Vanilla Fragrans Research Articles

Screening of Antifungal and Antibacterial Activity of 90 Commercial Essential Oils against 10 Pathogens of Agronomical Importance.

Nowadays, the demand for a reduction of chemical pesticides use is growing. In parallel, the development of alternative methods to protect crops from pathogens and pests is also increasing. Essential oil (EO) properties against plant pathogens are well known, and they are recognized as having an interesting potential as alternative plant protection products. In this study, 90 commercially available essential oils have been screened in vitro for antifungal and antibacterial activity against 10 plant pathogens of agronomical importance. EOs have been tested at 500 and 1000 ppm, and measures have been made at three time points for fungi (24, 72 and 120 h of contact) and every two hours for 12 h for bacteria, using Elisa microplates. Among the EOs tested, the ones from Allium sativum, Corydothymus capitatus, Cinnamomum cassia, Cinnamomum zeylanicum, Cymbopogon citratus, Cymbopogon flexuosus, Eugenia caryophyllus, and Litsea citrata were particularly efficient and showed activity on a large panel of pathogens. Among the pathogens tested, Botrytis cinerea, Fusarium culmorum, and Fusarium graminearum were the most sensitive, while Colletotrichum lindemuthianum and Phytophthora infestans were the less sensitive. Some EOs, such as the ones from A. sativum, C. capitatus, C. cassia, C. zeylanicum, C. citratus, C. flexuosus, E. caryophyllus, and L. citrata, have a generalist effect, and are active on several pathogens (7 to 10). These oils are rich in phenols, phenylpropanoids, organosulfur compounds, and/or aldehydes. Others, such as EOs from Citrus sinensis, Melaleuca cajputii, and Vanilla fragrans, seem more specific, and are only active on one to three pathogens. These oils are rich in terpenes and aldehydes.

New Record of Association of Bean yellow mosaic virus with Mosaic Disease of Vicia faba in India

Faba bean (Vicia faba L.) is a species of bean belonging to the family Fabaceae extensively cultivated in Africa and Asia for its high nutritional values. In India and Nepal, it is called as “Bakulla”, its young pods are consumed as a green vegetable. It is used as human food in developing countries and as animal feed in industrialized countries. It is rich in l-dopa and used in medicine to treat Parkinson’s disease and to control hypertension (http://en.wikipedia.org/wiki/Vicia_faba). During a survey in February of 2011 severe mosaic symptoms were observed on about 35 % (38/109) V. faba plants growing in kitchen gardens at Lucknow. The symptoms were mild to severe (Fig. 1a) and infected plants were dwarf as compared to healthy plants. Since infection of Bean yellow mosaic virus (BYMV) is reported on V. faba [6], therefore, infection of potyvirus was suspected. Fig. 1 aBean yellow mosaic virus (BYMV) infected V. faba leaf showing mosaic symptoms. b Phylogenetic analysis of nucleotide sequence of Bean yellow mosaic virus isolate-1 ({type:entrez-nucleotide,attrs:{text:JF798580,term_id:341650451,term_text:JF798580}} ... For the detection of BYMV, total RNA was isolated from leaves of naturally infected V. faba plants from two locations using the RaFlex total RNA isolation kit (GeNei, India) and Reverse transcription-polymerase chain reaction (RT-PCR) were performed using a pair of degenerate primers (Pot 1 and Pot 2) capable of amplifying a 1.6–2.1 Kb fragment from 3′ end of the genome (virion protein gene and part of the NIb gene) of 17 species of the Potyviridae [4]. RT-PCR amplification resulted in the expected size (~1.6 Kb) amplicons from symptomatic V. faba (4/5) similar to a positive control (potyvirus infected gladiolus) but not from the healthy one. An additional specific band of ~0.8 Kb was also obtained in all the symptomatic samples. These results indicated presence of potyvirus infection in V. faba. For identification of potyvirus associated with mosaic symptoms of V. faba, the amplicons of ~1.6 Kb obtained from two symptomatic samples were sequenced directly and deposited in GenBank under Accession numbers: {type:entrez-nucleotide,attrs:{text:JF798580,term_id:341650451,term_text:JF798580}}JF798580 (isolate-1) and {type:entrez-nucleotide,attrs:{text:JN692500,term_id:779787813,term_text:JN692500}}JN692500 (isolate-2). BLASTn analysis of isolate-1 from V. faba ({type:entrez-nucleotide,attrs:{text:JF798580,term_id:341650451,term_text:JF798580}}JF798580) revealed highest sequence identities 87 % with polyprotein gene of various BYMV strains: from Japan ({type:entrez-nucleotide,attrs:{text:AB097089,term_id:40645273,term_text:AB097089}}AB097089, {type:entrez-nucleotide,attrs:{text:AB079782,term_id:32351753,term_text:AB079782}}AB079782, {type:entrez-nucleotide,attrs:{text:AB097090,term_id:40645275,term_text:AB097090}}AB097090); China ({type:entrez-nucleotide,attrs:{text:AJ311371,term_id:29134762,term_text:AJ311371}}AJ311371) and Taiwan ({type:entrez-nucleotide,attrs:{text:AM884180,term_id:198446713,term_text:AM884180}}AM884180). It also showed 86 % sequence identities with strains of BYMV: M11 ({type:entrez-nucleotide,attrs:{text:AB079886,term_id:109638255,term_text:AB079886}}AB079886) and IbG ({type:entrez-nucleotide,attrs:{text:AB079887,term_id:109638257,term_text:AB079887}}AB079887) from Japan and Vanilla ({type:entrez-nucleotide,attrs:{text:AY845011,term_id:61740434,term_text:AY845011}}AY845011, {type:entrez-nucleotide,attrs:{text:AY845012,term_id:61740436,term_text:AY845012}}AY845012) from India. The sequence identity of isolate-2 ({type:entrez-nucleotide,attrs:{text:JN692500,term_id:779787813,term_text:JN692500}}JN692500) was highest 86 % with isolates-1 ({type:entrez-nucleotide,attrs:{text:JF798580,term_id:341650451,term_text:JF798580}}JF798580) and 85–84 % with BYMV strains ({type:entrez-nucleotide,attrs:{text:AB079782,term_id:32351753,term_text:AB079782}}AB079782, {type:entrez-nucleotide,attrs:{text:AB097089,term_id:40645273,term_text:AB097089}}AB097089, {type:entrez-nucleotide,attrs:{text:AY845012,term_id:61740436,term_text:AY845012}}AY845012, {type:entrez-nucleotide,attrs:{text:AM884180,term_id:198446713,term_text:AM884180}}AM884180, {type:entrez-nucleotide,attrs:{text:AJ311371,term_id:29134762,term_text:AJ311371}}AJ311371, {type:entrez-nucleotide,attrs:{text:AY845011,term_id:61740434,term_text:AY845011}}AY845011, {type:entrez-nucleotide,attrs:{text:AB097090,term_id:40645275,term_text:AB097090}}AB097090, {type:entrez-nucleotide,attrs:{text:AB079886,term_id:109638255,term_text:AB079886}}AB079886, {type:entrez-nucleotide,attrs:{text:AB079887,term_id:109638257,term_text:AB079887}}AB079887). The 86–87 % sequence identities of isolates under study with other BYMV strains reported all over the world and in accordance with the species demarcation criteria and identification guidelines for potyviruses [1] suggested the virus isolates of V. faba under study as two isolates of BYMV. Phylogenetic analysis of nucleotide sequence of the BYMV isolates-1 and 2 of V. faba ({type:entrez-nucleotide,attrs:{text:JF798580,term_id:341650451,term_text:JF798580}}JF798580 and {type:entrez-nucleotide,attrs:{text:JN692500,term_id:779787813,term_text:JN692500}}JN692500) with the various BYMV strains of V. faba and other host species reported all over the world was performed using MEGA 4.0 tool. The BYMV strains clustered into two clusters, where the isolates under study clustered in cluster 1 along with Indian BYMV strains of Vanilla fragrans ({type:entrez-nucleotide,attrs:{text:AY845011,term_id:61740434,term_text:AY845011}}AY845011, {type:entrez-nucleotide,attrs:{text:AY845012,term_id:61740436,term_text:AY845012}}AY845012). The analysis of the BYMV isolate-1 ({type:entrez-nucleotide,attrs:{text:JF798580,term_id:341650451,term_text:JF798580}}JF798580) and isolate-2 ({type:entrez-nucleotide,attrs:{text:JN692500,term_id:779787813,term_text:JN692500}}JN692500) also revealed closest relationships with BYMV strains of Gentiana scabra ({type:entrez-nucleotide,attrs:{text:AB097089,term_id:40645273,term_text:AB097089}}AB097089, {type:entrez-nucleotide,attrs:{text:AB079782,term_id:32351753,term_text:AB079782}}AB079782, {type:entrez-nucleotide,attrs:{text:AB097090,term_id:40645275,term_text:AB097090}}AB097090) from Japan; close relationships with BYMV strains of G. scabra ({type:entrez-nucleotide,attrs:{text:AB079886,term_id:109638255,term_text:AB079886}}AB079886, {type:entrez-nucleotide,attrs:{text:AB079887,term_id:109638257,term_text:AB079887}}AB079887) from Japan, Eustoma russellianum ({type:entrez-nucleotide,attrs:{text:AM884180,term_id:198446713,term_text:AM884180}}AM884180) from Taiwan, {type:entrez-nucleotide,attrs:{text:AJ311371,term_id:29134762,term_text:AJ311371}}AJ311371 from China, V. fragrans ({type:entrez-nucleotide,attrs:{text:AY845011,term_id:61740434,term_text:AY845011}}AY845011, {type:entrez-nucleotide,attrs:{text:AY845012,term_id:61740436,term_text:AY845012}}AY845012) from India and distinct relationship with other BYMV strains of V. faba reported from Australia and Japan (Fig. 1b). Based on nucleotide sequence identities and phylogenetic relationships of the virus isolates of V. faba with various strains of BYMV, the virus isolates of V. faba under study were identified as two BYMV isolates. These isolates did not differ in symptoms on V. faba plant. The occurrence of BYMV on gladiolus in India [5], Japan [6], V. fragrans in India [2], V. faba in Australia [3], Egypt [7] and Iran [8] were reported previously. However, to our knowledge this is the first report of the natural occurrence of BYMV on V. faba in India.

The complete sequence of Cymbidium mosaic virus from Vanilla fragrans in Hainan, China

The complete nucleotide sequence of Cymbidium mosaic virus (CymMV) isolated from vanilla in Hainan province, China was determined for the first time. It comprised 6,224 nucleotides; sequence analysis suggested that the isolate we obtained was a member of the genus Potexvirus, and its sequence shared 86.67-96.61% identities with previously reported sequences. Phylogenetic analysis suggested that CymMV from vanilla fragrans was clustered into subgroup A and the isolates in this subgroup displayed little regional difference.

Preparation and Recognition Properties of Vanillin‐Imprinted Polymers

AbstractSome new molecularly imprinted polymers (MIPs) were prepared by different protocols involving vanillin as the imprinted molecule, methacrylic acid (= 2‐methylprop‐2‐enoic acid; MAA) as the functional monomer, and ethylene glycol dimethacrylate (EGDMA = 2‐methylprop‐2‐enoic acid ethane‐1,2‐diyl ester) as the cross‐linking agent. The adsorption property of the imprinted polymers was studied by UV spectrophotometry and HPLC. The results indicated that the porogen solvent had a certain influence on the adsorption performance of the polymer. The vanillin‐imprinted polymer MIP1 prepared with MeOH as porogen, exhibited advantageous characteristics, i.e., a high binding activity, a good selectivity, and a rapid adsorption equilibrium. The binding parameters studied by Scatchard analysis established that there are two types of binding sites in MIP1. Finally, by packing an SPE column (SPE = solid‐phase extraction) with the polymer MIP1, the vanillin was separated and enriched successfully by this sorbent from the samples of Vanilla fragrans and beer.

Bioactive aromatic compounds from leaves and stems of Vanilla fragrans.

Alcoholic extracts of leaves and stems of Vanilla fragrans were fractionated with ethyl acetate and aqueous butanol. All three fractions of ethyl acetate, butanol, and water were screened for toxic bioactivity against mosquito larvae. The results of these experiments showed that the fractions from the ethyl acetate and butanol phases were both active in the bioassay. Bioactivity of the ethyl acetate fraction was found to be much greater than that from the butanol fraction in mosquito larvae toxicity. The water phase appeared to contain no substances that impaired mosquito larval growth. Repeated column chromatography of the ethyl acetate fraction on silica gel led to the isolation of 4-ethoxymethylphenol (1), 4-butoxymethylphenol (2), vanillin (3), 4-hydroxy-2-methoxycinnamaldehyde (4), and 3,4-dihydroxyphenylacetic acid (5). Compounds 4 and 5 were isolated from Vanilla species for the first time and 2 has not been reported to have been found in a natural form. 4-Ethoxymethylphenol (1) was the predominant compound, but 4-butoxymethylphenol (2) showed the strongest toxicity to mosquito larvae. The structures of the compounds were determined on the basis of their mass spectra and (1)H or (13)C NMR data.

Coat Protein Gene ofCymbidiummosaic virus fromVanilla fragransin Reunion Island

Nucleotide and amino acid sequences of the coat protein (CP) of 12 isolates ofCymbidiummosaic virus fromVanilla fragransin Reunion Island (CyMV‐R) were compared with each other and with those of previously described Asian strains. Alignment revealed that CyMV‐R isolates were highly homologous, suggesting that one strain is prevalent in Reunion. This strain also showed high homology with the Korean CyMV‐K2 and Singapore CyMV‐S2 strains, but nucleotide additions resulted in the carboxy‐terminal ends of the CP sequences differing from those of the Korean CyMV‐K1 and Singapore CyMV‐K1 strains.

Long-Chain Aliphatic β-Diketones from Epicuticular Wax of Vanilla Bean Species. Synthesis of Nervonoylacetone

Analysis of the neutral lipids from Vanilla fragrans and Vanilla tahitensis (Orchidaceae) without saponification resulted in the isolation and identification of a new product family in this plant: beta-dicarbonyl compounds. The compound structures are composed of a long aliphatic chain with 2,4-dicarbonyl carbons and a cis double bond at the n-9 position. They represent approximately 28% of the neutral lipids, that is, 1.5%, in immature beans, and approximately 10% of the neutral lipids, that is, 0.9%, in mature beans. Using retention indices, gas chromatography-mass spectrometry, derivatization reactions, and chemical degradation, five beta-dicarbonyl compounds have been identified including 16-pentacosene-2,4-dione, 18-heptacosene-2,4-dione, 20-nonacosene-2, 4-dione, 22-hentriacontene-2,4-dione, and 24-tritriacontene-2, 4-dione. Among them (Z)-18-heptacosene-2,4-dione, or nervonoylacetone, has been synthesized in two steps starting from nervonic acid. The major constituent, nervonoylacetone, represented 74.5% of the beta-dicarbonyl fraction. The range of these compounds has been studied in relation with bean maturity for V. fragrans and V. tahitensis species. This compound family has not been found in the leaves or stems of any of the three vanilla species studied and is markedly absent in the beans of V. madagascariensis.

4‐Demethylsterols and triterpene alcohols from two Vanilla bean species: Vanilla fragrans and V. tahitensis

Abstract4‐Demethylsterol and triterpene alcohol compositions of two Vanilla bean species (V. fragrans and V. tahitensis) were investigated. From retention times and gas chromatography‐mass spectrometry, nine 4‐demethylsterols were identified in V. fragrans and seven in V. tahitensis. The 4‐demethylsterol fraction of V. fragrans was characterized by a high content of 24‐methylene cholesterol (27–40%) and of β‐sitosterol (35–46%). The 4‐demethylsterol fraction of V. tahitensis was characterized by a high content of stigmasterol (27%) and of β‐sitosterol (57.5%), and a lower amount of 24‐methylene cholesterol (5%). Vanilla tahitensis was also characterized by the presence of ergosta‐5,25‐dien‐3β‐ol (2%) and the absence of campesterol, stigmasta‐5,22,25‐trien‐3β‐ol, and ergosta‐7,24(28)‐dien‐3β‐ol. The beans’ age modified the ratio 24‐methylene cholesterol/β‐sitosterol in V. fragrans. Combining liquid chromatography and gas chromatography allowed the identification of four other demethylsterols in V. fragrans (brassicasterol, 0.02%; stigmasta‐5,23‐dien‐3β‐ol, 1.43%; stigmasten‐22‐ol, 0.1%; and fucosterol, 0.5%) from the 4‐demethylsterol fraction. 24‐Methylene cholesterol and β‐sitosterol were isolated, and their structures were confirmed by 1H and 13C nuclear magnetic resonance. Four triterpene alcohols were identified in V. fragrans, including cycloartenol (0.9–1.6%) from the triterpene alcohol fraction, 24‐dihydrotirucallol (17–23%) from the triterpene alcohol fraction, tirucall‐7‐en‐3β‐ol (6–7.5%) from the triterpene alcohol fraction, and in a higher content cyclosadol (66–69%) from the triterpene alcohol fraction. The content ranges were studied as a function of the beans’ age. Demethylsterol and triterpene alcohols profile could be used for origin differentiation.

Improved determination of vanillin and related phenolic components in vanilla (Vanilla fragrans (Salisb.) Ames) by high‐performance liquid chromatography

AbstractThis paper describes a reverse‐phase high‐performance liquid chromatographic (HPLC) method for the separation, identification and quantification of vanillin and related phenolic compounds in ethanolic extracts of cured vanilla. The compounds were separated on a silica column, bonded with pentafluorophenyl, using a mobile phase gradient of methanol–citric acid with diode‐array detection at 275 nm. Vanillin, vanillic acid, p‐hydroxybenzoic acid and p‐hydroxybenzaldehyde were identified by the spectral data and retention times given by reference compounds. Retention times and spectral data for a further 36 compounds are presented. The new HPLC technique allows a more accurate means of determining the vanillin content of vanilla than the ISO spectrophotometric assay.

Vanilla viruses in the South Pacific

Surveys for viruses affecting Vanilla fragrans and V. tahitensis carried out in Cook Islands, Fiji, Niue, Tonga and Vanuatu detected cymbidium mosaic potexvirus and odontoglossum ringspot tobamovirus in all countries; vanilla necrosis potyvirus in Fiji, Tonga and Vanuatu; and vanilla mosaic potyvirus in Cook Islands, Fiji and Vanuatu. For all these viruses the surveys extended the previously known distribution in the South Pacific. Other, unreported potyviruses and rhabdovirus‐like particles were detected from Fiji and Vanuatu. Symptoms associated with potyvirus infection (leaf distortion, sunken chlorotic patches, stem necrosis and vine die‐back) were found in a majority of plantings in Fiji, Tonga and Vanuatu; in Fiji and Tonga there were instances where up to 50% of the plants in farmers’ plots were affected.

The Isolation and Evaluation of Two Naturally Occurring Mild Strains of Vanilla Necrosis Potyvirus for Control by Cross‐Protection

AbstractIn an attempt to find mild virus strains that would cross‐protect sgainst vanilla necrosis potyvirus (VNPV), Vanilla fragrans plants in Tonga were surveyed for the presence of mild or symptomless potyvirus infections. Potyviruses were detected by indirect ELISA using a commercially available portyvirus group monoclonal anibody. From 28 plants with mild or symptomless infections two portyvirus isolates, designated V1 and V3, included systemic infections in Nicotiana benthamiana following mechanical inoculation. V1, which causes a mild mottle in N. benthamiana, is serologically related to VNPV, while V3 which causes mild vein banding is serologically unrelated to VNPV. Prior inoculation with V1 protected N. benthamiana against the severe mosaic symptoms of VNPV when challenge inoculated after 14 and 21 days, but not after 7 days. When V3 was used as the protecting strain, cross‐protection was observed in some, but not all plants, when chalenged with VNPV after 14 and 21 days.

Some Hosts and Properties of a Potyvirus Infecting Vanilla fragrans (Orchidaceae) in the Kingdom of Tonga

Abstract A potyvirus was isolated from Vanilla fragrans exhibiting leaf distortion, chlorotic and necrotic lesions and vine die‐back. The virus can be propagated in Nicotiana clevelandii and N. benthamiana and induces local lesions in Chenopodium amaranticolor and C. quinoa. The flexuous particles have a mean length of 765 nm, pinwheel inclusions are found in systemically infected N. benthamiana leaves and SDS‐polyacrylamide gel electrophoresis indicates a single polypeptide of M 32.7 × 103. The virus is transmissible from N. clevelandii to N. clevelandii by the aphid species Myzus persicae and Aphis gossypii. A polyclonal antiserum raised to the Tongan potyvirus failed to react, with the potyvirus associated with mosaic symptoms of V. tahitensis in French Polynesia and ISEM failed to detect any close relationship to 22 other potyviruses.

Synthesis, identification and determination of glucosides present in green vanilla beans (Vanilla fragrans Andrews)

AbstractA preliminary study showed the presence of gluocovanillin and other glucosides in green vanilla beans (Vanilla fragrans Andrews). The glucosides of vanillin, vanillic acid, ρ‐hydroxybenzaldehyde and ρ‐hydroxybenzoic acid have now been synthesized and their structures confirmed by mass spectrometry and 13C‐NMR spectroscopy. A reversed phase (C‐18) HPLC method, using a photodiode array detector, enabled separation and characterization of the authentic substances and this method was applied to the determination of these glucosides in green vanilla bean extracts of various origin.