Historical biogeography of Tetrastigma (Vitaceae): Insights into floristic exchange patterns between Asia and Australia.

Biodiversity exchanges across the Malesian region, linking the distinct biotas of Asia and Australia, have long attracted the curiosity of biologists. Tetrastigma (Vitaceae) has a wide distribution in Asia through the Sunda archipelago to Australia and provides a good case to elucidate floristic exchange between Asia and Australia. Tetrastigma species have fleshy fruits that are consumed by birds, representing a lineage with a predictable dispersal across island chains. We herein estimate the divergence times and reconstruct the biogeographic history of Tetrastigma with intensive taxon sampling (96 of approximately 120 species; >80%) using 10 chloroplast loci. The biogeographic history of Tetrastigma was reconstructed with 4-area and 6-area divisions by delineating the Sunda region into one or three areas of endemism based on a phylogenetic bioregionalization analysis and the geological history of Malesia. The 4-area division shows that Tetrastigma originated in continental Asia and diverged from the recently segregated genus Pseudocayratia in the early Eocene (49.43 Ma). Dispersal from continental Asia might have started in the late Eocene but mainly occurred in the last 10 Myr. Continental Asia is indicated to be the most important source area while Sunda is the biggest sink, with 16 of the 27 dispersal events inferred from continental Asia to Sunda. Only seven dispersal events are inferred arriving in the Sahul plate and one reverse dispersal from Sahul back to Asia. The 6-area division suggests that the Philippines have been an active junction between Asia and Australia. The biogeographic history of Tetrastigma illustrates an asymmetric floristic exchange between Asia and Australia in this genus, which has been facilitated by the formation of terrestrial connections in the late Miocene and the expansion of wet tropical forests across Wallace's Line and beyond.

Phylogenomic relationships and character evolution of the grape family (Vitaceae).

The grape family consists of 16 genera and ca. 950 species. It is best known for the economically important fruit crop - the grape Vitis vinifera. The deep phylogenetic relationships and character evolution of the grape family have attracted the attention of researchers in recent years. We herein reconstruct the phylogenomic relationships within Vitaceae using nuclear and plastid genes based on the Hyb-Seq approach and test the newly proposed classification system of the family. The five tribes of the grape family, including Ampelopsideae, Cayratieae, Cisseae, Parthenocisseae, and Viteae, are each robustly supported by both nuclear and chloroplast genomic data and the backbone relationships are congruent with previous reports. The cupular floral disc (raised above and free from ovary at the upper part) is an ancestral state of Vitaceae, with the inconspicuous floral disc as derived in the tribe Parthenocisseae, and the state of adnate to the ovary as derived in the tribe Viteae. The 5-merous floral pattern was inferred to be the ancestral in Vitaceae, with the 4-merous flowers evolved at least two times in the family. The compound dichasial cyme (cymose with two secondary axes) is ancestral in Vitaceae and the thyrse inflorescence (a combination of racemose and cymose branching) in tribe Viteae is derived. The ribbon-like trichome only evolved once in Vitaceae, as a synapomorphy for the tribe Viteae.

Discrimination of Tetrastigma hemsleyanum according to geographical origin by near-infrared spectroscopy combined with a deep learning approach.

Recently, deep learning has presented as a powerful approach to overcome the deficiencies of the conventional biochemical approaches. In this study, a method for discriminating medicinal plant Tetrastigma hemsleyanum from different origins was proposed using near-infrared spectroscopy (NIRS) and deep learning models. Support vector machine (SVM), self-adaptive evolutionary extreme learning machine (SAE-ELM), and convolutional neural network (CNN) were used to process the near-infrared spectral data (4000-5600 cm-1). The results indicated that the average recognition accuracy of SVM on the test set samples (n = 60) reached 90%. The average recognition accuracy of SAE-ELM was 98.3%, while CNN correctly discriminated 100% of T. hemsleyanum from different origins. Notably, CNN avoids tedious redundant data preprocessing and is also able to save the trained model for the next call to achieve rapid detection. As above, this study provides an effective deep learning-based method for discriminating the geographical origins of T. hemsleyanum as well as providing a convenient and satisfactory approach to ensure the famous-region of other medicinal plants.

New flavonoid with antidiabetic and antioxidant potential from Tetrastigma angustifolia (Roxb. ) Deb leaves

Ethnomedicinal survey documents the traditional practices of Tetrastigma angustifolia leaves in the management of diabetes in the North-eastern region of India. The present study was aimed at isolation of possible antidiabetic principle(s) from T. angustifolia leaves and evaluation of antidiabetic efficacy of isolated compound(s) in experimental animal model. The methanolic extract of T. angustifolia leaves was obtained by Soxhlet extraction method and subjected to silica gel column chromatography (100-200 mesh). Fraction 18-176 chloroform:methanol (70:30) yielded a pale yellow colored compound. The structure of pure compound was elucidated with the help of UV, IR, NMR and Mass spectrometric/techniques. The antioxidant activity of the isolated compound was evaluated in vitro by various radical scavenfing assay methods.. Oral acute toxicity study was carried out according to OECD guideline 423 in Wistar rats. The antidiabetic efficacy of the isolated compound was evaluated in STZ-induced diabetic rats at the dose of 5 mg/kg b.w. for duration of 21 days. The present study reports a new flavocnoid compound isolated from the methanolic extract of T. angustifolia leaves and identified as 8-hydroxyapigenin 7-O-D-glucopyranoside. The flavonoid compound exhibited potent antidiabetic (hypoglicemic) activity in STZ-induced diabetic rats with promising antioxidant (radical scavenging activity) potential in vitro.

Micromorphological investigation of leaf epidermis and seeds of Vitaceae from Pakistan using light microscopy and scanning electron microscopy.

This present study is the first report on the detailed foliar epidermal anatomy and micromorphology of seeds of five species of Vitaceae from Pakistan using light microscopy (LM) and scanning electron microscopy (SEM). The studied species occur at various localities particularly higher altitude. Qualitative and quantitative leaf micromorphological characters investigated here are related to shape and size of epidermal cells, anticlinal wall pattern, stomatal pore, guard cell, stomatal complex, subsidiary cells, and trichomes. Variations in size and shape were observed in the studied species. Irregular, polygonal and elongated epidermal cells with straight or undulate margins, stomata anomocytic type, and trichomes nonglandular, unicellular to multicellular in all the studied plants. Seed micromorphological qualitative characters studied are shape of margins, integument cell, apical notch, chalaza, anticlinal wall thickness, and quantitative characters are number of seeds per berry, length, and width of seed, length to width ratio. The detailed distribution and microscopic characteristics investigated of family Vitaceae has a distinguished role in identification at genus and species level and may provide evidence in the determination of taxonomic rank of the family in the phylogenetic tree. This study possesses potential for plant taxonomists to further evaluate the species for phytochemical studies and physiology. RESEARCH HIGHLIGHTS: Light microscopy and scanning electron microscopy was used for the micromorphological investigation of family Vitaceae Qualitative and quantitative characteristics were studied Variation in microscopic features of leaf epidermis and seeds Identification of species based on micromorphological characters.

Molecular authentication of Tetrastigma hemsleyanum from its adulterant species using ISSR, CAPS, and ITS2 barcode.

Tetrastigma hemsleyanum is a rare and endangered herb, which is commercialized as the resource of anti-cancer drugs. Wild T. hemsleyanum plants are on the verge of extinction recently, there are increasing numbers of counterfeits on the market. In the present study, inter-simple sequence repeat (ISSR), Cleaved amplified polymorphic sequence (CAPS), and the internal transcribed spacer region II (ITS2) barcode were used for the first time for the authentication of T. hemsleyanum from its commonly counterfeits. ISSR analysis suggested that it was a useful method for distinguishing T. hemsleyanum from its adulterants of different genus. However, it was insufficient to distinguish T. hemsleyanum from those adulterants of the same genus. ITS2 of T. hemsleyanum and the commonly counterfeits were amplified and sequenced. The Neighbor-Joining tree constructed from the ITS2 sequences showed that T. hemsleyanum was clearly differentiated from all counterfeits samples. A mutation site in the ITS2 region of T. hemsleyanum had been found which could be recognized by the restriction endonuclease NcoI. T. hemsleyanum could be readily distinguished from counterfeits as the PCR products from T. hemsleyanum could be digested sufficiently by NcoI, while the PCR products from counterfeits could not be digested. The results indicated that CAPS and ITS2 barcode methods provided effective and accurate identification of T. hemsleyanum from all its adulterants, while ISSR could only distinguish T. hemsleyanum from its adulterants of different genus. The CAPS method developed in the present study will serve as a reliable tool for safe and effective use of T. hemsleyanum in the clinic application. It will also play an important role for the identification, management and conservation of this endangered species.

Phylogeny of the Ampelocissus-Vitis clade in Vitaceae supports the New World origin of the grape genus.

The grapes and the close allies in Vitaceae are of great agronomic and economic importance. Our previous studies showed that the grape genus Vitis was closely related to three tropical genera, which formed the Ampelocissus-Vitis clade (including Vitis, Ampelocissus, Nothocissus and Pterisanthes). Yet the phylogenetic relationships of the four genera within this clade remain poorly resolved. Furthermore, the geographic origin of Vitis is still controversial, because the sampling of the close relatives of Vitis was too limited in the previous studies. This study reconstructs the phylogenetic relationships within the clade, and hypothesizes the origin of Vitis in a broader phylogenetic framework, using five plastid and two nuclear markers. The Ampelocissus-Vitis clade is supported to be composed of five main lineages. Vitis includes two described subgenera each as a monophyletic group. Ampelocissus is paraphyletic. The New World Ampelocissus does not form a clade and shows a complex phylogenetic relationship, with A. acapulcensis and A. javalensis forming a clade, and A. erdvendbergiana sister to Vitis. The majority of the Asian Ampelocissus species form a clade, within which Pterisanthes is nested. Pterisanthes is polyphyletic, suggesting that the lamellate inflorescence characteristic of the genus represents convergence. Nothocissus is sister to the clade of Asian Ampelocissus and Pterisanthes. The African Ampelocissus forms a clade with several Asian species. Based on the Bayesian dating and both the RASP and Lagrange analyses, Vitis is inferred to have originated in the New World during the late Eocene (39.4Ma, 95% HPD: 32.6-48.6Ma), then migrated to Eurasia in the late Eocene (37.3Ma, 95% HPD: 30.9-45.1Ma). The North Atlantic land bridges (NALB) are hypothesized to be the most plausible route for the Vitis migration from the New World to Eurasia, while intercontinental long distance dispersal (LDD) cannot be eliminated as a likely mechanism.

Orthology inference in nonmodel organisms using transcriptomes and low-coverage genomes: improving accuracy and matrix occupancy for phylogenomics.

Orthology inference is central to phylogenomic analyses. Phylogenomic data sets commonly include transcriptomes and low-coverage genomes that are incomplete and contain errors and isoforms. These properties can severely violate the underlying assumptions of orthology inference with existing heuristics. We present a procedure that uses phylogenies for both homology and orthology assignment. The procedure first uses similarity scores to infer putative homologs that are then aligned, constructed into phylogenies, and pruned of spurious branches caused by deep paralogs, misassembly, frameshifts, or recombination. These final homologs are then used to identify orthologs. We explore four alternative tree-based orthology inference approaches, of which two are new. These accommodate gene and genome duplications as well as gene tree discordance. We demonstrate these methods in three published data sets including the grape family, Hymenoptera, and millipedes with divergence times ranging from approximately 100 to over 400 Ma. The procedure significantly increased the completeness and accuracy of the inferred homologs and orthologs. We also found that data sets that are more recently diverged and/or include more high-coverage genomes had more complete sets of orthologs. To explicitly evaluate sources of conflicting phylogenetic signals, we applied serial jackknife analyses of gene regions keeping each locus intact. The methods described here can scale to over 100 taxa. They have been implemented in python with independent scripts for each step, making it easy to modify or incorporate them into existing pipelines. All scripts are available from https://bitbucket.org/yangya/phylogenomic_dataset_construction.

Lineage diversification and hybridization in the Cayratia japonica-Cayratia tenuifolia species complex.

The Cayratia japonica-Cayratia tenuifolia species complex (Vitaceae) is distributed from temperate to tropical East Asia, Southeast Asia, India, and Australia. The spatiotemporal diversification history of this complex was assessed through phylogenetic and biogeographic analyses. Maximum parsimony, neighbor-joining, and maximum likelihood methods were used to analyze sequences of one nuclear (AS1) and two plastid regions (trnL-F and trnC-petN). Bayesian dating analysis was conducted to estimate the divergence times of clades. The likelihood method LAGRANGE was used to infer ancestral areas. The Asian C. japonica and C. tenuifolia should be treated as an unresolved complex, and Australian C. japonica is distinct from the Asian C. japonica-C. tenuifolia species complex and should be treated as separate taxa. The Asian C. japonica-C. tenuifolia species complex was estimated to have diverged from its closest relatives during the Late Eocene (35.1 million years ago [Ma], 95% highest posterior densities [HPD]=23.3-47.3Ma) and most likely first diverged in mid-continental Asia. This complex was first divided into a northern clade and a southern clade during the middle Oligocene (27.3Ma; 95% HPD=17.4-38.1Ma), which is consistent with a large southeastward extrusion of the Indochina region relative to South China along the Red River. Each of the northern and southern clades then further diverged into multiple subclades through a series of dispersal and divergence events following significant geological and climatic changes in East and Southeast Asia during the Miocene. Multiple inter-lineage hybridizations among four lineages were inferred to have occurred following this diversification process, which caused some Asian lineages to be morphologically cryptic.

Oldest fruits of the grape family (Vitaceae) from the Late Cretaceous Deccan cherts of India.

PREMISE OF THE STUDY: Despite the inferred Cretaceous origin of the Vitaceae, fossils of the grape family are relatively young, with the oldest previously known examples limited to the Paleocene of Europe and North America. New fossil evidence indicates that the family was already present in India in the Late Cretaceous (Maastrichtian), about 10-15 million years before the tectonic collision of India with Eurasia. • METHODS: Fruits and seeds were investigated by serial sections and peels of chert from the Deccan Intertrappean beds of central India, and compared anatomically with those of extant genera. • KEY RESULTS: Indovitis chitaleyae gen. et sp. n. is described based on immature fruits bearing four to six seeds, and isolated mature seeds. The seeds possess paired ventral infolds and a dorsal chalaza, features diagnostic of the order Vitales. Characters of chalaza shape, infold morphology, and seed coat anatomy place I. chitaleyae within Vitaceae and favor a phylogenetic position either sister to the Vitis-Ampelocissus clade or sister to the Ampelopsis-Clematicissus-Rhoicissus clade. • CONCLUSIONS: Presence of the oldest known vitaceous fossils in the latest Cretaceous of India indicates a previously undocumented Gondwanan history and a possible southern hemisphere origin for the Vitales. An "out-of-India" scenario might explain the relatively sudden appearance of diverse Vitaceae in the Late Paleocene and Early Eocene of the Northern Hemisphere.

Phylogeny of the non-monophyletic Cayratia Juss. (Vitaceae) and implications for character evolution and biogeography.

Cayratia consists of ca. 60 species primarily distributed in the tropical and subtropical regions of Asia, Australia, and Africa. It is an excellent candidate for exploring the evolution of intercontinental disjunct distributions in the Old World. Previous phylogenetic work of Vitaceae with a few species of Cayratia sampled showed that Cayratia was not monophyletic and was closely related to Cyphostemma and Tetrastigma. We herein expanded taxon sampling of Cayratia (25/60 species) with its allied genera Cyphostemma (39/150 species), Tetrastigma (27/95 species), and other related genera from Vitaceae represented, employing five plastid markers (atpB-rbcL, rps16, trnC-petN, trnH-psbA, and trnL-F), to investigate the phylogeny, character evolution and biogeography of Cayratia. The phylogenetic analyses have confirmed the monophyly of the Cayratia-Cyphostemma-Tetrastigma (CCT) clade and resolved Cayratia into three lineages: the African Cayratia clade, subg. Cayratia, and subg. Discypharia. The African Cayratia was supported as the first diverging lineage within the CCT clade and Tetrastigma is resolved as sister to subg. Discypharia. Character optimizations suggest that the presence/absence of a membrane enclosing the ventral infolds in seeds is an important character for the taxonomy of Cayratia. The presence of bracts on the lower part of the inflorescence axis is inferred to have arisen only once in Cayratia, but this character evolved several times in Tetrastigma. Both the branching pattern of tendrils and the leaf architecture are suggested as important infrageneric characters, but should be used cautiously because some states evolved multiple times. Ancestral area reconstruction and molecular dating suggest that the CCT clade originated from continental Africa in the late Cretaceous, and it then reached Asia twice independently in the late Cretaceous and late Oligocene, respectively. Several dispersals are inferred from Asia to Australia since the Eocene.

Estudo de biocompatibilidade in vivo e in vitro do extrato hidroalcoólico de Cissus sicyoides L. - Vitaceae em ratos / Study of in vivo and in vitro biocompatibility of Cissus sicyoides L. - Vitaceae hydroalcoholic extracts in rats

Biocompatibilidade é a capacidade de um material exercer funções específicas quando aplicado em contato com tecidos vivos de determinado hospedeiro, sem, contudo, causar danos ou prejuízo ao mesmo. Este trabalho objetivou determinar a biocompatibilidade in vivo e in vitro do extrato hidroalcoólico do Cissus sicyoides L - Vitaceae. Foram utilizados 30 ratos (Rattus novergicus albinus wistar), com idade entre 45 e 90 dias e pesando entre 170 e 260 g. Os animais foram divididos em 3 grupos (A1, A2 e A3) de 6 animais cada para o teste in vivo, os quais foram sacrificados com 2, 4 e 6 dias, respectivamente. Para o teste in vitro, foram utilizados 12 animais para obtenção do índice de aderência e da capacidade fagocítica dos macrófagos de ratos do grupo controle e do grupo experimental. Nos resultados encontrados no teste in vivo, conclui-se que o extrato apresentou-se biocompatível, visto que não provocou alterações significativas no tecido. Já no teste in vitro, o mesmo não se apresentou biocompatível, pois o extrato puro apresentou índice de aderência baixo (7,1) e taxa de fagocitose elevada (35,7), indicando diferença significante quando comparado ao controle. Porém, quando diluído, o extrato se mostrou inócuo, devido ao aumento dos valores do índice de aderência nas concentrações de 1/10 (61,4) e 1/100 (74,3) nos ensaios, as quais não apresentaram diferença significante quando comparadas ao controle. Após a análise dos dados, concluiu-se que a solução diluída do extrato hidroalcoólico do Cissus sicyoides L. não causa danos ou prejuízos. Entretanto, como nem todos os efeitos farmacológicos foram testados no presente trabalho, não se pode inferir automaticamente que ele é biocompatível em todos os casos.

Biocompatibility is the ability of a material to perform specifictasks when applied to living tissues without causing damage or injuries to it. Thus, this study aimed at determining the in vivo and in vitro biocompatibility of Cissus sicyoides L. - Vitaceae hydroalcoholic extracts. A total of 30 rats (Rattus norvegicus Albinus Wistar), with ages ranging from 45 to 90 days and weighing between 170 and 260g were used. The animals were divided into 3 groups (A1, A2 and A3) with 6 animals each, for the in vivo test, which were sacrificed after 2, 4 and 6 days, respectively. For in vitro test, 12 animals were used to obtain the index of adherence and phagocytic ability of macrophages of rats from the control and the experimental groups. In results found for the in vivo test, it was concluded that the extract was biocompatible, whereas no significant changes were observed in the tissue. As to the in vitro test, the extract was not biocompatible, since the pure extract showed a low rate of adherence (7.1) and a high rate of phagocytosis (35.7), indicating a significant difference when compared to the control group. However, when diluted, the extract was shown to be harmless, due to an increase in the values of the adherence index at the following concentrations : 1/10 (61.4) and 1/100 (74.3) in the tests, which showed no significant differences when compared to the control group. After analyzing the data, it was concluded that since the infusion of the plant is a kind of dilution, its use does not cause any harm to the body. A new study is necessary at the moment to possibily demonstrate its effects on the long term.

Requerimientos germinativos de enredaderas características del Chaco Serrano de Córdoba, Argentina / Germinating requirements of common vines from Chaco Serrano of Córdoba, Argentina

Vines are conspicuous elements of floras in different ecosystems. Patterns of distribution and ecology of this group has been studied at regional scales, mainly in tropical areas, but less is known about factors affecting their distribution at smaller scales. In this study, the germinating requirements of common vines from two plant communities (open shrubland and woodland) in xerophytic mountain forests (Chaco serrano) were studied. A total of 21 species were selected and classified as typical of woodland, shrubland, or indifferent. Experimental treatments were three temperature regimes (15-5ºC, 25-15ºC and 35-20ºC), in light (12-12h daily photoperiod) and in continuous darkness. Total germination percentage, the relative light germination (RLG) and germination rate (T50) were recorded. Among results we found that the temperature was the main factor triggering the germination process. No differences in the mean RLG and T50 between woodland and shrubland patches were found. The patterns observed in this work support the idea that, independently of patch type, germination is associated with temperature of the time of the year when most of the rainfall occurs. Germination of the studied species would be responding to macroclimate’s factors (seasonality of climate and rainfall), rather than to a microclimate. This indicates that differences in species composition between patches would not be due to the factors studied, rather than to characteristics of the later stages of the cycle life.

Las enredaderas son elementos conspicuos de la flora de diferentes ecosistemas. Numerosos estudios basados en patrones de distribución y ecología de este grupo de plantas se han realizado a escala regional, se conoce poco sobre los factores que afectan su distribución a nivel local. Se estudiaron los requerimientos germinativos de 21 enredaderas del Bosque chaqueño serrano de Córdoba, las cuales fueron clasificadas en típicas de bosques, matorrales e indiferentes. Se llevaron a cabo experimentos de germinación en tres regímenes de temperatura (15-5ºC, 25-15ºC y 35-20ºC), en luz (12-12hr luz-oscuridad) y oscuridad permanente. Se registró el porcentaje final de germinación, el índice de germinación relativa a la luz (GRL) y la velocidad de germinación (T50). La temperatura fue el principal factor regulador del proceso germinativo. El GRL y el T50 no registraron diferencias entre fisonomías. Los patrones observados apoyan la idea de que la germinación de este grupo de plantas estaría respondiendo a factores macroclimáticos (estacionalidad del clima y las precipitaciones), más que a un microclima (factores asociados a la fisonomía del sitio). La composición diferencial de enredaderas en los parches de bosques y arbustos no estaría explicada por los factores estudiados, sino por características de las etapas posteriores a la semilla.

[Germinating requirements of common vines from Chaco Serrano of Córdoba, Argentina]. / Requerimientos germinativos de enredaderas características del Chaco Serrano de Córdoba, Argentina.

Vines are conspicuous elements of floras in different ecosystems. Patterns of distribution and ecology of this group has been studied at regional scales, mainly in tropical areas, but less is known about factors affecting their distribution at smaller scales. In this study, the germinating requirements of common vines from two plant communities (open shrubland and woodland) in xerophytic mountain forests (Chaco serrano) were studied. A total of 21 species were selected and classified as typical of woodland, shrubland, or indifferent. Experimental treatments were three temperature regimes (15-5 degrees C, 25-15 degrees C and 35-20 degrees C), in light (12-12h daily photoperiod) and in continuous darkness. Total germination percentage, the relative light germination (RLG) and germination rate (T50) were recorded. Among results we found that the temperature was the main factor triggering the germination process. No differences in the mean RLG and T50 between woodland and shrubland patches were found. The patterns observed in this work support the idea that, independently of patch type, germination is associated with temperature of the time of the year when most of the rainfall occurs. Germination of the studied species would be responding to macroclimate's factors (seasonality of climate and rainfall), rather than to a microclimate. This indicates that differences in species composition between patches would not be due to the factors studied, rather than to characteristics of the later stages of the cycle life. 1523. Epub 2012 December 01.

[Fingerprints identification of Gynostemma pentaphyllum by RAPD and cloning and analysis of its specific DNA fragment].

OBJECTIVE: To identify the resources of Gynostemma pentaphyllum and its spurious breed plant Cayratia japonica at level of DNA. METHODS: Two random primers ( WGS001, WGS004) screened were applied to do random amplification with genomic DNA extracted from Gynostemma pentaphyllum and Cayratia japonica which were collected from different habitats. After amplificated with WGS004, one characteristic fragment about 500 bp which was common to all Gynostemma pentaphyllum samples studied but not to Cayratia japonica was cloned and sequenced. Then these sequences obtained were analyzed for identity and compared by Blastn program in GenBank. RESULTS: There were obvious different bands amplified by above two primers in their fingerprints of genomic DNA. On the basis of these different bands of DNA fingerprints, they could distinguish Gynostemma pentaphyllum and Cayratia japonica obviously. Sequence alignment of seven cloned bands showed that their identities ranged from 45.7% - 94.5%. There was no similar genome sequences searched in GenBank. This indicated that these seven DNA fragments had not been reported before and they should be new sequences. CONCLUSION: RAPD technique can be used for the accurate identification of Gynostemma pentaphyllum and its counterfeit goods Cayratia japonica. Besides, these specific DNA sequences for Gynostemmna pentaphyllum in this study are useful for the further research on identification of species and assisted selection breeding in Gynostemma pentaphyllum.

[Identification of Gynostemma from their adulterant Cayratia japonica by PCR-RFLP].

OBJECTIVE: To establish a convenient and effective method for the identification of Gynostemma and Cayratia japonica. METHOD: Eight species, including Gynostemm pentaphyllum, G. pentagynum, G. cardiospermum, G. longipe, G. yixingense, G. laxiflorum, G. guangxiense and C. japonica were investigated through PCR - RFLP of six chloroplast DNA fragments. The six gene fragments were digested by six restriction endonuclease respectively, including Taq I, Hpa II, EcoR I, Rsa I, Hha I, Hind III. RESULT: Seven species of Gynostemma and their adulterant could be identified by trnK1f-trnK2r and Rsa. CONCLUSION: PCR - RFLP provides a quick, reliable molecular marker technique for identification of Cynostemma and their adulterant Cayratia japonica.

Host-to-parasite gene transfer in flowering plants: phylogenetic evidence from Malpighiales.

Horizontal gene transfer (HGT) between sexually unrelated species has recently been documented for higher plants, but mechanistic explanations for HGTs have remained speculative. We show that a parasitic relationship may facilitate HGT between flowering plants. The endophytic parasites Rafflesiaceae are placed in the diverse order Malpighiales. Our multigene phylogenetic analyses of Malpighiales show that mitochondrial (matR) and nuclear loci (18S ribosomal DNA and PHYC) place Rafflesiaceae in Malpighiales, perhaps near Ochnaceae/Clusiaceae. Mitochondrial nad1B-C, however, groups them within Vitaceae, near their obligate host Tetrastigma. These discordant phylogenetic hypotheses strongly suggest that part of the mitochondrial genome in Rafflesiaceae was acquired via HGT from their hosts.