The reaction to Tomato spotted wilt virus (TSWV) was evaluated in 94 accessions of Nicotiana, originating from the Institute of Soil Science and Plant Cultivation tobacco germplasm collection in Puławy, Poland. Tests for resistance were conducted under greenhouse conditions using single TSWV isolate collected from tobacco plantation in Lublin district, Poland. The presence of the virus was verified using DAS-ELISA. SCAR markers associated with TSWV resistance gene were applied. The members of the section Alatae, the genus Nicotiana: N. alata, N. forgetiana, and Nicotiana x sanderae as well as N. tabacum cultivars: 'Polalta' and 'Wiktoria' with the TSWV resistance gene introduced from N. alata, displayed the hypersensitive reaction (HR) against TSWV (grade 0 on symptom intensity scale). In some of those accessions, the virus spread from the initially infected areas eliciting systemic hypersensitive reaction (SHR). Five accessions of N. alata and three of Nicotianaxsanderae were composed of 6.3-50.0 % of plants in which SHR symptoms appeared. In all of N. forgetiana plants HR reaction was followed by systemic infection (SHR). In N. tabacum 'Wiktoria' 21.1 % of plants showed HR reaction, while the remaining were susceptible (S). All of the genotypes which responded with HR or SHR reaction to TSWV infection demonstrated the presence of SCAR markers linked to the resistance gene. The remaining eighty tested accessions were identified as being susceptible upon exposure to TSWV.
Nicotiana tabacum cv. ÔPolaltaÕ is reported as having a hypersensitive type of Tomato spotted wilt virus (TSWV) resistance. In ÔPolaltaÕ hybrids with susceptible cultivars, morphological deformations (thickened and ribbon-shaped leaves, irregular venation, tumours) appear. Acceptable DH lines derived from hybrids of ÔPolaltaÕ with ÔWis´licaÕ were obtained by a method of in vitro culture. Those DH lines were evaluated for TSWV resistance. Tests for resistance were conducted under greenhouse conditions using one TSWV isolate from a local source. The presence of the virus was verified using DAS-ELISA. The DH lines PW-833, PW-834, and PW-900 showed a near correct morphology and stably inherited dominant monogenic resistance to TSWV.
Black root rot (BRR) caused by Thielaviopsis basicola as well as Tomato spotted wilt virus (TSWV) are the most serious problems in tobacco growing regions. We crossed the breeding line WGL 3 carrying BRR resistance derived from N.glauca with the line PW-834 the resistance of which to TSWV was transferred from cultivar Polalta. Anthers obtained from F1 hybrid plants were cultured to induce haploids combining resistance to Th. basicola and TSWV. Flow cytometry analysis revealed 242 haploids and 2 spontaneous doubled haploids among regenerants. All haploids were cloned and then evaluated for BRR as well as TSWV resistance. The presence of pathogens was detected by microscopic evaluation of roots or using DAS-ELISA test. Microscopic assessment showed that, 132 haploids had no symptoms of Th. basicola which, together with the absence of symptoms in the F1 hybrids, indicated a dominant monogenic mode of inheritance. At the same time only 30 haploids demonstrated resistance to TSWV. SCAR markers associated with TSWV resistance gene detection was applied. The results indicate that small proportion of TSWV-resistant haploids is probably due to the influence of deleterious genes flanking the resistance factor that reduced vitality of gametophytes. Altogether, 24 haploids showed multiple resistance to Th. basicola and TSWV.
SUMMARYFour cytoplasmic male sterile (cms) F 1 flue-cured hybrids cv. Wi³lica × cv. Virginia Golta (VG), the male fertile analogue and the parental varieties were tested at two locations in Poland in a replicated field trial. The cms sources in the hybrids were N. suaveolens, N. amplexicaulis, N. bigelovii and a N. tabacum cms mutant. Under the slight to moderate pressure from black root rot present at the trial sites the hybrids showed a moderate tolerance of the disease characteristic of VG as opposed to medium strong susceptibility of Wi³lica. Apart from the effect of black root rot tolerance the vegetative vigor of the hybrids (plant height, leaf size, earliness) was affected by cytoplasm source. The F 1 hybrid with N. suaveolens cytoplasm flowered approximately three days later than the remaining hybrids. Of the cms hybrids tested cms N. bigelovii produced the tallest plants with largest mid-position leaves. Yields of cured leaves were largely influenced by black root rot and were generally higher in VG and in the hybrids than in Wi³lica. Leaf yields and curability were generally little affected by cms source under low pressure from black root rot. At the site with a relatively high level of black root rot infestation the yields of cms N.
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