Sporadic outbreaks of potato yellow vein disease (PYVD) were first observed in the early 1940s by potato growers in Antioquia, Colombia. Long known to be transmitted by the greenhouse whitefly (Trialeurodes vaporariorum), the precise identity of its causal agent (presumably viral in nature) has remained obscure. Here, we present evidence that a closterovirus with a bipartite genome, potato yellow vein virus (PYVV), is associated with PYVD. Electrophoretic analysis revealed that diseased tissue contains 4-5 disease-specific dsRNAs ranging in size from c. 9 000-1 800 bp. RT-PCR reactions containing pairs of degenerate primers directed against conserved motifs in the closterovirus heat-shock protein homologue produced products of the expected sizes. Comparison of the corresponding amino acid sequences revealed striking similarities between PYVV and two bipartite, whitefly-transmitted criniviruses, Cucurbit yellow stunting disorder and Tomato chlorosis viruses. Epidemiological surveys carried out in Rionegro, Colombia identified Polygonum mepalense, Polygonum spp., Rumex obtusifolium, Tagetes spp., and Catharanthus roseus as potential viral reservoirs. PYVV is transmitted through tubers, and visual symptoms alone cannot be used to determine infection status. A sensitive hybridisation-based assay for PYVV has been developed for use in seed certification programmes.

Double-stranded RNA preparations produced from potato plants graft-inoculated with a Peruvian isolate of Potato yellow vein virus (PYVV; genus Crinivirus, family Closteroviridae) contain five RNA species denoted RNA 1, RNA 2, RNA 3, x and y of approximately 8, 5·3, 3·8, 2·0 and 1·8 kbp, respectively. The complete nucleotide sequences of PYVV RNAs 1, 2 and 3 and Northern hybridization analysis showed that PYVV RNA 1 contained the replication module and an additional open reading frame (p7), while two distinct species, RNAs 2 and 3, contain the Closteroviridae hallmark gene array. Pairwise comparisons and phylogeny of genome-encoded proteins showed that PYVV shares significant homology with other criniviruses but is most closely related to the Trialeurodes vaporariorum-vectored Cucumber yellows virus. Secondary structure prediction of the 3′-untranslated regions of all three PYVV RNAs revealed four conserved stem–loop structures and a 3′-terminal pseudoknot structure, also predicted for all fully characterized members of the genus Crinivirus and some members of the genera Closterovirus and Ampelovirus.

The inherent ability of seeds (orthodox, intermediate, and recalcitrant seeds and vegetative propagules) to serve as carriers of pests and pathogens (hereafter referred to as pests) and the risk of transboundary spread along with the seed movement present a high-risk factor for international germplasm distribution activities. Quarantine and phytosanitary procedures have been established by many countries around the world to minimize seed-borne pest spread by screening export and import consignments of germplasm. The effectiveness of these time-consuming and cost-intensive procedures depends on the knowledge of pest distribution, availability of diagnostic tools for seed health testing, qualified operators, procedures for inspection, and seed phytosanitation. This review describes a unique multidisciplinary approach used by the CGIAR Germplasm Health Units (GHUs) in ensuring phytosanitary protection for the safe conservation and global movement of germplasm from the 11 CGIAR genebanks and breeding programs that acquire and distribute germplasm to and from all parts of the world for agricultural research and food security. We also present the challenges, lessons learned, and recommendations stemming from the experience of GHUs, which collaborate with the national quarantine systems to export and distribute about 100,000 germplasm samples annually to partners located in about 90 to 100 countries. Furthermore, we describe how GHUs adjust their procedures to stay in alignment with evolving phytosanitary regulations and pest risk scenarios. In conclusion, we state the benefits of globally coordinated phytosanitary networks for the prevention of the intercontinental spread of pests that are transmissible through plant propagation materials.

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