N. Milosevic's Lab

Featured projects (1)

CherrySeRB is basically oriented towards the field of agro-biodiversity, aiming to add value to unexplored Serbian and Balkan fruit genetic resources through new protocols and approaches for their genotypic and phenotypic characterisation. Two fruit species of worldwide importance are targeted  diploid sweet cherry and tetraploid sour cherry. The main idea of CherrySeRB is connection of the production challenges arising from climate change and demands for quality and healthy fruits, and the richness of Serbian cherry germplasm that is still unused enough. Characterisation of the indigenous genotype potential for desirable reproductive properties and defining ʻgood reproductive behaviour cherry modelʼ that can face warmer temperature conditions during the flowering are in focus. CherrySeRB will also provide S-genotyping and the choice of suitable pollenisers that maximize fertilization efficacy for landraces, as well as their nutritional/phytochemical profiles that will enable obtaining of new high-added value nutraceutical products.Organized around four pillars that include six WPs, CherrySeRB combines interdisciplinary modern methodologies – molecular, for identification of S-haplotypes in definition of GSI-status (DNA diagnostic strategy); biological, for revealing suitable polleniser(s) (assessment of the pollen tube growth in vivo by fluorescence microscopy; the fertilization efficacy parameters); histological, for determining ovary function after exposing to different flowering temperatures (cyto-embryological and histochemical testing); nutritional, for characterisation of fruit quality and health-promoting compounds (texture and colorimetric analysis, HPLC, UV-Vis, etc.); genotype identification (S-genotyping; pollen grain characterisation by SEM); statistical (models in reaching new scientific conclusions). Data, protocols and novel cherry material will be made available to breeders, biotechnology scientists and cherry-sector stakeholders. Representing novelty in breeding approach that can be applicable to other fruit species, the ultimate goal of CherrySeRB is to foster the use of Serbian cherry germplasm as the material adapted to climate change in order to contribute to sustainable fruit production and ensure the availability of healthy food for the future.

Featured research (6)

A study was conducted to investigate the effect of different storage periods and temperatures on pollen viability in vitro and in vivo in plum genotypes ‘Valerija’, ‘Čačanska Lepotica’ and ‘Valjevka’. In vitro pollen viability was tested at day 0 (fresh dry pollen) and after 3, 6, 9 and 12 months of storage at four different temperatures (4, −20, −80 and −196 °C), and in vivo after 12 months of storage at distinct temperatures. In vitro germination and fluorescein diacetate (FDA) staining methods were used to test pollen viability, while aniline blue staining was used for observing in vivo pollen tube growth. Fresh pollen germination and viability ranged from 42.35 to 63.79% (‘Valjevka’ and ‘Čačanska Lepotica’, respectively) and 54.58 to 62.15%, (‘Valjevka’ and ‘Valerija’, respectively). With storage at 4 °C, pollen viability and germination decreased over the period, with the lowest value after 12 months of storage. Pollen germination and viability for the other storage temperatures (−20, −80 and −196 °C) were higher than 30% by the end of the 12 months. Pollination using pollen stored at 4 °C showed that pollen tube growth mostly ended in the lower part of the style. With the other storage temperatures, pollen tube growth was similar, ranging between 50 and 100% of the pistils with pollen tubes penetrated into the nucellus of the ovule in the genotype ‘Čačanska Lepotica’. The results of these findings will have implications for plum pollen breeding and conservation.
Five plum hybrids (38/62/70, IV/63/81, 32/21/87, 34/41/87 and 22/17/87) and newly released cultivar ‘Nada’, obtained by planned hybridisation and singled out within breeding programme at Fruit Research Institute, Čačak, were assessed for the main physical (fruit and stone weight and flesh percentage), chemical (soluble solids content, total and inverted sugars content, sucrose content, total acids content, pH value of fruit juice, ratio of soluble solids and total acids content and ratio of total sugars and total acids content) and sensorial (attractiveness, taste, aroma and consistency) traits compared with the standard cultivar ʻČačanska Lepoticaʼ. Results showed that the studied plum genotypes differed significantly in all of the assessed traits. Regarding the physical features, the best results were shown by hybrid 38/62/70, which had the highest fruit weight (56.92 g) and flesh percentage (96.91%), as well as by the new cultivar ‘Nada’, for which a larger fruit was observed in comparison to the standard (45.54 g and 42.24 g, respectively). Also, ‘Nada’ had better sensorial properties such as attractiveness, taste, aroma and consistency in relation to the other promising hybrids and the standard cultivar. From the aspect of all the studied sensorial characteristics, in addition to ‘Nada’, only hybrid 38/62/70 was aligned with the standard cultivar. Out of the studied genotypes, late ripening hybrid 22/17/87 had the best values of parameters of fruit chemical composition such as the soluble solids content (17.01%), total and inverted sugars contents (12.31% and 8.96%, respectively). The highest sucrose content (3.39%), pH value of fruit juice (3.51), as well as the highest ratio between soluble solids and total acids content (43.72) and ratio between total sugars and total acids content (32.58) were found in cultivar ‘Nada’. The highest total acids content (1.42%) was recorded in hybrid 32/21/87. Compared to ʻČačanska Lepoticaʼ, the same or better results in terms of the fruit chemical composition were determined in cultivar ‘Nada’ and hybrids 34/41/87 and 22/17/87. The study revealed existence of significant correlations between individual studied parameters of the fruit physical, chemical and sensorial properties.
In 2018 and 2019, we evaluated the influence of seven clonal rootstocks (Colt, MaxMa 14, Krymsk 6, Adara, Cigančica, Gisela 5 and Gisela 6) with different vigour on the total phenolic content (TPC), total flavonoid content (TFC), total anthocyanin content (TAc), total antioxidant capacity (TAC) and main fruit physical properties of ‘Šumadinka’ sour cherry. In particular, ‘Šumadinka’ on Gisela 5, especially on Gisela 6 had the highest fruit and stone weight, fruit dimensions and ripening index (RI) value. MaxMa 14 as a semi-dwarf rootstock induced the best values of all phenolic compounds evaluated and TAC, whereas Gisela 5 as a dwarf rootstock promoted the poorest levels of these compounds and antioxidant activity. Krymsk 6 and Colt induced the highest and similar flesh rate, whereas Adara promoted the lowest. Cigančica induced the highest soluble solids content (SSC) and titratable acidity (TA), but poor phenolic compounds content and antioxidant activity. In general, the more vigorous rootstocks produced significantly lower fruit weight, fruit size, SSC and TA, but induced better content of phenolic compounds in comparison with semi-dwarf and dwarf rootstocks, respectively.
A newly released, late ripening plum cultivar ‘Pozna Plava’ sets fruit poorly, although it produces high quality fruit. This study aimed to evaluate which factors in the reproductive process could be related to the lack of fruit set. In two consecutive years, establishment of a suitable polleniser and the stage of ovule development at anthesis as well as initial and final fruit set have been studied. In addition to this, the impact made by temperature fluctuations on the interaction between male gametophytes and female sporophytes was also analysed. Growth of the pollen tubes in the style and penetration into the nucellus as well as fruit set were more effective in cross-pollination than in open and self-pollination. A relative delay in ovule development was observed, and most ovules had an embryo sac with eight nuclei. Considering the results of the quantitative parameter study of pollen tube growth in the ovary as well as the results of the stage of ovule development, a conclusion can be made that this cultivar is characterised by an extremely short effective period of pollination.

Lab head

Nebojša Milošević
  • Department of Pomology and Fruit Breeding
About Nebojša Milošević
  • Nebojša Milošević currently works at the Department of Pomology and Fruit Breeding, Fruit Research Institute, Čačak. Nebojša does research in Food Science, Agricultural Plant Science and Horticulture. Their current project is 'TR-31064'.

Members (4)

Radosav Cerović
  • University of Belgrade
Ivana Glišić
  • Fruit Research Institute
Milena Đorđević
  • Fruit Research Institute
Slađana Marić
  • Fruit Research Institute, Čačak, Republic of Serbia