“Cross-protection”, a nearly 100 years-old virological term, is suggested to be changed to “close protection”. Evidence for the need of such change has accumulated over the past six decades from the laboratory experiments and field tests conducted by plant pathologists and plant virologists working with different plant viruses, and, in particular, from research on Citrus tristeza virus (CTV). A direct confirmation of such close protection came with the finding that “pre-immunization” of citrus plants with the variants of the T36 strain of CTV but not with variants of other virus strains was providing protection against a fluorescent protein-tagged T36-based recombinant virus variant. Under natural conditions close protection is functional and is closely associated both with the conservation of the CTV genome sequence and prevention of superinfection by closely similar isolates. It is suggested that the mechanism is primarily directed to prevent the danger of virus population collapse that could be expected to result through quasispecies divergence of large RNA genomes of the CTV variants continuously replicating within long-living and highly voluminous fruit trees. This review article provides an overview of the CTV cross-protection research, along with a discussion of the phenomenon in the context of the CTV biology and genetics.

The family Closteroviridae consists of two genera, Closterovirus and Ampelovirus with monopartite genomes transmitted respectively by aphids and mealybugs and the Crinivirus with bipartite genomes transmitted by whiteflies. The Closteroviridae consists of more than thirty virus species, which differ considerably in their phytopathological significance. Some, like Beet yellows virus (BYV) and Citrus tristeza virus (CTV) were associated for many decades with their respective hosts, sugar beets and citrus. Others, like the grapevine leafroll-associated ampeloviruses 1, and 3 were also associated with their grapevine hosts for long periods; however difficulties in virus isolation hampered their molecular characterization. The majority of the recently identified Closteroviridae were probably associated with their vegetative propagated host plants for long periods and only detected through the considerable advances in dsRNA isolation and sequencing of PCR amplified replicons. Molecular characterization of CTV and several other Closteroviridae revealed that, in addition to genomic and subgenomic RNAs, infected plants contain several different subviral defective RNAs (dRNAs). The roles and biological functions of dRNAs associated with Closteroviridae remain terra incognita.

The control of tristeza quick decline (QD) of citrus is based on the use of rootstocks that are tolerant or resistant to the Citrus tristeza virus (CTV), but some of them show bio-agronomic limits. The application of cross-protection (CP) has been insufficiently explored. The present study examined the possibility of QD control by cross-protection (CP) following reports showing the dependence of the CP strategy on the close genetic relationships between the protective and challenging CTV isolates. Taking advantage of deep sequencing technologies, we located six naturally infected trees harboring no-seedling yellow (no-SY) and no QD decline (mild) VT isolates and used these for challenge inoculation with three QD VT isolates. Symptom monitoring showed that all six Sicilian mild no-SY isolates, based on their genomic relatedness and mild symptoms reactions, provide effective protection against the three severe local VT isolates. The differences between the six mild and three severe isolates were confined to just a few nucleotide variations conserved in eight positions of three CTV genes (p23, p33, and Orf1a). These results confirm that the superinfection exclusion (SIE mechanism) depends on close genetic relatedness between the protective and challenging severe VT strain isolates. Ten years of investigation suggest that CP could turn into an efficient strategy to contain CTV QD infections of sweet orange trees on SO rootstock.

Citrus production in the Mediterranean area is of considerable importance, in both cultural and economic terms, and the viability of the industry greatly depends on proper phytosanitary management. In this review, we focus on exotic and emerging dangerous citrus viruses that have still not been reported in the countries of the Mediterranean area, that are not yet regulated or that are restricted to certain small areas. We also discuss the contribution that old and new technologies may offer for valuable surveys aimed at promoting the adoption and sharing of better control measures and the production of pathogen-tested citrus trees and rootstocks.

The devastating tristeza epidemic swept through South American citrus groves in the 1930s and subsequently spread to most citrus-growing regions worldwide, causing varying degrees of damage and prompting significant changes in citrus cultivation practices. The causal agent of the disease, citrus tristeza virus (CTV), belongs to the genus Closterovirus in the family Closteroviridae. CTV virions are approximately two microns long and possess the largest known positive-strand RNA genome in plants, spanning 19.3 kb. The history of tristeza disease and CTV’s molecular biology and taxonomic relationships have been extensively reviewed in the scientific literature. This paper primarily focuses on the author’s personal experiences with tristeza disease and its causal agent over the past six decades. The journey began during a period when biological indexing was the primary diagnostic tool. It later progressed through the isolation of purified CTV particles, which served as a practical diagnostic tool for CTV suppression efforts in Israel during the 1970s. However, biological indexing was first replaced by electron microscopy, followed by ELISA procedures; both were eventually abandoned after it was discovered that many ELISA-positive infections were caused by symptomless CTV isolates, even on trees grafted onto sour orange rootstocks. In retrospect, my work on CTV can be categorized into three main phases. It began with the biological phase, inherited from earlier generations of citrus virologists, followed by the isolation and partial characterization of CTV virions, and culminated in the genomic era. While we live in an age of remarkable biotechnological achievements, my recommendation for future CTV research is to integrate both biological and genomic approaches rather than viewing them as mutually exclusive. This is particularly important for economically significant pathogens such as CTV, which should be studied continuously as both biological agents and molecular pathogens.