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Background. The micropropagation methods are used for the conservation of Pyrus L. phytodiversity, as well as for the creation of collections of the threatened species and pear cultivars including genotypes necessary for breeding and genetic studies. Materials and Methods. The two Pyrus species, P. communis, and P. salicifolia Pall. as well as four pear P. communis cultivars, ’Bere Desiatova’, ’Umans’ka Juvileina’, ’Kniahynia Olga’, and ’Sofia Umans’ka’, were chosen as the experimental plants. Young shoots of Pyrus species and cultivars with apical meristem from three-year-old plants were used as primary explants. The effectiveness of sterilization was determined by the rate of sterile and viable explants. Rooted in vitro test tube shoots with 3–4 roots 2–5 cm long were transplanted to adapt into nutrient disks Jiffy-7 and Ellepress. The statistical analysis of the results was performed according to Ronald Fisher (2017) and Peter Bruce with co-workers (Bruce et al., 2020) using Statistica ver.10 (StatSoft, Inc. STATISTICA version 10.0). Results and Discussion. According to our results, mercury dichloride (HgCl2) was the most effective of all tested sterilizers of P. communis. The P. communis explants sterilization technique was extremely difficult or unsuccessful for P. salicifolia, probably due to the dense pubescence on shoots and buds of this species. Therefore, we used the seeds as explants for micropropagation of P. salicifolia according to the standard protocol. In the best variant of Murashige–Skoog (MS) medium modification for P. communis, where MS-276 was modified by 2.0 mg/L 6-benzylaminopurine (6-BAP) and 0.01 mg/L indole-3-butyric acid (IBA), a value of 9.50 microclones per passage was obtained. However, the most active shoot formation of P. salicifolia in vitro was observed in the variant where the Driver and Kuniyuki (DKW) medium was modified by adding 2.5 mg/L 6-BAP. To induce rhizogenesis, the best medium for P. communis and P. salicifolia rhizogenesis was MS-302, modified by the addition of 0.5 mg/L α-naphthaleneacetic acid (NAA). In this variant, the first root in microclones developed in 8–10 days, and after a month, rooting reached 87.50% of P. communis and 98.54 % of P. salicifolia. The efficiencies of both Pyrus species adaptation were better with Ellepress peat plugs, than with Jiffy-7 peat pellets. Conclusion. It is recommended to cultivate explants of P. communis on MS medium with the addition of 2.0 mg/L 6-BAP and 0.01 mg/L IBA, and P. salicifolia on DKW medium with the addition of 2.5 mg/L 6-BAP and 0.1 mg/L IBA. It is also recommended that MS medium with the addition of 0.5 mg/L NAA be used for rooting of the microclones of both Pyrus species, and Ellepress peat plugs for the adaptation of regenerated plants.
Background. The micropropagation methods are used for the conservation of Pyrus L. phytodiversity, as well as for the creation of collections of the threatened species and pear cultivars including genotypes necessary for breeding and genetic studies. Materials and Methods. The two Pyrus species, P. communis, and P. salicifolia Pall. as well as four pear P. communis cultivars, ’Bere Desiatova’, ’Umans’ka Juvileina’, ’Kniahynia Olga’, and ’Sofia Umans’ka’, were chosen as the experimental plants. Young shoots of Pyrus species and cultivars with apical meristem from three-year-old plants were used as primary explants. The effectiveness of sterilization was determined by the rate of sterile and viable explants. Rooted in vitro test tube shoots with 3–4 roots 2–5 cm long were transplanted to adapt into nutrient disks Jiffy-7 and Ellepress. The statistical analysis of the results was performed according to Ronald Fisher (2017) and Peter Bruce with co-workers (Bruce et al., 2020) using Statistica ver.10 (StatSoft, Inc. STATISTICA version 10.0). Results and Discussion. According to our results, mercury dichloride (HgCl2) was the most effective of all tested sterilizers of P. communis. The P. communis explants sterilization technique was extremely difficult or unsuccessful for P. salicifolia, probably due to the dense pubescence on shoots and buds of this species. Therefore, we used the seeds as explants for micropropagation of P. salicifolia according to the standard protocol. In the best variant of Murashige–Skoog (MS) medium modification for P. communis, where MS-276 was modified by 2.0 mg/L 6-benzylaminopurine (6-BAP) and 0.01 mg/L indole-3-butyric acid (IBA), a value of 9.50 microclones per passage was obtained. However, the most active shoot formation of P. salicifolia in vitro was observed in the variant where the Driver and Kuniyuki (DKW) medium was modified by adding 2.5 mg/L 6-BAP. To induce rhizogenesis, the best medium for P. communis and P. salicifolia rhizogenesis was MS-302, modified by the addition of 0.5 mg/L α-naphthaleneacetic acid (NAA). In this variant, the first root in microclones developed in 8–10 days, and after a month, rooting reached 87.50% of P. communis and 98.54 % of P. salicifolia. The efficiencies of both Pyrus species adaptation were better with Ellepress peat plugs, than with Jiffy-7 peat pellets. Conclusion. It is recommended to cultivate explants of P. communis on MS medium with the addition of 2.0 mg/L 6-BAP and 0.01 mg/L IBA, and P. salicifolia on DKW medium with the addition of 2.5 mg/L 6-BAP and 0.1 mg/L IBA. It is also recommended that MS medium with the addition of 0.5 mg/L NAA be used for rooting of the microclones of both Pyrus species, and Ellepress peat plugs for the adaptation of regenerated plants.
Korla fragrant pear is a high-quality local pear variety native to Xinjiang, China. Currently, the low fruit-setting rate and low calyx shedding rate problems in Korla fragrant pears have been highlighted, which seriously affect the fruit yield and quality. It is of great significance to research the fruiting characteristics and molecular-assisted identification of Korla fragrant pear bud mutation materials for enriching the germplasm resources of Korla fragrant pear. In this research, a natural pollination group (YB) of Korla fragrant pear bud mutation materials and a natural pollination group (CK) of Korla fragrant pears were established. On the fruiting characteristics, the fruit-setting rate and calyx-removal rate of the two groups were investigated. In terms of fruit quality, the fruit shape index, fruit specific gravity, soluble solids content, sugar:acid ratio, soluble sugar content, and other indicators were measured. For the anatomical structure of the calyx tube, the detachment cells were observed. The formation time of the two groups of detached cells was compared. In the GBS simplified genome sequencing, a phylogenetic tree was constructed based on the obtained SNP sites. A principal component analysis, population genetic structure analysis, and genetic diversity index analysis were carried out. In the aspect of SSR molecular marker identification, the SSR types were counted. Polyacrylamide gel electrophoresis was performed. The results demonstrate the following: (1) the fruit-setting rate (30.87%) and calyx-removal rate (68.11%) in the YB group were significantly higher than those in the CK group (19.37%) and the calyx-removal rate (55.18%). (2) There was no significant difference in fruit quality indexes, such as average fruit weight (127.10–130.00 g) and soluble sugar content (9.47–9.56%) between the two groups. (3) Abscission-layer cells were observed at 2, 4, 6, 8, and 10 h after calyx tube discoloration in the YB group and at 48, 72, and 96 h after calyx tube discoloration in the CK group. (4) The genetic background of the YB group and the CK group was similar at the GBS level, but there were differences at the DNA level. This research finally shows that Korla fragrant pear bud mutation material is a good germplasm resource. This germplasm resource can promote the structural optimization of Korla fragrant pear varieties and the healthy development of the industry.
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