Development of 10 microsatellite loci for Rheum tanguticum (Polygonaceae)

Rhubarb is an important Traditional Chinese Medicine. However, the wild resource has been declining. In order to design appropriate conservation methods for the official species across their natural distributions, it is important to characterize their genetic diversity. Here, we describe the development of 10 new microsatellite loci for AC/TG/CCA in Rheum tanguticum. The microsatellites were enriched using the combined biotin capture method. The polymorphism of each locus was further assessed in 12 individuals from four geographically distinct populations of this species. The number of alleles ranged from three to seven and the expected heterozygosity ranged from 0.53 to 0.73. All markers have been checked in the other three species in the genus and two of them together comprise the official medicinal rhubarb resource, with R. tanguticum. These microsatellite markers could. provide a useful tool for genetic and conservation studies of the rhubarb species.

Molecular phylogeny, recent radiation and evolution of gross morphology of the rhubarb genus Rheum (Polygonaceae) inferred from chloroplast DNA trnL-F sequences

Background and Aims Rheum, a highly diversified genus with about 60 species, is mainly confined to the mountainous and desert regions of the Qinghai-Tibetan plateau and adjacent areas. This genus represents a good example of the extensive diversification of the temperate genera in the Qinghai-Tibetan plateau, in which the forces to drive diversification remain unknown. To date, the infrageneric classification of Rheum has been mainly based on morphological characters. However, it may have been subject to convergent evolution under habitat pressure, and the systematic position of some sections are unclear, especially Sect. Globulosa, which has globular inflorescences, and Sect. Nobilia, which has semi-translucent bracts. Recent palynological research has found substantial contradictions between exine patterns and the current classification of Rheum. Two specific objectives of this research were (1) to evaluate possible relationships of some ambiguous sections with a unique morphology, and (2) to examine possible occurrence of the radiative speciation with low genetic divergence across the total genus and the correlation between the extensive diversification time of Rheum and past geographical events, especially the recent large-scale uplifts of the Qinghai-Tibetan Plateau.Methods The chloroplast DNA trnL-F region of 29 individuals representing 26 species of Rheum, belonging to seven out of eight sections, was sequenced and compared. The phylogenetic relationships were further constructed based on the sequences obtained.Key Results Despite the highly diversified morphology, the genetic variation in this DNA fragment is relatively low. The molecular phylogeny is highly inconsistent with gross morphology, pollen exine patterns and traditional classifications, except for identifying all samples of Sect. Palmata, three species of Sect. Spiciformia and a few species of Sect. Rheum as corresponding monophyletic groups. The monotypic Sect. Globulosa showed a tentative position within the clade comprising five species of Sect. Rheum. All of the analyses revealed the paraphyly of R. nobile and R. alexandrae, the only two species of Sect. Nobilia circumscribed by the possession of large bracts. The crude calibration of lineages based on trnL-F sequence differentiation implied an extensive diversification of Rheum within approx. 7 million years.Conclusions Based on these results, it is suggested that the rich geological and ecological diversity caused by the recent large-scale uplifts of the Qinghai-Tibetan Plateau since the late Tertiary, coupled with the oscillating climate of the Quaternary stage, might have promoted rapid speciation in small and isolated populations, as well as allowing the fixation of unique or rare morphological characters in Rheum. Such a rapid radiation, combined with introgressive hybridization and reticulate evolution, may have caused the transfer of cpDNA haplotypes between morphologically dissimilar species, and might account for the inconsistency between morphological classification and molecular phylogeny reported here.

Pollen morphology and its in systematic and ecological significance in Rheum (Polygonaceae) from China

Pollen morphology of 40 species of Rheum, belonging to eight sections, was investigated under LM and SEM. Four new exine patterns were found in the species: a) microcchinate-foveolate, b) rugulate, c) verrucate-perforate, and d) verrucaterugulate ornamentation. In addition, two patterns, the Rheum-type pollens with microechinate-perforate and fine-reticulate, as previously described, were also confirmed in the present study. Based on above study the evolution trends of pollen morphology in the taxa involved were discussed phylogenetically as below. As microechinate-perforate exine pattern existed commonly, the pattern is, therefore, regarded as the most primitive among all the six types. The fine-reticulate type was thought as a derivative type, deriving from the basic micro echinate-foveolate-perforate pattern, and followed by the rugulate and verrucate-perforate ornamentation. The verrucate-rugulate ornamentation should be the most advanced. More than one pollen type often exist in most of the sections in Rheum. The pollen morphology of Rheum was strongly correlated with its geographical and ecological distribution. Three medicinally important species R. officinale, R. palmation and R. tanguticum can be palynologically distinguished by their ornamentations.

In Vitro Evaluation of the Genotoxic Activity and Apoptosis Induction of the Extracts of Roots and Leaves from the Medicinal Plant Coccoloba mollis (Polygonaceae)

Coccoloba mollis (Family Polygonaceae) is a medicinal plant popularly used in cases of memory loss, stress, insomnia, anemia, impaired vision, and sexual impotence, but the scientific literature, to date, lacks studies on the biological effects of this species, particularly with regard to cytotoxicity and induction of DNA damage. The aim of the present study was to assess in vitro (in hepatic HTC cells) ethanolic extracts of the roots and leaves of C. mollis for cytotoxicity, genotoxicity, and induction of apoptosis. For these evaluations the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cytotoxicity assay, comet assay, micronucleus test with cytokinesis block, and an in situ test for detection of apoptotic cells with acridine orange staining were used. The results showed that the extract obtained from the roots of C. mollis is more cytotoxic than that obtained from the leaves and that the reduction in cell viability observed in the MTT assay was a result, at least in part, from the induction of apoptosis. Both extracts induced DNA damage at a concentration of 20 mu g/mL in the comet assay, but no genotoxicity was detected with any of the treatments carried out in the micronucleus test.

Diversidade de ácaros em indivíduos masculinos e femininos de Triplaris americana L. (Polygonaceae) e eficiência de seu pólen como alimento para Euseius citrifolius Denmark & Muma (Acari: Phytoseiidae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Estudo fitoquímico e atividades biológicas preliminares de extratos de Polygonum Acre (Polygonaceae) H.B.K. e Synadenium Carinatum (Euphorbiaceae) Boiss

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Insectos asociados a la lechuga de agua (Pistia stratiotes) en la presa Rocas Morenas, Camoapa, 2010

Entre el período comprendido de Julio a Diciembre del 2010, se realizaron captura de insectos en la presa Rocas Morenas, Camoapa, Boaco, Nicaragua, el objetivo fue llevar a cabo una identificación de las plantas acuáticas presente en la presa y de los órdenes y familias de insectos asociadas a la lechuga de agua (Pistiastratiotes). Las capturas se realizaron en cuatro puntos de muestreo ubicados sobre las riveras de la presa, en la metodología se aplicó monitoreos cada 8 días, estos se realizaban a través de capturas manuales con el uso de red entomológica y trampas lumínicas para insectos nocturnos, realizando su debida identificación posteriormente. El análisis correspondió a una estadística descriptiva para los órdenes y familias de las plantas e insectos encontrados, se estimó el índice de biodiversidad de espécimenes de insectos. Los resultados obtenidos fueron: plantas acuáticas presentes Polygomun densiflorum (Caryophyllales: Polygonaceae), Panicum repens (Cyperales: Poaceae) y en mayor abundancia fue Pistia stratiotes (Arales: Arácea). El total de insectos colectados fue de 564, distribuidos en entre los órdenes Coleóptera, Homóptera y Díptera. Las familias más abundantes de insectos se distribuyen entre Chrysomelidae, Delphacidae, Cicadelidae y Curculionidae. Se registro daño foliar en hojas de P. stratiotes de 10 % provocado por insectos masticadores, el índice de Shannon fue de 3.35.

新疆天山中段北部空中花粉数量的时空变化及其气候与植被指示意义

空中花粉是植被类型、气候状况与大气环境质量的指示物之一。空中花粉雨的取样实验对化石花粉谱的解释和古植被的恢复具有指导意义，因此空中花粉分析成为第四纪孢粉学和古生态学研究的有效辅助手段。本研究在新疆天山中段北坡进行了短期的空中花粉监测（2001年7月~2006年7月），目的是获得局域性和区域性植被所散发的花粉类型及其数量信息，并探讨花粉数量的植被与气候指示意义、花粉传播以及受环境条件影响的程度，同时分析和检测花粉浓度受气候条件的影响情况。空中花粉的收集使用Cour-type捕捉器，分别在3个不同海拔梯度进行5年连续的空中花粉收集（夏秋季节每周一个样品，冬春季节每两周一个样品）。取样地点分别是新疆天山中段北部的天池气象观测站（43°53'58.38"N, 88°07'15.75"E , 海拔1942.5m）、中科院阜康荒漠生态系统定位站（44°17'27.41"N, 87°55'52.65"E, 海拔477m）和准噶尔盆地古尔班通古特沙漠南缘的北沙窝试验站(44°22'40.74"N, 87°55'9.74"E, 海拔443m），取样点的植被带跨度天山北坡的森林草原到低海拔的典型荒漠。 光学显微镜鉴定的空中花粉主要有43种类型（24科30属），而蕨类、苔藓与真菌孢子数量较少。孢粉类型多样性指数为2.45（Shannon Index）。显然，从样品中所鉴定的空中花粉种类与所观察的植物种类有很大的差距。花粉类型与数量在三个不同海拔梯度的取样点上也有明显差异。天池取样点的花粉类型较丰富，与天山北坡垂直带植被分布以及丰富的植被成分相对应，花粉的优势种类为雪岭云杉(Picea schrenkiana)、蒿属(Artemisia)、藜科(Chenopodiaceae)、柽柳科(Tamaricaceae)和麻黄属(Ephedra)，高代表性的外源荒漠花粉类型“削弱了”本地花粉类型的代表性;阜康与北沙窝取样点的花粉类型主要来自荒漠成分的藜科、蒿属、麻黄属、蒺藜科(Zygophyllaceae, 白刺属Nitraria为主）、柽柳科（柽柳属为主）、蓼科(Polygonaceae, 沙拐枣属Calligonum为主）等耐旱植物。花粉数量与植被数量并非简单的线性关系。空中和表土花粉的数量分析表明，藜科、蒿属、麻黄属花粉具有超代表性;雪岭云杉花粉的代表性适中;禾本科(Gramineae)、榆属(Ulmus)、白刺属、柽柳科、沙拐枣属花粉代表性较差。 5年平均花粉浓度大小依次是阜康（81.668 grains/100m3）>天池（51.726 grains/100m3）>北沙窝（45.7685 grains/100m3）。蒿属花粉的峰值出现在秋季，并且出现双峰现象（可能与不同蒿类花期分布不均有关），而其它木本和草本类植物花粉浓度的峰值为单峰，具有明显的季节性，春夏之交开花的桦木属、榆属等类型较为明显。藜科花粉的峰值出现在夏季，麻黄和柽柳的峰值出现在盛夏，桦木属、柳属Salix、雪岭云杉等花粉浓度峰值出现在初夏，与其花期物候相对应。木本植物花粉以雪岭云杉为主，其次为桦木属(Betula)、榆属、柳属、胡颓子科(Elaeagnaceae)花粉，柳属花粉百分比在三个取样点分配比较均匀，桦木属花粉在空间分布上差异明显，与植物分布数量相关。胡颓子科的花粉出现时间短，数量也较少。榆属花粉出现的季节性明显，代表性也较差。因此解释地层中出现类似的低代表性和高代表性的花粉类型时，需要注意其百分比数量的适当校正。木本植物花粉百分比随海拔高度变化成正比关系，对植被的指示性较好。雪岭云杉的年平均花粉浓度在天池取样点明显高于其它两个取样点，5年的平均花粉浓度和百分比与海拔高度呈正比。天池取样点的百分比最高为28.85%，平均为21.15%，而北沙窝与阜康两地5年平均百分比含量分别为0.69%和1.57%，这种时空变化规律与植被数量的关系密切。藜科花粉百分比在阜康和北沙窝取样点占绝对优势，5年的平均花粉浓度与海拔高度呈反比。蒿属花粉年平均浓度与海拔高度关系微弱（R2=0.04，p =0.46），而花粉百分比与海拔高度有显著的线性正比关系（R2=0.72，p<0.0001）。但在低海拔的两个取样点之间差异不明显。 年平均花粉百分比和花粉浓度随海拔梯度而变化，主要花粉类型的变化规律存在差异。暗针叶林雪岭云杉花粉的年平均百分比在低海拔的阜康和北沙窝地区低于3%，在盛花期的6月初，低海拔的取样点也未见超过5%，雪岭云杉花粉的传播在研究区范围内数量分布变化较大，再次表明该类型的花粉在原地沉积效率很高。空中花粉数量能够较好地指示主要的植被带，盛花期内50%以上的雪岭云杉花粉含量则指示了雪岭云杉森林植被带，而高含量的蒿属植物花粉指示了天山山前冲积平原上的蒿类荒漠，相反，高含量的藜科花粉代表了低海拔地区的荒漠。雪岭云杉花粉百分比与浓度均与海拔高度呈显著的线性正比关系，表明雪岭云杉花粉传播的局限性。主要的非木本植物藜科花粉的浓度与海拔高度成反比，蒿属则相反，表明藜科与蒿属花粉在研究区具有很好的植被与气候指示意义。 花粉绝对数量、百分比、浓度具有明显的年际变化，其原因与气候状况的变化有关，年平均花粉浓度、主要类型的花粉浓度与气候参数（降水量、平均温度、最高和最低温度、相对湿度等）的相关分析表明，降水量和湿度与花粉浓度呈负相关程度较高，与其它气候参数则呈显著的正相关。干旱指数、花粉比值、花粉百分比（AP，NAP）与七月份平均气温，年平均降水量的相关系数（Pearson correlation），Ar/Ch与七月平均温度在0.01水平上呈显著的正相关（R2=0.894）, 与年平均降水量在0.01水平上呈显著的负相关（R2=0.874）。AP花粉百分比与七月份平均温度呈显著的负相关（R2=0.71, p<0.0001），与年降水量呈正相关（R2=0.43, p=0.01），而NAP花粉正相反。利用短期监测的空中花粉雨数据计算得到的花粉比值以及干旱指数在研究区具有明确的指示意义，尤其是干旱指数、Ar/Ch、Ar/Ep，这些都可为表土花粉、植被与气候关系模型的建立提供重要的参考信息。 气候参数在很大程度上影响大气花粉的浓度，而风速和风向对空中花粉的传播、散布影响较大，花粉的来源与传播受风向和风速的控制。不同取样点的空中花粉数量差异受地形条件影响很明显，这种差异也可利用地形空气动力学特征的差异来解释，天池监测站因“逆坡”（upslope）的气流促进了低海拔的花粉流向高海拔传播。 研究区的空中花粉数量信息是研究表土花粉、植被和气候关系以及第四纪古生态学的重要基础。本研究提高了对研究区空中孢粉的类型及其传播规律的认识和理解，增加了解空中花粉受环境影响的主要因素及其影响程度，对地层孢粉类型鉴定和花粉数据的解释提出指导性建议，对本地区的古植被与古环境研究增加现代孢粉学的参考依据和信息，并可为建立区域性的现代孢粉－植被－气候关系模型提供可靠依据。 同时，本报告也讨论了存在问题和不足，并提出了今后工作的可能完善方案。

Anti-HIV Activities of the Compounds Isolated from Polygonum cuspidatum and Polygonum multiflorum

The 70% EtOH extract of Polygonum cuspidatum showed inhibitory action against HIV-1-induced syncytium formation at non-cytotoxic concentrations in vitro with a 50% effective concentration (EC50) of 13.94 +/- 3.41 mu g/mL. Through bioactivity-guided fractionation, 20 phenolic compounds, including eight stilbenoids, were isolated from the roots of Polygonum cuspidatum, and their anti-HIV-1 activities were evaluated. Results showed that compounds 1, 13, 14, and 16 demonstrated fairly strong antiviral activity against HIV-1-induced cytopathic effects in C8166 lymphocytes at non-cytotoxic concentrations, with EC50 values of 4.37 +/- 1.96 mu g/mL, 19.97 +/- 5.09, 14.4 +/- 1.34 mu g/mL, and 11.29 +/- 6.26 mu g/mL and therapeutic index (TI) values of 8.12, > 10.02, > 13.89, and > 17.71, respectively. Other compounds showed either weak or no effects. Compound 6 also showed weak inhibition (153.42 +/- 19.25 mu g/mL); however, it possesses very good water solubility and showed almost no cytotoxicity (> 2000 mu g/mL), therefore achieving a fairly good TI (13.04). The activities of the two compounds (3 and 18) from Polygonum multiflorum were also assayed. The relationship between molecular structures and their bioactivities was also discussed.

果洛唐古特大黄染色体观察

对蓼科(Polygonaceae)植物唐古特大黄(Rheum tanguticum Maxim.ex Balf.)染色体核型进行了研究.结果表明,唐古特大黄体细胞染色体数目为2n=22;核型公式为K(2n)=22=20 m+2 sm,核型为"1A"型,少数细胞中发现有随体存在.

唐古特大黄染色体的核型分析

目的对蓼科（Polygonaceae）植物唐古特大黄（Rheum tanguticum Maxim、ex Balf．）染色体核型进行研究。方法采用常规压片方法，并结合显微摄影对染色体进行检测分析。结果唐古特大黄体细胞染色体数目为2n=22；核型公式为K（2n）=22=20m＋2sm，核型为“1A”型。结论唐古特大黄染色体的核型属于较原始的类型。

Cipó-guaraná (Ruprechtia obidensis Huber).

O cipó-guaraná é uma planta trepadeira lenhosa (liana) perene, pertencente à família Polygonaceae, nativa da Amazônia, de ocorrência natural em florestas de terra firme no Acre, Amazonas, Pará e Rondônia (MELO, 2015). O cipó-guaraná forma moitas densas que competem com o pasto por luz, embora não seja uma planta de propagação muito agressiva. Trata-se de uma erva daninha de difícil controle, com rebrotação superior a 90% em plantas roçadas rente ao solo. Mesmo o controle químico tradicional, com pulverização no toco cortado com herbicidas à base de 2,4-D e picloram, apresenta baixa eficácia (SILVA; ANDRADE, 2008). Não há relato de causar toxidez para o gado.

Modern pollen samples from alpine vegetation on the Tibetan Plateau.

1  A set of 316 modern surface pollen samples, sampling all the alpine vegetation types that occur on the Tibetan Plateau, has been compiled and analysed. Between 82 and 92% of the pollen present in these samples is derived from only 28 major taxa. These 28 taxa include examples of both tree (AP) and herb (NAP) pollen types. 2  Most of the modern surface pollen samples accurately reflect the composition of the modern vegetation in the sampling region. However, airborne dust-trap pollen samples do not provide a reliable assessment of the modern vegetation. Dust-trap samples contain much higher percentages of tree pollen than non-dust-trap samples, and many of the taxa present are exotic. In the extremely windy environments of the Tibetan Plateau, contamination of dust-trap samples by long-distance transport of exotic pollen is a serious problem. 3  The most characteristic vegetation types present on the Tibetan Plateau are alpine meadows, steppe and desert. Non-arboreal pollen (NAP) therefore dominates the pollen samples in most regions. Percentages of arboreal pollen (AP) are high in samples from the southern and eastern Tibetan Plateau, where alpine forests are an important component of the vegetation. The relative importance of forest and non-forest vegetation across the Plateau clearly follows climatic gradients: forests occur on the southern and eastern margins of the Plateau, supported by the penetration of moisture-bearing airmasses associated with the Indian and Pacific summer monsoons; open, treeless vegetation is dominant in the interior and northern margins of the Plateau, far from these moisture sources. 4  The different types of non-forest vegetation are characterized by different modern pollen assemblages. Thus, alpine deserts are characterized by high percentages of Chenopodiaceae and Artemisia, with Ephedra and Nitraria. Alpine meadows are characterized by high percentages of Cyperaceae and Artemisia, with Ranunculaceae and Polygonaceae. Alpine steppe is characterized by high abundances of Artemisia, with Compositae, Cruciferae and Chenopodiaceae. Although Artemisia is a common component of all non-forest vegetation types on the Tibetan Plateau, the presence of other taxa makes it possible to discriminate between the different vegetation types. 5  The good agreement between modern vegetation and modern surface pollen samples across the Tibetan Plateau provides a measure of the reliability of using pollen data to reconstruct past vegetation patterns in non-forested areas.

Fungos ectomicorrízicos em áreas de Mata Atlântica do Nordeste do Brasil

Ectomycorrhizal associations are poorly known from tropical lowlands of South America. Recent field trips to the reserve Parque Estadual das Dunas in Natal, in Rio Grande do Norte state, Brazil, revealed a undocumented community of ectomycorrhizal fungi. This type of Mycorrhizal association is frequently in the north hemisphere in temperate and boreal forests. The aim of this work is to analyze the occurrence of ectotrophic areas in atlantic rainforest. Collections along and around the trails in the reserve revealed six genera of putatively ECM fungi which belong to the basidiomycete, Amanitaceae, Boletaceae, Russulaceae, Entolomataceae, and Sclerodermataceae family which are poorly documented in Brazil. Plants belonging to Myrtaceae, Polygonaceae, Leguminosae/Caesalpinioideae, Erythroxylaceae, Malphigiaceae, Bromeliaceae, Loganiaceae, Sapotaceae e Celastraceae were found living next to the species of fungi analized. Our results suggest that the area studied is an ectotrophic environment which shows high diversity of putatively ECM fungi and some plants probably host ECM. The tropical lands are a potential focus to study reinforced by the new records of Scleroderma in Brazil and Northwest of Brazil

Evaluation of extracts from Coccoloba mollis using the Salmonella/microsome system and in vivo tests

The common everyday use of medicinal plants is an ancient, and still very widespread practice, whereby the need for studies on their possible toxicity and mutagenic properties. The species Coccoloba mollis has been much used in phytotherapy, mainly in cases involving loss of memory and stress. In order to investigate its genotoxic and mutagenic potential, ethanolic extracts from the leaves and roots underwent Salmonella/microsome assaying (TA98 and TA100 strains, with and without exogenous metabolism - S9), besides comet and micronucleus tests in vivo.There was no significant increase in the number of revertants/plate of Salmonella strains in any of the analyzed root-extract concentrations, although the extract itself was extremely toxic to the Salmonella TA98 strain in the tests carried out with S9 (doses varying from 0.005 to 0.5 µg/plate). on the other hand, the leaf-extract induced mutations in the TA98 strain in the absence of S9 in the highest concentration evaluated, although at very low mutagenic potency (0.004 rev/µg). Furthermore, there was no statistically significant increase in the number of comets and micronuclei, in treatments involving Swiss mice. It was obvious that extracts of Coccoloba mollis, under the described experimental conditions, are not mutagenic.