过表达AtchyB基因提高洋桔梗光胁迫耐受性的研究

类胡萝卜素尤其是叶黄素循环类物质在植物抵抗由强光照引起的非生物胁迫中发挥着重要的作用,为了提高洋桔梗对强光照的抗性,从拟南芥中克隆了类胡萝卜素生物合成途径中参与叶黄素循环关键酶——β-胡萝卜素羟化酶基因(AtchyB),利用农杆菌介导法将其转入洋桔梗中,最终得到遗传转化植株2个株系,研究发现,转基因植株中总类胡萝卜素含量高于对照组,且叶黄素循环池被不同程度地放大。在不同光照强度下,转基因洋桔梗植株对光照耐受性明显强于对照组,且转基因植株生物量也明显提高。表明过表达AtchyB基因使洋桔梗光胁迫耐受性有所增强。     

过表达IbOr基因甘薯增强抗旱性的生理机制

以超表达甘薯橙色基因(IbOr)的转基因甘薯(TS)以及非转基因甘薯(NT)为实验材料,通过15%聚乙二醇6000(PEG-6000)模拟干旱条件,研究转基因与非转基因甘薯幼苗在水分胁迫不同时间的光合系统,膜脂过氧化及抗氧化防御系统中主要指标的变化情况,探讨转基因甘薯耐旱性的生理机制。结果显示:(1)随PEG-6000胁迫时间延长,甘薯叶片的叶绿素、类胡萝卜素含量及其叶片净光合速率、气孔导度、胞间CO2浓度、蒸腾速率都显著降低,但转基因株系降低幅度小于非转基因植株。(2)在正常供水和水分胁迫下,超表达IbOr基因甘薯叶片中O-·2、MDA含量均低于非转基因甘薯,即转基因甘薯具有较低的活性氧水平且脂膜受损伤较小。(3)PEG-6000胁迫24h后,甘薯叶片中SOD、POD酶活性均增加,48h达到最大值,且转基因甘薯中2种酶活性显著高于非转基因甘薯。研究表明,过表达IbOr基因可以有效减轻甘薯在水分胁迫条件下受损害的程度,且可能主要通过提高甘薯的抗氧化胁迫能力来完成。     

干旱胁迫下荒漠植物红砂叶片抗氧化特性

以荒漠半荒漠地区优势种超旱生灌木红砂两年生植株为研究对象,测定了不同程度干旱胁迫下叶片膜脂过氧化和抗氧化特征参数,以探讨荒漠植物抗氧化系统对荒漠极端环境的适应机制.结果显示:红砂叶片过氧化氢(H2O2)含量随干旱胁迫加剧而显著增加,丙二醛(MDA)含量在不同程度干旱胁迫下均有不同程度的增加,可溶性蛋白含量随干旱胁迫的增加而显著降低;类胡萝卜素和脯氨酸含量在不同程度干旱胁迫下均有不同程度增加;超氧化物歧化酶(SOD)活性随干旱胁迫加剧而显著提高,过氧化氢酶(CAT)活性在不同程度干旱胁迫下均有不同程度增加;过氧化物酶(POD)活性在中度干旱胁迫下显著增加而在重度干旱胁迫下显著降低.研究表明,荒漠植物红砂叶片中胡萝卜素、脯氨酸、SOD和CAT在防御干旱引起的氧化胁迫中起到了显著保护作用, 而POD在重度干旱胁迫下没有起到积极保护作用.     

外源钙对干旱胁迫下桔梗幼苗生理特性和桔梗品质的影响

【目的】干旱胁迫抑制药用植物光合色素合成,降低植株光合活性、光合碳同化效率,阻碍养分运输,严重影响药材的产量和品质。钙是植物生理活动中多种酶的活化剂,同时Ca2+作为细胞内第二信使与植物非生物胁迫密切相关。揭示外源钙缓解桔梗干旱胁迫伤害的潜在生理机制,对利用外源钙提高干旱、半干旱地区桔梗耐旱性和药用品质具有重要意义。【方法】以桔梗幼苗为材料,采用盆栽试验在干旱条件下叶面喷施10 mmol/L CaCl2,研究外源钙对桔梗幼苗生长、光合气体交换参数、抗氧化酶活性及药用部位品质等的影响。【结果】（1）外源钙处理能促进干旱胁迫下桔梗根长和干鲜生物量显著增加。（2）外源钙使干旱胁迫下桔梗叶片气孔开度、叶绿素a含量、总叶绿素含量、类胡萝卜素含量、净光合速率（Pn）、气孔导度（Gs）和蒸腾速率（Tr）分别显著增加30.28%、57.67%、44.44%、100.33%、89.53%、60.00%和83.11%。（3）外源钙使干旱胁迫下桔梗叶片丙二醛（MDA）、过氧化氢（H2O2）含量分别显著降低13.82%和18.66%,同时使干旱胁迫下桔梗叶片中过氧化物酶（POD）、超氧化物歧化酶（SOD）和过氧化氢酶（CAT）活性分别增加了25.43%、7.90%和33.92%。（4）外源钙不仅促进桔梗根中药用成分皂苷D、多糖、总黄酮、游离氨基酸在正常条件下分别显著增加35.34%,34.87%,4.19%和6.52%,还使它们在干旱胁迫处理下分别显著增加10.94%、7.53%、6.07%和5.78%。（5）外源钙提高了干旱胁迫下桔梗根中可溶性蛋白含量,以及地上部和地下部矿质元素含量。【结论】喷施10 mmol/L CaCl2能进一步激发干旱环境中桔梗叶片抗氧化系统的保护作用,通过渗透调节提高光合色素含量和光合性能,协同促进次生代谢产物和矿质元素累积,改善药用部位品质,从而缓解干旱对桔梗幼苗的伤害。     

外源水杨酸对低温胁迫下圆柏属植物幼苗生理特性的影响

以3年生圆柏和祁连圆柏幼苗为材料,采用不同浓度水杨酸(SA)预处理两种圆柏属植物幼苗,测定-4℃低温胁迫处理第6天叶片相对含水量(RWC)、相对电导率(REC)、丙二醛(MDA)、可溶性糖(SS)、可溶性蛋白(SP)、脯氨酸(Pro)和类胡萝卜素(Car)含量及超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)、抗坏血酸氧化酶(APX)和谷胱甘肽还原酶(GR)活性,分析外源 SA 对低温胁迫下圆柏属植物叶片膜脂过氧化、渗透调节物质及抗氧化酶系统的影响,为培育较多品种的抗冷冻常绿植物提供理论依据。结果表明：低温胁迫下,一定浓度的 SA 预处理能有效保护幼苗叶片膜系统的稳定性、增加渗透调节物质含量和提高抗氧化酶活性,其中200 mg/L SA 预处理对提高圆柏抗寒性效果最好,30 mg/L SA 预处理对提高祁连圆柏抗寒性效果最好,且在外源 SA 预处理下,祁连圆柏的抗寒性比圆柏的强。因此,施用适合浓度的 SA 在提高圆柏属植物抗寒性方面具有较好的应用价值。     

异源过表达胡杨PeDWF4基因提高烟草对非生物胁迫的耐性

本研究以烟草为材料异源过表达了胡杨PeDWF4基因,选取T1代的转基因阳性植株和野生型烟草幼苗水培培养50 d后对其分别施加100 mmol/L Na Cl、100 mmol/L甘露醇和100μmol/L Cu SO4进行处理,通过测定其植株生物量、叶绿素含量、抗氧化酶系统酶SOD和POD活性,渗透调节物质(脯氨酸,可溶性糖和可溶性蛋白)含量等,研究在不同非生物胁迫下PeDWF4转基因烟草对胁迫的生理生化响应及过量表达PeDWF4基因对其抗逆方面的影响和作用。结果表明,(1)异源过表达PeDWF4基因可显著提高转基因烟草的植株鲜重和生物量;(2)在3种胁迫下,转基因烟草通过减少叶片内叶绿素的降解,增加抗氧化系统酶SOD和POD活性以及提高渗透调节物脯氨酸等的含量等方式来减少非生物胁迫对自身的伤害。以上结果表明,异源表达PeDWF4基因可能是通过提高了转基因烟草內源的BR含量,从而提高了烟草对于以上3种非生物胁迫的耐性。     

外源钙对汞胁迫下菹草(Potamogeton crispus L.)叶片抗氧化系统及脯氨酸代谢的调节效应

以组织培养的菹草无菌苗为实验材料,研究了外源Ca(NO3)2对汞(Hg)胁迫下菹草无菌苗抗氧化酶系统、可溶性蛋白、总抗氧化能力(T-AOC)、非蛋白巯基(NP-SH)、植物络合素(PCs)以及脯氨酸代谢的调节效应。结果表明:(1)Hg胁迫使菹草体内过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)及谷胱甘肽还原酶(GR)活性明显降低,但超氧化物歧化酶(SOD)及过氧化物酶(POD)的活性并没有明显变化。外源添加25 mmol·L-1Ca(NO3)2后,明显提升了SOD、POD、APX及GR的活性;(2)Hg胁迫使菹草体内可溶性蛋白含量及T-AOC大幅降低,并引起NP-SH和PCs大量积累。外添Ca(NO3)2则有效减缓可溶性蛋白含量及T-AOC的下降,显著降低NP-SH和PCs水平;(3)脯氨酸是植物体内重要的渗透调节物质,外源Ca添加后,脯氨酸代谢关键酶吡咯啉-5-羧酸合成酶(P5CS)及δ-鸟氨酸转氨酶(OAT)活性显著增加,脯氨酸含量显著高于对照组。以上结果表明,Hg胁迫打破了菹草无菌苗抗氧化酶系统的内在平衡,而外添Ca(NO3)2有效促进了可溶性蛋白合成,提高了抗氧化酶活性和抗氧化物质含量,使菹草无菌苗维持较高的总抗氧化能力,并通过调节脯氨酸代谢来减轻Hg胁迫对菹草的伤害,增强植物对重金属Hg的耐受能力。     

低温胁迫对转新疆雪莲sikRbcs2基因烟草光合作用的影响

研究分布于新疆的雪莲(Saussurea involucrata)中的sikRbcs2基因在低温条件下对植物光合作用的影响,以16、10、6、4℃温度梯度处理非转基因型和转sikRbcs2基因型烟草,每个温度处理72 h,比较研究其叶绿素荧光特性和光合特性。实验分析结果:低温胁迫下,转基因型烟草叶片叶绿素a(Chla)、叶绿素b(Chlb)、叶绿素a+b(Chla+Chlb)、类胡萝卜素(Car)含量都显著高于非转基因型烟草。叶绿素荧光参数分析:低温胁迫下,转基因烟草PSⅡ最大光化学效率(Fv/Fm)、q P(光化学猝灭系数)、ETR(电子传递效率)都显著或极显著高于非转基因型,光合参数测定:随着低温胁迫程度的加剧,转基因烟草和非转基因型烟草净光合速率(Pn)、气孔导度(Gs)、蒸腾速率(Tr)、胞间CO_2浓度(Ci)总体都呈下降趋势,但转基因烟草的净光合速率有一个明显的动态变化,先急剧下降后有一个稳定上升的趋势。通过对生长指标的测定发现:在低温处理后和恢复后转基因烟草生物量的积累都显著高于非转基因型烟草。研究结果表明:转新疆雪莲sikRbcs2基因的烟草在低温条件下具有较高的Fv/Fm、q P、ETR,对光合机构的损伤小,降低了低温胁迫效应,提高了烟草在低温胁迫条件下的耐受性。     

放线菌对干旱胁迫下黑麦草生长及抗氧化特性的影响

采用菌剂接种及盆栽生物实验,研究了干旱缺水条件下放线菌对黑麦草生长和抗氧化特性的影响。结果显示:(1)干旱胁迫下,土壤中接种放线菌显著促进黑麦草的生长,其中根分蘖及根部生物量分别显著增加35.00%和37.47%;(2)接种放线菌后,黑麦草叶片的叶绿素总量和类胡萝卜素含量分别显著增加了12.02%和10.38%;(3)加菌后3种主要的抗氧化酶超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)较不加菌对照均增加,其增率分别为12.72%、2.38%和24.83%,其中SOD和CAT的活性增加显著。研究表明,干旱胁迫下放线菌接种土壤后,提高了黑麦草叶绿素含量和类胡萝卜素含量,增强了黑麦草抗干旱胁迫的抗氧化酶活性,可显著促进黑麦草的生长,增加其生物量。     

铜胁迫条件下AMF对海州香薷光合色素含量、抗氧化能力和膜脂过氧化的影响

以盆栽海州香薷为研究对象,模拟Cu胁迫条件下,接种丛枝菌根真菌(AMF)对海州香薷叶片光合色素含量、抗氧化酶活性、抗氧化剂含量、膜脂过氧化程度的影响。结果表明:(1)与对照相比,Cu胁迫使海州香薷叶片叶绿素a(Chl a)、叶绿素b(Chl b)、总叶绿素(Chl(a+b))、类胡萝卜素(Car)含量以及叶绿素a/b(Chl a/b)均显著降低,抗氧化酶活性和抗氧化剂含量也显著下降,质膜相对透性(MRP)和丙二醛(MDA)含量显著增大。(2)与Cu胁迫相比,Cu胁迫下接种AMF可使海州香薷叶片叶绿素含量显著增加;超氧化物歧化酶(SOD)、抗坏血酸过氧化物酶(APX)活性显著提高;还原型谷胱甘肽(GSH)、抗坏血酸(As A)含量显著增加;MDA含量、MRP显著下降。总之,接种AMF可提高Cu胁迫下海州香薷叶片光合色素含量和抗氧化能力,降低膜脂过氧化水平,从而缓解Cu胁迫对植株造成的伤害,增强海州香薷对Cu胁迫的适应性,提高了植株的生物量。     

激光对花生诱变效果的研究

Ｈｅ－ＮｅＹＡＧ和ＣＯ２激光对花生Ｌ１代幼苗期有刺激生长的作用,ＣＯ２激光对花生Ｌ１代的株高有较明显的抑制作用,ＣＯ２,Ｈｅ－Ｎｅ和ＹＡＧ激光对花生Ｌ１代植株的总果数,饱果数有一定的增多作用,且前者最明显,但Ｎ２激光对花生Ｌ１代植株的总果数,饱果数均有明显的减少作用;Ｈｅ－ＮｅＹＡＧ,Ｎ２和ＣＯ２激光均能诱发药生根尖细胞染色体畸变,且畸变率随辐照剂量的增大而上升,以上四种激光所诱发发生的变异性状是     

Cross-resistance to antibiotics of Escherichia coli adapted to benzalkonium chloride or exposed to stress-inducers

AIMS: To study the effects of adaptation and stress on the resistance to benzalkonium chloride (BC) and cross-resistance to antibiotics in Escherichia coli. METHODS AND RESULTS: Precultivation of E. coli ATCC 11775 and E. coli DSM 682 in the presence of subinhibitory concentrations of BC or stress inducers (salicylate, chenodeoxycholate and methyl viologen) resulted in higher minimum inhibitory concentration (MIC) of BC and chloramphenicol (CHL). Adaptation to growth in sixfold of the initial MIC of BC resulted in stable BC resistance and enhanced tolerance to several antibiotics and ethidium bromide (EtBr). The MIC of CHL increased more than 10-fold for both strains. Enhanced efflux of EtBr in adapted E. coli ATCC 11775 indicated that the observed resistance was due to efflux. Changes in outer membrane protein profiles were detected in the BC-adapted cells. There were no indications of lower membrane permeability to BC. CONCLUSIONS: Induction of stress response or gradual adaptation to BC or CHL results in acquired cross-tolerance between BC and antibiotics in E. coli. Enhanced efflux was one of the observed differences in adapted cells. SIGNIFICANCE AND IMPACT OF THE STUDY: Provided not taking due precautions, extensive use of disinfectants could lead to emergence of antibiotic-resistant isolates.     

Ability of human T lymphocytes to adhere to vascular endothelial cells and to augment endothelial permeability to macromolecules is linked to their state of post-thymic maturation

The accumulation of mononuclear cells at sites of chronic inflammation is dependent on a number of factors including localized adherence of lymphocytes to vascular endothelial cells (EC), cytokine-mediated increased adhesiveness of endothelium, chemotactic factors and endothelial permeability. The present study investigates two of the above attributes of lymphocyte-EC interaction: namely, the ability of maturationally distinct subpopulations of human T lymphocytes to adhere to vascular EC and to increase vascular endothelial permeability to macromolecules in an in vitro model. Thus, human T lymphocytes were separated into CD4+ CD8-helper/inducer, CD4- CD8+ cytotoxic/suppressor, CD29+ CD45RA- CD45RO+ memory, and CD29- CD45RA+ CD45RO- naive/virgin T subpopulations, were activated with PHA and PMA, and then examined for their adherence to EC and also for their effect on endothelial permeability. Upon activation, cells within each of the above four subpopulations exhibited increased adherence to EC. In contrast, resting CD29+ CD45RA- CD45RO+ memory T lymphocytes exhibited two to three times greater ability to adhere to EC than their CD29- CD45RA+ CD45RO- naive/virgin counterparts. Consistent with their increased adherence to EC, CD29+ CD45RO+ memory T lymphocytes, when activated, significantly increased endothelial permeability to albumin. Although activated CD45RA+ naive T lymphocytes exhibited increased adherence to EC, these cells failed to increase significantly endothelial permeability. Similar to their polyclonal counterparts, Ag-specific CD4+ CD29+ CD45RO+ T cell clones, but not their actively released mediators, also increased endothelial permeability via a noncytolytic mechanism(s). This ability of CD29+ CD45RO+ memory T lymphocytes to augment endothelial permeability may facilitate their transendothelial migration into extravascular space. These observations may provide additional insights into molecular mechanism(s) underlying pathophysiology of localized chronic inflammatory responses in general and more specifically selective accumulation of CD29+/CD45RO+ memory T lymphocytes at sites of chronic inflammation such as rheumatoid synovium.     

Capacity of Old Trees to Respond to Environmental Change

Atmospheric carbon dioxide [CO2] has increased dramatically within the current life spans of long-lived trees and old forests. Consider that a 500-year-old tree in the early twenty-first century has spent 70% of its life growing under preIndustrial levels of [CO2], which were 30% lower than current levels. Here we address the question of whether old trees have already responded to the rapid rise in [CO2] occurring over the past 150 years. In spite of limited data, aging trees have been shown to possess a substantial capacity for increased net growth after a period of post-maturity growth decline.Observations of renewed growth and physiological function in old trees have, in some instances, coincided with Industrial Age increases in key environmental resources, including [CO2], suggesting the potential for continued growth in old trees as a function of continued global climate change.     

Failure of CD4 T-cells to respond to liver-derived antigen and to provide help to CD8 T-cells

CD4 T-cell help is required for the induction of efficient CD8 T-cells responses and the generation of memory cells. Lack of CD4 T-cell help may contribute to an exhausted CD8 phenotype and viral persistence. Little is known about priming of CD4 T-cells by liver-derived antigen. We used TF-OVA mice expressing ovalbumin in hepatocytes to investigate CD4 T-cell priming by liver-derived antigen and the impact of CD4 T-cell help on CD8 T-cell function. Naïve and effector CD4 T-cells specific for ovalbumin were transferred into TF-OVA mice alone or together with naïve ovalbumin-specific CD8 T-cells. T-cell activation and function were analyzed. CD4 T-cells ignored antigen presented by liver antigen-presenting cells (APCs) in vitro and in vivo but were primed in the liver-draining lymph node and the spleen. No priming occurred in the absence of bone-marrow derived APCs capable of presenting ovalbumin in vivo. CD4 T-cells primed in TF-OVA mice displayed defective Th1-effector function and caused no liver damage. CD4 T-cells were not required for the induction of hepatitis by CD8 T-cells. Th1-effector but not naïve CD4 T-cells augmented the severity of liver injury caused by CD8 T-cells. Our data demonstrate that CD4 T-cells fail to respond to liver-derived antigen presented by liver APCs and develop defective effector function after priming in lymph nodes and spleen. The lack of CD4 T-cell help may be responsible for insufficient CD8 T-cell function against hepatic antigens.     

Adhesion of cellulose to cell walls to Trichoderma reesei

Carbohydrate-binding components were shown to be present at the surface of Listeria monocytogenes by means of a panel of neoglycoproteins using direct agglutination. These lectin-like components bind on neoglycoproteins bearing D-glucosamine, L-fucosylamine, or para-amino-phenyl-alpha-D-mannopyrannoside residues. The interactions were inhibited by the carbohydrate moieties specific to the neoglycoproteins. The protein nature of the lectin-like components of L. monocytogenes was ascertained by the loss of carbohydrate-binding capacity following protease treatment.