染色体加倍对忍冬生物学性状及其生态适应性的影响

忍冬（Lonicera japonica Thunb.）属忍冬科忍冬属，是一种重要的药用植物，其花蕾称为金银花（Flos Lonicerae），在我国已有1000多年的药用历史，具有清热解毒、凉散风热等功效。我们利用秋水仙素诱导二倍体“大毛花”品种茎尖选育出同源四倍体“九丰一号”品种，并在生产中发现花蕾产量显著提高，而药效成份含量是否发生变化了呢？四倍体忍冬表现出典型的器官巨大性，这些变化是否与其较强生态适应性之间存在联系呢？染色体加倍增强了忍冬的生态适应性，其生态修复功能如何呢？在本研究中，主要从以下几个方面进行了探讨：（1）染色体加倍对其叶片，茎、花蕾和花蕾产量及其药效成份等生物学性状的影响;（2）染色体加倍后植物对水分胁迫的响应;（3）染色体加倍后植物对热胁迫的响应;（4）除上述两个品种外，增加一个变异品种“红银花”，探讨3个忍冬品种对退化生态系统的修复功能。主要结果如下： 1. 通过测定根尖细胞染色体数目和使用流式细胞仪分析茎尖细胞DNA含量，表明四倍体忍冬（2n = 4x = 36）确实来自二倍体忍冬（2n = 2x = 18）的染色体加倍。四倍体忍冬气孔细胞大小显著大于二倍体，而气孔密度显著低于二倍体。四倍体忍冬没有光合“午休”，而二倍体存在明显的光合“午休”，这可能与其存在抗高温的叶片解剖结构特性有关。四倍体忍冬叶片较二倍体变大、变厚、变浓绿（较高叶绿素含量）。与二倍体相比，四倍体忍冬单位叶面积重量显著增高，表明其具有较强的生态适应性。四倍体忍冬单个花蕾的鲜重和干重均显著大于二倍体。连续3年的花蕾产量调查表明，四倍体显著高于二倍体。染色体加倍使其茎干粗壮、节间变短、新梢上着生花蕾数目增多及单个花蕾变大，这是其花蕾高产的生物学基础。对金银花的药效成份而言，染色体加倍不影响绿原酸的含量，但增加了木犀草苷的含量。结果表明，染色体加倍能增加金银花的产量和药效成份的含量，建议四倍体忍冬在药材生产中推广应用。 2. 水分胁迫显著降低二倍体和四倍体忍冬叶片的净光合速率、气孔导度和蒸腾速率。水分胁迫也降低电子传递速率、光系统II实际量子产量和光化学猝灭，而增加非光化学猝灭、总可溶性糖、脯氨酸和丙二醛的含量。四倍体忍冬对水分胁迫的响应表现为其叶片水势、气体交换、叶绿素荧光和有关代谢物含量的变化程度低于二倍体，并且复水后其恢复能力快于二倍体，表明染色体加倍增强了忍冬的抗旱能力。分析其主要原因，可能是由于四倍体植株总叶面积减少、单位叶面积重量增加、叶片表皮细胞和栅栏组织增大以及叶片表皮毛较浓密等形态解剖结构特性有关。结果表明，染色体加倍能增加忍冬植物的抗旱能力，而使其具有较强的生态适应性。 3. 二倍体和四倍体忍冬受48 ºC热胁迫处理6 h和恢复10 h，以及离体叶片45 ºC，50 ºC，55 ºC 水浴热胁迫3 min，应用叶绿素荧光成像系统研究了它们对热胁迫的响应。热胁迫显著降低了两个品种叶片的最大光化学效率、电子传递速率、光系统II实际量子产量和光化学猝灭，降低了四倍体的非光化学猝灭，而增加了二倍体的非光化学猝灭。热胁迫增加了两个品种叶片的总可溶性糖、脯氨酸和丙二醛的含量。四倍体受热胁迫的叶绿素荧光参数和有关代谢物的响应程度低于二倍体，以及其恢复程度快于二倍体，表明染色体加倍提高了抗热性。此外，叶绿素荧光成像的异质性也表明，四倍体的抗热能力大于二倍体。进一步的叶片解剖结构分析表明，四倍体叶片的表皮细胞变大、栅栏组织增厚、表皮毛较浓密等特点，是其抗热性强的主要原因。 4.根据以上的研究结果，通过四倍体忍冬生态修复功能的野外试验证明染色体加倍后其生态适应性变化。在本研究中，针对北京市门头沟区大面积不同类型的废弃地急需恢复植被和景观的问题，在恢复生态学理念的指导下，综合运用集成技术，将3个忍冬品种植物用于这些退化生态系统的恢复。在王平镇的公路下边坡（以碎石和矿渣为主）、煤矿、石灰窑和采石场4种类型废弃地建立生态修复的试验示范区。结果表明，在4个废弃地类型上引进的3个金银花品种，具有使示范区快速复绿、当年成景和群落快速形成的潜力，并具有对不同退化迹地的适应能力和恢复效果，其中四倍体忍冬效果更好些，这主要与其形态解剖结构和较强的生态适应性有关。

Comparative studies on photosynthesis and phosphate metabolism of Cylindrospermopsis raciborskii with Microcystis aeruginosa and Aphanizomenon flos-aquae

The physiological differences for three bloom-forming cyanobacteria (Cylindrospermopsis raciborskii, Microcystis aeruginosa, and Aphanizomenon flos-aquae) were investigated. In comparison with M. aeruginosa and A. flos-aquae, C. raciborskii exhibited a significantly higher concentration of carotenoids, higher values in maximum photosynthesis rate (P-m), apparent photosynthetic efficieny (a), and maximum electron transport rate (ETRmax) during the growth period. In addition, higher extracellular alkaline phosphatase activities and lower light compensation point (I-c) were also detected in C raciborskii (p < 0.05, ANOVA). Therefore, it is suggested that the higher photosynthetic activities, more effective uptake and utilization to phosphate, and low light requirements might play important roles in the occurrence and invasive behavior of C. raciborskii. Crown Copyright (C) 2009 Published by Elsevier B.V. All rights reserved.

Mouse Toxicity of Anabaena flos-aquae from Lake Dianchi, China

Some species of the genera Anabaena can produce various kinds of cyanotoxins, which may pose risks to environment and human health. Anabaena has frequently been observed in eutrophic freshwater of China in recent years, but its toxicity has been reported only in a few studies. In the present study, the toxicity of an Anabaena flos-aquae strain isolated from Lake Dianchi was investigated. Acute toxicity testing was performed by mouse bioassay using crude extracts from the lyophilized cultures. The mice exposed to crude extracts showed visible symptoms of toxicity and died within 10-24 h of the injection. Serum biochemical parameters were evaluated by the use of commercial diagnostic kits. Significant alterations were found in the serum biochemical parameters: alkaline phosphatase (AKP), gamma-glutamyl transpepticlase (gamma-GT), aspartate amino transferase (AST), alanine amino transferase (ALT), AST/ALT ratio, total protein content, albumin content, albumin/globulin (A/G) ratio, blood urea nitrogen (BUN), serum creatinine (Ssr), and total antioxidative capacity (T-AOC). Histopathological observations were carried out with hematoxylin and eosin (HE) stain under light microscope. Severe lesions were seen in the livers, kidneys, and lungs of the mice injected with crude extracts. The alterations of biochemical parameters were in a dose-dependent manner, and the severities of histological lesions were in the same manner. Based on biochemical and histological studies, this research firstly shows the presence of toxin-producing Anabaena species in Lake Dianchi and the toxic effects of its crude extracts on mammals. (C) 2008 Wiley Periodicals, Inc. Environ Toxicol 24: 10-18, 2009.

Lysis of Aphanizomenon flos-aquae (Cyanobacterium) by a bacterium Bacillus cereus

A phytoplankton-lytic (PL) bacterium, Bacillus cereus, capable of lysing the bloom-forming cyanobacterium. Aphanizomenon flos-aquae was isolated from Lake Dianchi of Yunnan province, China. This bacterium showed lytic activities against a wide range of cyanobacteria/algae, including A. flos-aquae, Microcystis viridis, Microcystis wesenbergi, Microcystis aeruginosa, Chlorella ellipsoidea, Oscillatoria tenuis, Nostoc punctiforme, Anabaena flos-aquae, Spirulina maxima, and Selenastrum capricornutum. Chlorophyll a contents, phycocyanin contents, and photosynthetic activities of the A. flos-aquae decreased evidently in an infected culture for a period. Bacterium B. cereus attacked rapidly A. flos-aquae cells by cell-to-cell contact mechanism. It was shown that the lysis of A. flos-aquae began with the breach of the cyanobacterial cell wall, and the cyanobacterial cell appeared abnormal in the presence of the PL bacterium. Moreover, transmission electron microscope examinations revealed that a close contact between the bacterium and the cyanobacterium was necessary for lysis. Some slime extrusions produced from B. cereus assisted the bacterial cells to be in close association with and lyse the cyanobacterial cells. These findings suggested that this bacterium could play an important role in controlling the Aphanizomenon blooms in freshwaters. (c) 2006 Elsevier Inc. All rights reserved.

First report of aphantoxins in China - waterblooms of toxigenic Aphanizomenon flos-aquae in Lake Dianchi

The oligohaline cyanobacterium Aphanizomenon flos-aquae (L.) Ralfs (A. flos-aquae) has been reported in several countries to produce paralytic shellfish poisons (PSPs) or protracted toxic effects. In the past years, A. flos-aquae blooms have occurred annually in the eutrophic Lake Dianchi (300 km(2) in area, located in southwestern China). Material from natural blooms dominated by A. flosaquae was collected and lyophilized. Acute toxicity testing was performed by mouse bioassay using extracts from the lyophilized material. Clear symptoms of PSPs, intoxications were observed. To confirm the production of PSPs, a strain of A. flos-aquae (DC-1) was isolated and maintained in culture. Histopathological effects were studied by examining the organ damages using transmission electron microscopy (TEM). Slight hepatocytic damage with swollen mitochondria was found. The ultrastructural pulmonary lesions were characterized by distortied nuclei and indenting of karyotheca, together with degeneration and tumefaction of mitochondria and endoplasmic reticulum. Control animals injected with acetic acid did not exhibit histopathological damage in any organ. Toxic effects of cultured algal cells on enzymatic systems in the mouse were studied using sublethal doses of extracts. Significant glutathione-S-transferase (GST) and lactate dehydrogenase (LDH) increases, together with decrease of the glutathione (GSH) level, were measured. These results indicated a potential role of PSPs intoxicating and metabolizing in the test animals. HPLC-FLD and LC/MS analysis of extracts from cultured material demonstrated the PSP toxins produced by A. flos-aquae bloom. To the best of our knowledge, this is the first study reporting chemically and toxicologically confirmed PSP toxins related to A. flosaquae in China. (c) 2005 Elsevier Inc. All rights reserved.

Taxonomic re-evaluation of Aphanizomenon flos-aquae NH-5 based on morphology and 16S rRNA gene sequences

The taxonomy of Aphanizomenon flos-aquae strain NH-5, a producer of cyanotoxins, was re-evaluated by comparison with six other Aphanizomenon strains using morphological characteristics and 16S rRNA gene sequences. Strain NH-5 was concluded to be improperly identified as Aph. flos-aquae based upon (1) lack of bundle formation in the trichomes, (2) location of akinetes next to heterocytes, (3) lower similarities (less than 97.5%) in the 16S rRNA gene sequences relative to Aph. flos-aquae strains, and (4) comparison within a phylogenetic tree constructed from 16S rRNA gene sequences. The Aphanizomenon strains investigated in this study are classified to four morphological groups as described by the classical taxonomy of Komarek & Kovacik (1989). This classification was supported from the phylogenetic results of 16S rRNA gene sequences. This study also discusses the generic boundaries between Aphanizomenon and Anabaena.

铬离子对藻类生长的影响

利用自然光照条件,在不同铬离子质量浓度下,对5种藻类的生长进行了实验研究。研究结果表明:5种藻类在不同的光波长下,其吸收光谱存在两个最强的吸收区:420～440nm的蓝紫光部分和660nm的红光部分;在不同铬离子质量浓度0～20.63mg·L-1范围内,5种藻类生长受到不同程度的抑制,其耐受性也尽不相同,经过分析和比较后得出5种藻类对铬的耐受性大小顺序为:微囊藻(Microcystis)>水华鱼腥藻(Anabaena flos-aquae)>细长聚球藻(Synechococcus elongatus)>斜

焦酚对蓝藻和绿藻生长、光合色素及微量元素的作用

研究了焦酚对4种常见蓝藻和绿藻的生长影响.结果表明,焦酚对几种藻的生长具有抑制作用.相同的处理浓度,4种藻生长率从小到大依次是:铜绿微囊藻(Microcystic aeruginosa)、羊角月牙藻(Selenastrum capri-cornutum)、蛋白核小球藻(Chlorella pyrenoidosa)、水华鱼腥藻(Anabaena flos-aquae).焦酚作用后铜绿微囊藻和羊角月牙藻的叶绿素a和胡萝卜素的含量有显著的变化,随着焦酚浓度的升高,铜绿微囊藻和羊角月牙藻叶绿素a和胡萝卜素含量减少

三种水华蓝藻对不同磷浓度生理响应的比较研究

本实验研究了铜绿微囊藻(Microcystis aeruginosa FACHB469)、水华鱼腥藻(Anabaena flos-aquae FACHB245)和浮游颤藻(Oscillatoria planctonicaFACHB708)对磷浓度变化的生理响应。结果表明,在缺磷条件下,A. flos-aquae对低磷环境的适应能力较强,O. planctonica其次,M. aeruginosa最差;在磷充足条件,微囊藻对磷过量吸收的能力明显高于其他两种蓝藻。三种蓝藻胞外碱性磷酸酶活性(APA)与培养基中

不同盐度下水华束丝藻对CO_2浓度倍增的生理响应

本文研究了在盐度分别为2‰和4‰的条件下,CO2浓度倍增对水华束丝藻(Aphanizomenon flos-aquae)生长速率、光合活性及光合色素比例与丙二醛含量的影响。结果表明随着盐度的增加水华束丝藻的生长速率和光合活性受到显著抑制,叶绿素a与藻蓝素比例大幅降低,丙二醛含量明显提高。在实验盐度范围内,CO2浓度倍增显著促进水华束丝藻的生长速率和最大光合电子传递速率(ETRmax)与色素比例,而且盐度越高促进效果越明显。此外CO2浓度倍增能显著降低丙二醛(MDA)含量。从而减少膜脂过氧化,缓解盐度胁迫。

中国淡水微囊藻属常见种类的分类学讨论——以滇池为例

根据中国云南滇池藻类样品的观察结果,对中国分布的淡水微囊藻属Microcystis10个常见种的形态特征进行了描述,同时对它们的分类学进行了讨论,并整理出分类检索表。这10种微囊藻是铜绿微囊藻M.aeruginosa、放射微囊藻M.botrys、坚实微囊藻M.firma、水华微囊藻M.flos-aquae、鱼害微囊藻M.ichthyoblabe、挪氏微囊藻M.novacekii、假丝微囊藻M.pseudofilamentosa、史密斯微囊藻M.smithii、绿色微囊藻M.viridis、惠氏微囊藻M.w

Mechanism study on the frequent variations of cell-bound microcystins in cyanobacterial blooms in Lake Taihu: Implications for water quality monitoring and assessments

Although Microcystis-based toxins have been intensively studied, previous studies using laboratory cultures of Microcystis strains are difficult to explain the phenomenon that microcystin concentrations and toxin variants in natural blooms differ widely and frequently within a short-term period. The present study was designed to unravel the mechanisms for the frequent variations of intracellular toxins related to the differences in cyanobacterial colonies during bloom seasons in Lake Taihu, China. Monitoring of Microcystis colonies during warm seasons indicated that the variations in microcystins in both concentrations and toxin species were associated with the frequent alteration of Microcystis colonies in Lake Taihu. High concentration of microcystins in the blooms was always associated with two Microcystis colonies, Microcystis flos-aquae and Microcystis aeruginosa, whereas when Microcystis wesenbergii was the dominant colonial type, the toxin production of the blooms was low. Additionally, environmental factors such as temperature and nutrition were also shown to have an effect on the toxin production of the blooms, and may also potentially influence the Microcystis species present. The results of the present study provides insight into a new consideration for quick water quality monitoring, assessment and risk alert in cyanobacterium- and toxin-contaminated freshwaters, which will be beneficial not only for water agencies but also for public health. (C) 2009 Elsevier Ltd. All rights reserved.

Reducing organic substances from anaerobic decomposition of hydrophytes

Oxidation-reduction properties of surface sediments are tightly associated with the geochemistry of substances, and reducing organic substances (ROS) from hydrophytes residues may play an important role in these processes. In this study, composition, dynamics, and properties of ROS from anaerobic decomposition of Eichhornia crassipes (Mart.) Solms, Potamogenton crispus Linn, Vallisneria natans (Lour.) Hara, Lemna trisulca Linn and Microcystis flos-aquae (Wittr) Kirch were investigated using differential pulse voltammetry (DPV). The type of hydrophytes determined both the reducibility and composition of ROS. At the peak time of ROS production, the anaerobic decomposition of M. flos-aquae produced 6 types of ROS, among which 3 belonged to strongly reducing organic substance (SROS), whereas there were only 3-4 types of ROS from the other hydrophytes, 2 of them exhibiting strong reducibility. The order of potential of hydrophytes to produce ROS was estimated to be: M. flos-aquae > E. crassipes > L. trisulca > P. crispus approximate to V. natans, based on the summation of SROS and weakly reducing organic substances (WROS). The dynamic pattern of SROS production was greatly different from WROS. The total SROS appeared periodic fluctuation with reducibility gradually weakening with incubation time, whereas the total WROS increased with incubation time. Reducibility of ROS from hydrophytes was readily affected by acid, base and ligands, suggesting that their properties were related to these aspects. In addition to the reducibility, we believe that more attention should be paid to the other behaviors of ROS in surface sediments.

Morphological characteristics and phylogenetic relationship of Anabaena species from Lakes Dianchi and Erhai, China

Although Anabaena is one of the most prevalent planktonic freshwater genus in China, there are few taxonomic reports of Anabaena strains by morphology and genetics. In this study, morphological characteristics and phylogenetic relationships of seven Anabaena strains isolated from two plateau lakes, Lakes Dianchi and Erhai, were investigated. Morphological characteristics such as morphology of filament, cellular shapes and sizes, relative position of heterocytes and akinetes, and presence or absence of aerotopes, were described for these seven strains. Phylogenetic relationships were determined by constructing 16S rRNA gene tree using the neighbor-joining algorithm. The seven strains were morphologically identified as three groups, and phylogenetic analysis based on 16S rRNA gene sequences also showed that these seven strains were in three groups. Strains EH-2, EH-3, and EH-4 were in group A belonging to the Anabaena circinalis and A. crassa group, and strains DC-1, DC-2, and EH-1 were in group B and identified as A. flos-aquae. Strain DC-3 without aerotopes was significantly different from the other isolated strains and was determined as A. cylindrica.

Taxonomic notes on water bloom forming Microcystis species (Cyanophyta) from China - An example from samples of the Dianchi Lake

Ten common species of Microcystis, based on the examination of water samples from the Dianchi Lake, Yunnan, China, were morphologically described, and their taxonomy was also discussed. They are Microcystis aeruginosa, M botrys, M firma, M flos-aquae, M ichthyoblabe, M novacekii, M pseudofilamentosa, M smithii, M viridis and M wesenbergii. Taxonomic status of other Microcystis species reported in China was also evaluated.