光和暗条件培养的眼虫藻内RuBP羧化酶的免疫定位

应用免疫金标记技术证明,在眼虫藻和其它藻类中ＲｕＢＰ羧化酶主要分布在蛋白核部位,这与高等植物中ＲｕＢＰ羧化酶分布不同,在眼虫藻叶绿体间质中有少量ＲｕＢＰ羧化酶存在,这与高等植物中ＲｕＢＰ羧化酶的分布也有相似之处。暗中培养的眼虫藻不能形成类囊体,无ＲｕＢＰ羧化酶,无光合能力,只能进行异养代谢。     

分光光度法与^14C标记法测定RuBP羧化酶的活性的比较

方法的原理分光光度酶偶联法是根据RuBP羧化酶催化RuBP产生的磷酸甘油酸与NADH的氧化作用相偶联的原理设计的。当加入ATP及NADH后,NADH在PGK和GAPDH的催化下氧化为NAD~ ,每羧化1molRuBP有2molNADH被氧化,根据在波长340nm处光密度的变化,可测知NADH     

不同生境两种生态型芦苇叶片质膜H+-ATPase的比较

利用两相法化纯化质膜微囊,研究了分布西北沙地区的两种生态型芦苇(Phragmites communis trih.)水生芦苇和重度盐化草甸芦苇,分别简称为水芒和盐芦)叶片质膜H - ATPase的部分性质.结果显示,与水芦相比,盐芦质膜H -ATPase的ATP水解活性升高,Km值由1.27mmol\l降至Vmax没有显著差异.并且该酶活性对温度的敏感必玫PH谱型也发生了变化.以对硝基苯磷酸盐为底物,低浓度时盐芦的的质膜H -ATPase水解活性有差异.钡酸盐抑制实验表明,两种生态的质膜H -ATPase磷酸-酶区的催化性质不同.胰酶对质膜H -ATPase活性的活化谱型也存在差异,说明该酶C末端的结构或性质发生了变化.此外,与水芦相比,盐芦质膜H -ATPase的质子泵活性的耦联程度也升高了.以上结果明,当芦苇从水生环境向盐渍环境过渡时,质膜H -ATPase的催化性质发生了变化,这些变化可能是由酶结构的修饰和不同的同工酬酶谱引起的.H -ATPase催化性质的变化可能是对盐渍生境的适应性反应.     

河西走廊不同生态型芦苇对干旱和盐渍胁迫的响应调节

长期稳定的自然变异植物种是研究植物适应逆境机制的理想实验体系。就河西走廊芦苇不同生态型的划分、4种生态型的光合响应调节、抗氧化保护机制、膜流动性修饰机制和渗透调节机制中的响应特征、胁迫诱导的特异表达成分等作了概述。     

苜蓿二磷酸核酮糖(RuBP)羧化酶体内活化作用的调节

苜蓿RuBP羧化酶的初活性和活化作用在不饱和光强下与光合速率一样随光强增加而增加。缺硫培养苜蓿叶片的光合速率和RuBP羧化酶的含量、初活性及总活性均比对照有不同程度的降低,其中酶的初活性与光合速率两者减少的趋势比较接近,说明RuBP羧化酶的初活性可能在光合CO_2固定作用中具有决定作用。然而,缺硫植株中酶的活化作用比对照明显增高。酶的活化作用与叶片中的叶绿素,6-PG,NADPH及ATP相对酶含量的比值成正比,与体内的酶量成反比。     

不同生境两种生态型芦苇叶片质膜H~ -ATPase的比较(英文)

利用两相法纯化质膜微囊,研究了分布于西北沙漠地区的两种生态型芦苇(Phragmites communis Trin.)(水生芦苇和重度盐化草甸芦苇,分别简称为水芦和盐芦)叶片质膜H -ATPase的部分性质。结果显示,与水芦相比,盐芦质膜H -ATPase的ATP水解活性升高,Km值由1.27 mmol/L降至0.30 mmol/L,但Vmax没有显著差异。并且该酶活性对温度的敏感性和pH谱型也发生了变化。以对硝基苯磷酸盐为底物,低浓度时盐芦的质膜H -ATPase水解活性高于水芦,高浓度时则没有差异。Km在水芦和盐芦中分别为3.61 mmol/L和1.92 mmol/L,但Vmax在两种生态型中没有差异。钒酸盐抑制实验表明,两种生态型的质膜H -ATPase磷酸-酶区的催化性质不同。胰酶对质膜H -ATPase活性的活化谱型也存在差异,说明该酶C末端的结构或性质发生了变化。此外,与水芦相比,盐芦质膜H -ATPase的质子泵活性及与水解活性的耦联程度也升高了。以上结果说明,当芦苇从水生环境向盐渍环境过渡时,质膜H -ATPase的催化性质发生了变化,这些变化可能是由酶结构的修饰和不同的同工酶谱引起的。H -ATPase催化性质的变化可能是对盐渍生境的适应性反应。     

河西走廊不同生态型芦苇核酸代谢季节动态研究

分布在甘肃河西走廊的４ 种生态型芦苇（Ｐｈｒａｇｍ ｉｔｅｓｃｏｍ ｍ ｕｎｉｓＴｒｉｎ．）的核酸代谢季节变化有差异。盐化草甸芦苇ＲＮＡ 含量持续增加,ＤＮＡ 含量相对稳定,其它３ 种生态型芦苇的ＲＮＡ 和ＤＮＡ 含量以５月份为最高。过渡带芦苇的ＲＮＡ 含量、沼泽芦苇及沙丘芦苇的ＤＮＡ含量９ 月份略有增高。盐化草甸芦苇与过渡带芦苇的ＤＮａｓｅ和ＲＮａｓｅ的活性７ 月份最高,沼泽芦苇与沙丘芦苇的ＤＮａｓｅ和ＲＮａｓｅ活性５—９ 月份呈增高趋势。盐化草甸芦苇的ＤＮＡ 和ＲＮＡ 合成活性不断升高,过渡带芦苇和沼泽芦苇的ＤＮＡ 和ＲＮＡ 合成活性及沙丘芦苇的ＲＮＡ 合成活性５—９月份均降低,仅沙丘芦苇的ＤＮＡ合成活性增强。ＲＮＡ 聚丙烯酰胺梯度凝胶电泳分析结果表明,４ 种生态型芦苇均含有２５Ｓ、２３Ｓ、１８Ｓ、１６Ｓ大分子量ｒＲＮＡ 和小分子量５．８Ｓ、５Ｓ、４．５ＳｒＲＮＡ 及４ＳｔＲＮＡ。大分子量ＲＮＡ 的含量高于小分子量ＲＮＡ 含量。不同生态型及同一生态型芦苇的不同发育时期,相同的ＲＮＡ 组分含量各不相同。且发育过程中２３Ｓ、１８Ｓ、１６ＳｒＲＮＡ 在不同月份发生不同程度的降解。由此,我们认为,核酸代谢的差异性是４ 种生态型由生长转入衰     

Variation of RuBP carboxylase activity among anther-derived plants of Solanum phureja

The effect of genotype and ploidy on RuBP carboxylase (EC 4.1.1.39) activity, chlorophyll content, leaf area, chloroplast ultrastructure and not photosynthesis among monoploid. diploid and tetraploid anther-derived plants of Solanum phureja Juz, and Buk. was studied. Within the monoploid group, RuBP carboxylase activity and concentration displayed a significant genotypic effect. For the diploids, variation among genotypes was significant for total protein content and maximum specific activity of RuBP carboxylase, and among the tetraploids for net photosynthesis and specific leaf weight. Ploidy effect was evident regarding net photosynthesis, leaf area and chlorophyll content. The different ploidy groups among the anther-derived plants surpassed the anther donor plant for all characteristics except maximum activity of RuBP carboxylase and net photosynthesis. For the latter only the tetra-ploid group was superior to the anther source plant. However, a monoploid genotype with an increase of 9% in maximum activity of RuBP carboxylase over the anther-donor plant was identified. Segregation of trails rind differential gene expression together with possible mutations during androgenesis are discussed as sources of variation.     

盘锦湿地芦苇(Phragmites communis)群落蒸发散主导影响因子

利用2005年全年盘锦芦苇湿地梯度观测和涡动相关系统的监测数据,对芦苇群落的蒸发散变化规律及其主导影响因子分析表明,不同月份芦苇群落蒸发散日变化呈现出相同的变化趋势,即早晚低、中午高的单峰型曲线;芦苇群落蒸发散主要受当地气象因素、植被生长状况和生理生态特征的影响.相关分析表明,芦苇湿地蒸发散与净辐射、气温、地表温度、相对湿度、风速、土壤含水量等环境因素的变化都有很好的响应.回归分析表明影响生长季主要因素有:净辐射、土壤含水量、相对湿度、气温和地表温度;影响非生长季主要有:净辐射、地表温度和风速.同时,植被生长状况和生理生态特征对蒸发散也有显著影响,其中叶面积指数与气孔导度是芦苇群落蒸发散的主导影响因子.     

Mutations in loop six of the large subunit of ribulose-1,5-bisphosphate carboxylase affect substrate specificity

Mutagenesis in vitro of the gene encoding the large subunit of ribulose-1,5-bisphosphate carboxylase/ oxygenase (EC 4.1.1.39) from Anacystis nidulans Synechococcus PCC 6301) was used to generate novel enzymes in Escherichia coli. Residues in C-terminal loop 6 of the / barrel structure of the large subunit were changed. Replacement of valine 331 with alanine caused a 90% reduction in V _max but did not alter the enzyme's relative specificity towards either of its gaseous substrates, CO₂ and O₂. However replacement of alanine 340 with glutamate decreased the enzyme's specificity for CO₂ but had no significant effect on either the K _m for ribulose-1,5-bisphosphate or CO₂ or on V _max. In contrast replacing a small cassette of residues 338-341 produced a small increase in the specificity factor.Abbreviations Rubisco ribulose-1,5-bisphosphate carboxylase/oxygenase - RuBP ribulose-1,5-bisphosphate - CABP 2-carbox-yarabinitol-1,5-bisphosphate We thank Karen Moore for the statistical analysis of the specificity factors. We acknowledge helpful discussions with Jim Pitts and Richard Pickersgill. This work was aided by the invaluable technical assistance of Iain Major.     

松嫩平原不同生境芦苇种群分株的生物量分配与生长分析

在松嫩平原8月中旬,4个旱地生境芦苇种群分株的生长和生产力以季节性临时积水的低洼地为最高,以碱斑地为最低,两个生境之间分株高度相差2.8倍,分株生物量相差4.4倍,其它2个生境沙地和草甸混生 少也达到了显著或极显著水平,不同生境芦苇种群分株生物量分配的差异,蕴涵着重要的生长调节和物质分配策略。在分株株较小而又有充分生长空间的沙地生境和三斑生境,以及具有种间竞争的草句生境,分株将50％以上的物质分配给叶的建造上,以保证充分的物质生产;在环境条件相对较好的低洼地生境,分株将近40％的物质分配给茎的伸长生长,以提高个体对空间和光资源的竞争力。4个生境芦苇种群在分株增高与增重,以及茎、叶与分株的相对增重均具有相同的幂函数异速生长规律。     

用分子标记技术分析不同生态型芦苇的遗传多样性

用分子标记技术对河西走廊4种不同生态型芦苇进行了遗传多态性分子标记分析。从30条ISSR引物和45条RAPD引物中分别筛选出适合4种不同生态型芦苇分析的9条和13条引物,前者共扩增出99条带,多态性位点数为51;后者共扩增出195条带,多态性位点数为87。两种分子标记分析所得的遗传相似系数呈显著正相关(r=0.845,P<0.05)。4种生态型芦苇表现出由水生芦苇经盐渍芦苇向沙丘芦苇逐渐演化的趋势。     

澳门路氹湿地芦苇氮磷含量的季节变化

为探讨芦苇（Phragmites communis）的生长规律及其在湿地生态系统中的作用,对澳门路毡湿地芦苇氮磷含量变化进行了6次调查。结果表明：芦苇地上部分和地下茎的氮、磷含量都在秋冬达到最大值,在春夏达到最低值,其中叶氮、磷含量最高值分别为37．5和4．0g·kg^-1,最低值为28．0和3．5g·kg^-1;茎中部氮、磷含量最高值分别为14．1和2．0g·kg^1,最低值为11．5和1．6g·kg^1;地下茎氮、磷含量最高值分别为10．2和0．9g·kg^1,最低值为6．6和0．8g·kg^-1。各季节芦苇不同部位的氮、磷含量大小顺序均为叶〉地上茎〉地下茎。氮、磷没有在芦苇体不同器官或部位之间发生转移。     

Proteomic characterization of Phragmites communis in ecotypes of swamp and desert dune

Phragmites communis Trin. (common reed) is a recognized model plant for studying its adaptation to contrasting and harsh environments. To understand the inherent molecular basis for its remarkable resistance to combined stresses, we performed a comprehensive proteomic analysis of the leaf proteins from two ecotypes, i.e. swamp and desert dune, naturally growing in the desert region of northwestern China. First, a proteome reference map of Phragmites was established based on the swamp ecotype. Proteins were resolved by 2‐D/SDS‐PAGE and identified by MALDI‐TOF/TOF MS. In total, 177 spots were identified corresponding to 51 proteins. The major proteins identified are proteins involved in photosynthesis, glutathione and ascorbic acid metabolism as well as protein synthesis and quality control. Second, the 2‐DE profiles of the two ecotypes were compared quantitatively via DIGE analysis. Compared with swamp ecotype, 51 proteins spots are higher‐expressed and 58 protein spots are lower‐expressed by twofold or more in desert dune ecotype. Major differences were found for the proteins involved in light reaction of photosynthesis, protein biosynthesis and quality control and antioxidative reactions. The physiological significance of such differences is discussed in the context of a flow of complex events in relation to plant adaptation to combined environmental stresses.