Cloning and Expressing of a Gene Encoding Cytosolic Copper/Zinc Superoxide Dismutase in the Upland Cotton

In this study, a gene encoding a superoxide dismutase （SOD） was cloned from senescent leaves of cotton （Gossypium hirsutum）, and its expressing profile was analyzed. The gene was cloned by rapid amplification of cDNA ends （RACE） method. Northern blotting was used to show the profile of the gene expression, and the enzyme activity was mensurated by NBT deoxidization method in different growth periods. The full length of a gene of cytosolic copper/zinc superoxide dismutase （Cu/Zn-SOD） was isolated from cotton （GenBank Accession Number： DQ445093）. The sequence of cDNA contained 682 bp, the opening reading frame 456 bp, and encoded polypeptide 152 amino acids with the predicted molecular mass of 15.03 kD and theoretical pI of 6.09. The amino acid sequence was similar with the other plants from 82 to 87%. Southern blotting showed that the gene had different number of copies in different cotton species. Northern blotting suggested that the gene had different expression in different tissues and development stages. The enzyme activity was the highest in peak flowering stage. The cotton cytosolic （Cu/Zn-SOD） had lower copies in the upland cotton. The copper/zinc superoxide dismutase mRNA expressing level showed regular changing in the whole development stages; it was lower in the former stages, higher in latter stages and the highest at the peak flowering stage. The curve of the copper/zinc superoxide dismutase mRNA expressing level was consistent with that of the Cu/Zn-SOD enzyme activity. The copper/zinc superoxide dismutase mRNA expressing levels of different organs showed that the gene was higher in the root, leaf, and lower in the flower.     

孑遗植物银杏(Ginkgo biloba L.)雌雄株水分生理特征研究(英文)

[Objective] The experiment aimed to study the difference of water physiology of male and female Ginkgo biloba L. for discussing the strategy of water utilization as well as the important role of this difference during evolution process. [Method] The stem sap flow, stomatal conductance(Gs), transpiration rate(Tr) and water use efficiency (WUE) of male and female Ginkgo biloba L. were comparatively studied. [Result] The day-night processes of flow on male and female Ginkgo biloba L. were similar. The flow on male and female Ginkgo biloba L. in day were almost same while the flow at night on male Ginkgo biloba L. was bigger than that on female Ginkgo biloba L. The Tr and Gs of male and female Ginkgo biloba L. were high in morning and at night but low at noon ,while Tr and Gs of female Ginkgo biloba L. in morning and at night were higher than these of male Ginkgo biloba L. at the same time point. However, these indexes of female plant were lower than these of male plant from 11:00 to 14:00. WUE changing trends of male and female Ginkgo biloba L. were similar, while average water utilization rate of female Ginkgo biloba L. was slightly lower than that of male Ginkgo biloba L. [Conclusion] Compared with other companion plants, water physiology of male and female Ginkgo biloba L. had strong homoplasy. The phenomenon might be a survival strategy of dioecious plants under long term evolutionary pressure.     

3种虾类超氧化物歧化酶同工酶比较(英文)

[Objective] The aim of the study is to compare the activities of superoxide dismutase(SOD) isoenzymes in three species of shrimps Penaeus japonicus, Procambarus clarkia and Litopenaeus vannamei. [Method] The experimental materials were used to measure SOD activities after pretreatment, meanwhile the differences in SOD isoenzymes from different materials were assayed by polyacrylamide gel electrophoresis (PAGE). [Result] There are specific and histological differences in SOD activities of shrimps. With a similar electrophoresis pattern and migration rate, Penaeus japonicus and Litopenaeus vannamei showed remarkable differences with that of Procambarus clarkia. [Conclusion] The result showed the differences of cognation and origin of three shrimps.     

Effects of Cinnamon Acid on Respiratory Rate and Its Related Enzymes Activity in Roots of Seedlings of Malus hupehensis Rehd

This paper studied the effects of cinnamon acid treatments on the respiratory rate and related enzymes activity in the seedling roots of Malus hupehensis Rehd.It would provide information for understanding the mechanisms of inhibition damage caused by continuous cultivation of apple tree.20 mL of solution containing different concentrations of cinnamon acid was added into container with the tested seedlings.After treatment,the samples were taken periodically and the respiratory rates were measured by OXY-LAB oxygen electrodes under 25°C stable temperature and then the activities of related enzymes were measured.The rates of total respiration and other 2 pathways [tricarboxylic acid cycle （TCA） and pentose phosphate pathway （PPP）] appeared initially an increasing treads and late （on the 3rd d） began to decline.However,they again appeared an increase trend at the end period,on the contrast,the respiratory rate of embden-meyer- hot-parnas （EMP） pathway appeared a stead decline tread but it had a recover on the last day.The respiratory rate of total and 3 pathways were decreased under 125 mg kg-1 （soil）.The dynamic trends of the enzymes activities of pyrophosphate-dependent phosphofructokinase （PFK）,glucose-6-phosphate dehydrogenase （G-6-PDH） and malate dehydrogenase （MDH） showed similarly.In conclusion,treatments of certain concentration of cinnamon acid would inhibit the respiratory rate and related enzymes activity of roots of M.hupehensis Rehd.And the inhibition degrees were positively related with concentration of cinnamon acid treatments.     

Method for Isolating Mitochondrial DNA from Etiolated Tissue of Cabbage

Isolation of high-quality mitochondrial DNA（mtDNA） is an important premise for researching molecular mechanisms in cytoplasmic male sterility of cabbage（Brassica oleracea L.var.capitata）. An efficient protocol for separation and purification of mitochondria and extraction of mitochondrial DNA（mtDNA） from etiolated tissues of cabbage was developed. We took a method combined mannitol density gradient with differential centrifugation, selected appropriate rotational speed, extended DNase I treating time and changed mitochondria cracking condition. The results showed that the extracted mitochondria in this protocol had complete structure, appeared to ellipsoid and had not been contaminated with other impurities under the Jannus Green B staining. The isolated mitochondrial DNA had high purity and yield through detecting the optical density, nuclear specific primer PCR and agarose gel electrophoresis. The results indicated that mitochondrial DNA extracted by this protocol had high quality and enabled to be used in futher genetic studies.     

Comparative Proteomic Analysis Shows an Elevation of Mdh1 Associated with Hepatotoxicity Induced by Copper Nanoparticle in Rats

Copper nanoparticle is a new material widely used in biological medicine, animal husbandry and industrial areas, but its potential toxicity to human health and environment remains unclear. In order to study the hepatotoxic mechanisms of nanoparticles copper, two-dimensional gel electrophoresis （2-DE） and matrix-assisted laser desorption/ionization time of flight mass spectrometry （MALDI/TOF MS） of proteomics technology were used to isolate and identify the differentially expressed proteins from liver, which associated with hepatotoxicity induced by copper nanoparticle in rats. In this study, we have screened 15 kinds of proteins related with hepatotoxicity, of which spot8212 was identified as Malate dehydrogenase （Mdhl）. The mRNA expression trend of Mdhl was consistent with the result of 2-DE by RT-PCR validation. Bioinformatics analysis showed that Mdhl was stable and no signal peptides, subcellular location was in endoplasmic reticulum; it contained many functional sites such as malate dehydrogenase activity signal sites 155LTRLDHNRAKSQI167; a helixes and random coils were the two main elements. Homologous analysis demonstrated high homologous of Mdhl in rats with mouse and human, and the phylogenetic tree of Mdhl was constructed. The result indicated that copper nanoparticle could regulate up the Mdhl protein expression so as to compensate the energy deficit. Energy metabolic disturbance may be a pathway for copper nanoparticle particles to exert the hepatotoxic effects in rats.     

Effects of Cinnamon Acid on Respiratory Rate and Its Related Enzymes Activity in Roots of Seedlings of Malus hupehensis Rehd.

This paper studied the effects of cinnamon acid treatments on the respiratory rate and related enzymes activity in the seedling roots of Malus hupehensis Rehd.It would provide information for understanding the mechanisms of inhibition damage caused by continuous cultivation of apple tree.20 mL of solution containing different concentrations of cinnamon acid was added into container with the tested seedlings.After treatment,the samples were taken periodically and the respiratory rates were measured by OXY-LAB oxygen electrodes under 25°C stable temperature and then the activities of related enzymes were measured.The rates of total respiration and other 2 pathways [tricarboxylic acid cycle (TCA) and pentose phosphate pathway (PPP)] appeared initially an increasing treads and late (on the 3rd d) began to decline.However,they again appeared an increase trend at the end period,on the contrast,the respiratory rate of embden-meyer- hot-parnas (EMP) pathway appeared a stead decline tread but it had a recover on the last day.The respiratory rate of total and 3 pathways were decreased under 125 mg kg-1 (soil).The dynamic trends of the enzymes activities of pyrophosphate-dependent phosphofructokinase (PFK),glucose-6-phosphate dehydrogenase (G-6-PDH) and malate dehydrogenase (MDH) showed similarly.In conclusion,treatments of certain concentration of cinnamon acid would inhibit the respiratory rate and related enzymes activity of roots of M.hupehensis Rehd.And the inhibition degrees were positively related with concentration of cinnamon acid treatments.     

Physiological traits of rice seedlings in response to inorganic arsenic

The aim of our study was to better understand the different responses of rice seedling to different species of inorganic arsenic As203 （As（Ill）） and Na2HAsO4 （As（V））. Our results indicate that the biomass of rice seedling decreased as arsenic concentration increased, with the decrease being more significant at higher arsenic concentrations. In addition, the analysis of superoxide dimutase （SOD）, peroxidase （POD）, and catalase （CAT） in rice roots and leaves showed that the activity of these three enzymes significantly decreased in rice tissues, especially in rice roots, as arsenic concentration was increased,. Further, the uptake and utilization efficiencies of N, P, and K were found to decrease as arsenic concentration was increased. However, the uptake and utilization efficiencies of P and K were mainly affected by As（IlI）, whereas those of N were mainly affected by As（V）. Inductively coupled plasma-mass spectrometry （ICP-MS） was used to assay arsenic accumulation in rice tissues; the results indicate that the arsenic content in rice tissues was enhanced when arsenic concentration was increased, especially in rice roots after arsenic treatment.     

斜纹夜蛾感受器超微形态特征

采用扫描电镜技术,系统研究了斜纹夜蛾触角、跗节和产卵器的感受器形态结构特征,发现触角感受器主要包括毛形、锥形、刺形、耳形和腔锥感受器.雌、雄虫触角锥形感受器形态特征区别明显,雄蛾锥形感受器长约10～15 μm,基部直径约4～5μm,显著比雌虫锥形感受器长、粗,雄蛾锥形感受器顶端有一小锥,雌蛾没有.雌成虫和雄成虫前足、中足、后足跗节均分布有毛形和刺形感受器,但数量明显少于触角同类感受器数量.雌成虫产卵器上主要为长毛形和短毛形感受器,这些感受器可能与其感受环境信息、产卵场所选择等行为有关.

Abstract：

The ultrastructures of the sensilla on the antennae, the tarsi and the ovipositor of Spodoptera litura were observed with scanning electron-microscopy. The antennae of both male and female moths were found to contain five kinds of antennal sensilla, namely, sensillum trichodeum, sensillum basiconicum,sensillum chaeticum, ear-shaped sensillum and sensillum coeloconicum. The characters of sensillums basiconicum on the antennae were different between the male and the female. The length was 10-15 μm, the diameter at the base was approximately 4-5 μm for the male antennae, which were significantly longer and bigger than the female antennae, and the male moths had onesmall bradawl at the bass of the antennae while the female moths did not. The tarsi of both male and female moths contained two kinds of sensilla:sensillum trichodeum and sensillum chaeticum; however, their numbers were significant less than those on the antennae. The long sensillun trichodeum and short sensillun trichodeum were mainly distributed on the ovipositor. These results provided basis for exploring the molecular mechanism of oviposition deterrent.     

泰山赤鳞鱼酪氨酸酶的活性和表达差异(英文)

[目的]对泰山赤鳞鱼中酪氨酸酶的活性和表达差异进行研究。[方法]以泰山赤鳞鱼为材料,运用聚丙烯酰胺凝胶电泳与生化染色技术、分光光度技术和酶组织化学技术对赤鳞鱼雌、雄个体9种组织中的酪氨酸酶进行了分析研究。[结果]酪氨酸酶在雌、雄赤鳞鱼的肝胰脏、肠和脾组织中均有存在;同时在雄性赤鳞鱼的胆中也有分布。赤鳞鱼各组织中酪氨酸酶的活性大小存在较大差异。其中,雌雄性赤鳞鱼的脾脏中活性最强,但其活性低于雌性鱼脾中的酪氨酸酶活性。酶组织化学结果显示赤鳞鱼的脾脏、肠、肝胰脏和胆中都存在较强的酪氨酸酶阳性信号,而且脾中的阳性信号最强。[结论]该研究是关于泰山赤鳞鱼酪氨酸酶组织定位研究的首次报道,为进一步了解赤鳞鱼酪氨酸酶的功能提供了理论依据。     

泰山赤鳞鱼苹果酸酶同工酶表型差异的研究

应用聚丙烯酰胺凝胶电泳结合生化染色方法分别对雌雄泰山赤鳞鱼10种组织中苹果酸酶同工酶酶谱进行分析。结果表明,在所取的10种组织中,雌性泰山赤鳞鱼在心脏、性腺、肾、眼、鳃和脾6种组织中存在苹果酸酶,而雄性泰山赤鳞鱼只在心、肌肉、性腺、眼和鳃5种组织中存在苹果酸酶。雌性鱼心脏组织中只有1条ME-2带,而雄性鱼心脏组织含有ME-1和ME-22条酶带;卵巢中含有ME—1和ME-22务酶带,而精巢中只含有ME-11条带;雌性鱼的跟组织含有ME-1带,而雄性鱼眼中含有ME-2带。肾和脾组织只在雌性鱼中含有苹果酸酶酶带,而肌肉组织只在雄性鱼中含有苹果酸酶。苹果酸酶的表型在泰山赤鳞鱼不同性别同一组织和同一性别不同组织之间都存在明显差异,但在同一性别的不同个体之间不存在差异。该研究结果为赤鳞鱼种群的发育遗传学、品种改良和定向育种提供了基础资料,为赤鳞鱼的开发和保护奠定基础。     

泰山赤鳞鱼苹果酸酶同工酶表型差异的研究

应用聚丙烯酰胺凝胶电泳结合生化染色方法分别对雌雄泰山赤鳞鱼10种组织中苹果酸酶同工酶酶谱进行分析。结果表明,在所取的10种组织中,雌性泰山赤鳞鱼在心脏、性腺、肾、眼、鳃和脾6种组织中存在苹果酸酶,而雄性泰山赤鳞鱼只在心、肌肉、性腺、眼和鳃5种组织中存在苹果酸酶。雌性鱼心脏组织中只有1条ME-2带,而雄性鱼心脏组织含有ME-1和ME-22条酶带;卵巢中含有ME-1和ME-22条酶带,而精巢中只含有ME-11条带;雌性鱼的眼组织含有ME-1带,而雄性鱼眼中含有ME-2带。肾和脾组织只在雌性鱼中含有苹果酸酶酶带,而肌肉组织只在雄性鱼中含有苹果酸酶。苹果酸酶的表型在泰山赤鳞鱼不同性别同一组织和同一性别不同组织之间都存在明显差异,但在同一性别的不同个体之间不存在差异。该研究结果为赤鳞鱼种群的发育遗传学、品种改良和定向育种提供了基础资料,为赤鳞鱼的开发和保护奠定基础。     

利用清蛋白PAGE技术鉴定甜玉米杂交种初报

利用改进的聚丙烯酰胺凝胶电泳技术对7个甜玉米杂交种的清蛋白进行分析。结果表明,甜玉米种子清蛋白存在一定的多态性,每个品种都有各自独特的清蛋白PAGE“指纹”。利用该技术对7个不同甜玉米杂交种的种子纯度进行了室内检验,同时利用田间种植鉴定法进行了测定,通过对其中5个杂交种的电泳纯度和田间纯度进行相关分析,二者达显著水平(r=0.9408),说明改进的聚丙烯酰胺凝胶电泳技术用于甜玉米种子纯度鉴定是可行的,可用于甜玉米杂交种纯度鉴定。     

泰山赤鳞鱼的核型研究(英文)

[目的]研究泰山赤鳞鱼的核型。[方法]以稀有物种泰山赤鳞鱼的头肾细胞为实验材料对其染色体数目和核型进行了分析。[结果]泰山赤鳞鱼染色体的数目是50,核型是2n=50,16m+14sm+20t,NF=80,没有发现性染色体。[结论]这是首次关于泰山赤磷鱼核型分析的报道。     

玉米蛋白质PAGE法与田间鉴定法测定杂交玉米种子纯度的相关性研究

利用玉米蛋白质聚丙烯酰胺凝胶电泳法（蛋白质PAGE法）和田间小区种植鉴定法对生产上广泛应用的8个品种10个样品杂交玉米种子进行纯度测定，并对两种方法所得数据进行了回归分析1，二者相关达极显著水平（r=0.9006），表明蛋白质PAGE法用于玉米种子纯度检验是比较可靠的，据此，用电泳纯度可推算出田间鉴定纯度。     

番茄POD与EST同工酶PAGE方法研究

对番茄种子芽期过氧化物酶（ＰＯＤ）同工酶与酯酶（ＥＳＴ）同工酶聚丙烯酰胺凝胶电泳（ＰＡＧＥ）方法进行了初步研究。结果表明,ＰＯＤ同工酶电泳酶样提取液以ｐＨ６．５￣０．１ｍｏｌ／Ｌ磷酸缓冲液＋０．１％巯基乙醇较适宜;ＥＳＴ同工酶酶样提取液以ｐＨ６．０￣７．０的０．１ｍｏｌ／ＬＴｒｉｓ－柠檬酸＋０．１％巯基乙醇或０．１ｍｏｌ／Ｌ磷酸缓冲液＋０．１％巯基乙醇较适宜。常用的７．５％分离胶浓度不适宜番茄种子     

银染聚丙烯酰胺凝胶电泳技术体系的优化

【目的】针对银染PAGE实验中常出现的背景颜色深、条带弥散、凝胶变形及条带不整齐等问题进行归纳总结并提出优化方法。【方法】以甘蓝型油菜基因组DNA为研究对象,通过PAGE电泳后银染并调整试剂配方、实验条件,筛选出适合实验的技术体系。【结果】甲醇浓度、电压稳定性、电泳温度、气泡等显著影响实验结果。甲醛浓度过高(1.25%)时,虽所需染色时间短,但凝胶背景深;甲醛浓度过低(0.001 25%)时,背景浅但条带淡,需要长时间染色;电压不稳定会造成条带弥散;电泳温度过高,导致凝胶变形,条带弯曲;凝胶中的气泡会扰乱DNA条带的形状;NaOH浓度在0.875%~1.375%,染色结果无明显差异。【结论】甲醛用量4 mL(1%)、保持稳定电压、在冰浴中电泳、制胶时及时赶走所有气泡,可得到凝胶背景浅、条带清晰且正常的结果。     

微芯片电泳与聚丙烯酰胺凝胶电泳的比较分析

聚丙烯酰胺凝胶电泳是目前实验室常用的分离寡核苷酸的电泳技术,微芯片电泳是近几年新兴起的DNA/RNA快速分析系统。对玉米自交系8112、黑糯等SSR分子标记的PCR扩增结果进行两种方法的比较,旨在分析检验微芯片电泳在重复性和分辨率上的可靠性。与聚丙烯酰胺凝胶电泳相比,微芯片电泳无毒性、操作更简单、灵敏度和准确性高,而且重复性良好,今后在科研工作中将拥有更加广阔的前景。     

3种鲟血清蛋白的比较研究

采用聚丙烯酰胺凝胶电泳及饱和硫酸铵分步沉淀法对健康的3龄中华鲟Acipenser sinensisG ray、2龄施氏鲟A.schrenckiiB randt和3龄达氏鳇Huso dauricusGeorgi的血清蛋白及其组成成分进行了研究。结果表明,3种鲟的血清电泳图谱具有各自的特有谱带,其电泳图谱的相似程度表明,中华鲟与施氏鲟的亲缘关系较达氏鳇与施氏鲟近。3种鲟血清蛋白的总浓度分别为18.70、21.25、24.60 mg/mL。