New record for Halophila decipiens Ostenfeld in Kenya based on morphological and molecular evidence

The seagrass Halophila decipiens Ostenfeld was recorded for the first time in Kenya in 2003. It was growing in subtidal mixed meadows with H. ovalis in protected bays at a depth of 3 m. DNA sequence analysis and morphological characteristics confirmed the identification. It is possible that this species is more widely distributed in the east African region. There was very little sequence differentiation between the Kenyan H. decipiens and the other samples in the Indo-West Pacific, and between this region and the Atlantic Ocean, further validating from an evolutionary perspective recent dispersal of this species.     

Wavelength-specific photosynthetic responses of Halophila johnsonii from marine-influenced versus river-influenced habitats

The seagrass Halophila johnsonii Eiseman grows from the upper intertidal to 3 m depths in habitats ranging from near-marine inlets to tidal riverine. These habitats have distinct optical characteristics, primarily due to variable concentrations of watershed-derived chromophoric dissolved organic matter (CDOM), which increases the attenuation of short-wavelength (blue and UV) light. H. johnsonii contains a suite of flavonoids that are thought to serve as UV-protective pigments (UVP). In this study, photosynthetic responses at specific wavelengths were compared between plants from a river-influenced high-CDOM habitat (Oleta River) and those from an adjacent marine, low-CDOM environment (Haulover Inlet) in Florida. Oxygen flux was used to measure dark respiration and photosynthesis under near-constant radiant energy at nine specific wavelengths from 340 to 730 nm. Riverine plants had higher gross photosynthetic rates and quantum efficiencies than inlet plants at the shortest wavelengths (350, 400 and 450 nm), while inlet plant photosynthetic rates and quantum efficiencies were higher at the two longest wavelengths measured (694 and 730 nm). Riverine plants also exhibited greater variation in photosynthetic responses across the spectrum and more variable pigment levels among replicates. Chlorophyll a and b concentrations were significantly greater in riverine plants suggesting that they were more shade-acclimated compared to the marine populations. Differences in wavelength-specific photosynthetic responses and chlorophyll levels indicate that the riverine plants were blue-shade acclimated. The higher and more variable UVP levels in the riverine population were not consistent with shade acclimation; however, these flavonoid pigments may protect chloroplasts from photodamage during short-term, high intensity irradiance conditions that occur over the course of tidal cycles at this highly fluctuating riverine site.     

Megagametogenesis in Halophila johnsonii, a threatened seagrass with no known seeds,and the seed-producing Halophila decipiens (Hydrocharitaceae)

Megagametogenesis, the development of a megaspore into an embryo sac, has been identified in the seagrass Halophila johnsonii, a threatened species with no known sexual reproduction or seeds. Megagametogenesis in H. johnsonii was compared with megagametophyte development in Halophila decipiens, a related species known to readily produce viable seeds. In both species, ovules were structurally similar, megaspore mother cells were seen in premeiotic ovules, and linear tetrads and megagametophytes with two to eight nuclei were present in postmeiotic ovules. However, H. decipiens postmeiotic ovules had a chalazal pouch that was absent in the postmeiotic ovules of H. johnsonii. Late-stage H. decipiens ovules also contained embryos, indicating that they had been fertilized, whereas all late-stage H. johnsonii ovules were degrading and showed no signs of fertilization. These observations suggest that meiosis does occur in H. johnsonii megasporocytes, leading to the formation of viable megagametophytes and egg cells that could be fertilized if pollination occurred. Thus, the lack of seed set is due to a lack of pollination rather than any loss of capacity to produce seeds in this species.     

Photophysiological responses of Halophila johnsonii to experimental hyposaline and hyper-CDOM conditions

The endemic seagrass Halophila johnsonii grows intertidally to 3 m deep, in both marine and riverine influenced habitats of eastern Florida. Salinity and chromophoric dissolved organic matter (CDOM) levels widely fluctuate across this broad habitat range, changing tidally and with variable influx of freshwater from watershed runoff, river discharge and stochastic storm events. CDOM exponentially absorbs light in the UV to blue wavelengths, affecting optical water quality. H. johnsonii produces 15 flavonoid compounds that maximally absorb in the UV range. These flavonoids are thought to function as UV-protectants (UVP) in high-light and UV-intense environments. This mesocosm study examined the photosynthetic capacity, quantum efficiency and pigment content of H. johnsonii under experimental treatments of three salinities (10, 20 and 30) with and without CDOM. Main treatment effects and possible interactive effects at both short- (1 day to 1 week) and longer-term (1 month) time scales were examined. There were no significant CDOM or CDOM x salinity effects over any of the experimental treatment durations. There was 100% mortality of plants at salinity 10 after 10 days regardless of water color. UVP content of leaves was not affected by CDOM in this study, but there was significant variation in UVP in response to salinity. Our results do not support the primary role of UVP in this species as a sunscreen, but indicate that different salinity environments contribute to changes in the levels of these flavonoids. The UVP response to salinity stress response was not mitigated by a decrease in UV-radiation (increased CDOM) as H. johnsonii continued to put energy into the production of the carbon-rich flavonoids regardless of potential UV-stress. The experimental results indicate that prolonged hypo-salinity conditions are an important environmental factor to manage in the limited geographic range of H. johnsonii.     

热带滨海植物红厚壳的抗逆生物学特性

为了解红厚壳(Calophyllum inophyllum)的抗逆特性,对西沙群岛自然生长的红厚壳叶片的形态解剖、生理生态、以及叶片和适生土壤的元素含量进行了研究。结果表明,红厚壳是阳生性植物,其上表皮厚,海绵组织发达且栅栏组织排列紧密,气孔排列松散且密度小(24.40 mm~(-2)),有利于叶片保水抵御干旱。叶片的叶绿素a、b含量低(分别为0.87和0.43 mg g~(-1)),表明红厚壳具适应强光环境的能力。叶片的MDA含量低(13.46 nmol g~(-1)),PRO含量高(127.89μg g~(-1)),SOD活性高(149.42 U g~(-1)),总抗氧化能力高(388.60 U g~(-1)),显示红厚壳能通过提高自身的抗氧化能力抵御膜脂过氧化伤害。红厚壳自然生长的珊瑚岛土壤较为贫瘠、营养元素含量低,但红厚壳植株体内具有较高的营养元素含量,表明红厚壳营养元素利用率高,对于贫瘠土壤具有很好的适应能力。因此,红厚壳具有较高的抗氧化胁迫能力和耐受干旱的能力,适宜生长在热带珊瑚岛等土壤贫瘠的生境,可以作为热带珊瑚岛防风固沙和植被恢复的工具种。     

Effects of quercetin and enhanced UV-B radiation on the soybean (Glycine max) leaves

The possible ameliorative effects of quercetin on soybean [Glycine max (L.) Merr.] leaves exposed to UV-B radiation were conducted in greenhouse. The symmetrical leaves supplied with quercetin solution (0.2%, 1%) were exposed to UV-B radiation (0, 3.5, 6.5 kJ m⁻² d⁻¹). 0.2% quercetin ameliorated leaf photosynthesis, improved leaf water content (LWC), and decreased lipid oxidation. The unfavorable effect on photosynthetic parameter was displayed in 1% quercetin treatment. The effect of quercetin on phenylalanine ammonia lyase (PAL) activity varied with the quercetin concentration, UV-B radiation intensity and leaf development. In the later development polyphenol oxidase (PPO) activity was increased significantly by quercetin treatments. We suggested that quercetin with suitable concentration could serve as UV-B protective agent partly due to its antioxidant capacity.     

Northern range extension of the seagrasses Halophila johnsonii and Halophila decipiens along the east coast of Florida,USA

The northern geographic limit for Halophila johnsonii and Halophila decipiens has been reported as Sebastian Inlet, within the Indian River Lagoon, Florida. Surveys conducted in August 2007 determined the new northern limit to be 21.5 km north of the previously known limit. This new northern limit is a 10% range extension for H. johnsonii, a federally threatened species. We conclude that these range extensions are recent, based on (1) the small size of patches; (2) unusually good water clarity conditions due to a recent drought; (3) recent mild/warmer winters; and (4) a recent mechanism for transporting propagules, the numerous hurricanes of 2004. Although this recent range extension is considered ephemeral, similar range extensions may have occurred in the past and may occur again in the future under favorable conditions given the high capacity of these two species for dispersal to favorable sites. The northern limits of these species should not be viewed as static locations; rather, they must be considered dynamic features.     

基于HPLC-ECD研究牡荆叶抗氧化的谱-效关系

为研究牡荆叶指纹图谱与抗氧化活性的谱-效关系，该研究首先建立了18批牡荆叶的高效液相色谱-电化学检测法(HPLC-ECD)指纹图谱，对不同来源牡荆叶药材进行聚类分析，鉴定主要酚类化合物且测定其含量，分析牡荆叶的总酚和总黄酮含量，并采用DPPH自由基清除法、ABTS自由基清除法、氧自由基吸收能力法及铁离子还原能力法考察其体外抗氧化活性，通过皮尔逊相关分析、灰度关联分析及偏最小二乘回归分析法研究牡荆叶的谱-效关系。结果表明：(1)牡荆叶的指纹图谱标定21个共有峰，共指认出10个峰，其含量顺序为绿原酸>异荭草苷>木犀草苷>异牡荆素>异绿原酸A>异绿原酸C>原儿茶酸>荭草苷>异绿原酸B>新绿原酸；不同产地样品间相似性较高，相似度结果为0.816～0.983。(2)系统聚类分析显示，样品含量对分类有一定影响，不同来源样品被分为3类，其中南北方样品存在一定差异。(3)牡荆叶中总酚和总黄酮的含量分别为15.82～61.83 mg·g^-1和27.85～157.65 mg·g^-1,样品均具不同程度的抗氧化活...     

The photosynthetic performance of the tropical seagrass Halophila ovalis in the upper intertidal

Some ecophysiological adaptation strategies of the tropical seagrass Halophila ovalis were investigated with respect to this plant's ability to grow in the upper intertidal in either monospecific pools (but not together with other intertidal species) or emergent and exposed to high temperatures and irradiances during several hours every day. It was found that Halophila ovalis could raise the pH in simulated pools to 8.6, while the two other major (biomass wise) intertidal seagrasses raised the pH to 8.8 (Cymodocea rotundata) and 9.2 (Thalassia hemprichii). In situ, midday pH values of 8.5, 8.7 and 9.0 were recorded in pools inhabiting the three species, respectively. It was further found that photosynthetic electron transport rates (ETR) measured continuously in situ for one leaf during a diurnal cycle followed the irradiance up to a daily maximal value of 2200 μmol photons m⁻² s⁻¹, but dropped by ca. 50% when the leaf became air exposed just before noon. This drop in ETR upon emergence was verified by “point measurements”, while ETRs of leaves that grew in small pools formed as the tide receded followed the irradiance more closely. Similarly, F_v/F_m measured after 15 min of dark adaptation decreased by ca. 50% in emergent leaves during midday, but maintained higher daily values in the submerged leaves.     

Antioxidant flavonoids from leaves of Polygonum hydropiper L

Ten flavonoid compounds were isolated from the dried leaves of Polygonum hydropiper L. (Laksa leaves), and identified as 3-O-alpha-L-rhamnopyranosyloxy-3',4',5,7-tetrahydroxyflavone; 3-O-beta-D-glucopyranosyloxy-4',5,7-trihydroxyflavone; 6-hydroxyapigenin; 6"-O-(3,4,5-trihydroxybenzoyl) 3-O-beta-D-glucopyranosyloxy-3', 4', 5, 7-tetrahydroxyflavone; scutillarein; 6-hydroxyluteolin; 3',4',5,6,7-pentahydroxyflavone; 6-hydroxyluteolin-7-O-beta-D-glucopyranoside; quercetin 3-O-beta-D-glucuronide; 2"-O-(3,4,5-trihydroxybenzoyl) quercitrin; quercetin. Evaluation of the antioxidative activity, conducted in vitro, by using electron spin resonance (ESR) and ultraviolet visible (UV-vis) spectrophotometric assays, showed that these isolated flavonoids possess strong antioxidative capabilities. Measurement of the Trolox equivalent antioxidant capacity (TEAC) values, against ABTS (2,2'-azinobis(3-ethyl-benzo-thiazoline-6-sulphonic acid) radicals and phenyl-tert-butyl nitrone (PBN) azo initiator (AI) also showed strong anti-oxidative activity. The most powerful of the antioxidants was 2"-O-(3,4,5-trihydroxybenzoyl) quercitrin (galloyl quercitrin). A combination of two flavonoid compounds was tested for synergistic anti-oxidative capacity, but no significant improvement was observed.     

Flavonoids of Helianthus series Microcephali

Ray flower and leaf flavonoids were investigated for the three species of Helianthus series Microcephali. Ray flowers of all species contain coreopsin, sulphurein, and quercetin 7-O-glucoside; those of H. microcephalus also contain quercetin 3-O-glucoside. A mixture of flavonoid aglycones, mostly methoxylated flavones, occurs in leaves of H. microcephalus, but not in H. glaucophyllus or H. laevigatus which also lack the glandular trichomes that in Helianthus are typically associated with flavonoid aglycones. The presence of compounds with the 6,8,4′ pattern of methoxylation in H. microcephalus suggests that the series is more similar in flavonoids to series Angustifolii than to series Corona-solis.     

红树植物秋茄类黄酮代谢及其抗氧化活性对高盐胁迫的响应

以红树植物秋茄为试验材料,设置不同浓度NaCl(0、200和500mmol·L-1)处理的砂培实验,应用qRTPCR分析秋茄叶片中类黄酮物质合成上游的4个关键酶基因——苯丙氨酸解氨酶基因(PAL)、肉桂酸羟化酶基因(C4 H)、4-香豆酰辅酶A连接酶基因(4CL)和查尔酮合成酶基因(CHS)的转录水平,并对关键酶活性进行了分析,同时测定了幼苗生物量、钾钠离子含量、类黄酮含量及其抗氧化活性,以探讨秋茄耐盐性与类黄酮物质的关系,为揭示木本植物耐盐机制奠定基础。结果显示:(1)在盐处理条件下,秋茄叶片中PAL、4CL、C4 H和CHS4个关键酶基因的转录水平显著上调,PAL、4CL、C4H酶活性和CHS含量随着盐浓度的增加而明显上升。(2)与对照相比,秋茄根、茎、叶干重在盐处理3d和15d后均无显著变化,而秋茄株高仅在200mmol·L-1盐处理15d后显著增加,其余浓度和时间均未发现有显著性变化。(3)随着盐浓度的升高,秋茄叶片类黄酮含量显著增加,K+/Na+明显下降,丙二醛含量显著降低,活性氧自由基清除率显著增加。研究表明,盐处理加强了秋茄叶片中类黄酮代谢过程中相关酶基因的表达,类黄酮物质的累积有助于其抗氧化能力的提高,进而提高秋茄的抗盐性,维持盐胁迫下秋茄的正常生长。     

祁连山东段高寒草甸植物总酚和黄酮含量及抗氧化活性研究

采用Folin-Ciocalteu法和铝盐显色法对祁连山东段12种高寒植物(灌木嫩枝和草本植物叶片)进行总酚、黄酮含量的测定,并用FRAP法和ABTS法测定其总抗氧化能力。结果表明:(1)金露梅、山生柳、高山绣线菊、西藏沙棘、珠芽蓼的总酚和总黄酮含量较高且总抗氧化能力较强,其中山生柳的总酚含量最高为19.58%,高山绣线菊的黄酮含量最高为13.23%。(2)采用FRAP法测得西藏沙棘的抗氧化能力最强为49.58mmol/L,而用ABTS法测得金露梅的抗氧化能力最强为32.46mmol/L。(3)总酚、总黄酮与总抗氧化能力(FRAP法)具有极显著的相关性(相关系数分别为0.887 1和0.915 7)。分析认为,12种高寒植物的植株体内总酚和总黄酮含量越高,其抗氧化能力越强。     

遮阴对浙江三叶青生理生化及总黄酮的影响

为探究抗癌药物浙江三叶青在不同遮阴处理下其有效成分总黄酮含量及生理生化响应,该研究设置5个光照梯度(全光照CK、遮阴30%、50%、70%、90%),以三叶青二年生扦插苗为材料,处理6个月后,测定三叶青在不同遮阴梯度条件下的生长指标、生化指标以及不同部位(叶片和块根)的总黄酮含量。结果表明:遮阴70%的处理中植株长势最佳,随着光照强度的减弱,三叶青生物量呈现先增高后降低趋势;比叶重呈降低趋势;叶片中的可溶性蛋白(SP)、游离脯氨酸(FP)和超氧化物歧化酶(SOD)含量均在全光照下最高,遮阴70%时最低,分别比全光照降低了33.36%、17.22%、46.88%,表现出光胁迫特点;总黄酮含量为叶片块根,且均以遮阴70%下含量最高,从整体来看,总黄酮含量随着遮阴度的增大而呈现先增加后降低的趋势。这表明光照强度是影响浙江三叶青生长及有效成分积累的重要因素,且高光强在一定程度上抑制三叶青植株的生长,适当遮阴对三叶青植株的生长起促进作用,但块根和叶片中的总黄酮含量对光强的响应却不同,这为今后科学栽培种植及合理有效开发三叶青提供了科学依据。     

Seasonality in nutrient concentrations and stable isotope ratios of Halophila ovalis growing on the intertidal flat of SW Thailand

Halophila ovalis on the intertidal flat of SW Thailand maintains its presence under heavy grazing by dugongs; nevertheless, the site provides lower availability of nutrients in the water column to the seagrasses due to emersion during ebb tide. To examine the seasonality of the nutrient status of fast-growing H. ovalis in the intertidal flat, and to assess whether emersion at low tide affects the availability of nutrients and inorganic carbon, monthly changes in carbon (C), nitrogen (N), and phosphorus (P) concentrations of H. ovalis were determined. Carbon and nitrogen stable isotope ratios were also determined to examine the carbon and nitrogen sources. CNP concentrations of the leaves and petioles of H. ovalis at the intertidal flat were considerably lower than those of the same species previously reported from a subtidal environment throughout the observation period of 8 months. The δ¹³C of the above-ground parts of H. ovalis was lower than in previous reports, suggesting that CO₂ was sufficient for its growth. The CNP concentrations of H. ovalis on the intertidal flat increased after a decrease in grazing pressure by dugongs. It is suggested that H. ovalis sustained rapid growth on the intertidal flat by lowering its nutrient concentrations under heavy grazing by dugongs to maintain its presence. Received: March 2, 2001 / Accepted: June 6, 2001     

基于广泛靶向代谢组学分析黑老虎不同部位成分差异

为提高黑老虎(Kadsura coccinea)资源的综合利用率，采用广泛靶向代谢组学技术鉴定并分析了根、茎、叶代谢组分差异及高度富集成分。结果表明，在根、茎和叶中分别鉴定出642、650和619个代谢物，以酚酸、脂质、类黄酮和有机酸为主；叶与根、茎与根的共有成分分别为566和650个，显著差异成分有442和393个，主要为酚酸、类黄酮和脂质，差异代谢物在苯丙烷生物合成、黄酮与黄酮醇生物合成通路中显著富集。代谢物总丰度和次生代谢物丰度均表现为叶>根>茎，叶中酚酸、类黄酮和脂质及茎中酚酸积累量显著高于根，而氨基酸及其衍生物、萜类、木脂素、香豆素、生物碱的丰度在根中显著上调。因此，黑老虎根、茎、叶有大量共有成分，叶和茎中酚酸、叶中类黄酮和脂质高度富集，含有新绿原酸、绿原酸、槲皮素等多个丰度较高且具有重要生物活性化合物，具有较高利用价值。     

构树叶性状分化的生态功能初步研究

叶性状分化在自然界中较为普遍,不同的叶性状特征与植物对资源获得及利用效率密切关联,反映了植物适应特定环境所形成的生存对策。叶性状分化的生态功能一直以来备受生态学家和进化生物学家的广泛关注。自然界构树(Broussonetia papyifera)在个体发育过程中出现全缘叶和裂缺叶的分化,但其生态功能尚不清楚,推测两者的叶型分化是构树对虫害规避的结果。为了探讨构树叶性状分化对应的可能生态功能,该研究采用野外监测和室内分析的方法,对构树全缘叶和裂缺叶的虫害发生率、叶面积、与抗虫有关的酚类物质(总酚、缩合单宁、黄酮)含量进行了比较。结果表明:(1)相对于裂缺叶,全缘叶虫害发生率显著增加,全缘叶虫害发生率是裂缺叶的两倍。(2)自然条件下,全缘叶叶面积显著高于裂缺叶,增加了约44个百分点。(3)自然条件下,裂缺叶中总酚、缩合单宁、黄酮含量均显著高于全缘叶,分别提高了6.0%、4.2%和16.2%。(4)除黄酮外,虫害处理下裂缺叶中总酚、缩合单宁含量显著高于全缘叶,均提高了约5.0%。(5)人为移除部分叶片,裂缺叶中总酚、缩合单宁、黄酮含量均显著高于全缘叶,分别提高8.0%、1.6%和25.4%。这说明构树全缘叶和裂缺叶中酚类物质含量对外来损伤响应不一致,裂缺叶虫害发生率较全缘叶低可能由于两种类型叶片中酚类物质含量存在差异所引起。     

Production of adventitious root biomass and secondary metabolites of Hypericum perforatum L. in a balloon type airlift reactor

The effects of inoculum density, aeration volume and culture period on accumulation of biomass and secondary metabolites in adventitious roots of Hypericum perforatum in balloon type airlift bioreactors (3 l capacity) were investigated. The greatest increment of biomass as well as metabolite content occurred at an inoculum density of 3 g l⁻¹ and an aeration volume of 0.1 vvm. After 6 weeks of culture, an approximately 50-fold increase in biomass was recorded containing 60.11 mg g⁻¹ dry weight (DW) of phenolics, 42.7 mg g⁻¹ DW of flavonoids and 0.80 mg g⁻¹ DW of chlorogenic acid. Liquid chromatography–mass spectroscopy/mass spectroscopy demonstrated that the presence of quercetin and hyperoside in adventitious roots at a level of 1.33 and 14.01 μg g⁻¹ DW, respectively after 6 weeks of culture. The results suggest scale-up of adventitious root culture of H. perforatum for the production of chlorogenic acid, quercetin and hyperoside is feasible.     

Studies on the Polyphenolic Composition and the Antioxidant Properties of the Leaves of Poplar (Populus spp.) Various Species and Hybrids

The chemical composition in terms of flavonoid and salicylic compounds of leaves from 6 species and 3 hybrids of poplars (Populus) was identified with the use of TLC and HPLC-DAD/ESI-MS methods. Chromatographic analyses were carried out with 21 standard compounds including salicylic compounds (2), phenolic acids (3) and flavonoids (16). Moreover, on the basis of the obtained chromatographic data from the HPLC-DAD/ESI-MS and TLC separations, the presence of salicortin, tremulacin and chlorogenic acid was confirmed, depending on the analyzed poplar species or hybrid. The content of salicylic compounds was determined by HPLC-UV method and expressed on salicin as free and total fraction. Total flavonoid content was determined by spectroscopic method as quercetin equivalent. Significant qualitative and quantitative differences in the chemical composition of the analyzed leaves were demonstrated. The highest concentration of flavonoids (8.02 mg/g) was found in the leaves of Populus nigra, while the highest content of salicylic compounds (47.14 mg/g) was found in the leaves of P.×berolinensis. The antioxidant and xanthine oxidase inhibition properties of extracts from poplar leaves were investigated by TLC bioautography. It has been shown that the richest set of compounds with antioxidant properties are present in the leaves of P. alba, P.×candicans and P. nigra.