Identification and characterization of Peach latent mosaic viroid and Hop stunt viroid in different peach cultivars showing dapple fruit, fruit yellow mosaic and cracked suture symptoms

From the early 1990s, a fruit peach syndrome characterized mainly by small discoloured spots (dapple fruit) and/or yellow areas on the skin (yellow mosaic), cracked suture and deformations was identified in most commercial orchards in the Emilia Romagna region (Northern Italy). In the past, Peach latent mosaic viroid (PLMVd) and Hop stunt viroid (HSVd) have been detected in trees with symptomatic fruits. In order to ascertain the presence and spread of these two viroids, symptomatic fruit samples were collected from five different cultivars: ‘Royal Glory’, ‘Crimson Lady’, ‘Grenat’, ‘Diamond Princess’ and ‘Laura’. Dapple fruit symptoms affected all cultivars, ‘Grenat’ samples also showed evident yellow mosaic and fruit deformation, and ‘Royal Glory’ severe cracked sutures. The results showed a large diffusion of the two viroids, mainly in mixed infections. Anvaluation of the role the viroids could play in symptom expression has been complicated by the high number of samples infected by both viroids (60%). Nonetheless, PLMVd was confirmed to be strictly associated with the yellow mosaic, cracked suture and fruit deformation symptoms. The aetiological origin of the dapple fruit disease, however, seems to be more complicated, since in the ‘Diamond Princess’, only PLMVd has been found to be associated with the symptoms, whereas in all other cultivars, the presence of HSVd could have influenced the symptom expression. Moreover, the molecular characterization of some PLMVd isolates does not show any correlation between nucleotide sequence and symptoms although new PLMVd variants were identified. Keywords: peach fruit symptoms, PLMVd, HSVd, mixed infectio

The molecular characterization of HSVd isolates associated with dapple fruit and fruit rugosity in plum seedlings suggests a possible role of breeding in viroid dissemination

In a wide range of hosts, the infection caused by Hop stunt viroid (HSVd) appears to be latent, whereas in some others it is frequently pathogenic. In this work, the presence of HSVd has been found to be associated with symptoms of dapple fruit and fruit rugosity in plum seedlings obtained from cross breeding for quality. Symptomatic and symptomless plum seedling samples have been analyzed for the presence of the principal stone fruit viroids and viruses. HSVd was found in all symptomatic samples, whereas no other viruses or viroids were found in the analyzed samples with the exception of ACLSV, which was detected rarely. The RNAs of all HSVd isolates have been cloned and sequenced. The sequence analysis showed a high percentage of homology among the isolates, making it possible to hypothesize a potential unique origin of the infection. For this purpose, those plants used in breeding as pollen donors have been analyzed. The results showed that the same HSVd isolate was also present in the parental plants, both in the leaves and pollen, suggesting a possible role of breeding in the dissemination of the viroid.Keywords: Plum, seedlings, fruit rugosity, dapple fruit, HSVd, polle

Improvement of the reverse transcription loop mediated isothermal amplification (RTLAMP) method for the detection of Peach latent mosaic viroid (PLMVd)

Peach latent mosaic viroid (PLMVd) is the most known peach viroid. Among the diagnostic techniques used for its detection, the most recent described being the reverse transcription loop-mediated isothermal amplification method (RT-LAMP). Several modifications were done on the basic protocol proposed by Boubourakas et al. (2009), additional experiments were preformed in order to further evaluate the method. Namely, the reaction time was further reduced and traces of leaf tissue, taken by a sterile toothpick, instead of tRNA were used as the intial material. Moreover, the AMV reverse transcriptase proved to be more effective than Thermoscript, while restriction enzyme analysis was performed on the RT-LAMP products in order to confirm that products had the respective sequences of the selected target. Finally, the extremely high efficiency and sensitivity of RT-LAMP proved to be sufficient for the detection of PLMVd in hosts other than peach. Keywords: PLMVd, RT-LAMP, peach, reverse transcriptase

AB0901 PREVALENCE OF OSTEOPOROSIS IN ITALIAN POSTMENOPAUSAL WOMEN ACCORDING TO DEFRA ALGORITHM

Background:Osteoporosis is a recognized health problem and the burden of the disease is mostly associated with the occurrence of hip and vertebral fracture.Objectives:This study was aimed at evaluating the prevalence of osteoporosis in Italian postmenopausal women, defined by DeFRA calculation as a 10 years fracture risk equal or higher than 20%.Methods:This is a monocenter cohort study evaluating 1850 post-menopausal women aged 50 years and older. All the participants were evaluated as far as anthropometrics. Defra questionnaire was administered and calculated with bone mineral density (DXA) measured at lumbar spine and femoral neck.Results:The prevalence of osteoporosis as assessed by DeFRA was 29.8% in the whole population, according to literature. The frequency of a risk fracture equal or higher than 20% varied from 7.9% in the group aged 50-59 years to 35% in subjects aged >80. Among clinical risk factors for fracture, the presence of a previous fracture (spine primarily) was the most commonly observed.Conclusion:Our data showed that about one third of post-menopausal women aged 50 and older in Italy has osteoporosis on the basis of DeFRA algorithm, with a high 10 years fracture risk. A previous fracture is the most common risk factor. The data should be considered in relation to the need to increase prevention strategies and therapeutic intervention.Disclosure of Interests:None declare