Renate Koenig

Detection of beet necrotic yellow vein virus strains, variants and mixed infections by examining single-strand conformation polymorphisms of immunocapture RT-PCR products

Single-strand conformation polymorphism analysis was found to be a powerful tool for rapidly assigning large numbers of beet necrotic yellow vein virus (BNYVV) isolates to a known strain group as well as for detecting mixed infections, minor variants or new strain groups. The prevalence of the B-type in Germany and France and the A-type in most other countries was confirmed. Minor variants with a very restricted distribution were detected occasionally. New rhizomania outbreaks in Great Britain were caused either by the A- or B-type or mixtures of both suggesting introduction of BNYVV from several sites abroad. An entirely different BNYVV type (P-type) was identified in a small area in France. Evidence for further strain groups in China was also obtained.

Molecular analyses of European A, B and P type sources of Beet necrotic yellow vein virus and detection of the rare P type in Kazakhstan

Summary. Nucleotide sequence analyses revealed that the genomes of the various European types of Beet necrotic yellow vein virus (BNYVV), i.e. the A, B and P types, are strongly conserved. Almost identical sequences were found, for instance, for A types originating from The Netherlands, Italy and former Yugoslavia; these sequences were also almost identical to those determined c. 15 years ago by Bouzoubaa et al. (1985; 1986 and 1987). This sequence stability of BNYVV types is in contrast to a pronounced sequence variability observed with Beet soil-borne pomovirus, another Polymyxa-transmitted sugar beet virus with rod-shaped particles. Sequences of RNA 1, 2 and 4 of BNYVV sources from Kazakhstan were almost identical to those of the P type of BNYVV which so far had been found only in a small area around the French town of Pithiviers. RNA 5, which in Europe is found also only in the Pithiviers area, was detected in the bait plants grown in two out of three soil samples from different fields in Kazakhstan. It closely resembled RNA 5 from the Pithiviers area, but was very different from RNA 5 from various East Asian BNYVV sources.

Occurrence of two different types of RNA-5-containing beet necrotic yellow vein virus in the UK

Summary.Two types of RNA-5-containing beet necrotic yellow vein virus (BNYVV) have been detected in the UK at different sites in Norfolk. On the basis of nucleotide (nt) sequence comparisons, one virus source (UK-MH) was clearly identified as P type BNYVV, a virus type that had previously only been detected in two widely separated parts of the world, France and Kazakhstan. The other virus source (UK-FF) has a complex genome composition. The analysed portions of its RNAs 2 and 4 are closely related to the corresponding portions in the RNAs of the East Asian A type isolate S, whereas those of its RNAs 1 and 3 resemble P type RNA 1 from Kazakhstan and European A type RNA 3, respectively. Interestingly, the P25 encoded on its RNA 3 has an unique TYHG tetrad in the highly variable amino acid positions 67–70. RNA 5 of the UK-FF BNYVV source shares properties with P type RNA 5, but also with East Asian types of RNA 5. The possible origin and epidemiology of BNYVV types is discussed.

First record of A and B type Beet necrotic yellow vein virus in sugar beets in Sweden

Natural infections of sugar beet with Beet necrotic yellow vein virus (BNYVV), the causal agent of rhizomania, have been detected for the first time in Sweden in two small areas, one on the island of Öland and one in the Southeastern part of Scania. Single strand conformation polymorphism analyses of PCR products revealed that the infections on Öland were produced by A type BNYVV, whereas those in Scania were caused by the B type. This suggests that BNYVV has been introduced into Sweden at least twice. Alternatively, the virus may have invaded sugar beet from unknown native hosts. BNYVV RNA 5 was not detected in the samples investigated.

Distribution of various types and P25 subtypes of Beet necrotic yellow vein virus in Germany and other European countries

The distribution of various Beet necrotic yellow vein virus (BNYVV) genotypes was studied using beet samples received from Germany and neighbouring countries. Almost exclusively B type BNYVV was detected in Germany, whereas in neighbouring countries BNYVV A types with different compositions of the amino acid tetrad in positions 67–70 of the RNA-3-encoded P25 are widely distributed. Neither A types nor the P type have been able to become established in Germany in the past decades, although there must have been many opportunities for their introduction from neighbouring countries. In one field, however, an RNA-5-containing BNYVV genotype closely resembling the Chinese isolate Har4 was found.

Iranian beet necrotic yellow vein virus (BNYVV): pronounced diversity of the p25 coding region in A-type BNYVV and identification of P-type BNYVV lacking a fifth RNA species.

Beet necrotic yellow vein virus (BNYVV) was detected in 288 of the 392 samples collected in Iran. A-type BNYVV was detected most frequently. The p25 coding region on BNYVV RNA-3 was amplified by RT-PCR and sequenced. Nine different variants of the highly variable amino acid tetrad at positions 67–70 of p25 were identified, i.e. ACHG, AHHG, AYHG, ALHG, AFHR, AFHG, AHYG, VLHG and VHHG. These are more different tetrad variants than have been reported from any other country. The first three variants were found most commonly. In 23 out of the 288 BNYVV-positive samples, we detected P-type BNYVV that had previously been identified only in France, Kazakhstan and recently in the UK. Surprisingly, none of these samples contained the fifth RNA species usually associated with P-type BNYVV in other countries. As in other BNYVV P-type sources, the p25 amino acid tetrad in positions 67–70 of the Iranian P-type consists of SYHG.

The complete genome sequences of a Peruvian and a Colombian isolate of Andean potato latent virus and partial sequences of further isolates suggest the existence of two distinct potato-infecting tymovirus species

The complete genomic RNA sequences of the tymovirus isolates Hu and Col from potato which originally had been considered to be strains of the same virus species, i.e. Andean potato latent virus (APLV), were determined by siRNA sequencing and assembly, and found to share only c. 65% nt sequence identity. This result together with those of serological tests and comparisons of the coat protein gene sequences of additional tymovirus isolates from potato suggest that the species Andean potato latent virus should be subdivided into two species, i.e. APLV and Andean potato mild mosaic virus (APMMV). Primers were designed for the broad specificity detection of both viruses.

New isolates of carnation Italian ringspot virus differ from the original one by having replication-associated proteins with a typical tombusvirus-like N-terminus and by inducing peroxisome- rather than mitochondrion-derived multivesicular bodies

Five new isolates of carnation Italian ringspot virus (CIRV) from cherry trees, Gypsophila and surface water differ from the original carnation isolate (CIRV-car) and also from Pelargonium necrotic spot virus (PelNSV) by having an ORF 1/ORF1-RT with a typical tombusvirus-like 5′end and by inducing the formation of peroxisome- rather than mitochondrion-derived multivesicular bodies (MVBs). This supports with natural isolates earlier conclusions reached by others with artificially produced hybrid viruses that the 5′end of ORF 1 determines from which organelle the MBVs will be derived. CIRV-car might have resulted from a natural recombination event with genome elements of a PelNSV-like virus.

Molecular and serological characterization of an Iranian isolate of Beet black scorch virus

An isolate of Beet black scorch virus (BBSV) was obtained from Iranian sugar beet roots. Its genome organization closely resembles that of the previously described Chinese and North American isolates, but the nucleotide sequences of the three isolates differ considerably. Most of the nucleotide exchanges, however, are silent, and the Iranian and the Chinese isolates were serologically indistinguishable. Beets infected by the Iranian BBSV did not show black scorch symptoms, but severe root beardedness. This might have been caused by BBSV or the simultaneously present beet necrotic yellow vein virus, or both together.

Molecular characterization of isolates of anagyris vein yellowing virus, plantago mottle virus and scrophularia mottle virus – comparison of various approaches for tymovirus classification

Summary.The complete nucleotide sequences were determined for the genomic RNAs of three tymoviruses, i.e. isolates of anagyris vein yellowing virus (AVYV), plantago mottle virus (PlMoV) and scrophularia mottle virus (SrMV) which are all serologically closely related to ononis yellow mosaic virus (ibid) and to Nemesia ring necrosis virus (NeRNV), a recently described recombinant virus which is widely spread in commercially grown ornamental plant species belonging to the Scrophulariaceae. Total nucleotide and coat protein amino acid sequence identities revealed similar groupings in the genus tymovirus as serological studies did. The latter, however, tended to suggest much closer relationships than the molecular data and may fail to recognise the distinctiveness of new tymovirus species. The usefulness of various species demarcation criteria for the classification of tymoviruses is discussed.