槟榔黄化病组织RNA提取方法的比较

为了从发生黄化病的槟榔茎、叶、花中提取到完整的RNA,采用了CTAB法、Trizol法和异硫氰酸胍法提取槟榔黄化病组织RNA。从RNA的完整性、产率和纯度方面对这几种方法进行了比较,结果表明,异硫氰酸胍法所得RNA完整性较好,条带清晰无明显降解,OD260/OD280介于1.8-2.0之间,RNA得率较高,为120μg/g以上,并能成功进行反转录制备cDNA,表明该RNA可以进行后续分子生物学操作。     

东亚砂藓(Rhacomitrum canescens)RNA提取方法的比较和改进

方法:以东亚砂藓为材料,用CTAB法、热硼酸盐法和改良的SDS/酸酚法提取东亚砂藓总RNA,并采用3种方法进行沉淀,从提取RNA的纯度、完整性、产量、耗时和RT-PCR效果等分析确定适用于东亚砂藓总RNA提取的方法。结果:研究表明:CTAB法提取的RNA 28S rRNA明显缺失,泳道模糊不清,杂质多,热硼酸法和改良的SDS/酸酚法提取RNA 28S rRNA,18S rRNA条带清晰,OD260/OD280都在1.7以上,纯度高,但热硼酸法提取出的RNA有DNA污染,RNA的含量(15.68~35.2μg.g-1)低于SDS/酸酚法提取RNA的含量(46.5~51.9μg.g-1)3种沉淀方法比较认为无水乙醇配合LiCl沉淀RNA节省时间,反转录和RT-PCR效果好。结论:改良的SDS/酸酚法适合东亚砂藓RNA的提取。     

石斛总RNA提取方法的研究

采用TRIZOL法、异硫氰酸胍法、Tris-硼酸法和改良的RNA提取方法提取石斛的总RNA,并通过凝胶电泳、紫外分光光度法检测提取的RNA样品的品质。研究结果表明:改良的RNA提取方法提取的RNA具有28S rRNA和18S rRNA两条清晰的条带,且无降解。OD260nm/OD280nm接近2.0,具有较高的纯度。其它三种方法获得的RNA品质较差,有降解和弥散现象。将改良的RNA提取方法提取的RNA逆转录成cDNA,经RAPD扩增,出现清晰的条带,进一步证明改良的RNA提取方法提取的RNA具有很高的纯度,可以满足进一步分子生物学研究的要求。     

一种适合从柑橘果皮提取总RNA的方法

柑橘果皮由于富含果胶、酚类物质等干扰RNA分离的物质,较难提取到纯度高的RNA。本试验建立了一种适于从柑橘果皮提取RNA的方法,从脐橙和蕉柑两种柑橘的果皮提取总RNA,经凝胶电泳、紫外分光光度法检测所提RNA的品质。研究结果表明,该法所提RNA条带清晰、无降解。OD260/OD280接近2.0,具有较高的纯度。RT-PCR试验结果进一步表明,该法提取的RNA纯度高,完全能够用于后续的分子生物学研究。     

CTAB法提取野野村菌基因组DNA

针对用常规方法难以提取野野村菌基因组DNA的问题,通过选用添加甘氨酸的不同培养基和不同培养时间获得的菌丝体,采用液氮研磨结合CTAB法提取野野村菌DNA,电泳检测及计算OD260/OD280值。结果表明,在添加0.3%甘氨酸的麦芽汁-酵母膏(YE)培养基中振荡培养培养3d的菌丝体适合于DNA提取,用CTAB法获得的基因组DNA,长度约为20kb,且OD260/OD280在1.8左右,达到基因组DNA-DNA杂交的要求。     

麦冬根中总RNA的快速提取

目的:从富含多糖、多酚的麦冬根部组织中快速提取总RNA。方法:采用改进的苯酚法,提取液的配制为5%SDS、1mol/LNaAc(pH4.1)、20%HAC、0.1%PVP。结果:采用该方法提取的麦冬总RNA纯度高、完整性好,电泳条带清晰。通过琼脂糖凝胶电泳与紫外吸光度测定产量与纯度,麦冬根部组织总RNA的吸光值D260nm/D280nm值大于1.8,D260nm/D230nm值大于2.0,麦冬块根与不定根RNA的平均产量分别为79.716和76.144μg/g(鲜重)。结论:用本方法提取的RNA可用于后继的抑制消减杂交试验。     

富含RNA酶大鼠腺体组织总RNA提取方法改良

为了改良传统提取富含RNA酶大鼠胰腺和腮腺组织总RNA方法,本研究将组织样本分成对照组,0.5％,1.0％和2.0％β-巯基乙醇处理组,对照组采用Trizol法提取总RNA,β-巯基乙醇处理组分别于Trizol试剂中加入0.5％,1％,2％β-巯基乙醇.测定不同处理组单链核酸含量、完整性,A260/280和A260/230;采用实时荧光定量PCR检测管家基因β-actin,GAPDH,胰腺淀粉酶AMY2和腮腺水通道蛋白AQP-5表达情况,并通过计算扩增效率,验证β-巯基乙醇是否对后续PCR实验有影响.结果表明,不同处理组单链核酸含量、A260/280,A260/230无显著差异,β-巯基乙醇组RNA完整性显著高于对照组,1.0％,2.0％β-巯基乙醇组RNA完整性显著高于0.5％β-巯基乙醇组,1.0％β-巯基乙醇组RNA完整性与2.0％β-巯基乙醇组无显著差异;添加1.0％β-巯基乙醇对后续PCR试验无影响.综上所述,在传统Trizol法内添加β-巯基乙醇可显著提高胰腺和腮腺组织总RNA提取质量,其中添加1.0％β-巯基乙醇是最佳提取方案.     

富含RNA酶大鼠腺体组织总RNA提取方法改良

为了改良传统提取富含RNA酶大鼠胰腺和腮腺组织总RNA方法,本研究将组织样本分成对照组,0.5％,1.0％和2.0％β-巯基乙醇处理组,对照组采用Trizol法提取总RNA,β-巯基乙醇处理组分别于Trizol试剂中加入0.5％,1％,2％β-巯基乙醇.测定不同处理组单链核酸含量、完整性,A260/280和A260/230;采用实时荧光定量PCR检测管家基因β-actin,GAPDH,胰腺淀粉酶AMY2和腮腺水通道蛋白AQP-5表达情况,并通过计算扩增效率,验证β-巯基乙醇是否对后续PCR实验有影响.结果表明,不同处理组单链核酸含量、A260/280,A260/230无显著差异,β-巯基乙醇组RNA完整性显著高于对照组,1.0％,2.0％β-巯基乙醇组RNA完整性显著高于0.5％β-巯基乙醇组,1.0％β-巯基乙醇组RNA完整性与2.0％β-巯基乙醇组无显著差异;添加1.0％β-巯基乙醇对后续PCR试验无影响.综上所述,在传统Trizol法内添加β-巯基乙醇可显著提高胰腺和腮腺组织总RNA提取质量,其中添加1.0％β-巯基乙醇是最佳提取方案.     

富含RNA酶大鼠腺体组织总RNA提取方法改良

为了改良传统提取富含RNA酶大鼠胰腺和腮腺组织总RNA方法,本研究将组织样本分成对照组,0.5％,1.0％和2.0％β-巯基乙醇处理组,对照组采用Trizol法提取总RNA,β-巯基乙醇处理组分别于Trizol试剂中加入0.5％,1％,2％β-巯基乙醇.测定不同处理组单链核酸含量、完整性,A260/280和A260/230;采用实时荧光定量PCR检测管家基因β-actin,GAPDH,胰腺淀粉酶AMY2和腮腺水通道蛋白AQP-5表达情况,并通过计算扩增效率,验证β-巯基乙醇是否对后续PCR实验有影响.结果表明,不同处理组单链核酸含量、A260/280,A260/230无显著差异,β-巯基乙醇组RNA完整性显著高于对照组,1.0％,2.0％β-巯基乙醇组RNA完整性显著高于0.5％β-巯基乙醇组,1.0％β-巯基乙醇组RNA完整性与2.0％β-巯基乙醇组无显著差异;添加1.0％β-巯基乙醇对后续PCR试验无影响.综上所述,在传统Trizol法内添加β-巯基乙醇可显著提高胰腺和腮腺组织总RNA提取质量,其中添加1.0％β-巯基乙醇是最佳提取方案.     

胡椒叶片基因组DNA提取方法比较

目的：研究建立胡椒叶片中提取高质量DNA的方法。方法：采用各种CTAB法和SDS法的改良方法,提取胡椒叶片中的总DNA,并对DNA进行紫外和电泳检测。结果：改良CTAB法4和5提取的DNA经电泳检测,有一条明亮主带,且无拖尾现象,样品槽无荧光出现,说明抽出的DNA纯度较高,一致性好;测定其OD260和OD280值,并计算其比值,OD260/OD280值在1.8-2.0之间,提取率在51.667-60.000μg/g之间,获得的基因组DNA质量高。结论：改良CTAB法4和法5可从胡椒幼叶中提取高质量DNA。     

酶降解法精制肝素钠

肝素钠在哺乳动物的脏器和体液如：肝、肺、脾及胃肠粘膜等中含量很高．肝素钠作为一种有效的抗凝剂已广泛应用于临床,生物学功能在于阻止血液中凝血酶的作用．其衍生物还具有降血脂、消炎、抗过敏、利尿及调节免疫功能等多种用途．肝素钠属于多糖类化合物,作为溶栓类药...     

Osmotic and ionic regulation in the western rock lobster Panulirus longipes (Milne-Edwards)

The western rock lobster, Panulirus longipes (Milne-Edwards) is poikilosmotic over its tolerated salinity range, 25–45 ‰. Blood sodium is accumulated while chloride concentration is reduced. Sodium and chloride vary directly with the external salinity, although maintaining their differences in the same proportions as at normal salinity (36.0 ‰). Calcium is accumulated, ranging from over 150% at salinity 20 ‰ to about 117% at salinity 45 ‰. Potassium concentration is equivalent to the external at normal salinity, but is increased with lowered and decreased with raised salinity. Magnesium is reduced to about one-third that of the external concentration over the salinity range 20–40 ‰, but regulation begins to break down at 45 ‰. Individual ions exhibit, therefore, a range of regulation types, from poikilosmotic to homoiosmotic.Equilibrium for sodium, chloride, and calcium is attained in 10 h at salinities of 25, 30, 40 and 45 ‰ respectively. Rate constants for this exchange are linearly related to salinity differential, and rapid osmotic adjustment is by high permeability, equal in both directions, probably mainly via the gills. Muscle appears to act as a salt pool for sodium, chloride, and potassium but not for magnesium and calcium. Salt-loading causes a slight salt diuresis, the salts being excreted, probably via the gills. Except for calcium, there is no excretion of salt into the gut, but there is evidence of an exchange of chloride with another anion. Magnesium excretion is slow, and in the absence of osmotic stress possibly occurs via the antennal glands. All the ions examined appear to be regulated independently.     

Micellization of conjugated chenodeoxy- and ursodeoxycholates and solubilization of cholesterol into their micelles: comparison with other four conjugated bile salts species

Micelle formations of sodium glyco- and taurochenodeoxycholate (NaGCDC and NaTCDC) and sodium glyco- and tauroursodeoxycholates (NaGUDC and NaTUDC) was studied at 308.2 K for their critical micelle concentrations at various NaCl concentrations by pyrene fluorescence probe, and the degree of counterion binding to micelle was determined using the Corrin-Harkins plots. The degree of counterion binding was found to be 0.37-0.38 for chenodeoxycholate conjugates, while the determination of the degree was quite difficult for ursodeoxycholate conjugates. The change of I1/I3 values on the fluorescence spectrum with the conjugate bile salt concentration suggested two steps for their bile salt aggregation. The first step is a commencement of smaller aggregates, the first cmc, and the second one is a starting of stable aggregates, the second cmc. The aggregation number was determined at 308.2 K and 0.15 M NaCl concentration by static light scattering: 16.3 and 11.9 for sodium NaGCDC and NaTCDC, and 7.9 and 7.1 for NaGUDC and NaTUDC, respectively. The solubilization of cholesterol into the bile salt micelles in the presence of coexisting cholesterol phase and the maximum additive concentration (MAC) of cholesterol was determined against the bile salt concentration. The standard Gibbs energy change for the solubilization was evaluated, where the micelles were regarded as a chemical species. The solubilization was stabilized in the order of NaGUDC approximately = NaTUDC < NaTC < NaGC < NaTCDC < NaGCDC < NaTDC < NaGDC, where the preceding results were taken into the order.     

不同盐胁迫下小麦叶片渗透性调节和叶绿素荧光特性

盐胁迫对植物伤害机理受到普遍关注。本试验以‘西旱3号’小麦幼苗为材料,通过比较钠盐(150 mmol·L^-1)、钙盐(5、30 mmol·L^-1)单独及其复合胁迫对叶片渗透调节和光合特性的影响,揭示不同盐胁迫对小麦的伤害机理。结果表明: 钠盐或钙盐单独胁迫显著抑制了小麦幼苗根、茎的生长,使叶片可溶性糖和脯氨酸含量、调节性能量耗散电子产量、非光化学猝灭及玉米黄质相对含量均显著增加,而叶绿素a和叶绿素b含量、最大光化学效率、PSⅡ实际光化学效率、光化学猝灭及光合电子传递效率均显著下降。此外,钙盐对小麦幼苗生长的抑制作用更强,钠盐处理下叶片叶绿素含量减少和叶绿素荧光参数降低更显著。除了可溶性蛋白、叶黄素和玉米黄质相对含量以外,低浓度钙盐有效缓解了钠盐诱导其他各指标的变化,而高浓度钙盐进一步增大了钠盐处理小麦幼苗各参数的变化幅度。总之,钠盐和钙盐显著抑制了小麦幼苗的生长,低浓度钙盐能有效缓解钠盐对小麦幼苗的伤害,而高浓度钙盐加剧了钠盐的毒害作用。这均与叶片光合色素含量、光能捕获及光合电子传递的改变有关。此外,渗透调节物质在增强钠盐或钙盐环境中小麦幼苗的抗性方面发挥着重要作用。     

盐度对无瓣海桑幼苗的生长和某些生理生态特性的影响

本文研究了盐度(0‰～50‰)对无瓣海桑幼苗生长的影响。盐度对无瓣海桑幼苗长叶数、茎长、植株鲜重、主根长、根系鲜重等方面起抑制作用;随盐度的提高,无瓣海桑幼苗成活率下降;但盐度对无瓣海桑幼苗叶片面积存在一个低盐(0‰～10‰)促进、高盐(15‰～40‰)抑制的过程;盐度对叶绿素含量的影响总趋势是随盐度提高,低盐时叶绿素含量下降,而当盐度超过10‰时上升。因此认为：1)无瓣海桑幼苗在无盐存在下,也可正常生长;2)无瓣海桑具有较高的耐盐能力,在盐度0‰～25‰内可正常生长,超过25‰,其生长受到抑制。     

Accumulation and localisation of sodium ions within the shoots of rice (Oryza sativa) varieties differing in salinity resistance

Oryza sativa L. (rice) is a salt-sensitive crop species which is relatively ineffective in controlling the influx of sodium and chloride ions to the shoot. Nonetheless, there is considerable varietal and individual variability in salinity resistance, much of which must derive, therefore, from differences in the fates and subsequent effects of saline ions after they have entered the plant. The destination of sodium ions within the plant has been investigated, in saline conditions, by examining the time-course of sodium ion concentrations in different leaves of four varieties and breeding lines of rice of differing salinity resistance. Radionuclide tracers were employed to study short term effects and the degree of retranslocation of these sodium ions. Sodium was not distributed uniformly but accumulated in the older leaves before the younger ones. At least some leaves were maintained at sub-lethal salt concentrations in at least the more salt resistant varieties. Radionuclide tracer studies showed that the discontinuous distribution of sodium (from leaf to leaf) is constitutive, and cannot be explained by time of exposure or differential leaf growth rates, and that significant quantities of sodium were not subsequently retranslocated, either within the plant or to the root medium.     

真菌玫瑰红钠培养基平皿计数验证法的改进

为了避免玫瑰红钠培养基平皿法计数验证时细菌在培养基上生长而干扰真菌计数的准确性这一问题,采用在该培养基中选加了0.08g/L、0.10g/L和0.12g/L三种不同浓度的氯霉素的方法。结果证实,在玫瑰红钠培养基中加入浓度为0.10g/L的氯霉素可完全抑制供试大肠杆菌CMCC(B)44102株的生长,但又不影响供试真菌如白色念珠菌CMCC(F)98001、黑曲霉CMCC(F)98003的培养生长。因此,在微生物限度检查时的玫瑰红钠培养基中加入氯霉素最适宜的有效浓度为0.10g/L。     

The involvement of the transpirational bypass flow in sodium uptake by high- and low-sodium-transporting lines of rice developed through intravarietal selection

We report the characterization of high- and low-sodium-transporting lines developed by intravarietal selection within a cultivar, IR36, of rice (Oryza sativa L.). The purpose was to investigate the mechanistic basis of sodium uptake in material in which differences in salt uptake could be isolated from the many other morphological and physiological characteristics that affect the phenotypic expression of salt tolerance. The lines differed in mean sodium transport by a factor of 2. They differed in vigour and water use efficiency, which are characters that modify the effects of salt transport, by only 12% or 13%. The lines did not differ significantly in other physiological traits that are components of salt resistance: compartmentalization at the leaf and cellular levels. There was a strong correlation between the transport of sodium and a tracer for apoplastic pathways (trisodium, 3-hydroxy-5,8,10-pyrene trisulphonic acid, PTS) in both lines. The regression coefficient for sodium transport on PTS transport was the same in both lines. The individual variation in PTS transport was similar to that in sodium transport, and the variation in the transport of both was very much greater than the variation in any other character studied. The high-sodium-transporting line took up proportionately more PTS than the low-sodium-transporting line. It is concluded that the transpirational bypass flow is of major importance in sodium uptake by rice and that selection for differences in sodium transport has been brought about by selection for heritable differences in the bypass flow.     

Behavioral Adaptations of Moose to Roadside Salt Pools

Abstract: Sodium has many fundamental physiological functions in animals but is rare in boreal ecosystems where moose (Alces alces) thrive. In Québec (Canada), sodium is readily available in aquatic vegetation and in salt pools that form along highways. We do not know if moose are adopting specific behaviors to access sodium sources or if they simply use the sodium sources they encounter during their movements. We tested the hypothesis that moose modify both space and habitat use to gather sodium from salt pools. We expected moose to use salt pools mostly in spring and early summer, when needs are greatest and before aquatic vegetation has fully developed. We fitted 47 moose with Global Positioning System telemetry collars and collected data for 2 to 36 months between 2003 and 2006. We rarely located moose at salt pools (0.12% among the 95,007 locations collected). As we expected, use of salt pools was highest in late spring and in early summer, and we observed a time lag between peak use of salt pools compared to use of lakes and waterways, indicating moose fulfilled their sodium requirements in salt pools before aquatic vegetation was available. Moose selected salt pools over lakes and waterways when these 2 sodium sources were present in their home range and moved rapidly over large distances to reach them. Our results were consistent with moose using salt pools when they are likely to be sodium deficient. Salt pools were less accessible, required long-distance movements, and were located in habitually avoided areas along highways. Elimination of roadside salt pools should be considered among strategies to reduce cervid-vehicle collision risks in boreal environments.     

Salt drying: a low‐cost,simple and efficient method for storing plants in the field and preserving biological repositories for DNA diversity research

Although a variety of methods have been optimized for the collection and storage of plant specimens, most of these are not suited for field expeditions for a variety of logistic reasons. Drying specimens with silica gel in polyethylene bags is currently the standard for field‐sampling methods that are suitable for subsequent DNA extraction. However, silica‐gel repositories are not readily available in remote areas, and its use is not very cost‐effective for the long‐term storage of collections or in developing countries with limited research budgets. Salting is an ancient and traditional drying process that preserves food samples by dehydrating tissues and inhibiting water‐dependent cellular metabolism. We compared salt and silica‐gel drying methods with respect to dehydration rates overtime, DNA quality and polymerase chain reaction(PCR) success to assess whether dry salting can be used as an effective plant preservation method for DNA analysis. Specimens from eleven plant species covering a variety of leaf structures, leaf thicknesses and water contents were analysed. Experimental work indicated that (i) levels of dehydration in sodium chloride were usually comparable to those obtained when silica gel was used, (ii) no spoilage, fungal or bacterial growth was observed for any of the species with all drying treatments and (iii) good yields of quality genomic DNA suitable for PCR applications were obtained in the salt‐drying treatments. The preservation of plant tissues in commercial table salt appears to be a satisfactory, and versatile method that may be suitable in remote areas where cryogenic resources and silica repositories are not available.