Kenta Tomimura

The Dual Role of the Potyvirus P3 Protein of Turnip mosaic virus as a Symptom and Avirulence Determinant in Brassicas

Two isolates of the potyvirus Turnip mosaic virus (TuMV), UK 1 and CDN 1, differ both in their general symptoms on the susceptible propagation host Brassica juncea and in their ability to infect B. napus lines possessing a variety of dominant resistance genes. The isolate CDN 1 produces a more extreme mosaic in infected brassica leaves than UK 1 and is able to overcome the resistance genes TuRB01, TuRB04, and TuRB05. The resistance gene TuRB03, in the B. napus line 22S, is effective against CDN 1 but not UK 1. The nucleic acid sequences of the UK 1 and CDN 1 isolates were 90% identical. The C-terminal half of the P3 protein was identified as being responsible for the differences in symptoms in B. juncea. A single amino acid in the P3 protein was found to be the avirulence determinant for TuRB03. Previous work already has identified the P3 as an avirulence determinant for TuRB04. Our results increase the understanding of the basis of plant-virus recognition, show the importance of the potyviral P3 gene as a symptom determinant, and provide a role in planta for the poorly understood P3 protein in a normal infection cycle.

The phylogeny of Turnip mosaic virus ; comparisons of 38 genomic sequences reveal a Eurasian origin and a recent emergence in east Asia

The genomes of a representative world‐wide collection of 32 Turnip mosaic virus (TuMV) isolates were sequenced and these, together with six previously reported sequences, were analysed. At least one‐fifth of the sequences were recombinant. In phylogenetic analyses, using genomic sequences of Japanese yam mosaic virus as an outgroup, the TuMV sequences that did not show clear recombination formed a monophyletic group with four well‐supported lineages. These groupings correlated with differences in pathogenicity and provenance; the sister group to all others was of Eurasian B‐strain isolates from nonbrassicas, and probably represents the ancestral TuMV population, and the most recently ‘emerged’ branch of the population was probably that of the BR‐strain isolates found only in east Asia. Eight isolates, all from east Asia, were clear recombinants, probably the progeny of recent recombination events, whereas a similar number, from other parts of the world, were seemingly older recombinants. This difference indicates that the presence of clear recombinants in a subpopulation may be a molecular signature of a recent ‘emergence’.

Effect of the Timing of Fungicide Application on Fusarium Head Blight and Mycotoxin Contamination in Wheat

Fungicide application to control Fusarium head blight (FHB) and accompanying mycotoxin contamination in wheat is generally performed at anthesis because wheat is most susceptible to FHB around this stage. In this study, we evaluated the effect of the timing of fungicide application on FHB and mycotoxin (deoxynivalenol and nivalenol) accumulation in wheat based on our previous finding that the late period of grain development (beyond 20 days after anthesis [DAA]) is important to determine the final toxin contamination level in wheat. Thiophanate-methyl fungicide was tested under artificial inoculation conditions in which moisture and inoculum spores were provided throughout the testing period. Eight treatments differing in application timing (anthesis, 10, 20, and 30 DAA) and in the number of applications (0 to 2) were tested for 2 years. The results indicated that fungicide application timing differentially affects FHB (disease) and mycotoxin concentration. Fungicide application at 20 DAA reduced mycotoxin concentration in matured grain without reducing FHB severity, whereas application at anthesis was crucial for reducing FHB. These results and our previous findings suggest that around 20 DAA (late milk stage) is a potentially critical timing for mycotoxin control in wheat.

Differentiation of “Candidatus Liberibacter asiaticus” Isolates by Variable-Number Tandem-Repeat Analysis

ABSTRACT Four highly polymorphic simple sequence repeat (SSR) loci were selected and used to differentiate 84 Japanese isolates of “Candidatus Liberibacter asiaticus.” The Neis measure of genetic diversity values for these four SSRs ranged from 0.60 to 0.86. The four SSR loci were also highly polymorphic in four isolates from Taiwan and 12 isolates from Indonesia.

Evidence of a new Tomato yellow leaf curl virus in Japan and its detection using PCR

A new isolate of Tomato yellow leaf curl virus (TYLCV) has been identified from tomato plants in Kochi Prefecture in Japan and designated TYLCV-[Tosa]. The complete nucleotide sequence of the isolate was determined and found to consist of 2781 nt. In phylogenetic analyses of entire nucleotide sequences, TYLCV-[Tosa] was delineated as a single branch and was more closely related to TYLCV-[Almeria] than TYLCV isolates Ng, Sz, or Ai reported in Japan, which had spread since 1996. Isolate TYLCV-[Tosa] is suggested to be a newly introduced, novel isolate of TYLCV that dispersed into Kochi Prefecture. In addition, a rapid method using the polymerase chain reaction to separate TYLCV isolates into four genetic groups was established. This method would be useful for reliable diagnosis based on genetic differences among isolates of TYLCV.

Sequence homogeneity of the ψserA-trmU-tufB-secE-nusG-rplKAJL-rpoB gene cluster and the flanking regions of 'Candidatus Liberibacter asiaticus' isolates around Okinawa Main Island in Japan.

In Japan and Southeast Asia, ‘Candidatus Liberibacter asiaticus’ (Las) is the dominant causal agent of citrus greening (huanglongbing) disease. Using PCR techniques, we determined the 11168-nucleotide sequence of the ψserA-trmU-tufB-secE-nusG-rplKAJL-rpoB gene cluster and the flanking regions for 51 Japanese, four Taiwanese, four Indonesian, and three Vietnamese isolates of Las. The sequence is identical in 62 isolates collected from Japan, Taiwan, Indonesia, and Vietnam, except for nucleotide substitutions at 11 positions. Some Las isolates from Sakishima Islands near Taiwan had unique nucleotide mutations, but all Las isolates around Okinawa Main Island were homologous. On the basis of the pattern of single nucleotide polymorphisms (SNPs) of the 11 nucleotide substitutions, the 62 Las isolates from Japan, Taiwan, Indonesia, and Vietnam could be divided into 12 pattern groups, and the 51 Japanese isolates consisted of six patterns. The results suggested that one unique genetic group is dominant around Okinawa Main Island, whereas several different are commonly distributed around islands near Taiwan.

Evolutionary rates and genetic diversities of mixed potyviruses in Narcissus

There is no attempt to evaluate evolutionary rates, timescales and diversities of viruses collected from mixedly infected hosts in nature. Plants of the genus Narcissus are a monocotyledon and are susceptible to several viruses. In this study, narcissus plants (Narcissus tazetta var. chinensis) showing mosaic or striping leaves were collected in Japan, and these were investigated for potyvirus infections using potyvirus-specific primers. Individual narcissus plants were found frequently to be mixedly infected with different potyviruses, different isolates and quasispecies of same virus. The viruses were potyviruses and a macluravirus in the family Potyviridae, namely Narcissus late season yellows virus (NLSYV), Narcissus yellow stripe virus (NYSV), Narcissus degeneration virus (NDV), Cyrtanthus elatus virus A (CyEVA) and Narcissus latent virus (NLV). Genetic diversities of coat protein coding region of different virus species were different; NYSV and CyEVA were most diverse whereas NDV was least. Evolutionary rates of all five narcissus viruses were 1.33-7.15×10-3nt/site/year and were similar. The most recent common ancestors (TMRCAs) varied between virus species; NYSV and CyEVA were the oldest whereas NDV was the youngest. Thus, the oldness of TMRCAs of the viruses correlated well with the greatness of nucleotide diversities.

Development of simple sequence repeat markers for the Japanese population of the Asian citrus psyllid, Diaphorina citri (Hemiptera: Psyllidae)

We developed nine simple sequence repeat (SSR) markers for the Japanese population of Diaphorina citri Kuwayama, which is known as the vector for citrus greening disease or huanglongbing, for use in population genetic studies. Analysis of 50 individuals collected from Tokunoshima Island showed that allelic diversity ranged from 5 to 24 alleles per locus, and observed and expected heterozygosities ranged from 0.100 to 0.939 and from 0.222 to 0.926, respectively. These SSR markers can be used to understand the population genetics of D. citri, and the obtained information may be useful in the estimation of population genetic structure, gene flow and invasion routes of D. citri.

Corrigendum to “Comparisons of the genetic structure of populations of Turnip mosaic virus in West and East Eurasia” [Virology 330 (2004) 408-423]

Laboratory of Plant Virology, Faculty of Agriculture, Saga University, Saga 840-8502, Japan Institute of Plant Molecular Biology, Academy of Sciences of the Czech Republic, Ceske Budejovice, Czech Republic Plant Pathology Laboratory, Faculty of Agriculture, Aristotle University of Thessaloniki, 540 06 Thessaloniki, Greece Warwick HRI, University of Warwick, Wellesbourne, Warwick CV35 9EF, UK School of Botany and Zoology, Australian National University, Canberra, A. C. T. 0200, Australia