New isolate of sweet potato virus 2 from Ipomoea nil: molecular characterization, codon usage bias, and phylogenetic analysis based on complete genome.

BACKGROUND: Viral diseases of sweet potatoes are causing severe crop losses worldwide. More than 30 viruses have been identified to infect sweet potatoes among which the sweet potato latent virus (SPLV), sweet potato mild speckling virus (SPMSV), sweet potato virus G (SPVG) and sweet potato virus 2 (SPV2) have been recognized as distinct species of the genus Potyvirus in the family Potyviridae. The sweet potato virus 2 (SPV2) is a primary pathogen affecting sweet potato crops. METHODS: In this study, we detected an SPV2 isolate (named SPV2-LN) in Ipomoea nil in China. The complete genomic sequence of SPV2-LN was obtained using sequencing of small RNAs, RT-PCR, and RACE amplification. The codon usage, phylogeny, recombination analysis and selective pressure analysis were assessed on the SPV2-LN genome. RESULTS: The complete genome of SPV2-LN consisted of 10,606 nt (GenBank No. OR842902), encoding 3425 amino acids. There were 28 codons in the SPV2-LN genome with a relative synonymous codon usage (RSCU) value greater than 1, of which 21 end in A/U. Among the 12 proteins of SPV2, P3 and P3N-PIPO exhibited the highest variability in their amino acid sequences, while P1 was the most conserved, with an amino acid sequence identity of 87-95.3%. The phylogenetic analysis showed that 21 SPV2 isolates were clustered into four groups, and SPV2-LN was clustered together with isolate yu-17-47 (MK778808) in group IV. Recombination analysis indicated no major recombination sites in SPV2-LN. Selective pressure analysis showed dN/dS of the 12 proteins of SPV2 were less than 1, indicating that all were undergoing negative selection, except for P1N-PISPO. CONCLUSION: This study identified a sweet potato virus, SPV2-LN, in Ipomoea nil. Sequence identities and genome analysis showed high similarity between our isolate and a Chinese isolate, yu-17-47, isolated from sweet potato. These results will provide a theoretical basis for understanding the genetic evolution and viral spread of SPV2.

Complete genome sequence of a novel varicosavirus infecting tall morning glory (Ipomoea purpurea).

A novel varicosa-like virus was identified in a tall morning glory (Ipomoea purpurea) plant by high-throughput sequencing and tentatively named "morning glory varicosavirus" (MGVV). The complete genome of MGVV contains two segments of negative-sense single-stranded RNA of 6409 (RNA1) and 5288 (RNA2) nucleotides. RNA1 encodes a 224.3-kDa large protein (224K), and RNA2 encodes four putative proteins of 48.6 kDa (49K), 46.4 kDa (46K), 35.7 kDa (36K), and 36.8 kDa (37K), respectively. The 224K and 49K proteins show amino acid sequence similarity to the large protein (39.4%) and the 49K protein (22.6%), respectively, of red clover-associated varicosavirus, and the 36K protein shares 19.6% amino acid sequence similarity with protein 3 of lettuce big-vein associated virus. The 46K and 37K proteins share no significant sequence similarity to known functional viral sequences. Phylogenetic analysis based on the large protein of MGVV and other rhabdoviruses showed that MGVV clustered with the varicosaviruses. These analyses indicate that MGVV is a novel member of the genus Varicosavirus in the family Rhabdoviridae.

Stylet penetration behaviors of Myzus persicae (Hemiptera: Aphididae) on four Ipomoea spp. infected or noninfected with sweet potato potyviruses.

Myzus persicae (Sulzer) is an efficient vector of potyviruses in sweet potato, Ipomoea batatas (L.). These potyviruses also infect members of the morning glory family Ipomoea cordatotriloba Dennstedt and Ipomoea hederacea Jacqin commonly found within or around sweet potato fields. Infection of sweet potato with potyviruses increases the intrinsic rate of increase of M. persicae. Thus, from the epidemiological stand point, virus infection can modify vector population dynamics, and therefore increase virus spread. To better understand this, stylet penetration behaviors of M. persicae on virus infected and noninfected sweet potato cvs. 'Beauregard' and 'Evangeline', as well as morning glory plants I. cordatotriloba and I. hederacea were monitored. Stylet penetration behaviors associated with nonpersistent virus transmission such as time to first intracellular puncture (potential drop), number of potential drops, duration of potential drop, duration of potential drop subphase II-3, and number of potential drops with subphase II-3 pulses were significantly increased on virus-infected compared with noninfected Beauregard, but greatly reduced on virus-infected compared with noninfected I. hederacea plants. Stylet penetration behaviors associated with host acceptance such as reduced nonprobing duration and nonprobing events were greater on virus-infected compared with noninfected Beauregard plants. In contrast, on Evangeline, I. cordatotriloba and I. hederacea stylet penetration behaviors by M. persicae indicate it had less preference for virus-infected compared with noninfected plants.

Population dynamics of three aphid species (Hemiptera: Aphididae) on four Ipomoea spp. infected or noninfected with sweetpotato potyviruses.

Three aphid species, Aphis gossypii Glover and Myzus persicae (Sulzer) (efficient sweetpotato potyvirus vectors) and Rhopalosiphum padi (L.) (an inefficient vector), are commonly found in sweet potato, Ipomoea batatas (L.), in Louisiana. Field-grown sweet potatoes are naturally infected with several potyviruses: Sweet potato feathery mottle virus, Sweet potato virus G, and Sweet potato virus 2. Thus, these aphids commonly encounter virus-infected hosts. What is not known is how each of these aphids responds to sweet potato, either infected or virus-free. The objectives of this study were to 1) determine if these aphid species can colonize mixed virus-infected sweet potato 'Beauregard', and if so, 2) determine the effects of virus infection on the population dynamics of each aphid. A. gossypii failed to larviposit and R. padi deposited a single nymph that died within a day on mixed virus-infected Beauregard. M. persicae larviposited and colonized Beauregard and further life-table analyses were warranted. M. persicae had a significantly greater reproduction on sweet potato cultivars Beauregard and 'Evangeline' with mixed virus infection compared with noninfected plants. On morning glory species, Ipomoea cordatotriloba (Dennestedt) and Ipomoea hederacea (Jacquin), M. persicae had a significantly lower reproduction on Sweet potato feathery mottle virus-infected compared with noninfected plants.

Epidemiology of soybean vein necrosis-associated virus.

Soybean vein necrosis-associated virus has been linked to an emerging soybean disease in the United States and Canada. Virus distribution and population structure in major growing areas were evaluated. Data were employed to design and develop sensitive detection protocols, able to detect all virus isolates available in databases. The host range for the virus was assessed and several species were found to sustain virus replication, including ivyleaf morning glory, a common weed species in soybean-growing areas in the United States. Koch's postulates were fulfilled using soybean thrips and transmission efficiency was determined. This article provides significant insight into the biology of the most widespread soybean virus in the United States.

Natural association of two different betasatellites with Sweet potato leaf curl virus in wild morning glory (Ipomoea purpurea) in India.

Wild morning glory (Ipomoea purpurea) was observed to be affected by leaf curl and yellow vein diseases during summer-rainy season of 2009 in New Delhi, India. The virus was experimentally transmitted through whitefly, Bemisia tabaci to I. purpurea that reproduced the two distinct symptoms. Sequence analysis of multiple full-length clones obtained through rolling circle amplification from the leaf curl and yellow vein samples showed 91.8-95.3% sequence identity with Sweet potato leaf curl virus (SPLCV) and the isolates were phylogenetically distinct from those reported from Brazil, China, Japan and USA. Interestingly, two different betasatellites, croton yellow vein mosaic betasatellite and papaya leaf curl betasatellite were found with SPLCV in leaf curl and yellow vein diseases of I. purpurea, respectively. This study is the first report of occurrence of SPLCV in wild morning glory in India. SPLCV was known to infect other species of morning glory; our study revealed that I. purpurea, a new species of morning glory was a natural host of SPLCV. To date, betasatellite associated with SPLCV in Ipomoea spp. is not known. Our study provides evidence of natural association of two different betasatellites with SPLCV in leaf curl and yellow vein diseases of I. purpurea.

Sweepoviruses cause disease in sweet potato and related Ipomoea spp.: fulfilling Koch's postulates for a divergent group in the genus begomovirus.

Sweet potato (Ipomoea batatas) and related Ipomoea species are frequently infected by monopartite begomoviruses (genus Begomovirus, family Geminiviridae), known as sweepoviruses. Unlike other geminiviruses, the genomes of sweepoviruses have been recalcitrant to rendering infectious clones to date. Thus, Koch's postulates have not been fullfilled for any of the viruses in this group. Three novel species of sweepoviruses have recently been described in Spain: Sweet potato leaf curl Lanzarote virus (SPLCLaV), Sweet potato leaf curl Spain virus (SPLCSV) and Sweet potato leaf curl Canary virus (SPLCCaV). Here we describe the generation of the first infectious clone of an isolate (ES:MAL:BG30:06) of SPLCLaV. The clone consisted of a complete tandem dimeric viral genome in a binary vector. Successful infection by agroinoculation of several species of Ipomoea (including sweet potato) and Nicotiana benthamiana was confirmed by PCR, dot blot and Southern blot hybridization. Symptoms observed in infected plants consisted of leaf curl, yellowing, growth reduction and vein yellowing. Two varieties of sweet potato, 'Beauregard' and 'Promesa', were infected by agroinoculation, and symptoms of leaf curl and interveinal loss of purple colouration were observed, respectively. The virus present in agroinfected plants was readily transmitted by the whitefly Bemisia tabaci to I. setosa plants. The progeny virus population present in agroinfected I. setosa and sweet potato plants was isolated and identity to the original isolate was confirmed by sequencing. Therefore, Koch's postulates were fulfilled for the first time for a sweepovirus.

Distinct cavemoviruses interact synergistically with sweet potato chlorotic stunt virus (genus Crinivirus) in cultivated sweet potato.

Two serologically unrelated sweet potato viruses causing symptoms of vein clearing in the indicator plant Ipomoea setosa were isolated and their genomes have been sequenced. They are associated with symptomless infections in sweet potato but distinct vein-clearing symptoms and higher virus titres were observed when these viruses co-infected with sweet potato chlorotic stunt virus (SPCSV), a virus that is distributed worldwide and is a mediator of severe virus diseases in this crop. Molecular characterization and phylogenetic analysis revealed an overall nucleotide identity of 47.6 % and an arrangement of the movement protein and coat protein domains characteristic of members of the genus Cavemovirus, in the family Caulimoviridae. We detected both cavemoviruses in cultivated sweet potato from East Africa, Central America and the Caribbean islands, but not in samples from South America. One of the viruses characterized showed a similar genome organization as, and formed a phylogenetic sublineage with, tobacco vein clearing virus (TVCV), giving further support to the previously suggested separation of TVCV, and related viral sequences, into a new caulimovirid genus. Given their geographical distribution and previous reports of similar but yet unidentified viruses, sweet potato cavemoviruses may co-occur with SPCSV more often than previously thought and they could therefore contribute to the extensive yield losses and cultivar decline caused by mixed viral infections in sweet potato.

Novel begomovirus species of recombinant nature in sweet potato (Ipomoea batatas) and Ipomoea indica: taxonomic and phylogenetic implications.

Viral diseases occur wherever sweet potato (Ipomoea batatas) is cultivated and because this crop is vegetatively propagated, accumulation and perpetuation of viruses can become a major constraint for production. Up to 90% reductions in yield have been reported in association with viral infections. About 20 officially accepted or tentative virus species have been found in sweet potato and other Ipomoea species. They include three species of begomoviruses (genus Begomovirus, family Geminiviridae) whose genomes have been fully sequenced. In this investigation, we conducted a search for begomoviruses infecting sweet potato and Ipomoea indica in Spain and characterized the complete genome of 15 isolates. In addition to sweet potato leaf curl virus (SPLCV) and Ipomoea yellowing vein virus, we identified three new begomovirus species and a novel strain of SPLCV. Our analysis also demonstrated that extensive recombination events have shaped the populations of Ipomoea-infecting begomoviruses in Spain. The increased complexity of the unique Ipomoea-infecting begomovirus group, highlighted by our results, open new horizons to understand the phylogeny and evolution of the family Geminiviridae.