铜胁迫对不同基因型谷子幼苗基因组DNA多态性的影响

采用室内盆栽土培法,以4种基因型谷子(D2-8、安06、黄米、朝谷)为供试材料,研究了不同浓度Cu2+胁迫对谷子幼苗体内可溶性糖、脯氨酸、丙二醛(MDA)含量和基因组DNA多态性的影响.结果表明,经50~400 mg·kg-1Cu2+处理30 d后,4种谷子幼苗体内可溶性糖含量与对照相比呈现先上升后下降的趋势,在50 mg·kg-1时达到最大值.当浓度为200mg·kg-1以上时,4种谷子的可溶性糖含量的平均降幅为对照的32.44%~56.5%.脯氨酸则表现为低含量(≤50 mg·kg-1)的促进和高含量(≥100 mg·kg-1)的抑制效应,MDA含量均有增加且与对照差异显著(P<0.05).Cu2+胁迫下不同基因型谷子幼苗基因组DNA的RAPD图谱发生明显变化,表现为单条或多条RAPD谱带的增加和缺失或者荧光强度的改变,细胞中基因组模板DNA的稳定性下降,DNA多态性变化与Cu2+浓度之间存在剂量-效应关系.不同基因型谷子对Cu2+胁迫的生理和遗传损伤响应存在差异.利用RAPD技术获得的DNA多态性变化可作为检测Cu2+遗传毒性效应的生物标记物.     

细胞分裂素类物质对镉胁迫下玉米幼苗生长和抗氧化酶活性及脯氨酸含量的影响

采用水培试验,研究了镉胁迫下喷施细胞分裂素类物质6-苄氨基嘌呤(6-BA)和激动素(KT)对玉米幼苗生长、叶片叶绿素含量、脯氨酸含量、植株镉含量和活性氧清除酶活性的影响.结果表明,与单纯镉胁迫相比,喷施6-BA和KT提高了玉米幼苗的生物量、根和地上部镉含量、叶绿素和脯氨酸含量,降低了叶片丙二醛(MDA)含量,促进了叶片过氧化物酶(POD)和过氧化氢酶(CAT)活性的升高,但对超氧化物歧化酶(SOD)活性的影响不明显.说明细胞分裂素类物质6-BA和KT可缓解镉胁迫对玉米幼苗的伤害,其机制可能与细胞分裂素物质能维持叶绿素稳定、刺激细胞内H2O2清除酶类活性及促进脯氨酸积累有关.当6-BA浓度在0～20mg·L-1之间时,其对玉米幼苗镉胁迫的缓解效应随喷施浓度的提高而增强,而KT的最适喷施浓度为70mg·L-1.     

Cd胁迫对桐花树同工酶的影响

利用聚丙烯酰胺凝胶垂直板电泳方法,研究了不同浓度Cd胁迫对桐花树幼苗叶片过氧物酶(POD)、超氧化物歧化酶(SOD)、酯酶(EST)同工酶酶谱的影响.结果表明:Cd胁迫下POD和EST同工酶组成发生了较大的变化,酶带颜色深浅和条数均有变化.部分同工酶基因表达在镉胁迫下完全关闭而酶带消失,另一部分同工酶基因表达则被启动而有新的酶带产生.而SOD同工酶酶带的亮度强弱也有明显的变化.这说明桐花树体内POD,EST和SOD同工酶确实能够适应Cd胁迫而产生相应变化,从而调节细胞代谢,以抵抗逆境胁迫.     

AFLP法分析染料对黄孢原毛平革菌的损伤效应

利用AFLP对DNA碱基序列分辨率高的特性,研究纯培养条件下染料对黄孢原毛平革菌(P. chrysosporium)菌株DNA的损伤效应.在对黄孢原毛平革菌(P. chrysosporium)AFLP最佳引物筛选的基础上,通过分析最佳引物扩增结果中DNA相似性、染料浓度与DNA相似性关系以及构建UPGMA相似性树状图,评价染料对P. chrysosporium DNA损伤效应.结果表明, AFLP法可较好反映染料对DNA损伤效应,在优选的3种引物组合中, E-AAC/M-CAA引物组合AFLP扩增效果最好,共获得35条DNA扩增条带,其中有12条具有特异性,多态性比率34.3%;在进行DNA相似性分析和UPGMA法聚类分析后发现, AFLP技术较好地表达了DNA相似性与染料浓度的正相关关系,并且显示P. chrysosporium对染料损伤效应敏感,染料浓度不宜高于50mg/L,当染料浓度达到300mg/L即可对P. chrysosporium产生较大损伤作用,使之基本丧失原有的处理功能.     

水培条件下秋华柳对重金属Cd的富集特性及光合响应

采用水培试验的方法,研究了不同Cd胁迫浓度〔ρ(Cd2+)分别为0、1、5、10和20 mg/L〕下秋华柳的生长、光合耐性及其对Cd的富集特性和转移能力.结果表明:①随着Cd胁迫浓度的升高,秋华柳生长所受到的抑制程度有所增加.除20 mg/L处理组外,其余各处理组的耐性指数(TI)均大于70,表现出较高的耐性.②低Cd胁迫浓度(1 mg/L)下秋华柳具有较好的光合耐性,当Cd胁迫浓度达到20 mg/L时,秋华柳的光合系统才受到明显影响.③秋华柳对外源Cd的富集能力较强,随着Cd胁迫浓度的升高,秋华柳所积累的Cd显著增加且主要积累于根部,表现为叶<茎<根.当Cd胁迫浓度小于5 mg/L时,秋华柳根部吸收的Cd向地上部分转移的效果较好.试验显示,秋华柳对于较低Cd胁迫浓度(≤10 mg/L)具有较好的耐受性和富集能力,可用于三峡库区消落带表层土壤Cd污染严重区域进行植物修复.      

NO对Cd2+胁迫下绿豆幼苗DNA及光合作用的影响

以绿豆幼苗为材料,研究了外源一氧化氮(NO)处理对Cd~(2+)胁迫3天和8天时幼苗叶片和根系DNA含量、DNA链间交联程度以及叶片光合参数的影响。结果表明:单独Cd~(2+)胁迫导致叶片和根系DNA含量降低,DNA链间交联程度增加,叶片净光合速率、气孔导度和胞间隙CO2浓度降低,气孔限制值升高;与单独Cd~(2+)胁迫相比,NO和Cd~(2+)复合处理提高了叶片和根系DNA含量,降低了DNA链间交联程度,同时使叶片净光合速率和气孔导度升高,而使胞间隙CO2浓度和气孔限制值降低。结果说明,NO能缓解Cd~(2+)胁迫对绿豆幼苗DNA结构及其代谢的影响;Cd~(2+)胁迫抑制绿豆幼苗光合作用的主要原因是气孔因素,而NO主要通过提高幼苗叶片非气孔因素缓解了Cd~(2+)胁迫对幼苗光合作用的抑制。     

壳寡糖缓解小麦镉毒害的某些生理特性研究

利用不同浓度壳寡糖溶液对小麦种子浸种,种子萌发后,对小麦幼苗进行Cd胁迫,研究了不同浓度的壳寡糖对小麦幼苗生长、超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)等抗氧化酶活性的影响。结果表明:不同浓度壳寡糖浸种,对镉胁迫下小麦幼苗生长有促进作用,幼苗高度、根长、生物量均有增加;幼苗叶片中叶绿素含量增高;SOD、POD和CAT活性增加。随着镉胁迫时间的延长,SOD和CAT活性降低,但降低幅度小于对照;POD活性却表现增加,增加的幅度大于对照。壳寡糖不同浓度之间比较,低浓度壳寡糖对镉毒害的缓解效果优于高浓度。说明壳寡糖预处理对镉胁迫有缓解作用。     

重金属Cd、Cu对小麦(Triticum aestivum)幼苗生理生化过程的影响及其毒性机理研究

对Cd、Cu单因子处理下小麦幼苗叶片和根系的SOD、POD酶活、可溶性蛋白含量以及叶片中叶绿素含量的变化进行了研究.结果表明,Cd、Cu胁迫下,小麦幼苗受到损伤的明显症状之一是叶片叶绿素含量下降;小麦植株叶片POD、SOD酶活能够被诱导而升高;重金属Cd、Cu对小麦幼苗根系的损伤较叶片大;重金属对小麦幼苗的毒害机理之一是抑制了蛋白质的生物合成;重金属引起的各个生化指标随着处理浓度和处理时间的变化远比有机污染物(如豆磺隆)简单.     

蒌蒿(Artemisia selengensis)对土壤中镉的胁迫反应及修复潜力研究

以洞庭湖湿地新发现的Cd高富集植物——蒌蒿(Artemisia selengensis)为试验材料,采用盆栽方式,系统研究了蒌蒿在生长期内对Cd的性状反应及富集特征.结果表明,大于40mg.kg-1的Cd胁迫对蒌蒿叶片伤害明显,且导致生物量下降,但植株在100mg.kg-1胁迫下可完成生活史,对Cd有较强的耐受能力;土壤中有效态Cd的含量仅占土壤全Cd含量的15.3%～37.1%;相同胁迫浓度下,各生长阶段的植株体内Cd含量差别不大,但由于生物量的原因,幼苗期对Cd的提取量显著小于其他时期,其中40～60mg.kg-1的Cd处理可使蒌蒿地上部分Cd含量达492～588mg.kg-1(成株期),且植株对该浓度范围的Cd污染去除效果最好;蒌蒿可作为对湿地土壤Cd污染较理想的修复植物加以研究和利用.     

稀土镧(La)及La(POA)_3对镉污染绿豆幼苗生理影响

以水培的方式研究了稀土元素镧及其配合物La(POA)3(HPOA=苯氧乙酸)对绿豆幼苗的生理生态影响。结果表明,单一镉(10mg/L)胁迫时,NR、CAT及POD活性下降,而MDA、Pro含量上升;较低浓度(5～30mg/L)的镧及其配合物能明显地促进绿豆幼苗生长,并且可以缓解镉对绿豆幼苗的胁迫作用,经镧及其配合物作用后,上述生理指标皆比单一镉胁迫时有明显的改善,而高浓度(100mg/L)的镧及其配合物对镉污染的缓解作用都减弱;另外,较低浓度(10、15、30mg/L)的La(POA)3对镉胁迫的缓解作用较镧更明显;随着作用时间的延长,镧及其配合物对绿豆幼苗镉污染的缓解作用均逐渐下降。     

Heavy metal induced DNA changes in aquatic macrophytes： Random amplified polymorphic DNA analysis and identification of sequence characterized amplified region marker

Plants have been used as good bio-indicators and genetic toxicity of environmental pollution in recent years. In this study, aquatic plants Hydrilla verticillata and Ceratophyllum demersum treated with 10 μmol/L Cd, 5 μmol/L Hg, and 20 μmol/L Cu for 96 h, showed changes in chlorophyll, protein content, and in DNA profiles. The changes in DNA profiles included variation in band intensity, presence or absence of certain bands and even appearance of new bands. Genomic template stability test performed for the ...     

Oxidative stress and DNA damages induced by cadmium accumulation

Experimental evidence shows that cadmium （Cd） could induce oxidative stress and then causes DNA damage in animal cells, however, whether such effect exists in plants is still unclear. In the present study, Vicia faba plants was exposed to 5 and 10 mg/L Cd for 4 d to investigate the distribution of Cd in plant, the metal effects on the cell lipids, antioxidative enzymes and DNA damages in leaves. Cd induced an increase in Cd concentrations in plants. An enhanced level of lipid peroxidation in leaves and an enhanced concentration of H2O2 in root tissues suggested that Cd caused oxidative stress in Vicia faba. Compared with control, Cd-induced enhancement in superoxide dismutase activity was significant at 5 mg/L than at 10 mg/kg in leaves, by contrast, catalase and peroxidaseactivities were significantly suppressed by Cd addition. DNA damage was detected by neutral/neutral, alkaline/neutral and alkaline/alkaline Comet assay. Increased levels of DNA damages induced by Cd occurred with reference to oxidative stress in leaves, therefore, oxidative stress induced by Cd accumulation in plants contributed to DNA damages and was possibly an important mechanism of Cd-phytotoxicity in Vicia faba plants.     

Soluble protein and acid phosphatase exuded by ectomycorrhizal fungi and seedlings in response to excessive Cu and Cd

Fungi and their symbionts can alleviate heavy metal stress by exuding soluble proteins and enzymes. This study examined the role of soluble protein and acid phosphatase (APase) exuded by Xerocomus chrysenteron, an ectomycorrhizal fungus, and the seedlings of its symbiont, Chinese pine (Pinus tabulaeformis), under conditions of excessive Cu and Cd. The growth type showed that this poorly studied ectomycorrhizal fungus was capable of tolerating high concentrations of Cu, and may be useful in phytoremediation. X. chrysenteron grew well at 80 mg/L Cu, and the EC50 for Cd was 17.82 mg/L. X. chrysenteron also showed enhanced exudation of soluble protein in both isolated and inoculated cultivations under the influence of Cu and Cd. Soluble protein exudation, however, di ered under Cu and Cd stress in isolates. In mediums containing Cu, soluble protein exudation increased with concentration, but in mediums containing Cd the content of soluble protein increased to a comparable level at all concentrations. This study demonstrated that soluble protein was related to heavy metal tolerance, although the di erent ions played di erent roles. While APase activity in exudates of fungi and seedlings decreased under Cu and Cd stress in comparison to the control, the APase activity in seedlings was maintained by inoculation. Thus, X. chrysenteron facilitated the ability of plant to maintain a normal nutrient uptake, and therefore to protect it from heavy metal toxicity.     

铅污染对菱幼苗SOD、POD及叶细胞亚显微结构的影响

不同浓度Pb2+溶液处理菱(Trapa bicornis Osbeck)幼苗后,考察了菱幼苗的超氧化物歧化酶(SOD)、过氧化物酶(POD)以及叶细胞亚显微结构出现的变化,并初步探讨了Pb2+的毒性机理.结果表明,SOD的活性与铅离子浓度(5~20mg/L)表现出明显的剂量-效应关系, Pb2+浓度愈大,毒害程度愈深.POD活性在较低浓度(5,10mg/L)作用下,持续升高;在较高浓度(15,20mg/L)下,其活性出现先上升后下降的趋势.电子显微镜观察发现,在20mg/L Pb2+溶液处理7d后,叶细胞的叶绿体基粒数目减少,基粒片层解体,叶绿体双层膜断裂;线粒体、高尔基体、内质网等解体消失;细胞核的染色质与核质遭到破坏,核膜破裂.同样处理条件的叶片组织总DNA电泳显示,DNA完全断裂呈现弥散状,提示Pb2+毒害所致细胞死亡有别于细胞程序性死亡. Pb2+对菱的毒害是多途径综合作用的结果.     

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。     

不同作物种苗对水中重金属镉去除的比较研究

以玉米、向日葵和蓖麻为实验材料,研究了3种作物种苗对镉的去除效果以及种苗生理生化指标的变化情况.结果表明,①随着处理时间的延长,溶液中镉含量逐渐降低,去除过程主要集中在前48 h.在1 mg·L^-1和2 mg·L^-1 Cd溶液中,向日葵和蓖麻种苗的去除效果要好于玉米种苗的;而在5 mg·L^-1和10 mg·L^-1 Cd溶液中,去除效果顺序为：蓖麻>玉米>向日葵.②种苗从溶液中吸收的镉绝大部分集中在根部.3种作物种苗对镉的富集能力大小顺序为：向日葵>蓖麻>玉米.③在不同镉处理浓度下,3种作物种苗的叶片质膜透性、根系活力、叶绿素含量和游离脯氨酸含量等生理生化指标均有不同程度的变化.向日葵的根系活力和脯氨酸含量均与镉积累量呈现显著相关性;蓖麻叶片的质膜透性和脯氨酸含量均与镉含量呈现显著正相关.3种作物种苗对镉都有一定的耐受性,而向日葵和蓖麻种苗的耐受性比玉米种苗的强.     

旺盛期烟草对镉富集敏感性研究

采用盆栽试验方法,揭示了旺盛期烟草（云烟99）对镉的富集特点以及光合等生理指标对镉胁迫的响应.结果表明：当土壤镉含量分别为4.43,7.94,17.33和49.79mg/kg时,烟草茎、叶及地上部镉的富集系数（植物镉含量与土壤镉含量的比值）均大于1,转移系数（地上部镉含量与根镉含量的比值）也大于1,但镉含量未达到镉超富集植物的临界含量标准100mg/kg.当土壤镉含量为4.43mg/kg时,烟草的耐性较强.当土壤镉含量大于7.94mg/kg时,烟草的生物量、叶片光合色素含量、净光合速率、蒸腾速率、气孔导度和SOD活性均显著下降（P<0.05）,胞间CO₂浓度和MDA含量显著提高（P<0.05）.旺盛期烟草对镉富集比较敏感,建议烟草的种植要远离镉污染土壤或镉背景值较高的土壤.     

Multiple biomarkers response in maize (Zea mays L.) during exposure to copper

Biomarkers in higher plants played an important role to estimate exposure effects of pollutants in soil ecosystem and have received increasing attention in recent years. The qualitative and quantitative modifications arising in amplified fragment length polymorphism （AFLP） profiles as a measure of DNA effects were compared with a number of parameters, namely, the root length, total soluble protein content in root tips, chlorophylls content and shoot size to select the most sensitive biomarker responding to copper stress in the range of 0--600 mg/kg. The changes occurring in AFLP profiles of root tips following Cu treatment included loss of normal bands and appearance of new bands and variation in band intensity in comparison to that of the normal seedlings, A reduction in root length was observed at the 200 mg/kg of copper, which was accompanied with a decrease in total soluble protein content. According to their sensitivity to the copper toxicity, the above indicator rank in the following order： AFLP profiles 〉 total soluble protein content 〉 root length 〉 chlorophylls content 〉 shoot. We concluded that the AFLP offered a useful alternative hiomarker assay for the detection of genotoxic effects of environmental pollutants.