Genetic divergence among inbred lines in Cucurbita moschata from China

Knowledge of genetic divergence among inbred lines is essential for cross breeding. The objectives of this study were to (1) analyze the level and character of genetic diversity in C. moschata accessions, and (2) classify the genetic divergence among inbred lines in C. moschata to assist in selection of parent genotypes for genetic improvement. Twenty agronomic characters were investigated and rich diversities were demonstrated among 39 inbred lines of C. moschata from China. Various degree correlations among these characters made it possible to summarize the diversities of the twenty characters into 3 major principal components: leaf, fruit and flesh quality factor. Forty-one inbred lines of pumpkin were clustered into four groups based on principal component data, which is more distinct for classification that based on the original data of the 20 characters. However, parent inbred lines whose hybrids displayed significant heterosis in fruit weight, soluble solid and fruit shape were located in different clusters or sub-clusters based on standardized original data. It was suggested that genotypes in the same clusters may represent members of one heterotic group.     

Morphological and molecular analysis of genetic diversity in jackfruit

Summary

RAPD markers were used to estimate genetic diversity in 12 high-yielding jackfruit (Artocarpus heterophyllus Lamk.) accessions obtained from different locations in southern India. Marker data were compared with morphological data obtained over three successive seasons. PCR-amplifiable DNA was isolated using the CTAB method and 171 amplified fragments were obtained using 23 random primers. The genetic dissimilarity matrix was calculated based on Squared Euclidian Distances, which revealed a maximum genetic distance of 7.9% between a clone of ‘Mottavarica’ (‘M₀’), and ‘Chandrahalasu’ from distant locations, while the minimum genetic distance (5%) was between the genotypes (‘M₀’) and ‘Kerala’, indicating their similar geographical origin. Ward's method of cluster analysis grouped all individuals on the dendrogram into two major clusters according to their geographical location. The present study showed low-to-moderate genetic diversity among the 12 jackfruit accessions, which will assist in the identification and management of jackfruit germplasm for breeding purposes.     

Genetic divergence in seedling trees of Persian walnut (Juglans regia L.) for various metric nut and kernel characters in Himachal Pradesh

The nature and magnitude of genetic divergence was assessed using non-hierarchical Euclidean cluster analysis in 229 seedling trees of Persian walnut growing naturally in four districts of Himachal Pradesh for 15 nut and kernel characters. Minimum and maximum values of coefficients of variability were recorded for nut width and kernel weight, respectively. All genotypes were grouped into 16 different clusters. The clustering pattern of walnut genotypes belonging to the same location revealed their distribution in more than one cluster showing non-parallelism between geographic and genetic diversity. Genotypes of clusters 5 and 8 were highly diverse from each other. Mean nut weight and kernel weight were highest in clusters 8 and 14, respectively. The highest mean kernel percentage and minimum shell thickness were observed in cluster 13. The highest mean value for fat was recorded in cluster 9 whereas cluster 3 had the maximum value of protein.     

Genetic transformation of Prunus domestica L. using the hpt gene coding for hygromycin resistance as the selectable marker

The study and development of transformation technology with new selection schemes is important for various fundamental studies and for crop trait improvement via genetic engineering. Here we have shown that hygromycin resistance is an effective system for plum genetic transformation. Embryonic axes of mature seeds were co-cultivated with Agrobacterium strain LBA4404 containing the pC1381 plasmid carrying the hygromycin phosphotranferase gene (hpt) and β-glucuronidase (GUS) gene or with strain EHA105 containing the plasmid pC1301 carrying the same marker and reporter genes. The latter strain containing a pC2301 plasmid carrying the neomycin phosphotransferase gene (nptII) gene was used as a control. Infected explants were placed on shoot induction medium containing either 5 mg L⁻¹ hygromycin or 75 mg L⁻¹ kanamycin for selection. Green shoots developed from the explants under hygromycin pressure. These shoots showed continued and vigorous growth and development upon transfer onto fresh hygromycin medium. PCR using hpt sequence primers, and Southern blot analysis using a probe from the hpt gene, confirmed the presence of the transgenes and their stable integration in regenerated plants. Full transgenic plants were obtained in a greenhouse. Hygromycin selection was very effective and no escapes were observed. The study demonstrated that hygromycin resistance can be used as an effective selectable marker for plum transformation. The new system developed here is important and useful for multiple gene transformation in plum.     

Determination of the uptake and translocation of nitrogen applied at different growth stages of a melon crop (Cucumis melo L.) using N isotope

In order to establish a rational nitrogen (N) fertilisation and reduce groundwater contamination, a clearer understanding of the N distribution through the growing season and its dynamics inside the plant is crucial. In two successive years, a melon crop (Cucumis melo L. cv. Sancho) was grown under field conditions to determine the uptake of N fertiliser, applied by means of fertigation at different stages of plant growth, and to follow the translocation of N in the plant using ¹⁵N-labelled N. In 2006, two experiments were carried out. In the first experiment, labelled ¹⁵N fertiliser was supplied at the female-bloom stage and in the second, at the end of fruit ripening. Labelled ¹⁵N fertiliser was made from ¹⁵NH₄¹⁵NO₃ (10 at.% ¹⁵N) and 9.6 kg N ha⁻¹ were applied in each experiment over 6 days (1.6 kg N ha⁻¹ d⁻¹). In 2007, the ¹⁵N treatment consisted of applying 20.4 kg N ha⁻¹ as ¹⁵NH₄¹⁵NO₃ (10 at.% ¹⁵N) in the middle of fruit growth, over 6 days (3.4 kg N ha⁻¹ d⁻¹). In addition, 93 and 95 kg N ha⁻¹ were supplied daily by fertigation as ammonium nitrate in 2006 and 2007, respectively. The results obtained in 2006 suggest that the uptake of N derived from labelled fertiliser by the above-ground parts of the plants was not affected by the time of fertiliser application. At the female-flowering and fruit-ripening stages, the N content derived from ¹⁵N-labelled fertiliser was close to 0.435 g m⁻² (about 45% of the N applied), while in the middle of fruit growth it was 1.45 g m⁻² (71% of the N applied). The N application time affected the amount of N derived from labelled fertiliser that was translocated to the fruits. When the N was supplied later, the N translocation was lower, ranging between 54% at female flowering and 32% at the end of fruit ripening. Approximately 85% of the N translocated came from the leaf when the N was applied at female flowering or in the middle of fruit growth. This value decreased to 72% when the ¹⁵N application was at the end of fruit ripening. The ammonium nitrate became available to the plant between 2 and 2.5 weeks after its application. Although the leaf N uptake varied during the crop cycle, the N absorption rate in the whole plant was linear, suggesting that the melon crop could be fertilised with constant daily N amounts until 2–3 weeks before the last harvest.     

Molecular phylogeny in Indian Citrus L. (Rutaceae) inferred through PCR-RFLP and trnL-trnF sequence data of chloroplast DNA

The taxonomy and phylogeny of Indian Citrus is revisited using PCR-RFLP of the trnD-trnT and rbcL-ORF 106 regions as well as sequence data analysis of the trnL-trnF intergenic spacer region of cpDNA. The study was based on 50 accessions of Citrus genotypes, collected from wild, semi-wild and domesticated stocks. Of the 13 restriction enzymes (RE) used for restriction digestion of the polymerase chain reaction (PCR) amplicons, four (Hinf I, Msp I, Alu I, Hae III) generated 47 restriction fragments, of which 24 (51%) were polymorphic. PCR-RFLP data showed a genetic distance ranging from 0 to 0.79 among 50 accessions of Citrus, and a cluster analysis, based on Neighbor-Joining (NJ) method, placed all the accessions in eight major clusters. Analysis of trnL-trnF sequences from 23 representative accessions of Citrus showed a pair-wise sequence divergence rate in the range of 0–0.064. NJ, minimum evolution (ME) and maximum parsimony (MP) analyses of trnL-trnF sequences produced phylogenetic trees, which placed all the 23 accessions in five clusters. PCR-RFLP analysis resulted in a well resolved phylogenetic tree with branches supported by moderate to high bootstrap values, while the trnL-trnF sequence-based trees showed only moderate to low bootstrap support for the internal tree branches, indicating uncertain origin of some Citrus genotypes. This study shows that the trnL-trnF spacer sequence data can detect genetic variation in Indian Citrus genotypes, but the utility of the data in inferring phylogeny at intra and inter-specific levels is limited probably by factors such as hybridization, bud mutations, apomixis and polyploidy. However, PCR-RFLP and trnL-trnF data supported the recognition of C. maxima, C. medica, and C. reticulata as the basal species of edible Citrus.     

Genetic divergence among gerbera accessions evaluated by RAPD

Genetic diversity was evaluated by RAPD markers and morpho-agronomic characters for a total of 42 accessions of Barberton daisy (Gerbera jamesonii) consisting of 29 commercial and 13 wild accessions. A total of 74 polymorphic bands were obtained employing a set of 12 primer pairs. The average genetic similarity coefficient for the 42 accessions, evaluated by Jaccard index was 0.55 ranging from 0.28 to 1.00. The genetic structure found among Barberton daisy accessions was evaluated by hierarchic classification analyses and UPGMA modeling, revealing six clusters of genotypes where two of them include the wild accessions and the remaining four including commercial material, except for wild genotype number 9. Shannon (H′) index was calculated using the molecular markers to investigate the genetic variation among the Gerbera accessions and showed higher values for the commercial cluster in comparison to the values obtained for the individuals from the non-commercial cluster, namely 0.34 versus 0.27, respectively. Therefore, both calculated indices (Jaccard and Shannon) indicated the presence of higher genetic variation among commercial accessions in comparison to the cluster representing non-commercial accessions, suggesting that genetic breeding programs may focus on commercial accessions to recombine interesting genotypes with commercially important and marketing-desired characteristics.     

雷州半岛菠萝蜜种质遗传多样性的RAPD分析

 用RAPD标记方法对我国雷州半岛的65份菠萝蜜(Artocarpus heterophyllus Lam. ) 实生种质资源DNA的遗传多样性进行了检测, 16个引物共检测到78条带, 其中69条具多态性(占88.4% ) 。聚类分析表明, 65份菠萝蜜材料在遗传距离0.26处可分为3大类。供试种质虽具有丰富的形态性状变异, 但在DNA水平上平均相似性系数为0.7341。干、湿胞类型及引自马来西亚的种质均不能独立聚类。对菠萝蜜品种的干、湿胞分类法及菠萝蜜品种的引种区域进行了讨论。     

Effect of organic and inorganic fertilizers applied during successive crop seasons on growth and nitrate accumulation in lettuce

A romaine-type lettuce (Lactuca sativa L.) cv. Corsica was cultivated during three successive crop seasons (late-spring, late-autumn and late-winter) in the same soil of an experimental greenhouse in S.W. Peloponnese, Greece. Seven long-term fertilization treatments were tested for their effect on plant growth and nitrate concentration in the external lettuce leaves. Treatments included: three different doses of organic fertilization (composted sheep manure) applied at the start of each crop season, three different doses of inorganic N fertilization applied via fertigation during each crop season, and a control treatment in which no fertilizer was applied. A drip irrigation system was used to water all plants. The highest nitrate levels were observed in the medium and maximum inorganic fertilization treatments (572–664 mg kg⁻¹) in all crop seasons. They were significantly higher compared to the respective organic fertilization treatments (253–435 mg kg⁻¹) and all other fertilization treatments (148–435 mg kg⁻¹). Crop season affected lettuce growth more than nitrate accumulation in the lettuce leaves: lettuce biomass production was the smallest and most uniform in the late-autumn season and did not respond to the fertilization treatments tested (ranging from 409 to 439 g plant⁻¹), while in the late-spring season biomass production was the highest and most variable (561–841 g plant⁻¹), it correlated with nitrate concentration in the leaves and in the medium and maximum inorganic fertilizer doses it significantly exceeded production from all other fertilization treatments (827–841 g plant⁻¹). Following the three crop seasons the residual availability of N, P and K was clearly enhanced in the soil receiving the organic compared to the inorganic fertilization. Nitrate concentration in lettuce leaves was far below the upper limits set by the European Commission in all fertilization treatments throughout the three crop seasons, a result attributed mainly to the sufficient level of light intensity and duration throughout the year in Southern Greece.     

Fruit thinning in ‘Conference’ pear grown under deficit irrigation: Implications for fruit quality at harvest and after cold storage

Fruit thinning in pear is feasible for mitigation of water stress effects. However, it is not well known how fruit quality at harvest and after cold storage is affected by pre-harvest water stress. Even less is known about the effects of fruit thinning on quality under these circumstances. To elucidate these, we applied deficit irrigation (DI) and fruit thinning treatments to ‘Conference’ pear over the growing seasons of 2008 and 2009. At the onset of Stage II (80 and 67 days before harvest in 2008 and 2009, respectively), two irrigation treatments were applied: full irrigation (FI) and DI. FI trees received 100% of crop evapotranspiration (ETc). DI trees received no irrigation during the first three weeks of Stage II to induce water stress, but then received 20% of ETc to ensure tree survival. From bud-break until the onset of Stage II and during post-harvest, FI and DI trees received 100% of ETc. Each irrigation treatment received two thinning levels: no thinning leaving commercial crop load (∼180 fruits tree⁻¹), and hand-thinning at the onset of Stage II leaving a light crop load (∼85 fruits tree⁻¹). Under commercial crop loads, DI trees were moderately water-stressed and this had some positive effects on fruit quality. DI increased fruit firmness (FF), soluble solids concentrations (SSC) and acidity at harvest while no changes were observed in fruit maturity (based on ethylene production). Differences in FF and acidity at harvest between FI and DI fruit were maintained during cold storage. DI also reduced fruit weight loss during storage. But fruit size was reduced under DI. Fruit thinning under DI resulted in better fruit composition with no detrimental effect on fresh-market yield compared to un-thinned fruit. Fruit size at harvest and SSC values after five months of cold storage were higher in fruit from thinned trees than fruit from un-thinned trees. Fruit thinning increased fruit ethylene production, indicating advanced maturity. This may lead to earlier harvest which is desirable in years with impending drought. Fruit thinning is therefore a useful technique to enhance pear marketability under water shortage.     

Characterisation of cultivated breeding lines of eggplant (Solanum melongena L.) and related wild Solanum species from India

Immature fruit of cultivated species of eggplant (Solanum melongena L.) are commonly used as a summer vegetable in India. Rich morphological variation exists among the cultivated species of eggplant in different growing regions of the country. We have characterised 24 breeding lines of Solanum spp., including 20 eggplant cultivars and four wild species of eggplant, based on 13 morphological characters using Mahalonabis D² statistics. All 24 breeding lines of Solanum spp., including the 20 eggplant cultivars and four wild species, were grouped into four clusters by agglomerative clustering. Cluster II and Cluster IV contained the most accessions (eight each), while Cluster I and Cluster III had four accessions each. The highest inter-cluster (D²) distance (158.33) was observed between Cluster I and Cluster II, followed by Cluster I and Cluster III (108.48), and Cluster II and Cluster IV (102.96), which indicated that accessions in Cluster I and Cluster II were more divergent than those in the other clusters. The highest intra-cluster distance (5.80) was observed in Cluster IV, with eight genotypes, and the lowest intra-cluster distance (2.21) was observed in Cluster II, also with eight genotypes. The intra-cluster distances in all four clusters were lower than the inter-cluster distances, which indicated that genotypes within the same cluster were closely related. Genotypes in Cluster IV had the maximum number of flowers per cluster (3.63), the highest number of fruit per cluster (3.25), and number of fruit per plant (208.63), which revealed that genotypes could be selected from Cluster IV for these characters. The first three principal components (PC1, PC2, and PC3) accounted 73.99% of the total variation among the 24 genotypes. These phenotypic data increase the feasibility of prioritising breeding lines in a crossing programme based on the uniqueness of their desirable morphological traits.     

Direct and indirect effects of seed related characters on number of seed in guava (Psidium guajava L.) fruits

Dessert quality of guava fruit is considerably reduced by high seed content. Number of seeds in guava is associated with different seed and fruit characters. Influence of different seed characters on number of seeds per fruit (NSPF) was studied by analysing character association and direct and indirect effects on NSPF and seed related traits for 68 genotypes of guava collected from diverse sources and conserved at National Active Germplasm Site, CISH, Lucknow. Seed related characters were studied at the colour break stage of the fruits using image analysis software for counting number of seeds. The data was subjected to path analysis to find out direct and indirect effects of different characters on number of seeds in the fruits. At genotypic and phenotypic levels, NSPF was significantly and positively associated with seed weight per fruit (SWPF), number of seeds 100 g⁻¹ pulp (NSPHP) and fruit weight (FW). The genotypic (0.0029) and phenotypic (0.0563) residual values were fairly very low, which revealed that variables included in this study had significant contribution in determining NSPF. The NSPHP exhibited high positive direct effect on NSPF. The maximum direct response (P = 0.737, G = 1.004) of this component was attributed to the indirect positive effects of the SWPF and fruit:seed weight proportion (FSWP). SWPF also had a very high positive direct effect (P = 0.521, G = 0.694) on the NSPF. Correlation and path coefficient analysis revealed that SWPF, NSPHP and 100-seed weight (100SW) were deciding factors for realizing improvement for NSPF. The importance of small seeded genotypes in selection of less seeded varieties was emphasized.     

Phylogenetic analysis and genetic mapping of Chinese Hedychium using SRAP markers

In the present study, using the SRAP markers, we analyzed the interspecific phylogenetic relationship of 22 species of Chinese Hedychium to find out fine mapping population. Using the same technique, maps of two species selected by morphological characters were constructed. Phylogenetic analysis showed that 22 species were grouped into three clusters. The mapping parents selected were in one cluster, with a similarity coefficient of 0.31, which is lower than the mean value of 22 species. Two maps with medium-density were constructed. The maternal parent contained 139 loci and spanned 917.1 cM. These loci were distributed in 18 linkage groups. The paternal parent contained 203 loci and spanned 1386.8 cM, which spread over 23 linkage groups. Unfortunately, the number of loci with a segregation ratio of 3:1 was not sufficient for us to link two maps together. For parental selection, we suggested that the phylogenetic relationship as well as complements of characters should be considered. Moreover, our study showed that two species with similarity around or higher than the mean level tend to produce fine mapping population in Hedychium.     

Microsatellite markers reveal low breeding system efficacy and pollen contamination can limit production of full-sib avocado progeny

Phytophthora cinnamomi causes a severe root rot in avocado, Persea americana. Breeding tolerant rootstocks is thought to be the most promising method for phytophthora root rot disease control but breeding avocado is challenging. The avocado flowering syndrome (synchronous protogynous dichogamy), combined with high flowering and low fruit set, render controlled pollination exceedingly difficult. Juxtaposing complementary flowering types of elite parent cultivars (cultivars that produce progeny with tolerance to phytophthora root rot) was performed in an effort to increase the number of full-sib progeny for elite maternal parents and, hypothetically, the number of phytophthora root rot tolerant progeny. Although high outcrossing rates were achieved (estimated ∼93%), the majority of progeny had a non-elite paternal parent (56% of progeny were offtypes) implying maternal trees were pollinated by non-elite distant trees. Among progeny that could be confidently genotyped, a high number of cross types were detected (33). Contrary to our hypothesis, a significant portion of the progeny were the result of crosses between like, and not complementary, flowering types. The spatial distribution of productive trees and grafts helped to explain these data, as productive grafts were directly adjacent to grafts of the same flowering type more often than that of the complementary flowering type. Selfed progeny were significantly less tolerant to phytophthora root rot than outcrossed progeny. Progeny resulting from crosses between an elite maternal parent and non-elite pollen donor (offtypes) were less tolerant than full-sib progeny resulting from crosses between elite parents. Maternal effects may interfere with identifying truly disease tolerant selections. Thus, to reduce maternal effects and non-elite pollen donor contamination, removal of seedling cotyledons before screening for disease tolerance and better isolation of elite parent trees and windbreaks may improve breeding efficacy. This study also demonstrates the usefulness of microsatellite markers in parentage analysis where a high proportion of the putative parents are closely related.     

Genetic diversity for mineral accumulation in the foliage of Chenopodium spp.

Chenopodium spp. is being used as a leafy vegetable and subsidiary grain crop in different parts of the world due to its rich nutritional quality and its ability to grow in stress conditions. A field experiment was conducted in Lucknow (India) to assess the genetic diversity in 40 accessions of Chenopodium spp. based on mineral composition of the leaves. Principal Component Analysis (PCA) showed that the first 4 PCs (Principal Component) accounted for 74.70% of the total variance among the accessions. The first PC (PC1) accounted for 41.96% of the total qualitative variation and had nickel, zinc and chromium with high positive and copper with high negative coefficients. The most important loadings for PC2 were calcium and potassium. Cluster analysis grouped the accessions into 4 major clusters. The first cluster, which showed maximum diversity, had 17 accessions, all of C. quinoa having high content of most of the heavy metals viz. zinc, chromium, nickel and cadmium. Cluster II was the largest consisting of 18 accessions which had low content of nickel, cadmium and chromium. Cluster III contained 3 accessions that had lowest amount of calcium, iron, magnesium and zinc, while accessions in cluster IV were characterized by high levels of calcium, sodium, magnesium, nickel, chromium and cadmium. Significant genotypic differences existed in the heavy metal uptake by plants. Mineral uptake and concentration in Chenopodium spp. corresponds to the taxonomic categorization in the genus and could also be utilized as a supplementary taxonomic tool for grouping together of closely related taxa. Conclusively the diversity can be exploited in plant breeding programs for increasing nutritional value and for bioremediation purposes.     

Study of genetic diversity in Chamomile (Matricaria chamomilla) based on morphological traits and molecular markers

Chamomile is one of the most important medicinal plants in the world trade that has many applications in drug and sanitary industrials. In order to evaluate the genetic diversity of different chamomile landraces based on morphological and molecular markers, 20 landraces were collected from different area of Iran. In addition to that, five populations imported from European were examined. The augmented design with four blocks and five controls were used to assess morphological traits. The RAPD method was utilized for evaluating the genetic diversity. Results showed that the economical yield, the number of flowers in plant, and the essential oil content had maximum variance coefficient. The flower's diameter and height had minimum variance coefficient. According to the cluster analysis on both morphological and molecular markers, 25 populations were classified into 5 clusters, but the population intra-groups were different. From 29 reliable primers that were used, 369 bands were detected and from which 314 (85.44%) bands were polymorphic. Genetic Jaccard's similarity coefficient was estimated in the range of 0.15–0.63, and with a mean of 0.35. Results showed that the genetic diversity was not according to the geographical diversity.     

Variation in total phenolics and antioxidant capacity among European plum genotypes

Our objective was to determine the amount of variation in total phenolics and antioxidants present in European plum fruits, so that it can be utilized in breeding programs to enhance the health benefits. Total antioxidant capacity and total phenolic content of the fruit, and the fruit skin color were determined in 20 genotypes, comprising of released varieties and advanced selections of European plums. Among the 20 genotypes, the total antioxidant capacity ranged from 105 to 424 mg ascorbic acid equivalents (AAE)/100 g fresh weight (FW) while the total phenolic content was 86–413 mg gallic acid equivalents (GAE)/100 g FW. The two parameters had a strong correlation of r² = 0.96. A direct correlation between skin color intensity and total phenolic content could also be observed. This study demonstrates that there is adequate variation in total phenolic compounds and antioxidants within European plums and hence there is potential for improvement towards enhancing these health-promoting phytochemicals in this fruit.     

Isotopically-labelled nitrogen uptake and partitioning in sweet cherry as influenced by timing of fertilizer application

Sweet cherry (Prunus avium L.) is a fruit of increasing economic importance though it is less significant than other stone fruit species such as peach. Cherry has received little attention concerning nitrogen (N) uptake and dynamics in mature trees. The aim of this work was to determine N uptake and partitioning as influenced by the timing of fertilizer application in 7-year-old sweet cherry trees cultivated in a cold region (Los Antiguos, Santa Cruz, Argentina; 71°38′ W, 46°32′ S). Nitrogen (95 kg ha⁻¹) was applied as ammonium nitrate to a soil with ‘Bing’ sweet cherry trees grafted onto Prunusmahaleb rootstocks. Fertilization was split into two equal applications per treatment, involving either the commercial fertilizer ammonium nitrate or the same fertilizer labelled with ¹⁵N isotope (10% atom.). Treatments consisted of one early spring (full bloom, October 2005) or one summer (late January 2006, 15 days after harvest) application of ¹⁵N ammonium nitrate to three replicate trees. Fruit were harvested in early January and leaves were collected at both full canopy and leaf fall. All trees were excavated in winter (August, 2006). Trees were partitioned into their components: trunk, branches (current-season shoots, 1-year-old and over-1-year-old branches), buds of the same age, small roots (less than 1 mm thick), large roots, leaves (sampled in February and April), and fruit (collected at harvest). Those components were dried and analysed for total N and ¹⁵N content. Total N per tree and N content derived from the fertilizer did not differ between treatments. Summer postharvest ¹⁵N application partitioned not only to structural components (trunk and roots) but also to buds and leaves. Uptake efficiency was significantly (p = 0.0113) higher in the spring than in the summer application (65.7% vs. 37.44%). Nevertheless, 52.5% of N applied in spring was lost due to harvest and summer pruning. This emphasizes the importance of the postharvest N fertilization which increases N accumulation in both reserve organs and buds though, according to our data, it is less efficiently used. The extent of nitrogen uptake, efficiency of use and partitioning in the following growing seasons are still open questions that deserve further research.