Identification, detection and frequency of lily viruses in Northern India☆
Introduction
Lilies are in demand in the floriculture industry both as cut and potted flowers and ranked within the top 10 flowers in the export market. The genus Lilium includes 294 genera with 4500 species. The three commercially important divisions of lily include Easter lily (Lilium longiflorum), Asiatic and Oriental hybrids. In India, Asiatic and Oriental lilies are commonly grown in hilly areas and in mild climates. However, Easter lilies are grown with success in many parts of the country with tropical climate. Palampur is situated at latitude 32°6′N, longitude 76°5′E and altitude 1290 m, in district Kangra of Himachal Pradesh. It has a moderate climate area with temperature range of 10–30 °C. It receives 250 cm rainfall per year. The area is optimal for growing lilies commercially. Lilies can be grown from seed, bulb, scales, bulbets and bulbils. The main limiting factor in cultivation of lilies is susceptibility to virus diseases. Almost all lilies are propagated vegetatively, and infected bulbs used for forcing propagate the virus from one generation to another. These viruses affect the quality and yield of cut flowers resulting in great economic losses (Lawson, 1981, Raju and Olsen, 1985).
The most commonly occurring viruses of lily are Cucumber mosaic (CMV), Lily mottle (LMoV), Lily symptomless (LSV) and Lily X viruses (LVX) (Derks, 1995). In addition, Arabis mosaic (ArMV) (Mowat and Stefanac, 1974), Tobacco ringspot (Travis and Brierley, 1957), Tobacco rattle (McWhorter and Allenm, 1964), Tobacco mosaic, Lily virus X, Lily mild mosaic, Tomato ringspot, Narcissus mosaic (Bellardi et al., 1988, Stone, 1980, Memelink et al., 1990, Lee et al., 1996) and Strawberry latent ringspot viruses (SLRSV) (Cohen et al., 1995) have also been reported infecting lily. Ram et al. (1999) reported CMV infecting Asiatic hybrid lily cultivars on the basis of double stranded RNA (ds RNA) analysis, ELISA and host range studies.
Various lily cultivars grown in Himachal Pradesh, India, were collected and planted in the field of IHBT, Palampur, District Kangra. These plants were found to exhibit various types of symptoms, viz. mosaic, mottle, deformed leaves, deformed flowers, stunting of the plants and deformed bulbs.
The viruses were detected and characterized by bioassay, double antibody sandwich enzyme linked immunosorbent assay (DAS-ELISA), electron microscopy, immunosorbent electron microscopy, cytopathology, reverse transcription polymerase chain reaction (RT-PCR) and sequencing. Nucleotide sequences of Indian isolates were compared with other viruses reported from different parts of the world. In this way, the homologies were determined and nucleotide sequences of coat protein (cp) gene of viruses infecting lilies are presented.
Section snippets
Sample collection and establishment of virus(es) culture
The bulbs from infected plants showing symptoms were collected and planted in formalin sterilized plastic pots containing sterilized soil mixture (1 compost:2 sand:1 soil). The pots were kept in an insect proof polyhouse. The new leaves emerging from the infected bulbs were used for further analysis. Sap obtained by macerating the leaves in 0.1 M phosphate buffer, pH 7.2, followed by squeezing through double layered muslin cloth, was inoculated to Chenopodium amaranticolor, C. quinoa, Cucumis
Natural symptoms and disease incidence
Naturally infected plants of lily were found to contain either single virus or virus mixtures. When the plants were infected with a single virus, the symptoms were less pronounced and only mosaic, mottling and vein clearing was observed. In mixed infections, the plants showed severe mosaic, leaf deformation and stunting of the plants. Disease incidence on the basis of symptoms ranged from 83.7% in Asiatic hybrid cv. America to 40.7% and 40.9% in cvs. Alaska and Cavi, respectively (Table 2).
Discussion
Viruses are the major problem in lily growing fields. They often decrease the yield and quality of flowers. Lily growing fields in Himachal Pradesh, India, were found to have a high percentage of virus-infected plants. In our survey 16 cultivars of Asiatic and Oriental hybrids, L. longifolium and L. tigrinum were found to be infected with the virus diseases. The highest disease incidence on the basis of symptoms 83.7% was recorded in Asiatic hybrid cv. America and the lowest 40.7% was found in
Acknowledgements
We express gratitude to the Director and Mr. P. Navaghare, Institute of Himalayan Bioresource Technology, Palampur, for facilities and technical help, respectively. Financial grant (No. BT/PR/1421/SPD/11/099/98) from Department of Biotechnology, Govt. of India, for this study is gratefully acknowledged.
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