Microcystin-RR induces physiological stress and cell death in the cyanobacterium Aphanizomenon sp DC01 isolated from Lake Dianchi, China

The phytoplankton community in Lake Dianchi (Yunnan Province, Southwestern China) is dominated in April by a bloom of Aphanizomenon, that disappears Suddenly and is displaced by a Microcystis bloom in May. The reasons for the rapid bloom disappearance phenomenon and the temporal variability in the composition of phytoplankton assemblages are poorly understood. Cell growth, ultrastructure and physiological changes were examined in cultures of Aphanizomenon sp. DC01 isolated from Lake Dianchi exposed to different closes of rnicrocystin-RR (MC-RR) produced by the Microcystis bloom. MC-RR concentrations above 100 mu g L-1 markedly inhibited the pigment (chlorophyll-a, phycocyanin) synthesis and caused an increase of soluble carbohydrate and protein contents and nitrate reductase activity of toxin-treated blue-green algae. A drastic. reduction in photochemical efficiency of PSII (Fv/Fm) was also found. Morphological examinationn showed that the Aphanizomenon filaments disintegrated and file cells lysed gradually after 48 h Of toxin exposure. Transmission electron microscopy revealed that cellular inclusions of stressed cells almost leaked out completely and the cell membranes were grossly damaged. These findings demonstrate the allelopathic activity of Microcystis aeruginosa inducing physiological stress and cell death of Aphanizomenon sp. DC01 Although the active concentrations of microcystin were rather high, we propose that microcystin may function as allelopathic Substance due to inhomogeneous toxin concentrations close to Microcystis cells. Hence, it may play a role in species Succession of Aphanizomenon and Microcystis in Lake Dianchi.

Physiological and biochemical microcystin-RR toxicity to the Synechococcus elongatus

Freshwater Microcystis may form dense blooms in eutrophic lakes. It is known to produce a family of related cyclic hepatopeptides (microcystins, MC) that constitute a threat to aquatic ecosystems. Most toxicological studies of microcystins have focused on aquatic animals and plants, with few examining the possible effects of microcystins on phytoplankton. In this study we chose the unicellular Synechococcus elongatus (one of the most studied and geographically most widely distributed cyanobacteria in the picoplankton) as the test material and investigated the biological parameters: growth, pigment (chlorophyll-a, phycocyanin), photosynthetic activity, nitrate reductase activity, and protein and carbohydrate content. The results revealed that microcystin-RR concentrations above 100 mug (.) L-1 significantly inhibited the growth of Synechococcus elongatus. In addition, a change in color of the toxin-treated algae (chlorosis) was observed in the experiments. Furthermore, MC-RR markedly inhibited the synthesis of the pigments chlorophyll-a and phycocyanin. A drastic reduction in photochemical efficiency of PSII (F-v/F-m) was found after a 96-h incubation. Changes in protein and carbohydrate concentrations and in nitrate reductase activity also were observed during the exposure period. This study aimed to evaluate the mechanisms of microcystin toxicity on a cyanobacterium, according to the physiological and biochemical responses of Synechococcus elongatus to different doses of microcystin-RR. The ecological role of microcystins as an allelopathic substance also is discussed in the article. (C) 2004 Wiley Periodicals, Inc.

模拟酸雨对马尾松菌根影响研究

本实验表明：外生菌根真菌彩色豆马勃、劣味乳菇、丝膜菌对PH的适应范围较广，最适生长BH呈酸性。模拟酸雨对马尾松幼苗菌根的外部形态和内部结构有明显影响。在温室栽培中，模拟酸雨（PH2.0）显著抑制菌根侵染率，在田间实验中，对菌根侵染率有一定的影响。菌根PH和土壤PH值随模拟酸雨PH下降而逐渐降低，接种菌根菌可略提高菌根PH和土壤PH值。菌根真菌过氧化氢酶对培养基中PH的变化不敏感，模拟酸雨对菌根过氧化氢酶活性影响也不明显。但沙培中，模拟酸雨（PH2.0）显著抑制菌根过氧化氢酶活性。模拟酸雨（PH2.0）显著刺激菌根过氧化物酶活性，接种菌根菌可以降低菌根过氧化物酶活性。不同PH的培养基对菌体硝酸还原酶活性有明显影响，而且菌体生长速度与硝酸还原酶活性呈正相关。模拟酸雨（PH2.0）显著抑制菌根硝酸还原酶活性，而接种菌根菌明显提高根系硝酸还原酶活性。菌体酸性磷酸酶活性对培养基中PH变化不敏感，同样菌根酸性磷酸酶活性对模拟酸雨的影响也不明显，但是接种菌根菌可明显提高根系酸性磷酸酶活性。模拟酸雨对马尾松幼苗茎的高生长影响不显著。但是对幼苗茎、根系的干重和侧根总长度有显著抑制作用。轻度酸雨（PH4.5-3.0）对马尾松幼苗生长有促进作用，接种菌可提高幼苗生长。从菌根形态结构和生理活性上看，接种菌根菌可减轻模拟酸雨对马尾松幼苗根系的危害，增强对模拟酸雨的抗性。4dThe result of experiment showed that ectomycorrhizal fungi Pisolithus tinctorins. Lactarius insulsus. Cortinarius russus can be growth in broad PH rang in pure culture, the optimum growth PH is acidity. The external morphology and internal structure of ectomycorrhiza of P. massoniana are affected with simulated acid rain. In greenhouse, simulated acid rain (PH2.0) treatment caused significant decrease in the percent infection, but it's not marked in field. The PH of mycorrhizal and soil are reduced with reducing rainfall PH. These PH are slight higher for inoculation with ectomycorrhizal fungi. Catalase activity of ectomycorrhizal fungus is not sensitive to medium with different PH. Mycorrhiza catalase activiyt is not affected significantly with simulated acid rain, but it's inhibited significantly with simulated acid rain (PH2.0) in the sand culture. Peroxidase atcivity of mycorrhiza is enhanced significantly with simulated acid rain (PH2.0), but it's universally lower for inoculation with ectomycorrhizal fungus. Ectomycorrhizal fungus nitrate reductase activity is affected significantly to medium with differdnt PH, the rates of these fungi growth and nitrate reductase activity is significant correlation. Nitrate reductase activity of mycorrhiza is inhibited significantly with simulated acid rain (PH 2.0), but it's increased significantly for inocnlation with mycorrhizal fungi. Ectomycorrhizal fungas acid phosphatase activity is not affected to medium with different PH, Mycorrhiza acid phosphatase activity is not affected with simulated acid rain too, the acid phosphatase activity of roots inoculated with mycorrhizal fungas is increased significantly. The highest acidity level simulated rain reduced signhficantly root system biomass and the dry weight of stem. Iower acidity level simulated rain can stimulated the growth of P. massoniana, the growth of seedling inocnlated with mycorrhizal fungus can be increased.

花椒主效化感物质分离鉴定及化感作用研究

近年来，随着对作物重茬障碍原因的深入研究，植物的化感作用越来越受到国内外众多学者的重视。花椒（Zanthoxy piperitum.）为芸香科植物，是一种收益早、用途广、价值高的经济树种，是川西干旱河谷地区的重要经济作物，其连作障碍也倍受关注，系统研究花椒化感作用将有助于理解和最终解决花椒连作障碍问题。本文首先通过萃取、层析等方法分离花椒主效化感成分；通过外加不同浓度的花椒叶水浸液研究了对土壤氮素养分循环的影响；研究了花椒叶水浸液对苜蓿生理生化、光合作用、氮素养分吸收的影响，并对外施氮肥对这种化感影响的缓解作用做了研究；研究了花椒化感潜力对全球变化——UV-B增强辐射的响应。主要研究结果如下： 1．用不同极性的有机溶剂对花椒叶水浸液浓缩浸膏萃取、柱层析，结合生物活性检测，分离得到主效化感作用组分的一种化感物质——对甲氧基苯酚。采用该物质纯品进行生物活性检测，证明其具有化感作用。 2．花椒叶水浸液处理土壤30天后，土壤硝态氮、铵态氮、无机氮（硝态氮＋铵态氮）与对照相比，随着花椒叶水浸液浓度的增加呈现降低的趋势，其中土壤铵态氮含量显著降低，而硝态氮含量的变化则不显著，无机氮含量也显著降低。土壤脲酶和蛋白酶的活性与无机氮含量的变化趋势相同。随着花椒叶水浸液浓度的增加，氨化细菌数量显著降低，固氮菌的数量变化不显著，硝化细菌和反硝化细菌数量有减少的趋势。60天后，硝态氮含量、铵态氮含量、无机氮随水浸液浓度增加的变化趋势与30天时相似；随着花椒叶水浸液浓度的增加，氨化细菌、固氮菌的数量显著减少，硝化细菌数量、反硝化细菌数量仍呈减少趋势；土壤脲酶、蛋白酶活性与第30天的变化趋势相同。第60天与第30天的结果相比，相同水浸液浓度处理的硝态氮、铵态氮、无机氮均有下降的趋势，但除了25g.L－1水浸液处理的外，其它相同浓度的处理间差异均不显著；除了12.5 g.L的处理外土壤脲酶活性均呈增强的趋势；蛋白酶活性都有不同程度的增加；花椒叶水浸液处理的土壤硝化细菌和反硝化细菌数量呈增加趋势。 3．随着花椒叶水浸液浓度的增加，显著抑制了苜蓿根长、地上地下生物量、叶绿素含量、叶片中可溶性蛋白的含量，净光合速率。苜蓿体内四种抗氧化酶（POD、SOD、CAT、APX） 活性随着水浸液浓度的增加而降低，而丙二醛含量则增加。苜蓿氮初级同化相关酶硝酸还原酶（NR）、谷氨酰合成酶(GS)、谷氨酸脱氢酶（GDH）的活性随着水浸液浓度的增加受到不同程度的影响。总的来说，苜蓿硝酸还原酶、谷氨酰合成酶的活性受到抑制，而谷氨酸脱氢酶活性的变化则比较复杂，根呈先降低后增加的趋势，叶片则无显著变化。外施两种不同浓度的硝酸铵氮肥后，对12.5、25 g.L－1花椒叶水浸液处理的苜蓿化感作用有显著的缓解作用，表现在株高、生物量、光合作用等方面，大多达到与对照（0 g.L－1）未施氮肥无显著差异的水平，而对50 g.L－1水浸液处理的苜蓿幼苗，虽有一定的缓解作用，但这种作用均未达到与对照（0 g.L－1）未施氮肥时无显著差异的水平。 4． UV-B增强辐射处理花椒后，花椒的化感潜力显著增强。花椒叶片内UV-B吸收物质的含量和总酚含量均显著增加。 In recent years, with profound research on the reasons of continuous cropping obstacles, allelopathy received increasing attention to many scholars at home and abroad. Zanthoxy bungeanum as a Rutaceae plant is a high economic value species which gains early and uses widely. Zanthoxylum is an important economic crop in the arid valley of western Sichuan region, and its not even has received much concern for the continuous cropping obstacles. The systematic study of allelopathy of Zanthoxylum will contribute to the understanding and final settlement of this issue. The major allelopathic composition was separated through the extraction, chromatography combined with other methods. The impact on soil nutrient cycling was also studied through the addition of different concentrations of water extracts of Zanthoxylum. Furthermore, the effects of water extracts of Zanthoxylum leaves on alfalfa leaf physiological and biochemical indexes, photosynthesis, soil enzymes and nutrient uptake of nitrogen and the mitigation of allelopathy through using external fertilizer were studied to put forward scientific resolvent for Zanthoxylum continuous cropping obstacles .The response of allelopathic potential of Zanthoxylum to global change - UV-B enhanced radiation was studied . The main findings are as follows: 1. Through extraction with different polar organic solvents on concentrated water extract of Zanthoxylum leaf and then using column chromatography combined with detection of biological activity, one of the main allelopathic components- methoxy-phenol was isolated. The biological activity testing of the pure material of methoxy-phenol proved that it does have allelopathic potential. 2. Thirty days after treating soil with water extract of Zanthoxylum leaf, as compared with the control, the contents of soil nitrate, ammonium, nitrate plus ammonium nitrogen showed a trend of decrease with the increase of the concentration of water extract whereas the content of ammonium nitrogen showed a significant reduction, and the content of nitrate did not change significantly, the content of nitrate plus ammonium nitrogen also showed a significant (P <0.05) redction. The activity of soil urease and protease showed the same trend as the content of nitrate nitrogen plus ammonium nitrogen. With the increase in the concentration of water extract, the number of ammonification bacteria significantly reduced but nitrogen-fixing bacteria did not change significantly and there was a decreasing trend in the number of nitrifying bacteria and denitrifying bacteria. Sixty days after the treatment, with the increase in solution concentration of water extract of Zanthoxylum leaf, the content of nitrate、 ammonium nitrogen, nitrate plus ammonium nitrogen showed a similar change trend to 30 days’; the number of ammonification bacteria, nitrogen-fixing bacteria significantly reduced ; the number of nitrifying bacteria, denitrifying bacteria was still an downward trend; the activity of soil urease and protease showed the same trend as the 30th days’. Compared to the results of the 30th days’, the content of nitrate, ammonium, nitrate plus ammonium nitrogen showed a decrease trend between the treatment of same concentration, but there was no significant difference except the treatment of 25g.L-1 between the same concentration; the activity of soil urease showed enhanced trend except the treatment of 12.5 g.L-1; the activity of protease increased to varying degrees; the number of ammonification bacteria、 nitrifying bacteria and denitrifying bacteria were growing while nitrogen-fixing bacteria reduced.. 3. With the increase of the concentration of water extract of Zanthoxylum leaf, the water extract significantly inhibited the root length, aboveground biomass, content of chlorophyll and soluble protein in leaf and net photosynthetic rate. The activity of four antioxidant enzymes (POD, SOD, CAT, APX) reduced with the increase in concentration of the water extract but the content of MDA increased. The activity of enzymes related to primary nitrogen assimilation such nitrate reductase (NR), glutamyl synthetase (GS), glutamate dehydrogenase (GDH) were subject to different degrees with an increase in the concentration of water extracts. In general, the activity of nitrate reductase, glutamyl synthetase were inhibited, while change in the activity of glutamate dehydrogenase was more complex. The activity of glutamate dehydrogenase in leaf was first reduced and then increase,but did not change significantly in root. After using two external different concentrations of nitrogen fertilizer, there was a significant mitigation in inhibiton in plant height, biomass, photosynthesis, etc. in the treatment of 12.5,25 gL-1 of water extract of Zanthoxylum leaf, and most of these indexes showed no significant difference with the control (0 g.L-1, no external fertilizer was added) .Although there showed a certain degree of ease in the treatment of 50 g.L-1 , there was still a significant difference compared with the control (0 gL-1) in which no external fertilizer was used. 4.The allelopathic potential of Zanthoxylum positively responded to enhanced UV-B significantly. The content of UV-B absorbing compounds and the total phenol also significant increased.

在盐胁迫下外源维生素浸种对小麦发芽及幼苗生长的影响

维生素(Vitamin)又称维他命，为“万年青”产品，是维持人体生命健康必需的一类低分子有机化合物质。维生素对人体健康的作用人们研究很多, 维生素可以增强人体对感染的抵抗力，降低出生缺陷及降低癌症和心脏病发病率等，一旦缺乏，肌体代谢就会失去平衡，免疫力下降，各种疾病，病毒就会趁虚而入；而维生素对作物影响的研究却很少。目前为止，尚无对用维生素浸种的方法来研究外源维生素是否对小麦种子萌发及幼苗生长起调节作用的报道，且对其在小麦抗逆性方面影响的研究甚少，对盐的胁迫抗性研究尚未有人报道。小麦（Triticum aestivum L.）属于拒盐的淡土性作物。盐害不利于小麦生长，严重影响小麦的产量和品质。本研究采用4 种不同维生素VB1、VC、VB6、VPP，分别对供试小麦品种川育12（红皮）、川育16（白皮）小麦浸种后，在一般自然条件下和逆境（盐胁迫条件）下，进行试验。探讨在正常情况下与在不同盐浓度条件下，各维生素及盐浓度对小麦发芽及幼苗生长的影响，并且比较两种不同皮色的小麦在相同盐胁迫条件下的差异表现，同时研究维生素处理的特异性，且哪种维生素对盐害缓解作用最佳。研究结果表明：在无盐胁迫（自然）条件下，对用4 种不同维生素VB1、VC、VB6、VPP 浸种小麦川育12、川育16 后的种子萌发及幼苗生长（幼苗的根长、根重、苗高、苗鲜重）的研究结果表明：4 种外源维生素浸种均对小麦发芽有调节作用，都能提高其最终发芽率。但是提高幅度有所差异。用VB6 浸种后的小麦提高幅度最多，VC 次之，VPP 提高幅度最小。同时，4 种外源维生素浸种对小麦种子的出芽速度及芽后长势也有一定的影响。VB6、VC 处理的小麦种子出芽速度最快，萌发后长势最好；VB1 出芽速度相对较慢，VPP 最慢,但都大于对照；VB1 处理长势略高于对照，VPP 处理的小麦长势则低于对照。从整体来看，VB6、VC处理促进效应明显， VB1 次之，而VPP 在某些方面无效甚至产生负效应。此外，相同的维生素处理对不同的品种的种子萌发、生长效果也存在差异，各种维生素作用于川育12 的效应均强于对川育16。进一步对幼苗根系TTC 还原力及幼苗叶片中硝酸还原酶活性进行测定、分析。研究发现：并非所有种类的维生素对幼苗根系TTC 还原力及幼苗叶片中硝酸还原酶活性的提高都有帮助。幼苗根系TTC 还原力在不同维生素处理下存在显著差异，而与小麦品种关系甚微。经VB6、VC 处理后，根系TTC 还原力测定值均显著高于对照，VB1 不明显，VPP 则略低于对照。VB6、VC 处理的幼苗叶片中硝酸还原酶的含量大于对照，VB1 与对照相差无几，而VPP 处理的川育12 幼苗叶片中的硝酸还原酶活性比对照CK 略高，而在川育16 中则略比对照CK 有所下降，呈现出抑制效应。综上结果表明：VB6、VC 具有促进种子发芽，幼苗生长及根系生长的作用，是较好的促生长剂；VPP 具有抑制作用，是较好的抑制剂，可进一步研究、开发利用。在盐胁迫条件下，对用4 种不同维生素VB1、VC、VB6、VPP 浸种川育12、川育16 后的种子萌发及幼苗生长（幼苗的根长、根重、苗高、苗鲜重）的研究结果表明：在不同盐浓度胁迫条件下， 各处理的种子萌发及幼苗生长均受到不同程度的抑制。随着盐浓度的增加, 发芽率、发芽指数和活力指数成下降趋势；幼苗的根长、根重、苗高、苗鲜重不断降低。4 种维生素处理间也表现出较大差异。VB6、VC 在每个处理中均保持对盐害的缓解作用，VB6 较VC 更易于促进发芽及幼苗生长。最终发芽率高，根系多、长、重，苗高高、重。而VB1、VPP 则表现出抑制作用。在高盐浓度150mM 时，4 种维生素浸种后的种子，其最终发芽率均不能达到40％，但VB6、VC 处理最终发芽率、苗重、根重均高于对照，VPP 最终发芽率、苗重、根重均低于对照。进一步对幼苗根系TTC 还原力及幼苗叶片中脯氨酸含量进行测定、分析。研究发现：不同盐浓度，不同维生素处理、不同品种间存在差异。随着盐浓度的增加（75mM，100mM，150mM），幼苗根系TTC 还原力活性成下降趋势，幼苗叶片中脯氨酸的积累量成上升趋势。VB6 处理脯氨酸含量增加最为明显，VC 次之，VPP 与对照接近，其变化幅度最小。经VB6、VC 处理后的幼苗根系还原强度，在不同盐浓度下，测定值均显著高于对照，VB1 不明显，VPP 则低于对照，产生负效应。此外，品种间表现不尽相同，相同的维生素处理，相同的盐浓度对不同的品种的种子萌发、生长效果也存在差异， 4 种维生素对川育16 的作用均强于川育12，但其影响趋势是一致的。说明VB6、VC 具有耐（抗）盐性，可以促进种子发芽和幼苗生长，是较好的耐（抗）盐拌种剂。 Vitamin is one kind of necessary low molecular compound for humans tosustain health and life. Lots of Studies have been done on the effectc of the vitaminsfor people. Vitamin can help people improve the body's natural resistance to disease,Drop the rate of birth defects、cacers and the incidence of the heart diseases. Ifpeople have less of them, the metabolism of the organism may throw off balance,immunity may drop off, and catch disease; Though the effects for Vitamin to thecrops are limited. up to now, there’s no one use soking seeds of wheats with vitaminsas a method, to study on how the effects will happen on the wheat seed germinationand seedling growth, and there are only few reserches on antireversion force forwheats ,none for the antireversion force in Sault stress condition.Wheat（Triticum aestivum L.）is sensitive to the salt, so the salt damage will doharm to wheat’s growth, it will have an unfavorable impact on the output and thequality of wheat.On this reaserch, we Soaking CHY12(red)、CHY16 (white) wheat seeds withVitamin C, B1, PP, B6 (50mg/L) as a pretreatment first. Then under two condition: one is in the normal environment the other is in different Salinity, we begin ourexperiments. Then disscuss on if the vitamin and salinity affect the wheat seedgermination and seedling growth, and what is the different between the two of them,the result shows that:Under the normal condition, after soaking seeds with VB1、VC、VB6、Vpp，we study on the their seed germination and the seeding growth(the root length andweights, The seedling heights and weights), it shows that all of those four kinds ofvitamin can adjust the seed germination, but different in The growth rate. VB6 isbest for increase, VC comes second,VPP is the worst. Meanwhile, those four vitaminalso have effect on the speed of the sprouting of the wheat. VB6、Vc can faster theseed germination most, and the seedlings are all doing well; VB1 do little effects onthe budding, Vpp is the worst, but all treatments are better than CK; but in Vi, VB1some what above the CK, while VPP lower than that. On the whole, the acceleratingeffect of VB6、VC are obvious, VB1 takes second place, but VPP in some aspects arenoneffective even have negative effect. Furthermore, different kind of seeds with thesame vitamin may different in seed germination and seedling growth, four vitaminson CHY16 is better than CHY12.More studies on TTC reductive capacity of roots and the activity of nitratereductase in the leaves, the reasult shows not all the vitamin can help the seedlings toimprove the TTC reductive capacity and the activity of nitrate reductase. TTCreductive capacity in different treatments shows significant differences,but notcorrelate to the variety of the wheat. The TTC reductive capacity of VB6、Vctreatments are all higher than CK, VB1 is nearly the same as CK, VPP is a littlelower than CK. Through the study of acivity of nitrate reductase, it shows that,VB6、VC are higher than CK ,VB1 is nearly the same as CK also, VPP is a little higher inthe CK of CHY12 but lower in CHY16. Through all the results above: VB6、Vc helpthe wheat seed germination, seedling growth and the growth of roots, is theperfectable factor of stimulating the growth; Vpp is a inhibition, that’ll be furtherreserch,and well develop and utilize in the future.Under the different Salinity condition, after soaking seeds with VB1、VC、VB6、Vpp，we study on the their seed germination and the seeding growth(the root lengthand weights, The seedling heights and weights), it shows that: under differentsalinity, the seed germination and the seedling growth of any treatment are inhibited.With the increase of the concentration, the germination rate, Vi、Gi all had fallen; theroot length and weight, the seedling heights and weights steadily sank down. There are also have pronounced difference between all treatments with four differentvitamins.VB6、VC in all treatments are alleviative the salt damage, VB6 is easier tocause to put forth buds than VC, and it’s quantitative value is the highest in theultimate germination rate, in root and seedlings’ hight and weight. Though the VPP、VB1 are seems to inhibite its growth. Under the high concentration150mM Nacl, theultimate germination rate in all treatments are below the 40%, but VB6、VC’squantitative values in any experiments are higher than CK,while VPP lower thanCK.Then we study on the TTC reductive capacity of roots and the content of Polinein leaves, the result shows that between the different salinity, different vitamintreatments, different varieties of the wheat have discrepancy.along with theincreasing concentraion of the salinity（75mM，100mM，150mM），TTC reductivecapacity of roots decreases, the accumulation of the content of Poline in leaves havean upward trend. The increase of VB6’s treatment are obviously, VC comessecond,VPP is nearly come up with CK, changes a little. In TTC reductive capacity of roots’s reserch, VB6、VC are higher than CK at any time,VB1 is not palpable,VPP is lower than CK, makes negative affect on wheat. In addition, varieties of thewheats are remain different, no matter it shows promoting or inhibiting, all fourvitamins have moreobvious effects on CHY16 than CHY12, but the tendency of theeffection are the same. It is say that VB6、VC can help wheat to standwith the saultwell, and promot in growth，they are the better reagent to mix with the seed.

一种拟南芥突变体对高浓度CO_2反应的研究

The study on the response of a mutant and a wild type of Arabidopsis to 660 μl·L -1 CO 2 and ambient CO 2 showed that under elevated CO 2,the stomatal numbers of the mutant increased,while those of the wild type decreased. The chlorophyll content and NR (nitrate reductase) activity of the mutant increased,but those of the wild type had no obvious response. The mutant was not reproductively mature after the continuous exposure to increased CO 2 for five months. The results provided evidence of plant response to the changes of atmospheric CO 2 concentration,and the clues to related studies on other plants.

Viologen-thiol self-assembled monolayers for immobilized horseradish peroxidase at gold electrode surface

A stable, well-behaved self-assembled monolayer (SAM) of viologen-functionalized thiol was used to immobilize and electrically connect horseradish peroxidase (HRP) at gold electrode. Viologen groups in SAMs facilitated the electron transfer from the electrode to the protein active site so that HRP exhibited a quasi-reversible redox behavior. HRP adsorbed in the SAMs is very stable, and close to a monolayer with the surface coverage of 6.5 x 10(-11) mol/cm(2). The normal potential of HRP is -580 mV vs Ag/AgCl corresponding to ferri/ferro active center and the standard electron transfer rate constant is 3.41 s(-1) in 0.1 M phosphate buffer solution (pH 7.1). This approach shows a great promise for designing enzyme electrodes with other redox proteins and practical use in tailoring a variety of amperometric biosensor devices. Copyright (C) 1997 Elsevier Science Ltd.

Phosphorylation of plant actin-depolymerising factor by calmodulin-like domain protein kinase

actin-depolymerising factor (ADF)/cofilin group of proteins are stimulus-responsive actin-severing proteins, members of which are regulated by reversible phosphorylation. The phosphorylation site on the maize ADF, ZmADF3, is Ser-6 but the kinase responsible is unknown [Smertenko et al,, Plant J. 14 (1998) 187-193]. We have partially purified the ADF kinase(s) and found it to be calcium-regulated and inhibited by N-(6-aminohesyl)-[H-3]5-chloro-1-naphthalenesulphonamide. Immunoblotting reveals that calmodulin-like domain protein kinase(s) (CDPK) are enriched in the purified preparation and addition of anti-CDPK to in vitro phosphorylation assays results in the inhibition of ADF phosphorylation, These data strongly suggest that plant ADP is phosphorylation by CDPK(s), a class of protein kinases unique to plants and protozoa. (C) 2001 Published by Elsevier Science B.V. on behalf of the Federation of European Biochemical Societies.

Nitrogen Metabolism in leaves of a tank epiphytic bromeliad: Characterization of a spatial and functional division

The leaf is considered the most important vegetative organ of tank epiphytic bromeliads due to its ability to absorb and assimilate nutrients. However, little is known about the physiological characteristics of nutrient uptake and assimilation. In order to better understand the mechanisms utilized by some tank epiphytic bromeliads to optimize the nitrogen acquisition and assimilation, a study was proposed to verify the existence of a differential capacity to assimilate nitrogen in different leaf portions. The experiments were conducted using young plants of Vriesea gigantea. A nutrient solution containing NO(3)(-)/NH(4)(+) or urea as the sole nitrogen source was supplied to the tank of these plants and the activities of urease, nitrate reductase (NR), glutamine synthetase (GS) and glutamate dehydrogenase (NADH-GDH) were quantified in apical and basal leaf portions after 1, 3, 6, 9, 12, 24 and 48 h. The endogenous ammonium and urea contents were also analyzed. Independent of the nitrogen sources utilized, NR and urease activities were higher in the basal portions of leaves in all the period analyzed. On the contrary. GS and GDH activities were higher in apical part. It was also observed that the endogenous ammonium and urea had the highest contents detected in the basal region. These results suggest that the basal portion was preferentially involved in nitrate reduction and urea hydrolysis, while the apical region could be the main area responsible for ammonium assimilation through the action of GS and GDH activities. Moreover, it was possible to infer that ammonium may be transported from the base, to the apex of the leaves. In conclusion, it was suggested that a spatial and functional division in nitrogen absorption and NH(4)(+) assimilation between basal and apical leaf areas exists, ensuring that the majority of nitrogen available inside the tank is quickly used by bromeliad`s leaves. (C) 2011 Elsevier GmbH. All rights reserved.

Neisseria meningitidis and Neisseria gonorrhoeae are differently adapted in the regulation of denitrification: single nucleotide polymorphisms that enable species-specific tuning of the aerobic–anaerobic switch

The closely related pathogenic Neisseria species N. meningitidis and N. gonorrhoeae are able to respire in the absence of oxygen, using nitrite as an alternative electron acceptor. aniA (copper-containing nitrite reductase) is tightly regulated by four transcriptional regulators: FNR (fumarate and nitrate reductase), NarP, FUR (Ferric uptake regulator) and NsrR. The four regulators control expression of aniA in N. meningitidis by binding to specific and distinct regions of the promoter. We show in the present study that FUR and NarP are both required for the induction of expression of aniA in N. meningitidis, and that they bind adjacent to one another in a non-co-operative manner. Activation via FUR/NarP is dependent on their topological arrangement relative to the RNA polymerase-binding site. Analysis of the sequence of the aniA promoters from multiple N. meningitidis and N. gonorrhoeae strains indicates that there are species-specific single nucleotide polymorphisms, in regions predicted to be important for regulator binding. These sequence differences alter both the in vitro DNA binding and the promoter activation in intact cells by key activators FNR (oxygen sensor) and NarP (which is activated by nitrite in N. meningitidis). The weak relative binding of FNR to the N. gonorrhoeae aniA promoter (compared to N. meningitidis) is compensated for by a higher affinity of the gonococcal aniA promoter for NarP. Despite containing nearly identical genes for catalysing and regulating denitrification, variations in the promoter for the aniA gene appear to have been selected to enable the two pathogens to tune differentially their responses to environmental variables during the aerobic–anaerobic switch.

Respostas fisiológicas em mudas de banananeira tratadas com estrobilurinas

There are reports that strobilurin besides having a fungicide effect can promote physiologic benefits to the plants. However, this effect on banana plants was not studied yet. The objective of the present study was to evaluate the effect of strobirulins on the physiology of banana plantlets. For this purpose, cultivar Grand Naine banana plantlets were transferred to pots containing substrate and kept in a nursery with 50% shading. The experimental design was a completely randomized design with three treatments (water, azoxystrobin and pyraclostrobin) and five replications. The treatments were applied at 15, 30, 45, 60 and 75 days after transplanting at a dose 100 g a. i. ha(-1) with manual spray. Plant height, pseudostem diameter, shoot dry matter in strobilurin treated plants were higher than the untreated plants, however, the effect of fungicide treatment was different, being the most pronounced effect of pyraclostrobin compared to azoxystrobin. Plants treated with pyraclostrobin had higher leaf area, nitrate reductase activity and chlorophyll content of leaf total nitrogen than the plants treated with azoxystrobin and water, which did not differ. Strobilurins affect the physiology of the banana plantlets differently, the effect being more pronounced by pyraclostrobin.

Efeito simultâneo da deficiência hídrica e do alumínio tóxico no solo na cultivar IAC91-5155 de cana-de-açúcar

Sugarcane is a very important economic crop in Brazil. The effects of abiotic stresses cause negative reduction of the productivity in the sugarcane industry. In order to identify indicators of stresses tolerance, two physiological variables were evaluated, nitrate reductase activity and chlorophyll contents in young plants of sugarcane, cv. IAC91-5155. The simultaneous effect of abiotic stresses of high occurrence in Brazilian soils are, water deficiency and aluminum toxicity. The plants were submitted to three treatments of water availability (% field capacity, FC): no stress (70% FC), moderate stress (55% FC), and extreme stress (40% FC); and three acidity treatments in the soil (base saturation, V%): no acidity (V=55%), average acidity (V=33%), and high acidity (V=23%). The experiment was carried out in greenhouse, with 29.7 +/- 4.3 degrees C and 75 +/- 10% RH. The experimental design was in randomized blocks, in 3x3 factorial arrangement, with four replicates. After 60 days, nitrate reductase activity and chlorophyll contents were evaluated in the diagnostic leaf. The results demonstrate that the response of plants to a combination of drought and aluminum toxicity, similar to the conditions in many natural environments, is different from the response of plants to each of these stresses applied individually, as typically tested in the laboratory. The nitrate reductase activity can be used as a biochemical-physiological marker of water deficiency while chlorophyll contents can be used as a biochemical-physiological marker of both of them, water deficiency or aluminum toxicity in soil. Both parameters can not be as a biochemical-physiological marker for acclimation of young plants of sugarcane cv. IAC91-5155, under the combined stresses.

The contribution of genes required for anaerobic respiration to the virulence of Salmonella enterica serovar Gallinarum for chickens

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Nodulação e atividade da nitrato redutase em função da aplicação de molibdênio em soja

Among the nutrients that are essential for the biological nitrogen fixation by soybean plants, molybdenum stands out for being a cofactor of the nitrate reductase, affecting enzymatic activity and, consequently, the nodulation process. The research had as objective to evaluate the effects of molybdenum application on soybean nodulation and nitrate reductase activity. The experiment was conduced in greenhouse, sowing soybean in 12 L pots, with two plants per plot. The treatments consisted of two application via (with the seeds and leaf dressing) and two molybdenum doses (12 and 24 g ha(-1) with the seeds; 30 and 60 g ha(-1) leaf dressing) in ammonium molybdate form, plus the control. The number and dry mass of nodules and nitrogen content in soybean leaves were evaluated. Samples of leaves for the evaluation of nitrate reductase activity were taken at 10 a.m. and 10 p.m. It was concluded that soybean nodulation is affected by Mo dose and application via, resulting in higher number and weight of nodules when it is applied with the seeds. The enzymatic activity of the nitrate reductase is influenced by Mo fertilization and it is higher for leaf dressing with the double of the recommended dose.

METABOLISMO do NITROGÊNIO ASSOCIADO À DEFICIÊNCIA HÍDRICA E SUA RECUPERAÇÃO em GENÓTIPOS DE MILHO

O estresse hídrico afeta profundamente o metabolismo celular vegetal. Neste trabalho, objetivou-se quantificar os efeitos da deficiência hídrica e sua recuperação sobre a atividade das enzimas do metabolismo do nitrogênio: redutase do nitrato (RN), glutamina sintetase (GS) e glutamato sintase (GOGAT) e sobre o acúmulo de prolina em plantas dos genótipos de milho BR 2121 e BR 205. O experimento foi conduzido em casa de vegetação, sob o delineamento inteiramente casualizado, com quatro repetições, utilizando-se vasos que continham 14,3kg de solo. Os tratamentos consistiram da combinação dos dois genótipos e quatro intervalos entre irrigações (1, 3, 5 e 7 dias). No dia da avaliação (49 dias após emergência), os tratamentos com intervalos entre 3 e 7 dias, haviam sido irrigados no dia anterior, caracterizando-se portanto como recuperação da deficiência hídrica leve e severa, respectivamente. As extrações e análises foram realizadas utilizando-se a terceira folha basípeta completamente expandida. As atividades das enzimas estudadas não diferiram entre os tratamentos de estresse hídrico, controle e recuperação do estresse moderado, entretanto as plantas sob recuperação do estresse severo apresentaram atividade enzimática superior à das plantas controle. O acúmulo de prolina livre nas folhas aumentou com o estresse hídrico e respondeu à recuperação do estresse apresentando redução. de modo geral, a atividade enzimática e o acúmulo de prolina apresentaram respostas inversas dentro dos tratamentos.