D. Boscia et al., Production of monoclonal antibodies to Grapevine virus D and contribution to the study of its aetiological role in grapevine diseases, VITIS, 40(2), 2001, pp. 69-74
Six stable hybridoma cell lines secreting monoclonal antibodies (MAbs) to G
rapevine virus D (GVD) were obtained by fusing spleen cells of immunized BA
LB/c mice with mouse myeloma cell line Sp 2/0-Ag 14, In ELISA all MAbs dete
cted the virus in Nicotiana leaf extracts or cortical shavings from mature
grapevine canes, The use of a polyclonal antiserum for coating plates and o
f monoclonal antibodies and antimouse-conjugated antibodies for antigen det
ection, gave highly efficient and reproducible results for identification o
f GVD in field-grown grapevines. The reliability of the ELISA kit was confi
rmed by GVD-transmission tests to herbaceous hosts, using in vitro explants
as inoculum, 223 vines affected by one or more of the 4 syndroms of the ru
gose wood complex (Kober stem grooving, Corky bark, LN stem grooving and Ru
pestris stem pitting) were tested in ELISA for the detection of Grapevine v
irus A (GVA), Grapevine virus B (GVB) and GVD and by Western blot for the d
etection of Grapevine rupestris stem pitting associated virus (GRSPaV). The
possible cause-effect relationship between GVA and KSG, GVB and Co, and GR
SPaV and RSP was confirmed, but no consistent association was found between
GVD and any of the 4 above syndromes, Intriguingly, a reduction in the exp
ression of stem pitting symptoms in V. rupestris (from 90 % to 75 %) and of
stem grooving symptoms in Kober 5BB (from 95 % to 70 %) was observed when
vitiviruses and GRSPaV were contemporarily present in the same indicator. P
reliminary data of a survey involving 676 grapevine samples showed a high i
ncidence (31 %) of GVD, regardless of the geographical origin of samples.