsp. vulgare (GenBank accession number U40042) was used as a control gene. The result shows the transcription of the B-hordein genes in Z09 was found 7 days after flowering, while the transcription of the B-hordein genes in Z26 was found 4 days after flowering, but at a very low level, and it suggested that the B-hordein genes in Z26 probably expressed earlier than those in Z09, or the B-hordein genes in Z09 expressed at so a lower level than Z26 that it can not detected. In addition, B-hordein genes in Z26 accession showed higher expression levels than those in Z09 in four developing stages. Furthermore, a progressive increase in the expression levels of the B-hordein genes between 12 and 18 days after anthesis was observed in both Z09 and Z26. It implies that the B-hordein allelic variants encoded by Hor2 locus exist the differential expression in mRNA levels of during barley endosperm development.

~(19)F+~(27)Al耗散反应中双核系统的转动与形变

测量了19F+27Al耗散反应产物B,C,N,O,F和Ne的激发函数,入射束流的能量从110．25MeV 到118．75MeV,能量步长为250keV．从产物的能量自关联函数中提取了反应中所形成的中间双核系统的转动惯量,与相粘模型计算的刚体转动惯量相比较,结果表明形成的双核系统有大的形变．

用~(12)C离子模拟质子引起的单粒子效应

在理论分析的基础上 ,提出了一种利用兰州重离子加速器提供的高能 12 C离子模拟质子引起单粒子效应的途径 .在保证核反应机制是引起单粒子效应主要机制的前提下 ,用高能 12 C离子可以模拟质子在功率金属 -氧化物 -半导体场效应晶体管中引起的单粒子烧毁以及单粒子栅极击穿 ,获得质子单粒子效应的饱和截面 ,定性研究质子单粒子效应的角度效应 ,还可以作为高能质子单粒子效应实验前的预备实验 .该方法拓展了兰州重离子加速器加速的轻的重离子在单粒子效应实验研究方面的应用 ,对现阶段国内开展质子单粒子效应实验研究具有重要意义

纯金属Bi中高能重离子辐照效应

利用高能重离子（Ｋｒ， Ｘｅ， Ｓｎ和Ｐb ）辐照处于 １６或 １００ Ｋ的低熔点纯金属 Ｂｉ，通过测量分析辐照引起的样品电阻率增量及其变化速率随辐照剂量的变化，研究了入射离子在Ｂｉ中引起的辐照损伤效应如辐照损伤效率及复杂缺陷的产生等．结果表明，强的电子能损可在纯金属Ｂｉ中引起附加缺陷的产生，从而使得辐照损伤效率＞１．电子能损值大时，入射离子在Ｂi中引起的辐照效应主要是电子能损效应．辐照温度和入射离子速度对辐照效应的强弱有一定的影响．

C离子注入Si中Si_(1-x)C_x合金的形成及其稳定性

利用离子注入和高温退火的方法在Si中生长了C含量为0.6%~1.0%的Si1?xCx合金, 研究了不同注入剂量下Si1?xCx合金的形成及其在退火过程中的稳定性. 如果注入剂量小于引起Si非晶化的剂量, 850℃退火后, 注入产生的损伤缺陷容易与C原子结合形成缺陷团簇, 难于形成Si1?xCx合金. 随着注入C离子剂量的增大, 注入产生的损伤增强, 容易形成Si1?xCx合金, 但注入的剂量增大到一定程度, Si1?xCx合金的应变将趋于饱和, 即只有部分C原子进入晶格位置形成合金相. Si1?xCx合金一旦形成, 在950℃仍比较稳定, 而温度高于1 000℃, 合金的应力将部分释放. 随着合金中C原子浓度的升高, 合金的稳定性变差.

Rotation and deformation of the dinuclear system formed in dissipative reaction of F-19+Al-27

Excitation functions of the reaction products B, C, N, O, F and Ne emitted from the dissipative reaction of (19) F+(27) Al have been measured at incident energies from 110.25MeV to 118.75MeV in steps of 250keV. The moments of inertia of the intermediate dinuclear system formed in the reaction are extracted from the energy autocorrelation functions of the products. Comparing the moment of inertia extracted from the experimental data with the calculated one by using the sticking limit, it indicates that the formed dinuclear system has a large deformation in the reaction process.

Observation of an Antimatter Hypernucleus

Nuclear collisions recreate conditions in the universe microseconds after the Big Bang. Only a very small fraction of the emitted fragments are light nuclei, but these states are of fundamental interest. We report the observation of antihypertritons-comprising an antiproton, an antineutron, and an antilambda hyperon-produced by colliding gold nuclei at high energy. Our analysis yields 70 +/- 17 antihypertritons (3/Lambda(H) over bar) and 157 +/- 30 hypertritons (H-3(Lambda)). The measured yields of H-3(Lambda) (3/Lambda(H) over bar) and He-3 ((3)(He) over bar) are similar, suggesting an equilibrium in coordinate and momentum space populations of up, down, and strange quarks and antiquarks, unlike the pattern observed at lower collision energies. The production and properties of antinuclei, and of nuclei containing strange quarks, have implications spanning nuclear and particle physics, astrophysics, and cosmology.

同中子素核9C、8B、7Be和6Li在中能区与28Si反应总截面的测量

放射性束流(RIB)装置拓广了实验核物理在同位旋(T_z)自由度上从稳定核直到滴线核的广袤空间。通常，位于β-稳定线及其附近的核，N／Z在1－1.5范围变化，其分离能E_s无论对于质子还是中子，总是在6-8 MeV之间；对于远离稳定线的非稳定核，N／Z可在0.5-4范围变化，如~9C的N／Z = 0.5，~(10)He的N／Z = 4，而且分离能E_s是在0-40 MeV之间变化的，开展对这些远离β-稳定线非稳定核性质、结构的研究是目前核物理的前沿之一。核反应总截面σ_R是表征原子核性质特征的一个基本物理量，从实验测得的核反应总截面中可以得到有关核结构和核内核子分布的信息。在由放射性束流所产生奇异核的结构与各种反应机制研究中，反应总截面的测量更是有其特殊的重要性，具有奇异核结构如晕核的一个典型的物理特征就是其反应总截面要比稳定核大得多，Tanihata等人最早就是通过对放射性束流的相互作用截面的测量发现了具有奇异结构的核，即中子晕核。由于反应总截面的测量对探测器的要求不高，而且数据分析过程相对较为简单，因而反应总截面的测量已经成为放射性束核物理的研究的一个非常重要实验手段。中子晕核以及中子皮核的发现促使人们去寻找质子晕核和质子皮核，由于最后一个质子的结合能非常小只有136，keV，并且有较大的电四极矩，因而使得~8B成为质子晕的最大热门候选核，有关~8B是否具有质子晕核结构的问题，许多实验科学家得出了相互矛盾的回答；而目前有关另外一个质子晕候选核~9C的实验数据非常少，目前还没有人从实验上对~9C是否为质子晕核这一问题进行肯定或否定的回答。因此非常有必要测量~9C和~8B的反应总截面。对反应总截面进行研究的一个非常有用理论就是Glauber模型，该模型考虑了库伦效应的多次散射理论。它是一种基于自由核子-核子(N-N)碰撞的与核物质密度相关的理论，因而能够从实验测量到的反应总截面中提取核物质分布的信息。该理论对中低能区的反应总截面描述却有一个缺憾：理论值比实验值都要小。本论文主要描述了利用透射法测量了中能区同中子素核~9C、~8B、~7Be及~6Li与~(28)Si的反应总截面，并介绍了重离子碰撞以及描述重离子性质的几种常用理论。在论文里对实验测量得到的结果进行了理论分析，这些理论包括半经验的Shen公式、Glauber模型、BUU模型以及SHF理论。如果将~9C和~8B当成具有正常核结构来处理，半经验的Shen公式和Glauber模型(HO密度分布)的理论计算值总是比实验值要小得多；对于Glauber模型的理论计算值和实验值的差异，Ozawa等定义了一个差值因子d，方德清等人对轻核系统的d值进行了详细的分析。一般认为，正常核的d值在20％以内，而对于具有晕或皮奇异结构的核，其d值则超过30％，甚至可达50％，因此可根据一个核的d值是否超过30％而且比相邻核的d值明显大这种半经验的方法来判断一个核是否具有奇异结构；利用d值的分析结果，我们认为：~9C和~8B都具有奇异核结构；对于BUU模型用同样的方法引进差值因子d值，对于~9C和~8B有相同的结论。用SHF理论计算得到B和C同位素的密度分布结果显示，~9C和~8B的密度分布比相邻的同位素扩展都要大得多。为减小Glauber模型计算的反应截面与实验值的差别，本论文还对Glauber模型的输入密度形式进行了修改，在原单一HO分布基础上加一个高斯分布的尾巴，并对丰质子的同中子素核~9C、~8B、~7Be及~6Li与~(28)Si靶以及~(12)C和丰中子的C同位素核~(13-16)C与~(12)C靶的反应截面重新进行了计算，结果显示在中能区的计算值比原来单一密度分布的计算结构有明显改善。

基于3S技术的深圳市水土保持管理信息系统研究

深圳市水土保持管理信息系统采用B/W/D三层体系结构,实现GIS和MIS的紧密结合,克服了C/S结构的客户端维护量大的弱点,发挥B/W/D结构良好的数据交互能力和便于集中控制、维护简单的优点,充分利用了GIS优越的空间分析、直观的信息表达功能和MIS强大的数据存储、处理功能。系统通过对水土保持工作必需的信息进行分析和检索,利用地图、文字、表格、影像等多种手段进行信息表达。通过开发的水土流失估算模型,可对将要进行开发的建设项目的水土流失量进行估算,同时可以载入遥感影像图,为水土流失状况的对比、分析提供更为直观、清晰的效果,也可利用GPS接口转入GPS设备采集的数据,进行地块数据自动、批量录入、更新,实现系统专题图等数据的动态更新。

脱落酸对UV-C胁迫下小麦幼苗光合特性及抗氧化酶活性的影响

研究了UV-C辐射下短期和长期脱落酸(ABA)处理对小麦幼苗CO2同化作用、羧化效率、光合CO2响应以及抗氧化酶活性等的影响.结果表明,在无UV-C辐射情况下,短期和长期ABA处理能提高光合速率,比对照增加14·69%和20·46%,降低气孔导度,比对照降低14·74%和17·31%,但对胞间CO2浓度和羧化效率影响不大.当受到UV-C辐射时,光合速率、羧化效率、气孔导度和胞间CO2浓度逐渐降低.长期ABA处理变化最小,其次为ABA短期处理,对照降低最大.ABA处理能够提高小麦光合对CO2的响应,UV-C辐射抑制光合对CO2的响应.ABA处理能够提高小麦抗氧化酶(CAT、SOD、POD)活性而降低MDA含量.在UV-C辐射下,CAT活性先升高随后降低,在辐射处理1h时活性达最大值,ABA处理的SOD和POD活性先升高后降低,且ABA长期处理比短期处理增加明显,对照则逐渐降低.ABA处理可能通过提高小麦CO2同化作用和抗氧化酶活性增强对UV-C胁迫的抗性,且ABA长期处理比短期处理效果更明显.

增强UV-C辐射作用下小麦和豌豆幼苗光合响应及其相对抗性

以小麦和豌豆为材料,研究了UV-C辐射(波长<280nm)对叶片光合特性及抗氧化酶活性的影响.结果表明:UV-C辐射增强,可使豌豆叶片光合速率减弱,气孔导度、胞间CO2浓度、蒸腾速率和羧化效率明显降低,而对小麦叶片上述各项指标的影响则是先增加、后降低;在UV-C辐射下,豌豆的CO2补偿点逐渐升高,而小麦的CO2补偿点先降低、后升高.UV-C辐射除了使豌豆的POD活性和小麦的SOD活性逐渐降低外,其他抗氧化酶活性则呈先升高、后降低的变化趋势.小麦对短时间UV-C辐射的抗性比豌豆强,但随着UV-C辐射时间的延长,小麦和豌豆的抗氧化酶活性均降低,光合作用减弱.

蛭弧菌J26发酵生产维生素C前体产物的鉴定

采用有机溶剂沉淀方法去除J26发酵液中杂蛋白，通过离子交换、活性炭吸附去除发酵液中杂质，最后蒸发浓缩获得无色透明结晶。通过对所获晶体的红外光谱分析、熔点测定、纸层析鉴定，证明所获晶体确为维生素C前体2－酮基－L－古龙酸（2－KGA）。

中国东北西部地区沙质荒漠化过程与植被动态关系的生态学研究——群落多样性与沙质荒漠化过程

依据东北西部沙质荒漠化地区植被的分类与排序,分析了不同演替梯度上植被群落多样性指数与DCA排序坐标值的相关关系,利用其探讨群落多样性与沙质荒漠化过程的关系;科尔沁沙地、呼伦贝尔沙地,1)植物群落的生态优势度λ、Renyi的均匀度E1与群落分布的地下水位有显著的相关关系;2)植物群落Hill的多样性指数H0,H1、Hill的均匀度Eh、Hill指数的均匀度E′1与海拔、湿润系数、Thornthwaite指数、降水量、地下水位、放牧干扰等有显著的相关关系;3)植物群落Renyi的多样性指数N0,N1、Heip修正的均匀度Ep、Alatalo修正的均匀度E′h与经度、温暖指数、冷暖指数、Thornthwaite指数、土壤有机质、地下水位、放牧等干扰有显著的相关关系。4)呼伦贝尔沙地的完工-海拉尔沙带,樟子松林、贝加尔针茅羊草草甸草原;科尔沁沙地的油松林、羊草草甸草原、丛生禾草草原具有较高的多样性,各群落类型随着沙质荒漠化过程的逐渐加剧,物种丰富度逐渐降低,其递减趋势分为旱生沙化系列、湿生沙化系列、盐水至水生系列。按各群落类型的物种丰富度递减顺序可分为6个级别,6个级别所含的群落类型分别对应着稳定沙地(A、B),固定沙丘(C),半固定沙丘(D),半流动沙丘(E),流动沙丘(F)。

Phenolphthalein immobilized membrane for an optical pH sensor

A phenolphthalein immobilized cellulose membrane for an optical pH sensor was described. The phenolphthalein was first reacted with the formaldehyde to produce a series of prepolymers with many hydroxymethyl groups. In this paper, the prepolymers was abbreviated to phenolphthalein-formaldehyde (PPF). Then the PPF was covalently immobilized to the diacetylcellulose membrane via hydroxymethyl groups. Finally the membrane was hydrolyzed in the 0.1 M NaOH solution for 24 h to reduce the response time. Advantageous features of the pH-sensitive membrane include (a) a large dynamic range from pH 8.0 to 12.50, or even broader, (b) rapid response time (2-30 s), (c) easy of fabrication, and (d) a promising material for determination of high pH values. The immobilized PPF has a broader dynamic range from 8.0 to 12.50 than the free phenolphthalein from pH 8.0 to 11.0, and this was due to the newly produced methylenes in our investigation.

Preparation of a carbon supported Pt catalyst using an improved organic sol method and its electrocatalytic activity for methanol oxidation

The organic sol method for preparing ultrafine transition metal colloid particles reported for the first time by Bonnemann et al. [H. Bonnemann, W Brijoux, R. Brinkmann, E. Dinjus, T. Jou beta en, B. Korall, Angew. Chem. Int. Ed. Engl., 30 (1991) 1312] has been improved in this paper. The improved organic sol method uses SnCl2 as the reductant and methanol as the organic solvent. Thus, this method is very simple and inexpensive. It was found that the average size of the Pt particles in the Pt/C catalysts can be controlled by adjusting the evaporating temperature of the solvent. Therefore, the Pt/C catalysts prepared by the same method are suitable for evaluating the size effect of the Pt particles on electrocatalytic performance for methanol oxidation. The results of the X-ray diffraction (XRD) and transmission electron microscopy (TEM) showed that when the evaporating temperatures of the solvent are 65, 60, 50, 40, and 30 degrees C, the average sizes of the Pt particles in the Pt/C catalysts prepared are: 2.2, 3.2, 3.8, 4.3, and 4.8 nm, respectively. The X-ray photoelectron spectroscopic (XPS) results demonstrated that the small Pt particles are easily oxidized and the decomposition/adsorption of methanol cannot proceed on the surfaces of Pt oxides.