高压消毒蒸锅在石蜡切片免疫组织化学反应抗原恢复中的应用

将石蜡切片浸泡于１ｍｍｏｌ／ＬＥＤＴＡ缓冲液（ｐＨ８．０）中,置入高压消毒蒸锅处理（１２０℃,１０３ｋＰａ）１０ｍｉｎ,以恢复组织中因甲醛固定而被封闭的抗原。结果表明,一些常规免疫组织化学反应呈阴性的组织抗原,经本法处理后呈阳性反应;对不经本法处理即呈阳性的组织抗原,可大大提高第一抗体的稀释度或缩短孵育时间。同微波方法相比,本法可一次处理数百张切片,且对切片和组织的形态无明显损害。因此是石蜡切片免疫组织化学反应组织抗原恢复的理想方法     

激光对珠子参愈伤组织诱导影响的研究

通过不同激光功率密度和处理时间处理珠子参植株,然后分别用珠子参的茎和叶做外殖体进行愈伤组织诱导。结果表明,不经激光处理,珠子参茎和叶的愈伤组织诱导率分别为45.3%和0.7%。经激光处理后,珠子参茎和叶愈伤组织诱导率分别最高达到了92.0%和6.7%。经t检验,激光处理后,外殖体的愈伤组织诱导率与对照相比达到了显著性差异,表明激光处理有助于珠子参愈伤组织的诱导。     

高压静电场(HVEF)对苜蓿叶片愈伤组织增殖的影响

苜蓿叶片来源的愈伤组织经一定的高压静电场处理后生长速率显著高于对照组,而且处理组愈伤组织的呼吸速率,元素吸收速率也高于对照组。进一步的测定结果比较表明：静电场处理组的愈伤组织中可溶性蛋白质含量、蔗糖酶活性高于对照组;而ＩＡＡ氧化酶和ＲＮＡ水解酶活性则低于对照组。显示静电场处理组愈伤组织有较高的代谢水平和旺盛的分裂生长状态。探讨了静电场处理剌激愈伤组织增殖的可能机理     

BA对大麦花药培养中药壁的衰退和植株再生频率的影响

用含20ppm 6-BA的0.1％吐温-80溶液喷施花粉为单核前期的大麦上部叶片和穗部,明显影响大麦花药培养效率。实验结果表明:1)BA处理可明显延缓培养花药的药壁衰退进程。2)BA处理后的花药,在培养期间,其死亡的花粉数比对照大大减少,相反其双核或多核的花粉数比对照明显增加。3)BA处理虽然没有促进大麦花粉愈伤组织的诱导率,但显著地促进愈伤组织的生长。提高愈伤组织成长率,增加可转入分化培养的愈伤组织块数。4)BA处理促进愈伤组织的再分化,尤其是绿苗的分化。     

蓝光对绿豆下胚轴愈伤组织生长和呼吸的影响

与白光和黑暗条件相比,蓝光明显促进绿豆(Vigna radiata (Linn.)Wilczek)下胚轴愈伤组织的形成和生长。培养到18d,蓝光下生长的愈伤组织鲜重分别是白光和黑暗下的120％和191％。三种条件下愈伤组织的呼吸速率有明显的不同,培养5d,呼吸速率以蓝光处理的最高,白光处理的次之,黑暗处理的最低;培养14d,与上述相反,呼吸速率由高到低则依次是黑暗、白光和蓝光。应用呼吸抑制剂碘乙酸、丙二酸和正磷酸钠试验及对6-磷酸-葡萄糖脱氢酶活性测定的结果表明,培养5d,蓝光和白光处理时磷酸已糖支路(HMP)明显增加,被碘乙酸 丙二酸抑制后的呼吸占总呼吸的34％～36％,黑暗处理的仅占总呼吸9％～11％ 绿豆下胚轴愈伤组织中亦存在抗氰交替途径,黑暗处理时,此种途径的最大容量明显高于蓝光和白光处理的,培养14d,黑暗处理占总呼吸的31％左右.而蓝光和白光处理的则占20％以下。     

白桦愈伤组织化学诱变

用不同浓度的甲基磺酸乙酯（ethyl methane sulfonate,EMS）对白桦（Betula platyphalla Sak.）愈伤组织进行化学诱变处理。结果表明：EMS诱变剂的浓度和处理时间对愈伤组织的存活率有很大影响。在高浓度EMS短时间处理和低浓度EMS长时间处理条件下得到叶柄、叶片愈伤组织的半致死剂量。通过观察半致死剂量下愈伤组织的染色体发现,诱变后细胞中单倍体、非整倍体及多倍体比例均高于对照,这说明EMS的诱变处理引起了愈伤组织细胞中染色体数量的变化。     

白桦愈伤组织化学诱变

用不同浓度的甲基磺酸乙酯(ethyl methane sulfonate, EMS)对白桦(Betula platyphalla Sak.)愈伤组织进行化学诱变处理。结果表明: EMS诱变剂的浓度和处理时间对愈伤组织的存活率有很大影响。在高浓度EMS短时间处理和低浓度EMS长时间处理条件下得到叶柄、叶片愈伤组织的半致死剂量。通过观察半致死剂量下愈伤组织的染色体发现, 诱变后细胞中单倍体、非整倍体及多倍体比例均高于对照, 这说明EMS的诱变处理引起了愈伤组织细胞中染色体数量的变化。     

镉对黑斑蛙精巢组织结构的毒性效应

为了研究镉对两栖动物精巢组织结构的影响,用不同浓度Cd2 溶液对黑斑蛙(Rana nigromaculata)成体隔日皮下注射染毒,光镜下观察精巢组织的显微结构变化。结果表明,镉具有毒性效应,可以导致处理后的黑斑蛙精巢组织中的各级生精细胞、支持细胞和间质细胞受到损伤。对处理组和对照组的精巢组织分别用金属硫蛋白(MT)进行免疫组化定位观察,MT阳性颗粒在处理组和对照组均有表达,表达随处理组镉浓度的增大而减弱。     

石蜡包埋前组织去黑色素法探讨

在研究含有黑色素的肿瘤时,其组织石蜡切片HE染色如未经去色素处理,显微镜下可见大量浓密的黑色素颗粒,遮盖组织结构无法正常观察,作出正确判断,必须进行去黑色素处理。传统的去黑色素方法是在石蜡切片上进行脱色素反应,常用脱色剂为高锰酸钾或过氧化氢,二者均为强氧化剂,在去除黑色素的同时,极易使组织片脱落。尤其在进行免疫组化染色时脱片更为严重。     

组织叶酸检测方法的建立及应用

目的:采用目前常用的化学发光法检测组织叶酸,探索不同叶酸前处理,寻求合适的检测组织叶酸的实验方法。并将优化好的方法应用于叶酸缺乏小鼠模型的检测。方法:通过化学发光法检测组织叶酸含量,首先将一定量的组织放入合适的buffer中,匀浆器匀浆后,通过超声或煮沸处理样本,离心后,得到上清样本,建立标曲和质控后,进行上机测定。该方法中我们通过探讨不同的buffer(包括PBS、Lysis和Tris-base盐溶液)处理组织叶酸,和不同的处理条件(包括是否煮沸及煮沸时间、样本处理时间等),探讨该方法检测组织叶酸最优化条件;并应用该方法进行小鼠叶酸缺乏模型中各组织叶酸含量的检测。结果:我们选择雄鼠肝组织进行叶酸检测,分别用PBS、Lysis和Tris-base盐溶液进行样本前处理,结果发现同样的处理条件下,如不煮沸条件下,Tris-base盐溶液(38.72 ng/mg)PBS(15.68 ng/mg)Lysis(11.9 ng/mg),提示Tris-base盐溶液可能有助于叶酸的稳定,防止降解。同时,我们探讨煮沸对叶酸检测结果的影响,结果发现同一种前处理下(如Tris-base),不煮沸(38.72 ng/mg)煮沸1分钟(36.36 ng/mg)煮沸3分钟(33.28 ng/mg)煮沸5分钟(30.72 ng/mg),说明叶酸含量随着煮沸时间的延长逐渐降低。此外,我们还对叶酸检测结果的重复性和稳定性进行了评估,结果发现不煮沸条件下,叶酸结果重复性很好,但随着时间延长稳定性会逐渐下降。因此,我们选择了Tris-base盐溶液进行前处理,条件选择为不煮沸和立即检测来减少因样本处理问题产生的误差。我们用该方法检测了叶酸缺乏小鼠模型的各组织叶酸含量,结果发现肝组织18.81 ng/mg降为8.46 ng/mg,脑组织从0.37 ng/mg下降为0.19 ng/mg,脾脏组织从1.53 ng/mg下降为0.26 ng/mg,肺组织从0.47 ng/mg下降为0.13 ng/mg,肾脏组织从2 ng/mg下降为0.9 ng/mg,心脏组织从0.33 ng/mg下降为0.06 ng/mg说明饮食叶酸是体内叶酸的主要来源,其缺乏可能导致多种组织叶酸代谢异常。结论:组织叶酸化学发光法检测条件优化为Tris-base盐溶液进行前处理,条件选择为不煮沸,并将样本立即进行检测。我们用该方法检测小鼠叶酸缺乏模型发现低叶酸饮食能显著降低各组织的叶酸含量,可能对生长发育造成潜在的影响。     

Understanding plant rooting patterns in semi‐arid systems: an integrated model analysis of climate,soil type and plant biomass

Aim A consistent set of root characteristics for herbaceous plants growing in water‐limited environments has been developed based on compilations of global root databases, but an overall analysis of why these characteristics occur is still missing. The central question in this study is whether an ecohydrological model which assumes that rooting strategies reflect maximization of transpiration can predict the variations in rooting strategies of plants in dry environments. Location Arid ecosystems across the globe. Methods A model was used to explore interactions between plant biomass, root–shoot allocation, root distribution, rainfall, soil type and water use by plants. Results Model analyses showed that the predicted shifts in rooting depth and root–shoot allocation due to changes in rainfall, soil type and plant biomass were quite similar to observed shifts. The model predicted that soil type, annual rainfall and plant biomass each had strong effects on the rooting strategies that optimize transpiration, but also that these factors have strong interactive effects. The process by which plants compete for water availability (soil evaporation or drainage) especially affected the depth distribution of roots in the soil, whereas the availability of rainfall mainly affected the optimal root–shoot allocation strategy. Main conclusions The empirically observed key patterns in rooting characteristics of herbaceous plant species in arid environments could be explained in this theoretical study by using the concept of hydrological optimality, represented here by the maximization of transpiration.     

Age-determination of Dicotyledonous Herbaceous Perennials by Means of Annual Rings: Exception or Rule?

Thirty-five herbaceous dicotyledonous perennial plant species,with permanent root systems, from 16 families, were examinedfor the presence of growth rings in the secondary root xylem.Most of the species surveyed showed ring zonations in the roots,and these could be verified as annual growth rings in the tenspecies for which plants of known age were available. The potentialvalue of ‘herbchronology’ as a tool in ecologicalinvestigations of species and stands of perennial herbs of temperatezones is discussed. Annual rings; age-determination; dendroecology; herbaceous perennials; anatomical patterns; secondary root xylem     

Salt stress aggravates boron toxicity symptoms in banana leaves by impairing guttation

Boron (B) is known to accumulate in the leaf margins of different plant species, arguably a passive consequence of enhanced transpiration at the ends of the vascular system. However, transpiration rate is not the only factor affecting ion distribution. We examine an alternative hypothesis, suggesting the participation of the leaf bundle sheath in controlling radial water and solute transport from the xylem to the mesophyll in analogy to the root endodermis. In banana, excess B that remains confined to the vascular system is effectively disposed of via dissolution in the guttation fluid; therefore, impairing guttation should aggravate B damage to the leaf margins. Banana plants were subjected to increasing B concentrations. Guttation rates were manipulated by imposing a moderate osmotic stress. Guttation fluid was collected and analysed continuously. The distribution of ions across the lamina was determined. Impairing guttation indeed led to increased B damage to the leaf margins. The kinetics of ion concentration in guttation samples revealed major differences between ion species, corresponding to their distribution in the lamina dry matter. We provide evidence that the distribution pattern of B and other ions across banana leaves depends on active filtration of the transpiration stream and on guttation.     

New approaches for shoot production and establishment of in vitro root cultures of Cleome rosea Vahl.

Alternative methods for in vitro shoot culture of Cleome rosea, a Brazilian herbaceous species with ornamental value and medicinal potential, were evaluated. A protocol for rapid in vitro multiplication of roots, a valuable source of medicinal compounds, was also developed. Stem explants were cultured in liquid media (continuous immersion and paper bridge), while root explants were cultivated in continuous immersion and on solidified media. The highest numbers of shoots, 20 ± 4.6 shoots/explant, were obtained from stem explants incubated in a continuous immersion system in a liquid medium supplemented with 2.2 μM BA. Root explants cultivated in liquid media produced only hyperhydrous adventitious shoots. However, these explants generated 5.8 ± 0.8 shoots/explant by indirect organogenesis when cultivated on solidified medium supplemented with 2.2 μM BA. In addition, root multiplication was achieved in liquid medium in the presence of α-naphthaleneacetic acid. Adventitious shoots developed on newly formed roots when inoculated on solidified medium supplemented with 2.2 μM BA. Shoot microcuttings developed roots when transferred onto solidified MS medium without growth regulators. Rooted microcuttings were efficiently acclimatized when transferred ex vitro.     

虉草营养器官解剖结构与抗旱耐涝性及利用之间的关系

为了从显微结构上进一步探讨虉草(Phalaris arundinacea L.)的抗旱耐涝性及与利用的关系,于2011年采用常规石蜡切片技术,对其根、茎叶3种营养器官进行解剖观察。结果表明,虉草根的结构自外而内依次为表皮、皮层、维管束鞘、初生韧皮部和初生木质部;茎由表皮、基本组织和维管束构成;叶片内部结构可分为表皮、叶肉和叶脉3部分。根皮层大的细胞间隙和气腔,初生木质部的后生大导管和茎基本组织解体形成的髓腔都是虉草良好的通气组织,是其耐水淹的主要显微特征。茎、叶片角质化的表皮和叶表皮所含的丰富泡状细胞组是虉草具有抗旱性的主要解剖结构特征。叶肉细胞排列紧密且只有少量气孔分布于叶片下表皮,这样的结构可减少蒸腾;叶肉细胞富含叶绿体,增强光合作用,获得更多的同化产物,确保了植株在干旱条件下也有足够的光合产物来维持正常的生理活动。茎、叶维管束部分大量的木纤维起到支撑作用。虉草根的皮层和维管柱部分、茎的基本组织和维管束部分、叶的叶脉部分都含有大面积的厚壁细胞,厚壁细胞中含有丰富的粗纤维和木质素。丰富的粗纤维、木质素等成分则是虉草能成为新能源燃料植物的必备条件。     

A rapid method for generating sufficient amounts of uniform genotype-specific material from the woody perennial grapevine for ion transport studies

Experimentation with woody perennials may be difficult due to slow plant growth and a lack of sufficient amounts of uniform plant material. In this study, we sought to determine whether rooted leaves could be used as a substitute for whole plants in ion accumulation studies. Grapevine leaves are particularly amenable for rooting since their petioles are of sufficient length for dipping in rooting hormone and for holding the leaf above the soil surface. To determine whether rooted leaves would be useful for salinity experiments, we investigated the ion uptake characteristics of rooted leaves derived from a backcross population that differed in Cl^– accumulation. Long-term ion accumulation studies conducted over several weeks and short-term radioactive uptake studies conducted over several hours were performed. The data showed that the Cl^– content of rooted grapevine leaves from different genotypes grown at 25 and 50 mM NaCl was similar to data reported by others. Short-term radioactive uptake assays did not always reveal differences in uptake between the genotypes. Therefore, we suggest that rooted leaves under certain conditions may provide a space-efficient method for generating sufficient amounts of plant material. This material could be used for studying whole plant, molecular and electrophysiological aspects of ion transport and for conducting experiments where root material from specific genotypes is required.     

Rapid flood-induced stomatal closure accompanies xylem sap transportation of root-derived acetaldehyde and ethanol in Forsythia

Plants grown in containers frequently suffer from difficulties in managing their water status due to either insufficient or too much water. In the case of the latter, little information is available regarding how container-grown woody plants respond to anaerobic media. The aims of this work were therefore to use Forsythia as a model woody plant system to provide a mechanistic understanding of the physiological events and their timing during soil flooding. Exposure of pot-grown Forsythia to root hypoxia had a dramatic effect on leaf growth and stomatal conductance. Within 24 h of flooding a decline in leaf growth rate was detected along with a reduction in stomatal conductance. The effects of hypoxia appear initially with older leaves, but if flooding is prolonged (>2 days) younger expanding leaves are affected. These responses and their timing have not been described for woody perennial plants but appear comparable to those described for herbaceous plants such as tomato and castor bean. Measurements of stem and leaf tissue and during flooding showed large time dependent increases in the concentrations of acetaldehyde and ethanol; products associated with anaerobic respiration. Both of these chemicals were shown to be root-derived and are produced in a significant amount only as flooding time increases and the decline in leaf growth and conductance become apparent. Xylem sap was collected at a range of flow rates to measure whole root system conductance and determine changes in delivery of sap flux constituents. Calculated delivery rates of acetaldehyde and ethanol changed little with sap flow, particularly in well-drained control plants, while root hydraulic conductance declined when measured, 4 days after flooding. However, neither acetaldehyde nor ethanol, when used in a detached leaf transpiration bioassay, at physiologically realist concentrations (as determined from sap collection) failed to induce a dramatic reduction in leaf transpiration rates. The reasons for this discrepancy are discussed.     

Bt玉米光合作用和生长性状的变化

在华南地区种植美国先锋公司的Bt玉米34824与其非转基因近等基因系34823,比较它们的光合作用和部分重要的生理生长性状。结果表明,Bt玉米由于Bt基因的插入和表达使植株得到保护,受玉米螟危害的叶片显著减少;但Bt玉米植株的光合作用和部分生理生长性状也发生了一定的变化。与其非转基因近等基因系34823相比,虽然Bt玉米34824的胞问CO2浓度变化不大;并且在绝大多数生长时期,Bt玉米的蒸腾速率和气孔导度大于的非转基因玉米;但是从生长中期始,Bt玉米的净光合速率却一直小于非转基因玉米,在生长后期,这种差异甚至达到显著水平。在生长性状方面,各时期观察到的Bt玉米植株叶片数一直略少于非转基因玉米,表明Bt玉米的生长发育受到一定的延缓。同时,Bt玉米植株的株高变矮,单株叶面积变小,根系活力显著降低。在地下部,Bt玉米34824的植株根系总长度、根总表面积、根总体积和平均根直径均小于相对应的非转基因近等基因系34823。     

Suites of root traits differ between annual and perennial species growing in the field

Here, we tested whether root traits associated with resource acquisition and conservation differed between life histories (annuals, perennials) and families (Fabaceae, Asteraceae and Poaceae). Root topology, morphology, chemistry and mycorrhizal colonization were measured on whole root systems of 18 field-grown herbaceous species grown and harvested in central Argentina. Annuals differed from perennials in several root traits important in resource uptake and conservation. They exhibited higher specific root length (SRL), root nitrogen concentration (RNC) and mycorrhizal colonization but had lower root tissue density (RTD) than perennials. They did not differ in topology or construction cost. These differences were consistent among families. Families differed only in a few root traits known to be strongly associated with certain lineages such as topology and nitrogen concentration. There was a strong parallel between root traits and analogous leaf traits described in the literature for annuals and perennials. Our results suggest the existence at the root level of an acquisitive vs conservative syndrome consistent among families similar to that previously reported for above-ground traits.