Many viruses occur in apple (Malus domestica (Borkh.)), but no information is available on their seed transmissibility. Here, we report that six viruses infecting apple trees, namely, apple chlorotic leaf spot virus (ACLSV), apple green crinkle-associated virus (AGCaV), apple rubbery wood virus 2 (ARWV2), apple stem grooving virus (ASGV), apple stem pitting virus (ASPV), and citrus concave gum-associated virus (CCGaV) occur in seeds extracted from apple fruits produced by infected maternal trees. Reverse transcription polymerase chain reaction (RT-PCR) and quantitative RT-PCR (RT-qPCR) assays revealed the presence of these six viruses in untreated apple seeds with incidence rates ranging from 20% to 96%. Furthermore, ASPV was detected by RT-PCR in the flesh and peel of fruits produced by infected maternal trees, as well as from seeds extracted from apple fruits sold for fresh consumption. Finally, a large-scale seedling grow-out experiment failed to detect ACLSV, ASGV, or ASPV in over 1000 progeny derived from sodium hypochlorite surface sterilized seeds extracted from fruits produced by infected maternal trees, suggesting no detectable transmission via embryonic tissue. This is the first report on the seedborne nature of apple-infecting viruses.

Rapid apple decline is a phenomenon characterized by a weakening of young apple trees in high density orchards, often followed by their quick collapse. The nature of this phenomenon remains unclear. In this work, we investigated the root system architecture (RSA) of declining and non-declining apple trees in two orchards in New York State. High-density orchard A consisted of 4-year-old ‘Honeycrisp’ on ‘Malling 9 Nic29’, and conventional orchard B consisted of 8-year-old ‘Fuji’ on ‘Budagovsky 9’. In both orchards, a negative correlation (−0.4–−0.6) was observed between RSA traits and decline symptoms, suggesting that declining trees have weaker root systems. Scion trunk diameter at the graft union, total root length, and the length of fine and coarse roots were significantly (p < 0.05) reduced in declining trees in both orchards. Additionally, internal trunk necrosis at, above, and below the graft union was observed in declining trees in orchard A but not in orchard B. Finally, latent viruses were not associated with decline, as their occurrence was documented in declining and non-declining trees in orchard A, but not in orchard B. Together, these results showed weakened root systems of declining trees, suggesting that these trees may experience deficiencies in water and nutrient uptake, although distinct RSA and trunk health traits between the two orchards were noticeable.

Ceresini treehoppers are present in northern California vineyard ecosystems, including the closely related Spissistilus and Tortistilus (Hemiptera: Membracidae). These membracids are not direct pests of wine grapes, but S. festinus is a vector of grapevine red blotch virus (GRBV). No information is available on the ability of Tortistilus spp. to transmit GRBV. In this study, Tortistilus were collected on yellow panel cards across 102 vineyard sites and surrounding areas in Napa Valley, California, USA in 2021–2023. Specimens were morphotyped, sexed and tested for GRBV ingestion and acquisition by multiplex PCR or qPCR. Phylogenetic analysis of the partial sequence of mt-COI and ITS gene fragments of a subset of 40 Tortistilus specimens revealed clustering in a monophyletic clade with T. wickhami with the former barcode sequence. Only 6% (48/758) of the T. wickhami tested positive for GRBV, but none of the heads with salivary glands (0%, 0/50) of the dissected specimens tested positive for GRBV, indicating no virus acquisition. In contrast, half of the dissected heads with salivary glands of S. festinus (52%, 12/23), from the same collection vineyard sites, tested positive for GRBV. Together, our findings confirmed the presence of T. wickhami in northern California vineyards and suggested a dubious role of this treehopper as a vector of GRBV.

A sudden, unexplained decline and collapse of young apple trees on dwarfing and semi-dwarfing rootstocks has been reported across North America over the past decade. Although viruses have been detected in declining trees, no information is available on their potential causal role in the decline phenomenon. To this end, virus-inoculated apple trees were established in a high-density experimental orchard and monitored over five years. Tree decline was observed in year 4 (2022), resulting in 17% mortality, with declining trees exhibiting marked vascular tissue necrosis. However, none of the eight viruses and one viroid detected in the experimental orchard was significantly more prevalent in declining trees. Extreme temperature fluctuations in January 2022, followed by a severe water deficit in summer 2022, were recorded at the experimental orchard. Similar but distinct observations were made in a nearby commercial orchard with foliar nutrient imbalances documented in trees exhibiting symptoms of rapid decline. Together, our findings suggest that viruses are not primarily responsible for the rapid decline phenomenon and highlight the need for future work to investigate the roles of tree physiology and water stress in tree decline, as well as the potential efficacy of horticultural mitigation practices.

Viruses of cultivated apple ( Malus × domestica) are believed to be transmitted nearly exclusively via vegetative propagation, with few known biotic vectors and limited evidence of vertical transmission. To evaluate the seed transmission capabilities of six viruses and one viroid of apple, a large-scale seedling grow-out experiment was conducted using seeds harvested from 51 trees infected by several combinations of six viruses and one viroid. Virus detection via multiplex PCR-based amplicon sequencing followed by RT-qPCR validation demonstrated that citrus concave gum-associated virus and apple stem grooving virus were transmitted to seedlings at rates of 4.0% (32/792) and 0.3% (3/908), respectively. No evidence of seed transmission was obtained for apple chlorotic leaf spot virus, apple green crinkle-associated virus, apple hammerhead viroid, apple rubbery wood virus 2, or apple stem pitting virus. These findings document a previously unknown mode of transmission for two widely distributed apple viruses with direct implications for breeding programs, the selection of virus-free trees, and the exchange of germplasm.

The impacts of viruses on horticultural and fruit traits of apple trees were first characterized half a century ago, yet the responses to viruses of contemporary apple cultivars and rootstocks in high-density planting systems remains largely unknown. The aim of our study was to evaluate the effects of apple chlorotic leaf spot virus (ACLSV), apple stem pitting virus (ASPV), apple green crinkle-associated virus (AGCaV), and citrus concave gum-associated virus (CCGaV) on horticultural and fruit traits of young apple trees representing six commercially relevant scion-rootstock combinations in a high-density experimental orchard in New York. Analyses of horticultural traits showed statistically significant differences between infected and uninfected trees in trunk diameter (CCGaV) and terminal shoot growth (CCGaV, AGCaV); however, differences in means were small and inconsistent. Analyses of fruit traits revealed a statistically significant yield reduction of ‘Baigent’ Gala trees on ‘Geneva 935’ (G.935) rootstocks associated with ACLSV (-27%), ASPV (-16%) and CCGaV (-24%) in 2021, but not in 2022. Additionally, the number of concurrent virus infections per tree, as determined by multiplex PCR-based amplicon sequencing, was significantly negatively correlated with the yield of ‘Baigent’ Gala trees on G.935 in 2021 and significantly positively correlated with the firmness of fruits collected from trees on G.935 in both years. Together, these highly variable effects suggest that apple cultivars, rootstock genotypes, or combinations thereof, as well as variable environmental conditions, may influence the impacts of viruses in a high-density production system.