In many cases the diagnosis of a viral disease in perennial woody plants requires indexing by grafting on indicator plants. In the case of grapevine leafroll and corky-bark diseases, indexing requires 2 to 3 years for symptom recording. Both diseases are found in all grapevine-growing countries. It would therefore be advantageous to develop a sensitive, quick, and reliable diagnostic technique. Explant shoots infected with corky-bark were micrografted onto healthy indicator rootstocks and maintained in vitro. Typical corky-bark symptoms appeared on the indicator within 8 to 12 weeks. Osmotic stress, in vitro, induced by sorbitol, enhanced leafroll symptoms. Explants expressed symptoms after 2 to 3 months of growth on these media. The advantages of these techniques are: Rapid indexing, saving of space and labor, could be performed year-round. Further experiments are underway for adaptation of the micrografting to leafroll disease and the stress method for corky-bark disease.
N. Shlamovitz, P. Spiegel-Roy, and E. Tanne
Edna Tanne, N. Shlamovitz, and P. Spiegel-Roy
Grapevine (Vitis vinifera L.) explant shoots indexed forcorky-bark and rootstocks from healthy LN33 indicator plants were sterilized and maintained in vitro. When infected shoot tips were micrografted onto LN33 shoots, typical corky-bark symptoms appeared in 8 to 12 weeks. We suggest developing this method further to replace the regular, 2-year indexing procedure.
M.J. Striem, G. Ben-Hayyim, and P. Spiegel-Roy
Excluding seeded offspring at an early stage could be of great value to the breeder concerned with the development of seedless grapes (Vitis vinifera L.). We used the random amplified polymorphic DNA (RAPD) technique to identify molecular genetic markers, analyzing 82 individuals of a progeny resulting from a cross between `Early Muscat' (seeded) and `Flame Seedless'. Seven variables representing the traits of seedlessness were analyzed: mean fresh weight of one seed, total fresh weight of seeds per berry, perception of seed content, seed size categories evaluated visually, degree of hardness of the seed coat, degree of development of the endosperm, and degree of development of the embryo. Among 160 10mer primers, 110 gave distinct band patterns. Twelve markers yielded significant correlations with several subtraits of seedlessness, mainly with the mean fresh weight of one seed and the total fresh weight of seeds per berry. Multiple linear regression analysis resulted in high coefficients, such as R = 0.779 for fresh weight of seeds per berry, when the seven markers were included as independent variables in the model. Most of the seeded individuals, about 44% of the progeny, could be excluded using a two-step process of marker assisted selection.
D.W. Ramming, R.L. Emershad, P. Spiegel-Roy, N. Sahar, and I. Baron
Immature grape embryos from early ripening genotypes of Vitis vinifera were successfully cultured in vitro on Difco orchid agar or a modified White's agar medium. Germination was increased in vitro for five genotypes from 0%, 7%, 11%, 12%, and 16% in vivo to 15%, 24%, 23%, 34%, and 24%, respectively. Subculturing embryos onto liquid culture from seeds that failed to germinate on agar also was possible. Differences in germination rates, as affected by pollen, were significant. This method will allow accelerated development of early ripening cultivars by allowing breeders to use such genotypes as females, as well as males.