Studies on the diagnosis of hop stunt viroid in fruit trees: Identification of new hosts and application of a nucleic acid extraction procedure based on non-organic solvents

Abstract

A non-radioactive digoxigenin-labelled RNA probe specific for hop stunt viroid (HSVd) diagnosis has been developed. The high sensitivity and specificity of this RNA probe in dot blot hybridizations to nucleic acids from field samples, allowed the confirmation of the presence of HSVd in apricot (Prunus armeniaca L.) and its detection in two fruit tree species not previously described as hosts of this pathogen, almond (Prunus dulcis Miller) and pomegranate (Punica granatum L.). This result supports and extends the notion of the world wide distribution of HSVd, infecting cultivated fruit trees. HSVd was also found to accumulate to much higher levels in mature apricot fruits than in leaves. Additionally, a sample processing procedure which does not involve the use of organic solvents was demonstrated to render faithful results when used for viroid detection. The combined reliability and facility of use of both this extraction procedure and the non-radioactive probe will benefit agronomic investigations addressing the detection and eradication of HSVd. Other applications of the work described here, as the study of possible causal relations between specific disorders and HSVd infection, are also discussed.

Molecular sampling of hop stunt viroid (HSVd) from grapevines in hop production areas in the Czech Republic and hop protection

“…Two viroid species, hop stunt viroid (HSVd) and hop latent viroid HLVd (Puchta et al 1988a), both belonging to the family Pospiviroidae, have been described as hop (Humulus lupulus L.) pathogens. HSVd apparently has a wide host range and besides hop it is able to propagate in cucumber, grapevine, citrus, plum, peach, pear (Shikata 1990), apricot and almond plants (Astruc et al 1996, Canizares et al 1999. Currently, eighty-four HSVd sequences are present in the subviral RNA database (Pelchat et al 2000), but as many as 120 HSVd sequence entries are in common biological databases, and about eleven of them are designated as hop-adapted.…”

A Viroid Resembling Hop Stunt Viroid in Infected Apple Trees with Apple Scar Skin Disease in the Northeast of Iran

“…HSVd belongs to the genus Hostuviroid and has the broadest host-range known for any viroid. HSVd was first described as the causal agent of the stunt disease of hop in Japan but since then it has been found in several plant species including some fruit trees like citrus, pear, peach, plum, apricot, almond, cherry, pomegranate and grapevine (8,9). Some of these plants have shown specific abnormalities or appeared symptomless.…”

Sensitive Detection of Apple Stem Grooving and Apple Stem Pitting Viruses From Infected Apple Trees by Rt-PCR

“…These degenerate primers allowed reliable amplification of virus specific sequences from transcripts of total RNA preparations from herbaceous host plants growll in the greenhouse and from leaves of some virus-infected apple-trees from the orchard, but only during spring. Meanwhile (Astruc et al, 1996: Spiegel et al, 1996 (Kummert et a1.,1997), have been cloned and sequenced. The alignment of these sequences, and those published for ASGV (Yoshikawa et al, 1992) or ASPV (Jelkmann, 1994) …”