土壤类过氧化物酶活性对长期施肥的响应

为探究不同施肥处理对土壤类过氧化物酶活性的影响，以灰漠土和红壤长期定位试验站施肥处理土壤为研究对象，借助纳米矿物颗粒的类酶颜色反应原理，研究了不施肥(CK)、施用化肥(NPK)、施用化肥和有机肥(NPKM)和施用有机肥(M)土壤类过氧化物酶活性的变化。结果表明：与天然辣根过氧化物酶(HRP)相似，土壤也具有经典的催化颜色反应，灰漠土和红壤类过氧化物酶活性范围为0.4～1.0 U/kg。长期施肥显著改变了土壤类过氧化物酶活性，其中以M处理土壤类过氧化物酶活性为最高，可达0.7 U/kg，比CK处理增加了70%以上。不同施肥年限下，CK和NPK处理土壤类过氧化物酶活性并无显著差异，而M和NPKM处理土壤类过氧化物酶活性则随施肥时间线性增加(R²=0.71,P<0.01)。高温灭菌处理后，土壤类过氧化物酶活性大多没有改变，表明其活性是土壤本身固有的氧化还原特性；但M处理红壤类过氧化物酶活性比CK和NPK处理红壤稳定性更高。皮尔森相关性分析表明，类过氧化物酶活性可能影响土壤可溶性矿质元素(如硅和镁)水平。因此，施肥措施可以通过改变土壤固有类过氧化物酶活性而影响养分周...     

MTBE和Cd复合作用对土壤酶活性和小麦吸收重金属的影响

采用盆栽试验研究不同浓度(0,5,10,50,100,200,300,400,500mg/kg)的MTBE和相同浓度系列MTBE与Cd(5 mg/kg)复合作用对土壤酶活性的影响以及复合作用对小麦吸收重金属的影响.结果表明:MTBE作用下土壤脲酶、蔗糖酶、过氧化氢酶活性表现出先增高后降低的趋势,在MTBE浓度为50mg/kg时,3种土壤酶活性达到最高值,与对照相比分别提高了14.7％,4.2％,10.1％;复合作用下3种土壤酶活性表现出先降低然后升高再降低的趋势,在MTBE浓度为10mg/kg时与对照相比分别降低了17.2％,8.7％,14.0％;在MTBE浓度为50 mg/kg时,可以缓解重金属对3种土壤酶活性的抑制作用,但酶活性仍低于空白组;在MTBE浓度为200,300,400,500mg/kg时,2种物质的复合作用抑制了3种酶的活性;复合作用对土壤酶活性的抑制比MTBE作用要明显;复合作用下小麦中重金属含量表现出增高的趋势,这说明MTBE增加了Cd的可迁移性,从而使土壤中的Cd含量降低.     

仿刺参体腔液补体类似物化学发光免疫检测

首次应用酶联化学发光免疫检测(Chemiluminesent Immunoassay,CLIA)技术测定仿刺参体腔液补体类似物AjC3和AjC4。羊抗人C3、C4抗体吸附到经过紫外线处理的聚苯乙烯管内,采用辣根过氧化物酶(HRP)标记抗体。过氧化氢和鲁米诺为辣根过氧化物酶的底物。捕获抗体包被最适浓度为1μg/ml,免疫反应20℃孵育2h达到平衡。HRP-IgC3、IgC4抗体复合物适宜稀释度为1:2000,HRP-IgC3I、gC4抗体复合物4℃下保存8d性能稳定,室温下5d内性能稳定。标准品浓度在0.1～10ng/ml范围内时与化学发光值之间具有良好的线性相关性,检测灵敏度为0.1ng/ml。结果表明应用酶联化学发光免疫检测技术能够检测到仿刺参体腔液中含有补体类似物,AjC3含量为6.58±1.4μg/ml,AjC4含量为0.67±0.3μg/ml。     

土霉素及镉污染对土壤呼吸及酶活性的影响

随着饲料工业以及畜禽养殖业的规模化发展,抗生素和重金属在土壤环境中同时存在的几率不断增大。为了分析抗生素和重金属对土壤微生物生态系统的影响,以土霉素(OTC)与镉(Cd)为污染物,采用室内培养法,研究了土霉素(OTC)与镉(Cd)单一处理及复合污染对土壤呼吸和酶活性的影响。结果表明,10mg/kg重金属镉单独污染对土壤微生物呼吸表现为先抑制后激活作用,且显著抑制了土壤蔗糖酶、脲酶、磷酸酶活性,对3种酶活性平均抑制率从大到小依次为:蔗糖酶磷酸酶脲酶;1mg/kg土霉素显著激活土壤微生物呼吸,50和200mg/kg土霉素对土壤微生物呼吸的影响呈现出先抑制后激活的规律。各处理浓度下的土霉素对蔗糖酶和脲酶活性均主要表现为抑制作用,对磷酸酶活性的影响呈现出一定的波动性;当土霉素的浓度为1和200mg/kg时,其与10mg/kg镉的复合污染对土壤微生物呼吸及3种酶活性的影响主要为拮抗作用,但当土霉素的浓度为50mg/kg时,与10mg/kg镉的复合污染对土壤微生物呼吸及3种酶活性的影响则主要为协同作用。微生物呼吸对土霉素与镉胁迫更为敏感,最高抑制率和激活率分别可达98.98%和300.82%,土壤酶活性受土霉素和镉污染的影响要弱于它们对土壤微生物呼吸的影响。     

稀土配合物农用对土壤微生物活性的影响

研究了稀土(La,Nd)与唑类二元和三元配合物及其稀土盐对耕作土壤中细菌、霉菌及土壤酶活性的影响作用;结果表明,各供试稀土盐配合物在浓度小于1.5 mg/g时对土壤生物活性影响不大。当浓度大于3mg/g时,对土壤中细菌、霉菌及脱氢酶、过氧化氢酶活性均具有一定抑制作用。在化合物浓度为15mg/g土样中:细菌生长的抑制率达47％-63％,霉菌为23％-34％,过氧化氢酶抑制率为9.5％-38.7％,脱氢酶活性明显降低。稀土化合物农用对土壤微生态环境的影响不容忽视。     

硅对Cd胁迫下黄瓜苗期光合及抗氧化酶系统的影响

为了解石灰性土壤硅对Cd胁迫下黄瓜苗期叶片的光合特性和抗氧化酶系统及对实际土壤Cd毒害植物的防治,研究了硅介入下Cd污染土壤中黄瓜苗期的叶片净光合速率(P_n)、气孔导度(G_s)、蒸腾速率(T_r)、胞间CO_2(C_i)等光合作用指标,叶绿素含量,丙二醛(MDA)、超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)等抗氧化酶系统等相关参数。结果表明:与空白相比较,单施Cd处理下,P_n、G_s、T_r和叶绿素含量下降,Ci提高。硅介入后,能明显提高P_n、G_s、T_r和叶绿素总量,降低C_i含量。当C_d含量为5mg/kg时,硅施加量100~200mg/kg情况下,与空白相比,P_n、G_s、T_r和叶绿素上升,C_i下降,但100mg/kg与200mg/kg的硅处理差异不明显,300mg/kg的硅添加量效果最好。对于抗氧化酶系统,与空白相比较,土壤Cd含量为3~5mg/kg时,3类抗氧化酶的活性均受到显著抑制,200~300mg/kg硅的引入可明显提高SOD、POD、CAT活性,降低MDA值。可见,硅的引入可明显改善Cd胁迫下黄瓜叶片的光合特性和抗氧化酶系统。施用少量硅肥在石灰性土壤中对缓解植物Cd毒害具有一定的应用前景。     

模拟酸雨和镍复合污染红壤中莴笋的生长与抗氧化反应

采用盆栽实验,研究了在模拟酸雨、镍单一或复合污染的红壤中,莴笋的生长和叶片中抗氧化酶活性、活性氧和丙二醛含量的变化。结果表明,单一酸雨污染或酸雨与25 mg/kg镍的复合污染对植株生长无明显影响;酸雨增强了50-100 mg/kg镍对莴笋生长的抑制效应。复合污染下,酸雨为pH4.5时,莴笋叶片超氧化物歧化酶（SOD）、过氧化氢酶（CAT）的总体活性较单一镍污染高,而酸雨为pH3.5时则两者活性降低。复合污染下,叶片过氧化物酶（POD）活性升高,抗坏血酸过氧化物酶（APX）活性降低,且随酸雨pH降低,变化的幅度增大;同时,O2^＊ˉ的产生加快,H2O2和MDA含量升高,表明叶片组织遭受了氧化胁迫和膜脂过氧化损伤,这可能是复合污染下莴笋生长受阻的重要原因。     

钙对大豆幼苗镍毒害的缓解效应及对抗氧化酶活性的影响

采用盆栽方法,研究了镍毒害条件下外源钙对大豆镍毒害的缓解效应以及对其叶片组织中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)和抗坏血酸过氧化物酶(APX)活性的影响,以探讨钙缓解植物镍毒性的生理机制.研究结果表明:Ni2+60 mg/kg添加量对大豆幼苗生长表现出明显的抑制作用,但是当加入不同钙量后,大豆的镍毒害症状得到不同程度的缓解,Ca2+ 200 mg/kg处理的缓解作用好于Ca2+ 100 mg/kg.与不施镍处理相比,单施Ni2 60 mg/kg处理的大豆叶片组织SOD、CAT和APX活性明显下降,而POD活性明显上升.当配施Ca2以后,大豆叶片组织中SOD、CAT、POD和APX活性明显提高,且Ca2+ 200 mg/kg处理的上升程度比Ca2+ 100 mg/kg处理的大.因此,Ca2+是通过提高叶片组织抗氧化酶活性,增强活性氧清除能力,而使大豆的镍中毒症状得到缓解,这是Ca2+减轻大豆镍毒害的生理机制之一.     

施磷对镉胁迫下黄瓜苗期光合作用及抗氧化酶系统的影响

  目的  探究石灰性土壤中施磷对镉胁迫下黄瓜苗期光合作用及抗氧化酶系统的影响,为合理施用磷肥、缓解镉毒害提供依据。  方法  采用盆栽完全组合设计方案,研究了不同土壤镉添加水平（0、1、3和5 mg kg?1土）下施加不同浓度磷（0、50、100、150和200 mg P₂O₅ kg?1土）对黄瓜苗期生物量、叶片光合作用及抗氧化酶系统的影响。  结果  随着施Cd量的增加,黄瓜生物量先增后降,黄瓜叶片的净光合速率（Pn）、气孔导度（Gs）、蒸腾速率（Tr）显著降低,胞间CO₂浓度（Ci）呈上升趋势。施加适量磷（≤ 150 mg P₂O₅ kg?1）能够显著提高黄瓜生物量、Pn、Gs、Tr、叶绿素总量和类胡萝卜素含量（Car）,降低叶片中Ci。对于抗氧化酶系统来说,随着施Cd量的增加黄瓜叶片中过氧化物酶（POD）、过氧化氢酶（CAT）活性显著降低,丙二醛（MDA）含量明显增加。施磷后,能够明显增强抗氧化酶系统。施Cd量为1 mg kg?1,施磷量为50和100 mg P₂O₅ kg?1时,CAT活性增加了1.9% ~ 9.6%。施Cd量为3和5 mg kg?1时,随着施磷量的增加,POD和CAT活性出现不同程度增加,与不施磷相比,POD活性分别增加了149.1% ~ 197.8%和150.4% ~ 252.0%,CAT活性分别增加了5.8% ~ 16.6%和0.55% ~ 19.4%,其中施磷量为100 mg P₂O₅ kg?1时黄瓜叶片POD和CAT活性达到最大。同时,施P能够降低叶片中MDA含量,且随着施磷量的增加而降低。  结论  在石灰性土壤中,施磷量为100和150 mg P₂O₅ kg?1时,能够明显缓解镉毒害,增强黄瓜叶片的光合作用及抗氧化酶系统,而施磷量达到200 mg P₂O₅ kg?1时则对黄瓜生长产生抑制作用。     

LaCl3对桑树组培苗生长与分化的影响

本试验目的是通过研究不同浓度的稀土化合物(LaCl3)对桑树组培苗生长分化以及生根的影响,确定培养基中加入LaCl3的浓度。在前人研究稀土元素对植物生理生化影响的基础上,本试验将不同浓度稀土化合物(LaCl3)添加于桑树组培苗分化与生根培养基中,研究其对桑树组培苗生长与分化的影响。结果表明,低浓度的LaCl3（40 mg/L）能促进桑树组培苗的生长与分化,株高、生芽数、生根率、根长和根重都显著高于对照及其他浓度处理;分化芽数、株高及生根率分别比对照高84.2%、63.5%、12.5%。同时还能提高叶绿素含量,其中叶绿素a含量为对照的2.06倍;叶绿素b含量为对照的1.64倍;以及改变光合色素比例,提高过氧化物酶活性。高浓度LaCl3（60～80 mg/L）的促进作用减小甚至抑制桑树组培苗的生长。说明本试验条件下,有利于桑树组培苗分化及生根的LaCl3最佳浓度为40 mg/L。     

Broccoli processing wastes as a source of peroxidase

A peroxidase isozyme (BP) was purified to homogeneity from broccoli stems ( Brassica oleraceae var. maraton) discarded from industrial processing wastes. BP specific activity was 1216 ABTS [2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)] units/mg, representing 466-fold that of crude extract. BP is a monomeric glycoprotein containing 16% carbohydrates, with a molecular mass of 49 kDa and an isoelectric point close to 4.2. From kinetic data it showed a two-substrate ping-pong mechanism, and the catalytic efficiency measured as the rate-limiting step of free BP regeneration was 3.4 x 10(6) M(-1) s(-1). The ABTS K m value was 0.2 mM, which was about 20 times lower than that reported for acidic commercial horseradish peroxidase (HRP). Assessment of BP secondary structure showed 30% helical character, similar to HRP and cytochrome c peroxidase. BP lost only 25% activity after 10 min of heating at 55 degrees C and pH 6; it was stable in the pH range from 4 to 9 and showed an optimum pH of 4.6 using ABTS as substrate. BP was active on substrates normally involved in lignin biosynthesis, such as caffeic and ferulic acids, and also displayed good catechol oxidation activity in the presence of hydrogen peroxide. Reverse micellar extraction was successfully used as potential large-scale prepurification of broccoli peroxidase, achieving a purification factor of 7, with 60% activity yield. Stems from the broccoli processing industry have a high potential as an alternative for peroxidase purification.     

Purification and properties of a neutral peroxidase isozyme from turnip (Brassica napus L. Var. Purple Top White Globe) roots

A neutral peroxidase isozyme (pI 7.2) from turnip roots (TNP) was purified to homogeneity and partially characterized. TNP is a monomeric glycoprotein with 9.1% carbohydrate content and a molecular weight of 36 kDa. Optimum pH values for activity using 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid (ABTS) and guaiacol as H donors were 4.5 and 5.5, whereas the K(m) values were 0.7 and 3.7 mM, respectively. The ABTS K(m) was approximately 7 times higher than that reported for basic commercial horseradish peroxidase (HRP-C). TNP retained approximately 70% activity after 11 min of heating at 65 degrees C, whereas the activation energy for inactivation (132 kJ/mol) was higher than or comparable to that of other peroxidases. The low ABTS K(m) and high specific activity (1930 units/mg) gave a high catalytic efficiency (500 M(-1) s(-1)). These properties make TNP an enzyme with a high potential as an alternative to HRP in various applications.     

Enzymatic oxidative treatments of wheat bran layers: effects on ferulic acid composition and mechanical properties

Enzymatic treatments known to induce the gelation of feruloylated arabinoxylans solutions were applied to tissue strips isolated from peripheral layers of wheat grain to tentatively produce in situ arabinoxylan reticulation. The treatments by horseradish peroxidase (HRP) and manganese dependent peroxidase (MnP) induced a dimerization of ferulic acid (FA) in wheat bran with concomitant decrease of arabinoxylan solubility. Similar results were obtained, but to a lesser extent, by simple incubation of bran strips in water, suggesting the action of endogenous peroxidases. The fact that these treatments proved to be ineffective on the isolated aleurone layer and pericarp suggested that dimerization occurred mostly at the aleurone-pericarp interface. In addition, the MnP system generated a consumption of monomer and dimer of ferulic acid in the pericarp, perhaps due to their incorporation into lignin. Micro-mechanical tests using DMTA were performed on isolated tissue strips and showed that oxidation of wheat bran increased their mechanical strength (increase of stress and strain to rupture).     

Purification and partial characterization of three turnip (Brassicanapus L. var. esculenta D.C.) peroxidases

Three turnip peroxidases (fractions C1, C2, and C3) were partially purified and characterized, to permit study of their feasibility for use in clinical and enzyme immunoassays. These fractions represented 20% of the initial activity, and fractions C1 and C2 were purified to homogeneity. The optimum pH was between 5.0 and 5.5, while optimum temperature ranged from 40 to 55 degrees C. The ABTS K(m) values for the two acidic fractions (C2 and C3) were 0.70 and 0.42 mM, respectively; about 5 times lower than that reported for the acidic commercial horseradish peroxidase (HRP). Fraction C3 had 4 times higher K(m) value than commercial cationic HRP. The molecular weights determined by SDS-PAGE ranged from 39.2 to 42.5 kDa. Activation energies for inactivation were 113 (C1), 130 (C2), and 172 kJ/mol (C3) which are higher or comparable to other peroxidase isoenzymes reported. Fractions C1 and C3 represent an alternative source of peroxidase because of their higher purification yield and specific activity, when compared to fraction C2.     

施肥和外源砷对小白菜生长和抗性生理的影响

通过盆栽试验,研究了施用有机肥料与氮、 磷、 钾肥以及不同浓度砷对小白菜（Brassica campestris L. ssp. Chinensis L.）株高、 根长、 单株鲜重、 砷含量以及小白菜植株脯氨酸（Pro）、 丙二醛（MDA）、 超氧化物歧化酶（SOD）、 过氧化物酶（POD）等的影响。结果表明: 在施肥条件下外源砷10 mg/kg促进小白菜生长,鲜重最高,株高、 根长增加,外源砷继续提高则抑制小白菜生长; 小白菜砷含量随外源砷提高而增加,施肥则促进砷吸收; 小白菜Pro和MDA含量随外源砷提高而增加,施肥则有助于降低Pro和MDA含量; 小白菜POD活性在不施肥处理下随砷浓度的提高呈上升下降的变化趋势,且以砷 50 mg/kg时最高; 在施肥条件下随砷浓度的增加而增强; 小白菜SOD活性在有机肥处理下随砷浓度的增加而增强,在不施肥、 施氮磷钾肥以及氮磷钾肥和有机肥混施处理下随砷浓度的升高呈上升下降的变化趋势,均以砷 50 mg/kg处理最高。     

赤霉素对盐害下碱茅的萌发和幼苗生长的影响

以碱茅为试材,在100 mmol/L的NaCl盐胁迫下,通过种子培养、室内盆栽试验,研究盐胁迫下不同浓度赤霉素（8,15,20,25,50 mg/L）对碱茅种子萌发、幼苗生长的影响。结果表明:（1）100 mmol/L的NaCl严重抑制碱茅的种子萌发和幼苗生长,碱茅种子的发芽率、发芽势和发芽指数、根茎长显著降低,根系活力、过氧化物酶活力受到严重抑制;（2）在外源赤霉素溶液为8～20 mg/L的范围内,随着浓度的升高,碱茅种子的发芽率、发芽势和发芽指数、根茎长逐渐升高,根系活力、过氧化物酶活力、耐盐指数明显提高;（3）高浓度（ > 20 mg/L）的外源赤霉素浸种处理,对种子的促进作用逐渐下降,当赤霉素浓度为50 mg/L时,碱茅的发芽率和根茎比低于对照,产生了抑制效应。结论:不同浓度赤霉素对提高碱茅种子耐盐性有不同程度的促进作用,以赤霉素浓度为20 mg/L时作用效果最佳。     

Development of a new method, based on a bioreactor coupled with an L-lactate biosensor, toward the determination of a nonspecific inhibition of L-lactic acid production during milk fermentation

The development and characteristics of a bioreactor employing bacteria (Streptococcus thermophilus) encapsulated in Ca-alginate beads coupled with an L-lactate biosensor are reported. The biosensor comprises a carbon paste electrode modified with enzymes HRP (horseradish peroxidase), LOD (lactate oxidase), and FcH (ferrocene) as redox mediator. The measurement of L-lactate is based on the signal produced by H(2)O(2), the product of the enzymatic oxidation of L-lactate by LOD. The detection of H(2)O(2) is performed at the electrode surface via HRP/FcH at low operating potential (-100mV vs Ag/AgCl). Optimization studies were performed using the bioreactor in conjunction with an L-lactate electrode operating in a flow injection system to assess the ability of encapsulated bacteria to ferment carbohydrate solutions. The possibility of using the developed method to assess the fermentation capability of milk samples was evaluated. Bronopol (2-bromo-2-nitro propane-1,3-diol) was chosen to simulate the effect of an inhibitory agent of milk fermentation. The obtained results indicated that the evaluation of the amount of L-lactate amount produced through the bioreactor could be used as a measure of inhibition of lactic acid production in milk samples.     

Toxic reactivity of wheat (Triticum aestivum) plants to herbicide isoproturon

The herbicide isoproturon is widely used for controlling weed/grass in agricultural practice. However, the side effect of isoproturon as contaminants on crops is unknown. In this study, we investigated isoproturon-induced oxidative stress in wheat ( Triticum aestivum). The plants were grown in soils with isoproturon at 0-20 mg/kg and showed negative biological responses. The growth of wheat seedlings with isoproturon was inhibited. Chlorophyll content significantly decreased at the low concentration of isoproturon (2 mg/kg), suggesting that chlorophyll was rather sensitive to isoproturon exposure. The level of thiobarbituric acid reactive substances (TBARS), an indicator of cellular peroxidation, showed an increase, indicating oxidative damage to plants. The isoproturon-induced oxidative stress resulted in a substantial change in activities of the majority of antioxidant enzymes, including superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX). Activities of the antioxidant enzymes showed a general increase at low isoproturon concentrations and a decrease at high isoproturon concentrations. Activities of CAT in leaves showed progressive suppression under the isoproturon exposure. Analysis of nondenaturing polyacrylamide gel electrophoresis (PAGE) confirmed these results. We also tested the activity of glutathione S-transferase (GST) and observed the activity stimulated by isoproturon at 2-10 mg/kg.     

棉花氮素营养状况的诊断研究

为进行棉花氮素营养诊断指标和追肥推荐,应用反射仪开展了不同氮肥用量对棉花倒四叶叶柄的硝酸盐浓度影响的研究。田间试验在新疆阿瓦提县丰收三场二连进行。设5个氮肥用量,分别为N.0、180、240、300、360kg/hm2。结果表明,畦灌条件下的陆地棉花期、花铃期和铃期的叶柄硝酸盐浓度和施氮量之间有显著正相关,用一元二次模型模拟氮肥与产量之间的关系,得到最佳经济施氮量为304.kg/hm2,对应的经济产量为2420.kg/hm2;初步确立花期和铃期叶柄硝酸盐诊断临界值分别为10463和6901mg/L。花期、花铃期和铃期时棉株叶柄硝酸盐浓度与产量间有极显著相关性。以此为依据,建立了棉花植株硝酸盐诊断的氮肥推荐模型,并根据该模型计算出棉花各生育期不同硝酸盐测试值所对应的氮肥追肥用量。