Characterization of the trnL-trnF spacer sequence of the chloroplast tRNA genes in Spirodela species

The trnL-trnF spacer region of the chloroplast tRNA genes was sequenced and characterized in 14 accessions of the genus Spirodela (Lemnaceae). Only a low intraspecific variation of the spacer was observed in geographically isolated and morphologically different accessions of S. polyrrhiza, the most widespread Spirodela species. Five haplotypes of the spacer were identified, differing in mono-and oligonucleotide repeats and extended indels, specific to S. polyrrhiza, Landoltia punctata, and Lemna sp. The result supported the isolation of Landoltia from Spirodela.     

RAPD analysis of genome polymorphism in the family Lemnaceae

The multilocus RAPD analysis of intergeneric, inter- and intraspecific nuclear genome polymorphism was used for the first time to assess intergeneric, interspecific, and intraspecific polymorphism in Lemnaceae growing on the territory of Russia. The origin of the chosen accessions overlapped with the natural range of duckweeds in Russia. Seventy-five Lemnaceae accessions representing eight species (L. minor, L. gibba, L. turionifera, L. japonica. L. trisulca, L. aequinoctialis, S. polyrhiza, and L. punctata) from three genera (Lemna, Spirodela, and Landoltia), were analyzed. The highest variability levels were revealed in L. minor accessions (0.03-0.20). Species L. trisulca and S. polyrhiza were characterized by values of genetic distance 0.01-0.18 and 0.03-0.16, respectively. The lowest polymorphism levels were detected for L. turionifera (0.01-0.11). The dendrogram based on RAPD data showed that L. aequinoctialis was the most genetically distant species of the genus Lemna. Accessions of species L. turionifera and L. japonica, as well as L. minor and L. gibba, did not form separate species-specific subclusters; rather, they fell into clusters with L. japonica/L. turionifera and L. minor/L. gibba. Accessions of the genera Spirodela and Landoltia formed two separate clusters combined into one group.     

不同光温条件下紫背浮萍叶肉细胞和叶绿体超微结构的研究

以浮萍科紫萍属紫背浮萍为试验材料,利用透射电子显微镜研究了紫背浮萍叶肉细胞和叶绿体的超微结构。结果发现:强光下,紫背浮萍叶肉细胞内的叶绿体数明显增加;弱光时虽然叶绿体数减少,但叶绿体基粒片层结构变厚,增加了光合作用反应面积。与25℃适温条件相比,10℃低温和35℃高温下,紫背浮萍细胞均出现一定程度的逆境胁迫作用。表现在低温时部分叶绿体皱缩成带状,叶肉细胞之间出现较大空隙;高温时叶绿体外膜溶解,基质外渗,脂质小球数量增多。紫背浮萍通过改变细胞内部形态结构的方式来适应不同光强和温度环境条件,对逆境光温条件具有一定的耐受性。     

贵州省浮萍形态学鉴定与遗传多样性分析

为调查贵州省浮萍种间亲缘关系及变异情况, 研究在贵州省辖9个地级行政区的多处水环境中共采集到 41份浮萍种质, 通过形态鉴定这些种质中33个属于绿萍属(Lemna), 6个属于紫萍属(Spirodela), 2个属于斑萍属/兰氏萍属(Landoltia)。利用叶绿体atpF-atpH间隔序列和rpS16内含子序列进行分子生物学分析和鉴定, 41份浮萍种质聚类到Lemna aequinoctialis、Lemna minor、Spirodela polyrhiza和Landoltia punctata 4个种。atpF-atpH间隔序列和rpS16内含子序列的单倍型多样性分别为0.98700和0.64700, 群体突变率分别为0.15380和0.14334, 平均每kb核苷酸差异数为47.20000和61.72200, 核苷酸多态性分别为0.08725和0.09158。研究讨论了中国亚热带温润季风地区浮萍的遗传多样性, 为浮萍亲缘关系分析提供了依据, 也为亚热带湿润季风气候地区植物的种属鉴定及资源化利用奠定了基础。     

响应面优化紫萍黄酮提取工艺及其抗氧化活性

采用响应面法优化微波提取紫萍黄酮的条件。在单因素实验的基础上,选取乙醇体积分数、微波功率、液料比为影响因子,应用Box-Behnken中心组合法进行三因素三水平的试验设计,以紫萍黄酮得率为响应值,进行响应面分析(RSM),结果表明:微波提取紫萍黄酮的最佳提取条件为乙醇体积分数62%、液料比17∶1(mL/g)、微波功率500 W、微波时间9 min;在优化工艺下,紫萍黄酮的得率达到4.14%。对微波提取的紫萍黄酮进行抗氧化性研究发现紫萍黄酮对DPPH.清除作用较好,其抗氧化活性要优于相同浓度的抗坏血酸(VC)和2,6-二叔丁基对甲酚(BHT)。     

Molecular phylogenetic relationships among Lemnaceae and Araceae using the chloroplast trnL-trnF intergenic spacer

We test competing hypotheses of relationships among Aroids (Araceae) and duckweeds (Lemnaceae) using sequences of the trnL-trnF spacer region of the chloroplast genome. Included in the analysis were 22 aroid genera including Pistia and five genera of Lemnaceae including the recently segregated genus Landoltia. Aponogeton was used as an outgroup to root the tree. A data set of 522 aligned nucleotides yielded maximum parsimony and maximum likelihood trees similar to those previously derived from restriction site data. Pistia and the Lemnaceae are placed in two separate and well-supported clades, suggesting at least two independent origins of the floating aquatic growth form within the aroid clade. Within the Lemnaceae there is only partial support for the paradigm of sequential morphological reduction, given that Wolffia is sister to Wolffiella+Lemna. As in the results of the restriction site analysis, pantropical Pistia is placed with Colocasia and Typhonium of southeastern Asia, indicative of Old World affinities. Branch lengths leading to duckweed terminal taxa are much longer relative to other ingroup taxa (including Pistia), evidently as a result of higher rates of nucleotide substitutions and insertion/deletion events. Morphological reduction within the duckweeds roughly correlates with accelerated chloroplast genome evolution.     

紫萍磷转运蛋白1;1 cDNA序列的克隆及表达模式

紫萍Spirodela polyrrhiza是一种在生物质资源开发利用和水体生物修复中广泛使用的漂浮植物。随着水体富营养化日益严重,紫萍在水面随处可见。有文献报道紫萍对水体中的氮和磷有良好的净化作用。为了深入研究紫萍对水体中无机磷的吸收和转运,以紫萍为材料提取RNA反转录为cDNA,以此为模板,扩增得到1条特异性片段,其开放读码框 (ORF) 全长序列为1 620 bp,编码539个氨基酸,命名为SpPHT1;1,在GenBank中登录号为MN720003。生物信息学分析表明,SpPHT1;1没有内含子,编码的蛋白是一种稳定的、疏水的、具有11个跨膜结构域的蛋白,其结构具有主要协助转运蛋白超家族 (Major facilitator superfamily,MFS) 保守的结构域,聚类分析显示紫萍SpPHT1;1与玉米ZmPHT2和高粱SbPHT1-8有较近的亲缘关系,推测其属于植物磷转运蛋白家族PHT1成员。正常磷条件,SpPHT1;1在紫萍叶中的表达明显多于根;低磷条件促进该基因的表达,相对表达量在紫萍根和叶生长48 h时均达到峰值;丰磷条件会抑制基因表达,说明SpPHT1;1的表达会受到外界磷浓度的影响。研究结果有助于紫萍磷转运蛋白基因功能的进一步深入研究,为紫萍的进一步开发和利用提供理论依据。     

不同浓度铜对紫背萍和青萍色素含量及抗氧化酶系统的影响

通过水培试验研究了不同浓度的重金属Cu对紫背萍(Spirodela polyrrhiza)和青萍(Lemna minor)的色素含量和抗氧化酶系统的影响.结果表明,低浓度Cu(0.056 mg·L^-1)的处理下,紫背萍和青萍的叶绿素a、叶绿素b和叶绿素(a+b)的含量均出现不同程度的增加,分别高出其对照11％、46％、22％和8％、15％、11％,而在高浓度Cu(0.18～5.60 mg·L^-1)的处理下,上述色素含量均显著下降,平均下降幅度分别达63％、62％、65%和46％、44％、45％.紫背萍体内丙二醛(MDA)含量为青萍的2.57倍.两种浮萍的超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和过氧化物酶(POD)的活性均随Cu浓度的增加而呈先升后降的趋势,紫背萍体内3种酶活性在Cu浓度仅为0.18 mg·L^-1时即达峰值,随后则大幅下降,而青萍体内3种酶活性却在Cu浓度分别升高到0.56、1.0和1.0 mg·L^-1时才达到峰值.可见,在Cu胁迫下,紫背萍受Cu毒害较青萍深,且其体内抗氧化酶系统也较青萍敏感.     

Phytoremediation of Cr(VI) by Spirodela polyrrhiza (L.) Schleiden employing reducing and chelating agents

Phytoremediation of Cr(VI) by Spirodela polyrrhiza in binary combinations with low molecular weight organic compounds (LMWOCs) with a reducing or chelating potential, viz., ascorbic acid, citric acid, tartaric acid, oxalic acid, lactic acid, and glycerol was studied in Cr(VI) containing hydroponic media. Significant increase in the relative dry weight of plants with respect to Cr(VI) treated controls was observed with ascorbic acid and glycerol. The uptake of chromium by S. polyrrhiza followed Michaelis-Menten kinetics of active ion uptake. Interaction between Cr and ascorbic acid, oxalic acid, and lactic acid decreased Cr uptake, whereas citric acid, glycerol, and tartaric acid increased it. Supplementation of LMWOCs to Cr(VI) containing media decreased the MDA content of the plants. Multiple regression models revealed that LMWOCs decrease lipid peroxidation independently, as well as that induced by Cr(VI). It was found that superoxide dismutase (SOD), guaiacol peroxidase (GPX), and catalase (CAT) activities were increased significantly in plants growing in media containing Cr(VI). The study established that lactic acid, citric acid, ascorbic acid, and glycerol were most effective in increasing the Cr(VI) phytoremediating potential of S. polyrrhiza and LMWOCs with reducing or chelating properties decrease Cr(VI) stress in S. polyrrhiza.     

基于人工培养的浮萍快速无性繁殖营养条件研究

为探究并优化浮萍人工培养技术, 研究以广布种紫萍(Spirodela polyrrhiza)和青萍(Lemna minor)为主要研究对象, 探索两种浮萍植物在不同营养水平的Hoagland和Hunter培养液中的鲜重、叶状体数的生长变化状况。结果表明: (1)紫萍和青萍鲜重在Hoagland培养液的不同营养水平下先增加后减少, 而在Hunter营养液中呈持续增长趋势; 两种浮萍的鲜重最大相对增长率(RGR)分别为0.11和0.18, 鲜重受浮萍品种和培养基类型及其不同营养水平影响显著(P<0.05), 以青萍在Hunter原液培养基下的无性繁殖产生的生物量最高。(2)紫萍和青萍叶状体数在Hoagland培养液的不同营养水平下先增加后减少, 而同鲜重一样在Hunter营养液中呈不断增长趋势; 两种浮萍的叶状体最大相对增长率分别为0.14和0.19, 叶状体生长的RGR变化同样受浮萍品种和培养基类型及营养水平影响显著(P<0.05), 以青萍在Hunter原液的营养环境下收获的叶状体数最高。(3)两种浮萍在Hoagland和Hunter营养液的不同营养水平下鲜重/叶状体比呈下降趋势, 表明两种浮萍在适应不同营养时优先繁殖子代叶状体, 以扩大种的适合度。研究认为Hunter原液可作为广布种青萍的最优培养条件, 可实现短时间内收获较大浮萍鲜生物量和叶状体数, 为进一步资源化利用提供原材料。     

The development of someLemnaceae under different nutrient conditions

Plants ofSpirodela polyrrhiza, Lemna gibba, L. minor andL. minuscula were grown experimentally in nutrient media under various nutrient conditions involving different concentrations and forms of nitrogen and different concentrations of phosphorus. The treatments affected the multiplication rate of the plants, their root length and formation of anthocyanin and of turions.     

nrDNA ITS和cpDNA rpl16基因测序法对六种鼠尾草的检测

为从鼠尾草属植物中鉴别丹参品种,采用基因测序方法,用核糖体核酸内转录间隔区基因(nrDNA ITS),编码核蛋白体大亚基多肽L16的基因(rpl16)及叶绿体DNA上包含trnL以及trnL和trnF间隔区的区域基因(trnL-trnF)的序列,检测六种鼠尾草属新鲜植物.由于nrDNA ITS和rpl16突变率较高,可以做为6种鼠尾草的基源鉴定标记,依此设计了两对特异引物,从6种鼠尾草中鉴定出丹参(Salvia miltiorrhiza)和云南鼠尾草(S.yunnanensis).但trnL-trnF突变率太低,未能用于鉴别.商品干燥中药材因加工和储藏的方式致使DNA降解严重,基因测序法难于应用.     

The nucleotide sequence of the 4.5S and 5S rRNA genes and flanking regions from Spirodela oligorhiza chloroplasts

The base sequence of Spirodela oligorhiza chloroplast DNA coding for 4.5S and 5S ribosomal RNA, the flanking regions and the spacer between these two genes has been determined. We have compared these sequences with the corresponding ones in other higher plants. Besides a high degree of homology, some interesting differences are found.     

pH及无机氮化合物对细脉浮萍生长的影响

研究以细脉浮萍 ( Lemna aequinoctialis)为研究对象 ,在完全培养液的基础上模拟生活污水的氨氮、硝酸氮浓度和 p H范围 ,研究了培养条件对其生长的影响。结果表明 ,这个浮萍种对 p H耐受的低限在 4～ 5之间 ,最适 p H范围为 5～ 6。非离子态的氨会对其生长产生抑制作用 ,在氨氮为唯一氮源的条件下 ,非离子态氨的浓度大于 0 .1 mg/L时会对此种的生长产生明显的抑制作用 ,接近 2 mg/L时 ,基本上致死。在研究的氨氮和硝酸氮浓度范围内 ( <40 mg N /L) ,铵根离子和硝酸根离子浓度增加可促进细脉浮萍的生长。氨氮中生长的植株过氧化物酶活性高于硝酸氮中的活性。     

The phylogenetic utility of nucleotide sequences of sorbitol 6-phosphate dehydrogenase in Prunus (Rosaceae)

Sequences from s6pdh, a gene that encodes sorbitol-6-phosphate dehydrogenase in the Rosaceae, are used to reconstruct the phylogeny of 22 species of Prunus. The s6pdh sequences alone and in combination with previously published sequences of the internal transcribed spacer (ITS) and the cpDNA trnL-trnF spacer are analyzed using parsimony and maximum likelihood methods. Both methods reconstructed the same phylogeny when s6pdh sequences are used alone and in combination with ITS and trnL-trnF, and the topology is in agreement with previous studies that used a larger sample size. The s6pdh sequences have about twice as many informative sites as ITS. A molecular clock is rejected for s6pdh, most likely due to greater rates of evolution in subgenera Padus and Laurocerasus than in the rest of the genus. Phylogenetic reconstruction of Prunus as determined by analysis of the combined data set suggests an early split into two clades. One is composed of subgenera Cerasus, Laurocerasus, and Padus. The second includes subgenera Amygdalus, Emplectocladus, and Prunus. Species of section Microcerasus (formerly in subgenus Cerasus) are nested within subgenus Prunus. The order of branching and relationships among early diverging lineages is weakly supported, as a result of very short branches that may indicate rapid radiation.     

Concurrent removal and accumulation of heavy metals by the three aquatic macrophytes

Under the present investigation effectiveness of three aquatic macrophytes Pistia stratiotes L. (water lettuce), Spirodela polyrrhiza W. Koch (duckweed) and Eichhornia crassipes were tested for the removal of five heavy metals (Fe, Zn, Cu, Cr and Cd). These plants were grown at three different concentrations (1.0, 2.0 and 5.0mgl(-1)) of metals in laboratory experiment. Result revealed high removal (>90%) of different metals during 15 days experiment. Highest removal was observed on 12th day of experiment, thereafter it decreased. Results revealed E. crassipes as the most efficient for the removal of selected heavy metals followed by P. stratiotes and S. polyrrhiza. Results from analysis confirmed the accumulation of different metals within the plant and a corresponding decrease of metals in the water. Significant correlations between metal concentration in final water and macrophytes were obtained. Plants have accumulated heavy metals in its body without the production of any toxicity or reduction in growth. Selected plants shown a wide range of tolerance to all of the selected metals and therefore can be used for large scale removal of heavy metals from waste water.     

基于形态和cpDNA序列探讨柽柳和甘蒙柽柳的亲缘关系

柽柳科植物的系统学关系已初步明确,但部分种间的系统进化关系仍有待澄清。为明确中国黄河流域广泛分布的柽柳(Tamarix chinensis)和甘蒙柽柳(T. austromongolica)的亲缘关系,利用13个稳定的形态学性状和3个cpDNA间隔区片段(rpl20-5′-rps12, rps16, trn L-trn F)对柽柳科3属9种植物进行研究,结果表明3条序列的位点变异程度依次为trn L-trn F>rps16>rpl20-5′-rps12,柽柳和甘蒙柽柳群体共享有同一单倍型,表型和分子数据均支持柽柳和甘蒙柽柳为单系分支的近缘种,推测柽柳和甘蒙柽柳的分化大致发生在0.69Ma (0-1.93) Ma,与同属内其它物种相比,分化时间相对较晚。     

Phylogenetic relationship in Fritillaria spp. of Iran inferred from ribosomal ITS and chloroplast trnL-trnF sequence data

The genus Fritillaria embraces up to 165 taxa in the family Liliaceae, most of which are of high medicinal and ornamental value and importance. In this study, 44 specimens of the genus representing 9 species were collected from their natural habitats located in 10 provinces of Iran. Phylogenetic analysis was performed based on DNA sequences of the internal transcribed spacer (ITS) of the nuclear ribosomal cistron and the trnL-trnF regions. The phylogeny was constructed using the neighbor joining inference method. Results indicate that the examined samples were evidently diverged into 2 distinct clades. Members of the subgenera Fritillaria and Rhinopetalum formed one clade while the other clade contained the subgenera Theresia and Petilium. There can be seen a high degree of similarity between the only yellow-colored crown imperial specimen and the red-colored specimens. The endemic species of Fritillaria straussii, Fritillaria zagrica and Fritillaria kotschyana which their status within the subgenera known in the genus Fritillaria has been remained undefined, fell into the subgenus Fritillaria. The clades also had relatively reasonable distribution patterns based on the genetic structure, geographical conditions and climate specifications. This study revealed the feasibility of the ITS and trnL-trnF DNA sequence for phylogeny of the genus Fritillaria. This is the first phylogenetic analysis of Fritillaria spp. in Iran.