镉引起蚕豆(Vicia faba)叶片DNA损伤和细胞凋亡研究

以蚕豆为研究对象,采用碱性、半碱性和中性彗星实验方法研究Cd对植物DNA的损伤,同时还采用DAPI染色法从细胞形态学入手研究Cd诱导的蚕豆叶片的细胞凋亡.用3种彗星实验研究Cd处理蚕豆叶片DNA损伤,结果表明Cd能够引起蚕豆叶片DNA损伤,但是不同Cd浓度引起DNA损伤的种类不同:5mg·L^-1Cd处理主要引起单链DNA损伤和碱性不稳定位点的形成;10mg·L^-1Cd处理时开始检测到DNA双链断裂;20mg·L^-1Cd处理下,各种类型的DNA损伤均明显增加,且DNA双链断裂尤其明显.DAPI染色法通过细胞形态学变化来检测蚕豆叶片细胞凋亡的结果表明,蚕豆叶片细胞在Cd处理下发生凋亡的过程与DNA损伤过程有很大的相关性.结果表明,Cd是一种基因毒性物质,同时证明Cd引起的DNA损伤是引起细胞发生凋亡的机制之一.     

Effects of Glomus mosseae on the toxicity of heavy metals to Vicia faba

A glasshouse pot experiment was conducted to investigate effects of the arbuscular mycorrhizal fungus Glomus mosseae on the growth of Vicia faba and toxicity induced by heavy metals （HMs） （Cu, Zn, Pb and Cd） in a field soil contaminated by a mixture of these metals. There was also uninoculation treatment （NM） simultaneously. Mycorrhizal （GM） plants have significantly increased growth and tolerance to toxicity induced by heavy metals compared with NM plants. P uptake was significantly increased in GM plants. Mycorrhizal symbiosis reduced the transportation of HMs fi＇om root to shoot by immobilizing HMs in the mycorrhizal, shown by increasing the ratios of HMs from root to shoot. Oxidative stress, which can induce DNA damage, is an important mechanism of heavy metal toxicity. GM treatment decreased oxidative stress by intricating antioxidative systems such as peroxidases and non-enzymic systems including soluble protein. The DNA damage induced by heavy metals was detected using comet assay, which showed DNA damage in the plants was decreased by the GM treatment.     

镉胁迫对菜豆幼苗基因组DNA多态性的影响

采用随机扩增多态性DNA (RAPD)等技术,检测不同浓度镉(Cd)胁迫对菜豆幼苗叶片DNA损伤及相关生理特性的影响.结果表明,随着Cd处理浓度(0,20,40,80mg/L)的增加,菜豆幼苗中的Cd含量及叶片中丙二醛(MDA)含量升高;叶片中可溶性蛋白、脯氨酸、还原型谷胱甘肽在低浓度(20mg/L)Cd处理下含量增加,中、高浓度(40,80mg/L)Cd处理下减少.选取8条寡核苷酸引物(10bp),利用RAPD随机扩增技术,对菜豆幼苗叶片细胞基因组DNA进行扩增,8条引物均产生特异性谱带.正常叶片基因组DNA的RAPD图谱得到46条谱带.与正常组相比,Cd处理下利用不同引物扩增得到的RAPD图谱发生改变,呈现不同程度的谱带强度增强或减弱、谱带缺失或新增,与对照的RAPD图谱存在明显差异.Cd胁迫下基因组模板稳定性下降,并随Cd处理浓度的增加,稳定性下降幅度增大.利用RAPD技术获得的DNA多态性变化,可指示镉毒害生物遗传效应,并为镉污染地区蔬菜品种的选育及镉毒性的评价提供依据.     

Interactions of zinc and cadmium toxicity in their effects on growth and in antioxidative systems in tomato plants （Solanum lycopersicum）

The interaction between zinc and cadmium was investigated in tomato plants (Solanum lycopersicum). Ten-day-old seedlings were treated with 10 mol/L CdCl2 associated to di erent concentrations of ZnCl2 (10, 50, 100, and 150 mol/L). Zn supply clearly reduced Cd accumulation in leaves and simultaneously increased Zn concentration. Cd induced oxidative stress in leaves as indicated by an increase in thiobarbituric acid-reactive substances (TBARS) level and chlorophyll breakdown. Furthermore, compared with control, Cdtreated plants had significantly higher activities of superoxide dismutase (SOD, EC 1.15.1.1), whereas, catalase (CAT, EC 1.111.1.6), ascorbate peroxidase (APX, EC 1.11.1.11), and glutathione reductase (GR, EC 1.6.4.2) activities were significantly suppressed by Cd addition. Zn supplementation, at low level, restored and enhanced the functional activity of these enzymes (SOD, CAT, APX and GR) as compared to Cd-alone-treated plants. The beneficial e ect of adequate Zn level on Cd toxicity was confirmed by a significant decrease in TBARS level and restoration of chlorophyll content. However, when Zn was added at high level in combination with Cd there was an accumulation of oxidative stress, which was higher than that for Cd or excess Zn alone treatments. These results suggested that higher Zn concentrations and Cd are synergistic in their e ect on plant growth parameters and oxidative stress.     

Interactions of zinc and cadmium toxicity in their effects on growth and in antioxidative systems in tomato plants (Solanum lycopersicum)

The interaction between zinc and cadmium was investigated in tomato plants (Solanum lycopersicum). Ten-day-old seedlings were treated with 10 μmol/L CDC12 associated to different concentrations of ZnC12 (10, 50, 100, and 150 μmol/L). Zn supply clearly reduced Cd accumulation in leaves and simultaneously increased Zn concentration. Cd induced oxidative stress in leaves as indicated by an increase in thiobarbituric acid-reactive substances (TBARS) level and chlorophyll breakdown. Furthermore, compared with control, Cdtreate plants had significantly higher activities of superoxide dismutase (SOD, EC 1.15.1.1), whereas, catalase (CAT, EC 1.111.1.6),ascorbate peroxidase (APX, EC 1.11.1.11), and glutathione reductase (GR, EC 1.6.4.2) activities were significantly suppressed by Cd addition. Zn supplementation, at low level, restored and enhanced the functional activity of these enzymes (SOD, CAT, APX and GR) as compared to Cd-alone-treated plants. The beneficial effect of adequate Zn level on Cd toxicity was confirmed by a significant decrease in TBARS level and restoration of chlorophyll content. However, when Zn was added at high level in combination with Cd there was an accumulation of oxidative stress, which was higher than that for Cd or excess Zn alone treatments. These results suggested that higher Zn concentrations and Cd are synergistic in their effect on plant growth parameters and oxidative stress.     

模拟酸雨和Zn复合污染对蚕豆生长及其生理生化特性的影响

通过盆栽试验研究了模拟酸雨和Zn复合污染对蚕豆生长及其生理生化特性的影响．结果表明,与对照相比,在酸雨和Zn复合污染下,蚕豆株高和叶绿素均有不同程度的下降;蚕豆叶和根中的MDA含量上升,SOD活性下降,POD活性则是先升后降．MDA含量和SOD活性变化能较好地反映蚕豆受酸雨和Zn污染的状况．同时,酸雨和Zn的复合污染比其中任何一种单一污染对蚕豆造成的伤害更严重．     

商陆耐重金属Cd关键酶抗氧化酶的研究

研究商陆(Phytolacca americana L.)对Cd胁迫的抗氧化酶响应,对于揭示其耐Cd机制具有重要意义.商陆幼苗在200μmol/L或400 μmol/L CdCl2营养液中处理4 d,叶片活性氧自由基和MDA含量升高,膜透性增加,光合速率降低,表明Cd胁迫诱导产生氧化胁迫.随着Cd处理含量的提高和时间的...     

Cd胁迫对大豆各发育阶段生长及生理指标的影响

采用土壤盆栽试验方法,研究了不同浓度Cd胁迫下,大豆幼苗期、花荚期和成熟期植株的生长及叶片叶绿素含量、超氧化物歧化酶(SOD)活性、过氧化物酶(POD)活性和丙二醛(MDA)含量的变化.结果表明,大豆植株的生长在3个生长期均表现为低浓度Cd(￡0.50mg/kg)下的刺激效应和高浓度Cd(31.00mg/kg)下的抑制效应;随着Cd胁迫浓度的累积,大豆生长整体上受到抑制.在幼苗期,大豆植株的叶绿素含量下降、SOD活性受到抑制、POD活性迅速激活,共同缓解Cd毒性;在花荚期,大豆植株的防御系统得到有效激发,保护性酶POD、SOD的活性急剧升高,叶绿素含量呈上升趋势;在成熟期,因为长时间的Cd毒害,尤其是Cd浓度较高的情况下,大豆植株的SOD、POD活性和叶绿素含量急剧下降.在Cd胁迫下,大豆植株的生长发育以及生理生化指标在大豆的3个生长期的变化模式差异显著,且大豆植株的生物量和株高与SOD活性在整个生长期均表现出显著的相关性.     

不同浓度铵态氮对苦草的生理影响

研究了苦草在不同浓度(0.02,0.05,0.10,0.30,0.60,1.00,3.00mg/L)铵态氮中暴露14d后,其生物量的变化、叶片游离氨基酸态氮、叶绿素、可溶性蛋白含量以及O2-×信号强度、抗氧化酶活性和丙二醛(MDA)含量的响应.结果表明,各浓度组苦草的生物量无显著变化,但是各生理指标变化显著.当铵态氮浓度为0.30mg/L时,苦草叶片中游离氨基酸态氮的含量即开始显著升高.当铵态氮浓度达到0.60mg/L时,超氧化物歧化酶(SOD)活性显著升高,表明苦草诱导产生氧化应激但未受到氧化损伤.当铵态氮浓度高于1.00mg/L时,SOD和过氧化物酶(POD)活性显著升高,O2-×信号强度显著增强,叶绿素、可溶性蛋白含量显著降低.当铵态氮浓度为0.02mg/L时,O2-×信号强度显著增强.综上,铵态氮浓度低于0.60mg/L苦草生长良好,浓度31.00mg/L苦草光合能力受到抑制、代谢受到干扰.苦草对铵态氮最敏感的生理生化指标是叶片中游离氨基酸态氮含量.铵态氮作为沉水植物的一种营养物质,当其含量较低时,植物由于营养缺乏诱导产生自由基.     

Cd、Cu单一污染对小麦体内活性氧消除系统影响

采用溶液培养法,研究不同浓度Cd、Cu单一污染对小麦SOD、POD、CAT活性及MDA含量的影响。结果表明:(1)SOD和CAT活性对Cd胁迫反应较敏感,而POD活性变化不大,叶片中有效的活性氧自由基清除系统的平衡被打破,MDA含量增加;(2)Cu胁迫下,小麦叶片中MDA含量的变化趋势与根部的相反,在Cu处理浓度为25、50mg/L时,叶片中有效的活性氧自由基清除系统的平衡未被打破,MDA含量保持在较低的水平;而根部恰好相反,该系统出现了失衡现象,MDA含量偏高。     

喜旱莲子草对Pb、Cd胁迫响应的研究

采用营养液培养法,以喜旱莲子草为对象,选择重金属Pb在浓度梯度0,0.1,10,50,100和200 mg/L条件下,Cd在浓度梯度0,0.01,1,10,20和40 mg/L条件下,研究了不同浓度Pb、Cd单一及复合胁迫对喜旱莲子草生长的影响。结果表明,在试验条件下,对单一Pb及Cd对喜旱莲子草生长及生理影响而言,与对照相比,当Pb处理浓度为0.1 mg/L时,植物最高株高和最长根长分别增加了3.8%和15%。随着Pb处理浓度增加到200 mg/L时,植物生长受到明显抑制,茎叶株高呈显著下降。当Cd处理浓度为1 mg/L时,与对照相比,植物最高株高和最长根长分别增加了9.0%和10.0%,随着Cd处理浓度增加(20~40 mg/L),植物生长受到明显抑制,茎叶株高呈显著下降。随Pb和Cd处理浓度增加,喜旱莲子草叶绿素含量呈下降趋势,POD活性先升后降,MDA含量呈上升趋势,植株体内Pb、Cd积累量呈上升趋势。在研究Pb、Cd对喜旱莲子草生长及生理胁迫的基础上,选择Pb、Cd不同配比浓度,探讨其对喜旱莲子草的复合胁迫作用,研究植物对重金属Pb、Cd富集特性,结果表明,低浓度Pb、Cd处理表现出相互促进吸收富集,高浓度处理表现出相互抑制吸收富集的特性。     

La对Cd伤害大豆幼苗的生态生理作用

以盆载法研究了Cd对大豆幼苗的伤害及La对Cd伤害的缓解效应。结果表明,500mg/L CdCl₂严重抑制大豆幼苗的代谢与生长,叶面喷布30mg/LLaCl³一次,能缓解Cd毒造成的伤害。实验证明,这与La能增加大豆幼苗光合速率,提高叶绿素含量与硝酸还原酶活性、降低细胞质膜透性与植株体内Cd含量、维持TTC还原力等多重作用相关。     

久效磷农药对金鱼肝细胞DNA的损伤及其机制研究

以金鱼(Carassius auratus)为模式生物,研究了久效磷农药暴露导致的DNA损伤类型及其作用机制.结果表明:0.01,0.10,1.00mg/L的久效磷农药暴露24,48,96,168h均导致金鱼肝细胞DNA损伤程度显著升高,48h损伤最严重;暴露48h采用碱性、pH值12.1和中性彗星电泳发现DNA损伤类型主要为碱不稳定位点形成,其次为单/双链断裂;采用Endo Ⅲ酶和FPG酶处理的碱性彗星电泳,发现DNA出现氧化损伤;暴露24h肝脏谷胱甘肽过氧化物酶(GSH-Px)活性显著降低,丙二醛(MDA)含量显著升高达到峰值,96~168h超氧化物歧化酶(SOD)和GSH-Px活性显著升高,MDA含量与24h相比有所降低,表明活性氧自由基(ROS)含量在短期暴露升高(24h)、在96~168h暴露逐渐降低.影响抗氧化酶活性、干扰ROS清除过程是久效磷农药造成DNA损伤的主要机制.     

镉对小麦叶片DNA伤害的彗星实验研究

为了建立植物基因毒性的研究方法 ,以小麦为对象 ,进行了植物彗星实验方法的研究 .以 0 1mg·L- 1 的Cd2 + 溶液处理小麦叶片后 ,采用机械方法分离细胞核 ,制备玻片后变性 0 ,5 ,15 ,3 0min ,在 10 0 ,2 0 0 ,3 0 0mA下电泳 5 ,15 ,3 0min .实验结果表明 ,采用机械分离的方法可以获得大量的、能够进行彗星实验的细胞核 ,增加DNA的变性时间、电泳电流大小和电泳时间可以增加DNA片段在电场中的迁移 ,提高实验的灵敏度 ,但变性时间和电泳时间过长增加了对照处理DNA片段的迁移 .因此 ,本实验提出了小麦叶片彗星实验的最佳条件是DNA变性 15min ,3 0 0mA下电泳 15min ,并在该条件下研究了重金属镉对小麦的基因损伤 .结果表明 ,重金属镉能引起小麦基因的损伤 ,并随镉浓度的增加伤害程度加大     

焦化废水对蚕豆和大麦毒性的影响

以COD作为主要参照指标,研究了焦化废水在符合GB 13456-92<钢铁工业水污染物排放标准>焦化一级、二级排放标准限值要求时,对蚕豆和大麦幼根生长、根尖细胞遗传毒性的影响.结果表明:在实验周期内,焦化废水对蚕豆幼根根长、根重和有丝分裂指数的影响不大;对大麦幼根根重无明显影响,而对大麦根长和有丝分裂指数有促进作用.焦...     

球果蔊菜脯氨酸的积累及与Cd耐性的关系

采用盆栽方法,以风花菜(Rorippa islandica)为对照植物,研究Cd超积累植物球果蔊菜[Rorippa globosa (Turcz.) Thell.]脯氨酸的积累、丙二醛含量、生物量及叶绿素含量的变化,及其与Cd生物积累的关系.结果显示,Cd处理能明显促进2种植物体内脯氨酸的积累,当土壤中Cd浓度≤25μg/g时,球果蔊菜叶片脯氨酸的积累效应尤为显著;此时叶片生物量、叶绿素及丙二醛含量与对照相比均无显著差异,表明其叶片脯氨酸的积累在一定程度上可缓解Cd胁迫造成的氧化损伤.相关分析表明,球果蔊菜根系脯氨酸可指示球果蔊菜根系中Cd的积累.     

外源精胺对镉胁迫下苎麻生理代谢的影响

采用水培实验法,研究了不同浓度和不同施加方式下外源精胺对苎麻镉污染毒害的缓解效应.结果表明,镉胁迫(10mg/L)使苎麻叶绿素、类胡萝卜素和可溶性蛋白质含量均不同程度地下降,膜脂过氧化程度提高,丙二醛含量增加;加入外源精胺后(以10^-5mol/L效果最佳),叶绿素、类胡萝卜素和可溶性蛋白质含量均显著上升,膜脂过氧化产物丙二醛含量明显降低,说明苎麻对镉的抗性得到显著提高,其中叶面喷洒并同时向营养液中加入精胺能更有效地缓解叶片的生理伤害,而仅叶面喷洒外源精胺则更有利于缓解根中镉的毒害.进入苎麻体内的镉约73.2%积累在根部,2种方式加入精胺均使得镉在苎麻体内的累积量下降,同时镉向地上部分的转移率提高.     

外源脯氨酸对Cd胁迫下107杨光合与叶绿素荧光特性的影响

为揭示Cd胁迫下外源脯氨酸对107杨（Populus×euramericana cv.‘74/76’）光合与叶绿素荧光特性的影响,以一年生107杨苗木为试验材料,采用盆栽控水试验,研究了添加不同浓度（0、10和20 mmol/L）外源脯氨酸对Cd（0、100、200和300 mg/kg）胁迫下107杨叶片光合气体交换参数、光响应参数和叶绿素荧光参数的影响.结果表明:①Cd作为独立因子可显著影响107杨叶片的光合气体交换参数、光响应参数和叶绿素荧光参数,外源脯氨酸作为独立因子对这些参数无显著影响,Cd与脯氨酸的交互作用对这3类参数均有显著影响.②随着w（Cd）的增加,107杨叶片净光合速率（P_n）、蒸腾速率（T_r）、气孔导度（G_s）和水分利用效率（WUE）的变化趋势基本一致,均显著下降,而胞间CO₂浓度（C_i）呈显著上升趋势.与单独Cd处理组相比,在w（Cd）为200 mg/kg处理下,添加外源c（脯氨酸）为10 mmol/L时苗木叶片的P_n显著提高123.12%;WUE、PSⅡ原初反应最大量子效率（F_v/F_m）、实际光化学量子效率（φ_PSⅡ）、光化学淬灭系数（qP）和表观光合电子传递速率（ETR）分别显著提高43.73%、6.52%、27.99%、28.96%和63.26%;而非光化学淬灭系数（NPQ）显著降低19.97%.研究显示,w（Cd）低于200 mg/kg时,外源添加脯氨酸处理可增强107杨的PSⅡ光能转换效率和光强利用效率,防止过剩光能对光合机构的伤害,从而提高植株抗逆性与适应性,且以c（脯氨酸）为10 mmol/L时的处理效果较好.      

A proteomic analysis of rice seedlings responding to 1,2,4-trichlorobenzene stress

The proteomic analysis of rice （Oryza sativa L.） roots and leaves responding tol,2,4-trichlorobenzene （TCB） stress was carded out by two dimensional gel electrophoresis, mass spectrometric （MS）, and protein database analysis. The results showed that 5 mg/L TCB stress had a significant effect on global proteome in rice roots and leaves. The analysis of the category and function of TCB stress inducible proteins showed that different kinds of responses were produced in rice roots and leaves, when rice seedlings were exposed to 5 mg/L TCB stress. Most responses are essential for rice defending the damage of TCB stress. These responses include detoxication of toxic substances, expression of pathogenesis-related proteins, synthesis of cell wall substances and secondary compounds, regulation of protein and amino acid metabolism, activation of methionine salvage pathway, and also include osmotic regulation and phytohormone metabolism. Comparing the TCB stress inducible proteins between the two cultivars, the β-glucosidase and pathogenesis-related protein family 10 proteins were particularly induced by TCB stress in the roots of rice cultivar （Oryza sativa L.） Aizaizhan, and the glutathione S-transferase and aci-reductone dioxygenase 4 were induced in the roots of rice cultivar Shanyou 63. This may be one of the important mechanisms for Shanyou 63 having higher tolerance to TCB stress than Aizaizhan.     

Cd2+胁迫下中肋骨条藻细胞内多胺的生理响应

通过室内添加Cd2+培养中肋骨条藻(Skeletonema costatum),开展了Cd2+胁迫对中肋骨条藻生长的影响及藻细胞内超氧化物歧化酶(SOD)、二胺氧化酶(DAO)、多胺氧化酶(PAO)的活性以及丙二醛(MDA)和各形态多胺含量变化的实验,以探究多胺对于海洋微藻在重金属胁迫下的响应及生理作用.结果显示,Cd2+对中肋骨条藻的生长产生胁迫作用,且Cd2+浓度越大对藻细胞生长的抑制作用越强.Cd2+胁迫下中肋骨条藻细胞内MDA含量略有升高,SOD活性显著降低,DAO和PAO活性分别增强.随着Cd2+胁迫作用的增强,藻细胞内游离态腐胺(Put)、亚精胺(Spd)、精胺(Spm)含量先降低后升高;结合态Put、Spd和Spm含量先升高后降低;束缚态Put、Spm和Spd含量及Put总量和Spm总量升高.表明Cd2+胁迫会导致中肋骨条藻细胞产生氧化损害,同时细胞内DAO和PAO活性的增强以及游离态Put、束缚态Spd和Spm、Put总量和Spm总量的升高可能会增强藻细胞抵抗Cd2+胁迫的能力.