外源超氧物歧化酶基因转化巴西橡胶树的研究

本研究采用农杆菌侵染法成功实现玄参花叶病毒（FMV）34S启动子控制下的氧物歧化酶基因对巴西橡胶树（Hevea Brasiliensis Muell.Arg）的遗传转化。来自巴西橡胶无性系RRⅡ105未成熟花药的愈伤组织用含有双元质粒载体的根癌农杆菌菌株EHA 101进行转化。该双元载体带有用作选择标记的新霉素磷酸转移酶（nptⅡ）基因及β-葡糖醛酸酶（GUS）报告基因,同时含有带FMV 34S启动子的Mn-超氧物歧化酶（Mn-SOD）目的基因。试验中采用含4.5μM 2,4-D,2.2μM BAP,以及1.1μM NAA的MS培养基筛选并增殖抗卡那霉素的GUS阳性愈伤组织,并利用附加1.5μM BAP,1.4μM KIN,1.1μM NAA,以及2.0μM GA_3的MS培养基诱导高频体胚发生。用于胚胎成熟的培养基为附加1.2μM GA_3及1.5μM TDZ（噻苯隆）的改良MS培养基。试验结果显示,成熟的转化胚胎中有20%发育成小植株。这些小植株经健化之后被移植到营养袋中并置于温室中继续培育生长。最后,本研究通过用SOD及nptⅡ特异性引物作PCR扩增以及用nptⅡ特异性探针与分离自转化植株的DNA模板进行Southern杂交进一步证实了转化事件的发生。     

Responses of Pinus massoniana and Pinus taeda to freezing in temperate forests in central China

Abstract

Masson pine (Pinus massoniana Lamb.), a native species widely distributed in temperate forests in central China, and Loblolly pine (Pinus taeda L.), an exotic tree species introduced to China from southeastern United States, are dominant evergreen conifers that play a pivotal role in maintaining forest structure and functions for the region. We examined the effects of freezing on these species with chlorophyll fluorescence and electrolyte leakage using both field- and laboratory-based experiments in September 2009 and January 2010, respectively. We found that freezing could cause a greater impact on the Loblolly pine than the Masson pine. Although the two species showed similar values of F _v /F _m and electrolyte leakage before freezing, the Masson pine needles showed lower F _v /F _m and higher electrolyte leakage ratios than those of the Loblolly pine when treated in low temperatures (?15 to 0°C). We also found that cold-acclimation was crucial for both species to adapt to low temperatures with the F _v /F _m ratio decreased approximately by 80% in the first freezing hour for the non-acclimated needles of both species while the cold-acclimated needles showed little changes in the F _v /F _m ratio. This finding is also supported by our measurements of electrolyte leakage. These results suggest that the Loblolly pine could be more susceptible to freezing damages than the Masson pine in central China.     

青霉菌发酵液对大豆幼苗生长及生理特性的影响

采用室内生理检测及生物化学方法,研究了青霉菌发酵液对大豆幼苗生长及生理特性的影响。结果表明:用一定浓度的青霉菌发酵液对大豆浸种和茎叶处理,大豆幼苗的生长呈明显上升趋势,处理后大豆的株高、根长、株鲜重和根鲜重均明显得到了提高,且生长状态好于对照,不同浓度处理间具有明显差异,在发酵液浓度为5 g·L-1时,效果最佳。尤其是对根鲜重的影响,促进率最高可达56.21%,同时增强了幼苗根系活力,最大可以提高57.82%,浸种和茎叶处理后,分别提高了叶绿素含量41.25%和15.40%;超氧化物歧化酶活性12.20%和11.70%;过氧化物酶31.33%和29.95%;过氧化氢酶活性42.8%和20.0%,这为发酵液在大豆田推广应用、提高大豆产量提供了重要理论依据。     

饮用天然高钼水诱发的钼中毒耕牛主要生物酶测定

本研究对高钼饮水诱导钼中毒的耕牛及对照牛的血浆及组织（肝脏、肺脏、肾脏、脾脏、心脏、肌肉）中超氧化物歧化酶（ＳＯＤ）、黄嘌呤氧化酶（ＸＯＤ）及谷胱甘肽过氧化物酶（ＧＳＨ－ＰＸ）等３种生物酶活性进行了对比分析测定。结果表明,血浆和组织中ＳＯＤ、ＸＯＤ及ＧＳＨ－ＰＸ均随着钼中毒的加重其活性显著下降。这３种生物酶是钼中毒比较灵敏且相关较大的生化指标,对钼中毒具有一定诊断价值;测定ＳＯＤ。ＸＯＤ、ＧＳＨ－ＰＸ可作为钼中毒监测的重要手段。     

动物锰营养中含锰超氧化物歧化酶研究进展

锰超氧化物歧化酶(MnSOD)是一种催化超氧阴离子自由基发生歧化反应 ,生成氧和过氧化氢的重要含锰金属酶。本文对MnSOD的分布、结构、活性、基因表达以及动物饲粮锰及其它因素对MnSOD的调节等进行了综述 ,以期推动锰营养学的进一步深入研究     

Induction of Systemic Resistance in Cucumber against Several Diseases by Plant Growth-promoting Fungi: lignification and Superoxide Generation

Five fungal isolates (Trichoderma, Fusarium, Penicillium, Phoma and a sterile fungus) from zoysiagrass rhizosphere that promote plant growth were tested for their ability to induce systemic resistance in cucumber plants against Colletotrichum orbiculare. Roots of cucumber plants were treated with these fungal isolates using barley grain inocula (BGI), mycelial inocula (MI) or culture filtrate (CF). Most isolate/inoculum form combinations significantly reduced the disease except BGI of Trichoderma. These fungal isolates were also evaluated for induction of systemic resistance against bacterial angular leaf spot and Fusarium wilt by treatment with BGI. Penicillium, Phoma and the sterile fungus significantly reduced the disease incidence of bacterial angular leaf spot. Phoma and sterile fungus protected plants significantly against Fusarium wilt. Roots treated with CFs of these fungal isolates induced lignification at Colletotrichum penetration points indicating the presence of an elicitor in the CFs. The elicitor activity of CFs was evaluated by the chemiluminescence assay using tobacco callus and cucumber fruit disks. The CFs of all isolates elicited conspicuous superoxide generation. The chemiluminescence activity of the CF of Penicillium was extremely high, and its intensity was almost 100-fold higher than that of other isolates. The chemiluminescence activity was not lost following treatment with protease or autoclaving or after removal of lipid. The MW 12,000 dialyzed CF fraction was highly effective in eliciting chemiluminescence activity. Chemiluminescence emission from cucumber fruit disks treated with Penicillium was the same as that obtained from tobacco callus, except that the lipid fraction also showed a high activity. Both the MW 12,000 fraction and the lipid fraction induced lignification in the epidermal tissues of cucumber hypocotyls.     

柠檬酸对小鼠睾丸组织抗氧化酶活性的影响

为研究柠檬酸对小鼠睾丸组织抗氧化酶活性的影响,将40只健康昆明系小鼠随机分为对照组(生理盐水)、柠檬酸低(180mg/kg)、中(270mg/kg)、高(360mg/kg)剂量组,各组小鼠每周腹腔注射相应剂量试剂0.2mL,连续注射3周,第3次注射后的第7天时颈椎脱臼处死小鼠,取其睾丸组织制作匀浆液检测过氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、乳酸脱氢酶(LDH)、总一氧化氮合酶(T-NOS)活性及丙二醛(MDA)含量。结果表明,与对照组相比,中、高柠檬酸剂量组小鼠睾丸组织SOD活性显著降低(P<0.05);柠檬酸处理组GSH-Px活性、MDA含量均显著下降(P<0.05);随着柠檬酸剂量的增加,LDH、NOS活性分别呈下降、上升趋势。此结果提示,一定剂量的柠檬酸可以降低小鼠睾丸组织SOD、GSH-Px活性,但可能通过NO途径减少睾丸细胞膜脂质过氧化,提高细胞膜的稳定性。     

In UV‐B Irradiated Rice High Levels of Mn Limits Chloroplasts Injury Triggered by Oxidative Stress

The molecular mechanism to control the oxidative burst exerted by Mn accumulation in rice (Oryza sativa L.) plants, grown in hydroponics containing 0.5, 2 and 8 mg l^–1 Mn and irradiated with a total biological effective UV‐B irradiation of 20.825 kJ m^?2, was investigated in the chloroplasts at the 15th and 21st days after germination. In both experimental periods, Mn accumulation kinetics in the leaves and in the chloroplast lamellae displayed overall increases. Coupled to higher membrane selectivity, superoxide production and acyl lipids peroxidation in the thylakoids decreased, prompting upper rates of the Hill and Mehler reactions. Connected to UV‐B irradiation, higher accumulated Mn in thylakoids was found to be chelated in a 36.5 kDa protein, with Mn/protein ratio of about 1 and high content of Gln, Asp, Glu, Leu and Gly, being its EPR spectrum characteristic of high‐spin Mn(II), in a S = 5/2 ground state. As this protein exhibited enzymatic catalysis of superoxide dismutation, it was concluded that, under UV‐B irradiation, the high internal tolerance of Oryza sativa L. to Mn during the vegetative growth also triggers the synthesis of a manganese protein that mimics superoxide dismutase functioning, therefore furnishing an additional intimate protection against oxidative stress.     

罗非鱼活性肽分离及抗氧化能力研究

利用正交试验L9(34),以清除超氧自由基能力为指标,分别对木瓜蛋白酶、中性蛋白酶水解罗非鱼肉的水解条件进行优化,并对最佳清除率下的两种酶解物进行Sephadex G-50凝胶柱分离,检测了各组分对超氧自由基清除率及活性肽分子量分布情况.结果表明:中性蛋白酶在45 ℃、酶的质量分数2.0%、水解105 min及肉水比1∶3的水解条件下对超氧自由基有较好清除作用;木瓜蛋白酶在60 ℃、酶的质量分数2.0%、时间150 min及肉水比1∶2的水解条件下对超氧自由基有较好清除效果.木瓜蛋白酶酶解物在分子量为660 Da的多肽洗脱峰具有最大超氧自由基清除率,中性蛋白酶酶产物在分子量为1320 Da多肽洗脱峰具有最大超氧自由基清除率.     

低氧胁迫对河川沙塘鳢抗氧化酶及ATP酶活性的影响

研究了在急性低氧1.5 h、5 h和慢性低氧3 d下,河川沙塘鳢(Odontobutis potamophila)5种组织(心、脑、肝、鳃和肾)的超氧化物歧化酶(SOD)、过氧化氢酶(CAT)及谷胱甘肽过氧化物酶(GPX)3种抗氧化酶和ATP酶活性的变化规律。结果显示:在急性低氧暴露1.5 h时,河川沙塘鳢SOD和GPX的活力在各组织中与对照组相比均无显著差异,CAT活力在心、鳃和肝3种组织呈现显著升高(P0.05),ATP酶活力在心和肝组织极显著升高(P0.01);在急性低氧暴露5 h时,除肝组织SOD酶活性显著降低外(P0.05),其它4种组织的CAT、GPX和ATP酶均不同程度显著升高(P0.05);在慢性低氧处理3 d时,心、脑组织的抗氧化酶已基本恢复至与对照组无显著差异的水平,但鳃、肝和肾中酶活力仍较高(P0.05)。研究表明,河川沙塘鳢能通过自身调节抗氧化酶及ATP酶活性,改变代谢底物,提高机体适应低氧环境的能力。