Journal of Taiwan Agricultural Research

Author:Chin-Chih Chen*, Chin-An Chang*, and Fen-Lang Chiang

Abstract:

    A virus isolate (HM4), obtained from a symptomless blood lily (Haemanthus multiflorus Martyn) plant, was established on Chenopodium quinoa Willd. by three successive single lesion reisolation and inoculation. The original blood lily plant and inoculated C. quinoa leaf tissues of HM4 were reacted strongly with the antibody against the LY56 isolate of Narine latent virus (NeLV) from golden spider lily in indirect ELISA and western blotting. A 1.5-kbp DNA fragment was amplified by RT-PCR from HM4-infected blood lily and C. quinoa tissues using a primer pair specific to the 3'-end of genomic sequences of NeLV. Sequence analyses revealed that the amplicon comprises 1,560 nucleotides spanning the entire open reading frame of coat protein (CP) and the complete cysteinerich protein region of NeLV. Comparison of nucleotide and amino acid sequences indicated that the CP of HM4 shares 96–99% identities with those of the other six blood lily isolates and the known NeLV isolates documented in the GenBank. Results indicated that HM4 and the other six blood lily isolates are the isolates of NeLV. In phylogenetic analysis, these blood lily isolates and those NeLVs from golden spider lily (Lycoris aurea), amaryllis (Hippeastrum hybridum Hort.) and narcissus (Narcissus spp.) were grouped in the same clade evidently differing from that of Hippeastrum latent virus (HLV) and Narcissus common latent virus (NCLV). Phylogenetic analysis indicated that the blood lily isolates are grouped in the same sub-clade. Our studies also confirmed that all the NeLV isolates from blood lily, golden spider lily, amaryllis and narcissus can be detected by the same antiserum against NeLV-LY56 in indirect ELISA and western blotting. Moreover, a 552-bp DNA fragment can be amplified from all these NeLV isolates in RT-PCR using the primers designed from the genomic sequence of HM4. Field surveys were conducted during 2006–2014 by indirect ELISA and showed 18.4, 32, 79, and 13.3% of the incidences of NeLV in blood lilies, golden spider lilies, amaryllis and narcissus, respectively, in Taiwan. Based on the serological and molecular evidences, NeLV naturally infecting blood lilies without showing discernible symptoms is first reported.

Key words:Blood lily (Haemanthus multiflorus Martyn), Nerine latent virus (NeLV), Serological detection, RT-PCR detection, Phylogenetic analysis

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