Lily Mottle Virus Research Articles

Occurrence of Lily mottle virus on Lilium in Italy.

Lily mottle virus (LMoV), a member of the genus Potyvirus, is one of the main viruses infecting lily. Symptoms on lily differ according to the susceptibility and sensitivity of different cultivars and hybrids. They range from leaf mottle or mosaic, vein clearing, chlorotic and yellow streaking, leaf curling, and necrotic spots, to milder forms of leaf symptoms. Plants may even be symptomless at some stages of growth. A varietal collection of Lilium from the early 1990s is held in Pistoia Province (Tuscany, Italy) and is composed of Asian hybrids obtained from intraspecific breeding of commercial cultivars. During a survey conducted from May to June 2010, several plants showing vein clearing, leaf mottle, leaf mosaic, and reddish brownish necrotic spots were observed. Leaf samples from 60 symptomatic or symptomless lily plants, belonging to 20 cultivars, were collected and tested for the presence of LMoV. Samples were assayed by double-antibody sandwich (DAS)-ELISA and eight of them, belonging to four different cultivars, tested positive. Total RNA was extracted from 2 g of leaf tissue of every collected sample according to the protocol described earlier (3) and cDNA synthesis was performed with an iScript cDNA Synthesis Kit (Bio-Rad, Hercules, CA). Samples were tested by reverse transcription (RT)-PCR and real-time PCR assays using primers LMoV1 (5'-GCAAATGAGACACTCAATGCTG-3') and LMoV2 (5'-CGTGCGTGAAGTAACTTCATAG-3') designed to amplify 651 bp of the coat protein (CP) gene of LMoV (1). Results obtained with RT-PCR and real-time PCR exactly matched those achieved with ELISA assay, and the eight positive samples showed amplicons of the expected size. PCR products from five infected samples were directly sequenced from both directions and submitted in GenBank (Accessions Nos. JQ655106 to JQ655110). Our isolates share more than 99% nucleotide identity among each other. Comparison with other LMoV-CP gene sequences present in GenBank showed nucleotide identities ranging from 93 to 94% with LMoV isolates from South Korea (GenBank Accession Nos. GQ150683 to GQ150686), China (GenBank Accession Nos. EU348826, AJ748256, AJ564636, and AJ564637), Australia (GenBank Accession No. JN127341), and Japan (GenBank Accession No. AB570195). To our knowledge, this is the first report of LMoV on Lilium in Italy where this virus was already reported to infect escarole (2). Considering the economic importance of Lilium production as a flowering plant in Pistoia Province, and in several other areas of Italy, the report of LMoV present on lilies suggests the use of healthy propagation material and the adoption of preventive measures to avoid its diffusion.

ユリから検出されたレンブラントチューリップ斑入りウイルス（新称）

In 2009, Asiatic hybrid lily plants with mosaic symptoms were observed in Akita Prefecture, Japan. In a RT-PCR assay, using universal primers for the detection of potyviruses, a potyvirus different from Lily mottle virus was detected. The conserved area of the virus coat protein (CP) shared the highest amino acid (aa) sequence similarity (95.7%) to that of Rembrandt tulip breaking virus (ReTBV). As far as we know, this report is the first on the occurrence of ReTBV in Japan and on the full aa sequence of the CP for this virus.

Production of Virus-free Bulblets by Meristematic Tip Culture with Antiviral Chemical in Lilium brownii var. colchesteri

Lilium brownii var. colchesteri has unique and ornamental floral characteristics in graceful harmony with flower and anther colors, flower shape, elegant fragrance, and flower color change from yellowish cream to white during anthesis. There are, however, few accessions conserved in Japan up to the present, and they often show abnormally shaped flowers and mosaic leaves seemingly due to virus infection. Virus-free bulblets were established by combining meristematic tip culture and chemotherapy. At initial diagnosis with RT-PCR, Cucumber mosaic virus (CMV), Lily mottle virus (LMoV) and Lily symptomless virus (LSV) were detected from leaf tissues of the mother plants. All regenerated bulblets obtained by meristematic tip culture of the mother plants were still infected by at least one of the viruses. The meristematic tip of the bulblets infected with either LSV or LMoV was selected for subsequent culture with 2,4-dioxohexahydro-1,3,5-triazine (DHT), an antiviral chemical. LSV was eliminated successfully in mericlones from bulblets with LSV, whereas LMoV was not from those with LMoV. The virus-free bulblets were transferred to new medium without DHT, and multiplied by in vitro scaling. They were then acclimated in a phytotron glass room at 20°C. The plants were confirmed to be virus-free after 18-months’ acclimation. It was concluded that the combination of meristem tip culture and chemotherapy is practical for producing virus-free plants of L. brownii var. colchesteri.

Genetic mapping in Lilium: mapping of major genes and quantitative trait loci for several ornamental traits and disease resistances

With 1 figure and 3 tables AbstractConstruction of genetic linkage maps for lily was achieved using two populations, LA and AA that share one parent ‘Connecticut King’. Three different molecular marker systems (AFLP™, DArT and NBS profiling) were used in generating linkage maps for ‘Connecticut King’. The LA and the AA populations consist of 20 and 21 linkage groups (LGs), respectively. Average density between markers was 3.9 cM for the LA and 5 cM for the AA population. Several horticultural traits were mapped for the first time in Lilium and showed to be single gene based. We propose to name these genes as LFCc for flower colour, lfs for flower spots, LSC for stem colour, lal for antherless phenotype and lfd for flower direction whereby upper and lower case names refer to dominant and recessive genes, respectively. Additionally, resistance to Lily mottle virus (LMoV) was mapped as a locus on LG AA10. For Fusarium resistance, the Kruskal–Wallis test identified six putative quantitative trait loci (QTL) in the AA population of which one QTL (explaining 25% of the variation in resistance) could be confirmed by interval mapping.

Detection and Frequency of Lily Viruses in Argentina.

In a survey of lily growing fields in various regions of Argentina, three viruses, Lily symptomless virus (LSV), Lily mottle virus (LMoV), and Cucumber mosaic virus (CMV), were found in Longiflorum, Asiatic, Oriental, Longiflorum × Asiatic (LA), and Oriental × Trumpet (OT) hybrids. The areas surveyed were between latitude 26° 56' S and 43° 03' S, and longitude 65° 21' W and 71° 29' W. Virus detection was performed by double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) using polyclonal antiserum. In infected samples, viruses detected in decreasing order were LSV (60.5%), LMoV (51.0%), and CMV (28.7%) present in single or mixed infections. Virus infection varied among tested hybrids from 36.0% (Oriental Montecristo) to 94.7% (Lilium longiflorum Avita) in 2006 and from 38.9% (OT Yelloween) to 82.1% (LO Triumphator) in 2007, with an overall incidence of 64.1 and 70.7% in 2006 and 2007, respectively. A variation in virus incidence among localities was also observed. The highest virus incidence (89.6 and 87.6% in 2006 and 2007, respectively) was observed in Bahía Blanca (38° 44' S, 62° 16' W). The lowest virus incidences, detected in Trevellin (43° 03' S, 71° 29' W) and in Malargüe (35° 28' S, 69° 35' W), were 47.4 and 48.6% in 2006 and 2007, respectively. Moreover, a different distribution of each virus was observed between localities. The high occurrence of viruses infecting lily crops in Argentina could be due to both the use of infected bulbs for propagation and the lack of preventive virus vector control measures.

Production and Evaluation of Monoclonal Antibodies Against Recombinant Coat Protein of Lily mottle virus for Western Blotting and Immono-blot Analysis

Lily mottle virus (LMoV) causes flower quality reduction in Lilium spp. The coat protein gene was RT-PCR-amplified from total RNA extracted from infected lily leaves and the amplified fragment was cloned into the pRSET expression vector tagged with a His-MBP. The plasmid of recombinant coat protein was used to transform an Escherichia coli strain pLysS and was expressed. The coat protein was purified by affinity chromatography using a Ni-NTA resin. The identity of the purified protein was confirmed by SDS-PAGE. The in vitro-expressed protein was used for immunization of mice. The polyclonal and monoclonal antibodies reacted specifically for the detection of LMoV in lily extracts in Western blot. Moreover the monoclonal antibodies reacted with lily extracts in DAS-ELISA with no unspecific or heterologous reactions against other non-serologically related viruses, but the polyclonal antibodies revealed a weak reaction against both infected lily and healthy control.

Production of Antiserum to Recombinant Coat Protein for Detecting Lily mottle virus in Yunnan, China

Abstract Lily mottle virus (LMoV), Potyvirus genus, is very difficult to purify for the preparation of diagnostic antisera. The coat protein (CP) gene of LMoV was amplified by RT‐PCR from infected plants and cloned into the prokaryotic pET‐30a vector to generate the recombinant plasmid pET‐CP. The resulting carboxy‐terminal His‐tagged CP was over‐expressed in Escherichia coli BL 21 cells by Isopropyl‐β‐d‐thiogalactoside (IPTG) induction and purified over Ni‐NTA affinity columns. The purified CP was used to elicit a polyclonal antiserum in rabbits. The antiserum had a titre of 1 : 128 in double diffusion tests, and specifically recognized LMoV in Western blots, Enzyme‐Linked Immunosorbent Assays and Immuno‐Electron Microscopy. The CP antiserum was also used to evaluate LMoV infection of lily bulbs used for commercial production in Yunnan, China. Substantial levels of infection were found in both imported and native bulbs, and provide the basis to implement flexible, rapid and large‐scale virus indexing of lily plants for use in propagation and to meet virus‐free quarantine regulations.

THE CONTRIBUTION OF QUALITY INSPECTIONS TO THE IMPROVEMENT OF THE QUALITY OF THE DUTCH FLOWERBULBS AND ACCESS TO EXPORT MARKETS

THE CONTRIBUTION OF QUALITY INSPECTIONS TO THE IMPROVEMENT OF THE QUALITY OF THE DUTCH FLOWERBULBS AND ACCESS TO EXPORT MARKETS

Identification, detection and frequency of lily viruses in Northern India

In a survey of lily growing fields in various parts of Himachal Pradesh, India, three viruses, Cucumber mosaic (CMV), Lily symptomless (LSV) and Lily mottle virus (LMoV) lily strain, were found quite prevalent in Asiatic (12 cultivars) and Oriental hybrids (4 cultivars) of lily, Lilium longiflorum and L. tigrinum. Apart from these viruses, Strawberry latent ringspot virus (SLRSV) was also found to infect Oriental hybrid lily. The disease incidence on the basis of symptoms observed ranged between minimum (40.7%) in cv. Alaska and maximum (83.7%) in cv. America, both Asiatic hybrid lily types. These viruses were indexed by testing outer and inner scales of bulbs and the leaves at early and flowering stages using ELISA and reverse transcription polymerase chain reaction (RT-PCR). Mechanical and insect transmission, purification, electron microscopy and sequencing of the PCR fragments were carried out for these viruses. Cloning and sequence analysis confirmed the viruses as CMV-subgroup II, LSV and LMoV. There were 96–98% nucleotide and 93–97% amino acid homology with CMV subgroup II sequences, 97–99% nucleotide and 91–92% amino acid homology with LSV sequences and 92–99% nucleotide and 85–98% amino acid homology with LMoV sequences.

신나팔나리(Lilium x fomolongi) 바이러스 감염이 절화품질에 미치는 영향

신나팔나리(Lilium x fomolongi) 뇌산 품종 실생묘에 Cucumber mosaic virus(CMV), Lily mottle virus(LMoV) 및 Lily symptomless virus(LSV)를 인공적으로 접종하여 생육에 미치는 영향을 조사하였다. 또한 재배포장에서 자연감염된 바이러스 병징과 감염율을 조사하였다. 재배포장에서 가장 많이 관찰되는 병징은 LMoV 감염에 의한 잎 모틀 증상이었다. 바이러스 감염율은 6년 이상 재 사용된 구근으로 재배하는 포장의 경우 CMV, LSV, LMoV 바이러스 가운데 두 종류 이상의 바이러스에 의한 복합 감염율은 80% 가량이었다. 신나팔나리 (Lilium x fomolongi) 품종 뇌산에 CMV-Li1과 LMoV-Li 복합감염시 건전주에 비해 초장 14%, 생체중 38%, 꽃 길이가 15% 감소하였다. CMV-Li1과 LMoV-Li 단독감염시에도 생체중은 건전주에 비해 각각 21.8%, 28.4%의 유의성 있는 감소를 보였다. Damages caused by infection with Cucumber mosaic virus (CMV), Lily mottle virus (LMoV) and Lily symptomless virus (LSV) were assessed by comparing growth of plants produced from seeds of Lilium x fomolongi cultivar 'Noesan' both infected and free from infection with those viruses. Symptoms and infection rate were investigated in field-grown lily.Dominant viral infection symptom in the field was mottle on leaves, caused by natural infection with LMoV. Incidence of viral disease caused by mixed infection with two of LMoV, CMV and LSV in field-grown Lilium x fomolongi cultivars, reused for more than 6 years consecutively, was 80 percent. In comparison with healthy Lilium x fomolongi cultivar 'Noesan', plants doubly infected with CMV-Li1 and LMoV-Li diminished their plant height by 14 percent, fresh weight by 38 percent, and flower length by 15 percent. Lily plants singly infected with CMV-Li1 or LMoV-Li significantly reduced their freish weight by 21.8% and 28.4% compared to healthy plants, respectively.

Occurrence and sequences of Lily mottle virus and Lily symptomless virus in plants grown from imported bulbs in Zhejiang province, China.

Degenerate primers were used to amplify virus sequences from imported lilies in Zhejiang province, China. Two viruses, Lily mottle virus (LMoV, genus Potyvirus) and Lily symptomless virus (LSV, genus Carlavirus) were detected, purified and completely sequenced from a mixed infection in a plant raised from bulbs imported from the Netherlands. The sequence of LMoV was 9644 nt long and encoded a polyprotein of 3095 amino acids with a calculated M(r) of 351.0 kDa that had only 45.1-54.4% identity to other completely sequenced potyviruses. Phylogenetic analysis of the complete polyproteins of members of the genus demonstrated that LMoV was distantly grouped with LYSV, BYMV and ClYVV. Two partial LMoV sequences from different cultivars were identical to one another and very similar (98.3% identical nucleotides) to the corresponding region of the complete sequence. Analysis of the coat protein sequences of LMoV isolates revealed two subgroups, corresponding to the earlier "Tulip breaking virus lily strain" and "Tulip band breaking virus" isolates. Our newly-determined isolates showed an extremely close relationship to the first of these. The LSV sequence was 8393 nucleotides long and had the typical carlavirus genome organization. The ORF1 protein was most closely related to that of Blueberry scorch virus (57.2% identical amino acids). Sequences of 1796 nt at the 3'-end of three additional LSV isolates from different cultivars were very similar (>98% identical nucleotides) to the corresponding region of the complete sequence. This is the first report of complete sequences for LMoV and LSV.

First Report of Tulip band breaking virus in Mosaic Diseased Tulip in Japan.

Tulip (Tulipa spp.) is an ornamental plant of major economic importance in Japan. Regions in Toyama Prefecture are some of the most productive for producing tulip bulbs, shipping approximately 50 million bulbs annually. However, mosaic diseases caused by viruses such as Tulip breaking virus (TBV) currently limit bulb production in these areas. Only the potyviruses TBV and Lily mottle virus (LMoV) have been reported infecting tulip in Japan. A virus isolate from tulip with flower-breaking symptom in Toyama Prefecture was tentatively named OE4 and was presumed to be LMoV after detection by LMoV-specific reverse transcription-polymerase chain reaction (RT-PCR) (4). When OE4 was mechanically inoculated on test plants (13 species from six families), RT-PCR confirmed that it infected plants in the Liliaceae (Tulipa spp., Lilium formosanum, and L. concolor) with mosaic symptoms but did not induce any symptoms in Chenopodium quinoa, Tetragonia tetragonoides, and Nicotiana benthamiana. According to Dekker et al. (2) LMoV and Tulip band breaking virus (TBBV) infected Tulipa spp. and TBBV did not infect C. quinoa, T. tetragonoides, N. clevelandii, and N. benthamiana, species that were local or systemic hosts for LMoV. To analyze the genomic sequence of OE4, a primer set was designed for amplifying the coat protein (CP) gene of LMoV, with 5' -GCAAATGAGACACTCAATG-3' as a forward primer and 5'-TTACATAGAAATTCCAAGTAAG-3' as a reverse primer. The fragment obtained was cloned and sequenced (DDBJ/EMBL/GenBank Accession No. AB078007). The CP gene of OE4 consisted of 825 nucleotides and had 86.5% identity (90.6% identity in deduced amino acid sequence) with the CP gene of LMoV-Netherlands (S44147) (3). When it was compared with partial sequences (277 nucleotides) of LMoV (S60810) and TBBV (S60805) (2), the nucleotide sequence identities were 89.9 and 96.4%, respectively. Multiple alignment and a phylogenetic tree based on the 277 nt of the tulip potyviruses, including OE4, showed the close relationship between OE4 and TBBV. These results indicated that OE4 was an isolate of TBBV. However, the CP amino acid sequence identity between TBBV and LMoV was more than 80%, and it seemed logical that TBBV be assigned to a strain of LMoV, according to potyvirus species demarcating criteria (1). Another screening using RT-PCR based detection, and an inoculation test for 55 flower-breaking tulips collected from fields in Toyama Prefecture revealed that four tulip plants were infected with TBBV and 54 with TBV. This suggests that TBV is more prevalent than TBBV, and we need to produce an attenuated virus of TBV, which will be effective for managing tulip breaking disease in the prefecture.

Detection of cucumber mosaic virus, lily symptomless virus and lily mottle virus in Lilium species by RT-PCR technique

Cucumber mosaic virus (CMV), lily symptomless virus (LSV) and lily mottle virus (LMoV) were detected in three lily cultivars by reverse transcription-polymerase chain reaction (RT-PCR). Results were compared with those obtained in an enzyme-linked immunosorbent assay (ELISA). The amplified DNA fragments with expected size and specific bands for each virus were obtained by RT-PCR using degenerate primers. In 18 out of 22 plants tested at least one virus was detected with RT-PCR in the 18 (81.8%) plants compared to 4 (18.2%) plants with ELISA. The present study showed that the RT-PCR is a more sensitive method than ELISA to determine the presence of viruses in Lilium plants of various species.

Occurrence of Lily mottle virus in Escarole.

Lily mottle virus (LMoV), genus Potyvirus, an important virus of lily that also causes flower-breaking in tulip (1), is considered to have a natural host range restricted to the family Liliaceae. In 1996, escarole (Cichorium endivia L. var. latifolium LAM) plants growing in fields close to Torino, Italy, and showing mosaic and necrotic spots on outer leaves were infected by a potyvirus related to LMoV. The virus was identified by immunoelectron microscopy (IEM) done on experimentally infected Nicotiana benthamiana and Chenopodium quinoa. The virus isolated from escarole (LMoV-E) had an experimental host range similar to that of lily isolates of LMoV, although species within the Liliaceae were not tested. LMoV-E systemically infected all nine escarole cultivars and six of seven endive cultivars (C. endivia L. var. crispum LAM) but did not infect any of six lettuce and two chicory cultivars (C. intybus L. var. foliosum HEGI). Symptoms ranged from mild to severe mosaic and were generally more severe on escarole than on endive. Some of the same escarole, endive, and lettuce cultivars were inoculated with a typical LMoV isolate from lily (Le97/49, from A. F. L. M. Derks, the Netherlands), which induced mild systemic infections in only one escarole and one endive cultivar. Both cultivars were also susceptible to LMoV-E. LMoV-E was purified from N. benthamiana, and an antiserum was prepared. IEM decoration titer experiments were done with LMoV-E and four other LMoV isolates from Liliaceae and their homologous antisera. Heterologous titers ranging from identity to serological differentiation index values of 2 to 4 were obtained, confirming the identity of the escarole isolate as a LMoV strain and indicating considerable serological variability among LMoV isolates. In a field survey of endive and escarole crops in 1998, in the area where LMoV-E was first identified, the virus was again detected by enzyme-linked immunosorbent assay in 4 of 80 escarole plants tested. LMoV-E appears to be a LMoV strain particularly adapted to escarole. To our knowledge, this is the first report of LMoV identified in a naturally infected host outside monocotyledonous plants. Reference: (1) E. L. Dekker et al. J. Gen. Virol. 74:881, 1993.

A Comparison of Sensitive and Specific Methods for the Detection of Lily Mottle Virus in Lily Plants

The nucleotide sequence of the 3′‐terminal region of a potyvirus from lily in Miyagi, Japan, exhibited 96.5% homology in the CP gene and 98.9% homology in deduced amino acid sequence to an isolate of Lily mottle virus (LMoV) from the Netherlands. Dilution end‐points were compared for the detection of LMoV in lily by an indirect enzyme‐linked immunosorbent assay; an assay with immunocapture, reverse transcription (RT) and the polymerase chain reaction (PCR); and an assay with RT and PCR using total RNA extracted from infected plant material. The RT‐PCR assay with total RNA was the most sensitive assay which, with primers based on the nucleotide sequence information, was able to distinguish between infection by LMoV and Tulip breaking virus, another potyvirus that can infect lily.

Molecular evidence that a lily-infecting strain of Tulip breaking virus from Japan is a strain of lily mottle virus

The sequence of the 3′-terminal 2074 nucleotides (nts), excluding the 3′-poly (A) tail, of RNA of a potyvirus isolated from lily (Lilium Asiatic hybrid cv. Enchantment) in Japan, currently tentatively designated as Tulip breaking virus-li (TBV-li), was determined. The sequence started within a single open reading frame (ORF) that encoded the carboxyl terminus of the large nuclear inclusion protein (NIb) and the complete 275-amino-acid coat protein (CP), followed by a 3′-untranslated region (3′-UTR) of 204 nts. The CP of TBV-li shared 91% amino acid (aa) sequence identity with that of TBV lily strain Dutch isolate (TBV-lily). The nt sequences of their 3′-UTR were 94% identical. However both viruses shared only 60–65% sequence identities with TBV tulip strain Niigata isolate in the corresponding regions. The results suggest that TBV-li is closely related to TBV-lily, and that these two TBV lily strains should be classified into a species different from TBV tulip strains. We therefore support a proposal to rename TBV-lily Lily mottle virus (LMoV), and suggest that TBV-li is another strain of LMoV (LMoV-J).

Control of aphid-borne Lily symptomless virus and Lily mottle virus in Lilium in the Netherlands

An overview is presented on the management of viruses of Lilium crops in the Netherlands since the 1960s. This mainly concerns Lily symptomless virus (LSV) and Lily mottle virus (LMoV). Various factors which affect the efficiency of control are considered. The variable symptoms of LMoV in the many vegetatively propagated cultivars grown (c. 340) pose problems in the efficient roguing of diseased plants of some cultivars. Additionally the acceptable incidence of viruses such as LSV which are generally symptomless in field-grown plants may cause problems under the unfavourable light and growing conditions of year-round cut-flower production in greenhouses. The reduction in bulb yield and quality caused by viruses of lilies necessitates a further decrease in the already low virus levels still tolerated in lily bulb stocks. The routine detection of LSV and LMoV by ELISA has been developed extensively over the years. The impact of testing bulbs (‘bulb test’) during storage has been important in achieving an overall decrease in virus incidence in many stocks of lily cultivars, e.g. in Asiatic hybrids. The ‘leaf test’ used to assess many other cultivars, e.g. oriental hybrids, in which the bulb test for LMoV is not applicable, will be developed so as to eliminate at an early stage severely infected stocks that were initially intended for further propagation. The spread of the aphid-borne LSV and LMoV generally occurs very rapidly. The low virus incidence in the initially virus-tested stocks obtained by tissue culture procedures is effective in decreasing the access of vectors to virus-infected lilies. The viruses spread mainly in June and July, considerably less in May and least in August/September. Consequently the routine spraying of mixtures of mineral oil and pyrethroid insecticide is generally done weekly in May, June and July and fortnightly in August and September. There are differences in efficacy of the different brands of mineral oil. The rapid propagation of lilies, as done routinely by the scaling of bulbs and by tissue culture procedures, enables the rapid bulking of virus-tested and other stocks of high quality which have the health status required. The impact of the different factors in the management and control of viruses in Lilium crops in the Netherlands is discussed.

DETECTABILITY OF VIRUSES IN LILY BULBS DEPENDS ON VIRUS, HOST AND STORAGE CONDITIONS

Lily symptomless virus (LSV) and lily mottle virus (LMoV) are detected reliably with ELISA in bulbs of Asiatic hybrids stored at 0-2°C for at least 2-3 weeks after lifting prior to testing. If this protocol was applied to Lilium longiflorum-cultivars and Oriental hybrids, however, it generally failed in the detection of LMoV. This phenomenon was virus dependent since LSV, cucumber mosaic virus and lily virus X were easily detected. The detection of LMoV in L. longiflorum-cvltivais greatly improved after storage of scales at 20°C for 2-3 weeks under white fluorescent light (12-16 h/day, 13-16 W.m-2). This protocol was also applicable to LSV in L. longiflorum, but not for LMoV in Oriental hybrids.