Anemone Plants Research Articles

Complete genome sequences of anemone mosaic virus and ranunculus mild mosaic virus isolated from anemone imported from the Netherlands into Japan.

Anemone mosaic virus (AnMV) and ranunculus mild mosaic virus (RanMMV) infect anemone plants, which exhibit characteristic mosaic patterns on their leaves. Employing next-generation sequencing of plant material imported from the Netherlands, the complete genome sequences of these two viruses were determined for the first time. AnMV and RanMMV have 9698 and 9537 nucleotides (nt), respectively, excluding the poly(A) tail. They share 80.0%/82.0% and 98.0%/97.0% nt/amino acid (aa) sequence identity, which is above the species demarcation value, in the previously reported AnMV and RanMMV coat protein sequences, but they share 69.0%/70.0% nt/aa sequence identity or less with other potyviruses in all 10 mature protein coding regions of the genome. Additionally, phylogenetic analysis confirmed the relationship of the AnMV and RanMMV genome sequences to previously reported partial sequences and placed them within the genus Potyvirus. These results show that these two viruses represent separate species within the genus Potyvirus.

In Vitro Anti-proliferative Activity and Mechanism of Action of Anemone nemorosa.

Anemone nemorosa is part of the Ranunculaceae genus Anemone (order Ranunculales) which comprises more than 150 species. Various parts of the plant have been used for the treatment of numerous medical conditions such as headaches, tertian agues, rheumatic gout, leprosy, lethargy, eye inflammation as well as malignant and corroding ulcers. The Anemone plants have been found to contain various medicinal compounds with anti-cancer, immunomodulatory, anti-inflammatory, anti-oxidant and anti-microbial activities. To date there has been no reported evidence of its use in the treatment of cancer. However, due to the reported abundance of saponins which usually exert anti-cancer activity via cell cycle arrest and the induction of apoptosis, we investigated the mode of cell death induced by an aqueous A. nemorosa extract by using HeLa cervical cancer cells. Cisplatin was used as a positive control. With a 50% inhibitory concentration (IC50) of 20.33 ± 2.480 µg/mL, treatment with A. nemorosa yielded a delay in the early mitosis phase of the cell cycle. Apoptosis was confirmed through fluorescent staining with annexin V-FITC. Apoptosis was more evident with A. nemorosa treatment compared to the positive control after 24 and 48 h. Tetramethylrhodamine ethyl ester staining showed a decrease in mitochondrial membrane potential at 24 and 48 h. The results obtained imply that A. nemorosa may have potential anti-proliferative properties.

Ocena skuteczności działania niektórych substancji i preparatów pochodzenia naturalnego w zwalczaniu węgorka chryzantemowca – Aphelenchoides ritzemabosi (Schwartz, 1911) Steiner i Buhrer, 1932

In 2011–2012 the effects of azadirachtin (NeemAzal T/S), abamectin (Vertimec 018 EC), glycol-aqueous extract of Quillaja saponaria (Quillaja Extract), aqueous extracts of minced garlic, aqueous extracts of marigold roots and leaves, aqueous extract of salvia leaves in the control of chrysanthemum foliar nematode Aphelenchoides ritzemabosi (Schwartz, 1911) Steiner & Buhrer, 1932 were examined. Efficacy of these products was determined in the laboratory test on chrysanthemum leaves, and on clematis and anemone plants infested by A. ritzemabosi in greenhouse experiments. The highest mortality of chrysanthemum foliar nematode on chrysanthemum leaves was obtained after application of Q. saponaria extract and minced garlic extract. In greenhouse conditions the highest effectiveness had Q. saponaria extract (above 90%), statistically significant in trial on anemone plants. Unfortunately, application of this product resulted in phytotoxicity symptoms on plants. W latach 2011–2012 oceniano skuteczność działania azadirachtyny (NeemAzal T/S), abamektyny (Vertimec 018 EC), wodnoglikolowego ekstraktu z Quillaja saponaria (Quillaja Extract), wyciągu z miazgi czosnkowej oraz wyciągów z korzeni i liści aksamitki, i liści szałwii w warunkach laboratoryjnych na liściach chryzantemy oraz w szklarni na powojniku i zawilcu japońskim. W warunkach laboratoryjnych istotnie najwyższą śmiertelność węgorka chryzantemowca uzyskano po zastosowaniu ekstraktu z Q. saponaria i wyciągu z miazgi czosnkowej. W warunkach szklarniowych spośród testowanych substancji najwyższą śmiertelność nicieni (powyżej 90%) uzyskano po zastosowaniu ekstraktu z Q. saponaria, udowodnioną statystycznie w doświadczeniu na zawilcu. Niestety produkt ten wykazywał na badanych roślinach fitotoksyczność.

First Report of Cucumber mosaic virus in Anemone sp. in the United States

In the spring of 2008, more than a dozen, aphid-infested, anemone plants (Anemone sp.) grown at the campus of the University of Arkansas in Fayetteville showed stunting and mosaic, whereas only two were asymptomatic. Leaf homogenates from four symptomatic plants were inoculated onto Nicotiana benthamiana that became stunted and developed severe mosaic approximately 7 days postinoculation, whereas buffer-inoculated plants remained asymptomatic. Double-stranded RNA (dsRNA) extraction (4) from symptomatic anemone revealed the presence of four predominant bands of approximately 3.2, 2.9, 2.2, and 0.9 kbp, a pattern indicative of cucumovirus infection. Cucumber mosaic virus (CMV) is the only cucumovirus reported in anemone in Europe (2) and Israel (3), and for this reason, anemone and N. benthamiana plants were tested by Protein A ELISA with antisera against CMV developed by H. A. Scott. ELISA verified the presence of CMV in symptomatic anemone and inoculated N. benthamiana, while asymptomatic plants were free of the virus. Using cucumovirus degenerate primers, essentially as described by Choi et al. (1), a region of approximately 940 bases that includes the complete coat protein gene of the virus was amplified from symptomatic anemone and N. benthamiana but not asymptomatic plants of either species. This anemone isolate (GenBank Accession No. FJ375723) belongs to the IA subgroup of CMV because it shares 99% nucleotide and 100% amino acid sequence identities with the Fny isolates of the virus. To my knowledge, this is the first report of CMV infecting anemone in the United States and an important discovery for the ornamental industry since anemone is commonly grown together with several ornamental hosts of CMV in nursery and garden settings.

First Report of Orobanche crenata Parasitism on Ornamental Anemone (Anemone coronaria) in Israel.

Broomrapes (Orobanche spp.) are obligatory parasitic weeds that infect roots of vegetables and field crops worldwide, resulting in severe damage. Orobanche crenata Forsk is common in agricultural fields in the Mediterranean Basin, Southern Europe, and the Middle East and is known as an important scourge of grain and forage legumes and of some Apiaceous crops such as carrot (Daucus carota L.) and celery (Apium graveolens L.) (3,4). To our knowledge, in this note, we report for the first time on Anemone coronaria L. (Ranunculaceae) as a new host for O. crenata and this is also the first report of Orobanche parasitism on a geophytic crop. Anemone (Anemone coronaria L.) is a high-value ornamental crop, which is commercially grown for cut flowers. Four anemone cultivars (Meron Red, Galil White, Jerusalem Blue, and Jerusalem Pink) were planted in September 2006 in a 2-ha field in Israel. The previous crop, broad bean (Vicia faba L.), was heavily infected during 2005 by O. crenata. In February 2007, O. crenata parasitized the anemone plants and developed numerous fertile flowering stalks throughout the field. The four anemone cultivars were equally infected by the parasite. Additional flowering stalks were still emerging on anemone plants during July 2007. Washing the root system clearly verified direct connection between the parasite and anemone roots. The parasite species was identified morphologically after Flora Europea (1) and Flora Palaestina (2). In addition, the stem had the fragrance typical of O. crenata. Neither symptoms nor visible qualitative or quantitative damage could be detected on infected anemone plants compared with noninfected plants. However, anemone appears to be an alternate host on which O. crenata can produce additional seed for the parasite seed bank.

First Report of Anemone Plants Infected by Ranunculus white mottle virus.

Ranunculus white mottle virus (RWMV) (1), genus Ophiovirus, has been reported in crops of several cultivars of commercial ranunculus (Ranunculus asiaticus hybrids) during the 1990s in Liguria in Northwest Italy. Symptoms associated with RWMV in ranunculus are not clear-cut owing to the presence of mixed viral infections. During autumn 1999, a severe disease in commercial crops of anemone (Anemone coronaria) was noted in the same area. Plants appeared stunted with young leaves showing curling, deformation, and necrotic spotting. Disease incidence in some fields reached 40 to 50%. DAS- and TAS-enzyme-linked immunosorbent assays (ELISAs) for presence of RWMV and for the viruses most frequently infecting anemone in Italy were run on 24 field samples. Seven proved to be infected by RWMV in mixed infection with Cucumber mosaic virus subgroup II or with Tobacco necrosis virus. Ophiovirus-like particles were detected by negative staining and electron microscopy from sap extracts of field plants that were RWMV-positive by ELISA. Sap from these plants was also mechanically inoculated to indicator plants. Total RNAs were extracted from RWMV-infected field samples and from inoculated Nicotiana benthamiana and N. clevelandii and used in molecular tests. A DIG-DNA probe targeting the 1.8-kb RNA2 of RWMV was used in Northern blots and dot blots of total RNAs, confirming the infection in field samples and multiplication of the virus in test plants, unfortunately still in mixed infection. At present, it is difficult to evaluate RWMV symptomatology in anemone, but the presence of this virus in mixed infection seems to produce serious effects. This is the first report of RWMV in anemone. Reference: (1) A. M. Vaira et al. Arch. Virol. 142:2131, 1997.

First Report of Impatiens Necrotic Spot Tospovirus (INSV) in Israel.

In January 1999, Anemone coronaria L. imported from Europe and grown in open fields near Jerusalem in Israel developed foliar ringspots and foliar necrosis. Within a few weeks of the first appearance of these symptoms, further anemone plants in the surrounding area were affected and seriously damaged. Impatiens necrotic spot tospovirus (INSV) was detected in affected plants by enzyme-linked immunosorbent assay (ELISA; anti-INSV monoclonal antibodies were provided by H. T. Hsu, USDA, Beltsville, MD, and a polyclonal antibody to INSV was purchased from Loewe, Otterfing, Germany). Crude sap extracted from symptomatic tissue was mechanically transmitted to Emilia spp., Petunia hybrida, Nicotian glutinosa, N. benthamiana, and N. rustica plants that developed symptoms characteristic of INSV infection (1). ELISAs of leaf sap extracted from anemone plants and mechanically inoculated indicator plants gave a strong positive reaction to INSV. Leaf-dip preparations prepared from leaf samples of anemone plants were analyzed by transmission electron microscopy (TEM). Virus particles typical of a tospovirus were observed in samples taken only from symptomatic plants. TEM studies with ultrathin sections of infected anemone and Emilia spp. leaves revealed the presence of tospovirus-like particles. This first report of INSV interception in Israel brings the count of the Tospovirus members in Israel to three, including tomato spotted wilt virus (TSWV), which was found in the past to infect anemone and other ornamental crops, and the Iris yellow spot tospovirus, infecting onion (2). INSV is known to occur in Europe and in the U.S., mostly in flowers grown in greenhouses. The virus is transmitted by the Western flower thrips (WFT; Frankliniella occidentalis Pergande). Although all infected plants were destroyed, precautions to prevent further introduction of the virus must be made. INSV might spread by the WFT, which is abundant in Israel year round, and might also infect other greenhouse or field crops.

Anemone - an Additional Perennial Ornamental Host of Tobacco Rattle Virus in the U.S.

A previous report (1) drew attention to the occurrence in the U.S. of tobacco rattle tobravirus (TRV) in several perennial ornamentals that move freely in international trade. Here we report the occurrence of TRV in an additional host plant of this type. The virus was identified in anemone (Anemone × hybrida cv. Honorine Jobert) with leaf symptoms consisting of chlorotic blotches, chlorotic line patterns, and distortion. Characteristic tobravirus-like particles with modal lengths of 50, 80, 130, and 180 nm were observed by electron microscopy in partially purified extracts of symptomatic but not of asymptomatic plants. These particles reacted specifically with antibodies to TRV and pea early-browning (PEBV) tobraviruses in immunoelectron microscopic (IEM) assays, produced typical TRV-induced symptoms in Nicotiana benthamiana and N. clevelandii, in which similar particles were detected by IEM, but, like other TRV isolates, did not infect pea (2). No other viruslike particles were observed in partially purified extracts of symptomatic anemone plants. This is the first report of TRV infection in anemone. While TRV occurs widely in Europe, it had been identified previously in the U.S. only in Oregon, Washington, and California (1,2,3). This virus is an important pathogen of several crops, including potato. Several areas of the Midwest, including Minnesota and Wisconsin, have significant seed-potato industries, and further introduction and possible dissemination of TRV pose potential regulatory and quarantine issues. This example of the movement of an exotic pathogen of potential economic importance into new areas underlines the need for closer monitoring of plant material entering international trade in an era of increasing globalization.

Triterpenoid Saponins fromAnemone flaccida

From the rhizome of ANEMONE FLACCIDA Fr. Schmidt, used in Chinese folk medicine, four saponins of oleanolic acid-A,B,C,D were isolated. By means of spectroscopy and chemical transformation, saponins A, B, and D were identified as 3- O-[alpha- L-rhamnopyranosyl-(1-->2)-alpha- L-arabinopyrasyl]-oleanolic acid, 3- O-[alpha- L-rhamnopyranosyl-(1-->2)-beta- D-xylopyranosyl]-oleanolic acid, and 3- O-[alpha- L-rhamnopyranosyl-(1-->2)-beta- D-glucopyranosyl]-oleanolic acid. These three saponins are known compounds but isolated from ANEMONE plants for the first time. Saponin C was elucidated as 3- O-[beta- D-glucopyranosyl-(1-->2)-beta- D-xylopyranosy1]-oleanolic acid, which has not been reported in the literature before, and is named as flaccidoside I.

Protoanemonin , an antimutagen isolated from plants

Protoanemonin was identified as the factor responsible for the antimutagenicity of Ranunculus and Anemone plants against the strain E. coli B/r WP2 trp. The specific activities found were A D 50 = 30 and 47 μmg/plate, respectively, for UV- and N-methyl- N′-nitro- N-nitrosoguanidine-induced mutations. Quantitative analyses of protoanemonin in the crude extracts of Ranunculus and Anemone plants were performed by reversed-phase high-pressure liquid chromatography.

Anemone necrosis, a disease caused by a strain of tobacco ringspot virus

SUMMARYA strain of tobacco ringspot virus (TRSV) was isolated from severely necrotic Anemone coronaria plants in Somerset in 1957, and was named anemone necrosis virus (ANV). ANV infected, by sap‐rubbing inoculation, fifty‐eight of 103 plant species tested, in twenty‐one out of thirty‐two families. Experimental infection of anemone plants induced symptoms of necrosis and die‐back, but less virulent than those seen in the field. The virus was cultured and assayed in Nicotiana clevelandii or N. tabacum. The in vitro properties in crude sap were: maximum infective dilution 1/125 to 1/10,000 according to the source plant; thermal inactivation point (10 min.) 70°C.; survival in vitro 21 but not 28 days at 18°C., and at least 12 weeks at 0°C. Preparations preserved by vacuum sublimation (freeze‐drying) were still infective after 5 years. Some N. tabacum plants could be freed from ANV by growing at 37°C. for 4–6 weeks.Although less easily than TRSV, ANV was purified by extraction of N. clevelandii leaves macerated for 1–5 days at 0°C. in butanol + phosphate buffer, followed by differential centrifugation. Purified preparations were highly infective, serologically active, gave specific light‐scattering zones in density‐gradient centrifugation, and contained numerous angular particles (c. 29 mμ diam.) in electron micrographs. Antisera to ANV and to TRSV had specific titres up to 1/16,384. The two viruses were serologically closely related, very similar in host range and symptoms, in vitro properties, density‐gradient zones and particle form; they differed principally in the ways in which they could be purified.Anemone necrosis disease is apparently localized in Britain and its incidence small.