Influence of nitrogen availability on shoot development in young peach trees [Prunus persica (L.) Batsch]

It is commonly stated that nitrogen (N) influences biomass accumulation in plants. For trees, however, a precise characterisation of shoot response to N and its impact on architecture is lacking. We attempted to study on the phytomer scale the effects of N limitation on shoot growth components, i.e. leaf emergence rate, final internode length and branching on the main and secondary axes of 1-year-old peach trees [Prunus persica (L.) Batsch]. Trees were grown on recycled nutrient solutions in which N concentration was restored once a week. We used two hydroponic set-ups in which weekly N availability, i.e. amount of N per tree, differed being either low (N1) or high (N2). Net N availability was defined weekly as the relative amount of N remaining in each set-up before solution replenishment. It declined with time and three periods of contrasting net N availability were identified. During these periods, leaf emergence rate and final internode length were similar on the main axis of N1 and N2 trees, so too was the distribution of secondary axes along the main axis. Secondary axes responded to N limitation by decreasing their growth components according to their position along the main axis. Differences were most important for basal secondary axes. Leaf emergence rate and final internode length responded similarly to N availability depending on axis order and position in the tree. It was concluded that N availability, particularly during the period of maximum growth of axes, influenced the shoot growth components and thus tree architecture.     

Assessment of nutrient removal in bearing peach trees (Prunus persica L. Batsch) based on whole tree analysis

Background and Aims

The aim was to assess the amounts of macro- (N, P, K, Ca and Mg) and micro-elements (Fe, Mn, Cu and Zn) lost by peach trees (Prunus persica L. Batsch) in all the nutrient removal events (pruning, flower abscission, fruit thinning, fruit harvest and leaf fall), as well as those stored in the permanent structures of the tree (roots, trunk and main branches).

Methods

Three peach cultivars were used. The biomass and nutrient composition of materials lost by trees at the different events were measured during 3 years. The biomass and nutrient composition of permanent tree structures were also measured after full tree excavation.

Results

Winter pruning and leaf fall were the events where most nutrients were removed. Nutrient losses and total requirements are given as amounts of nutrients needed per tree and also as amounts necessary to produce a t of fresh fruit.

Conclusions

The allocation of all nutrients analyzed in the different plant parts was similar in different types of peach trees, with each element having a typical “fingerprint” allocation pattern. Peach tree materials removed at tree pruning and leaf fall include substantial amounts of nutrients that could be recycled to improve soil fertility and tree nutrition. Poorly known tree materials such as flowers and fruit stones contain measurable amounts of nutrients.     

Mapping QTLs controlling fruit quality in peach (Prunus persica (L.) Batsch)

 The organoleptic quality of fleshy fruits is in a large part defined by their composition of soluble sugars and organic acids. An F₂ population issuing from a cross between two peach varieties, ‘Ferjalou Jalousia’, a non-acid peach, and ‘Fantasia’, an acid nectarine, was analysed over 2 successive years for agronomic characters and for molecular-marker (isoenzymes, RFLPs, RAPDs, IMAs and AFLPs) segregations. Blooming and maturity dates, as well as productivity, were noted for each tree. Four fruits per tree were analysed at maturity for fresh weight, colour, pH, titratable acidity, soluble-solids content (SSC), acid (malic, citric and quinic acids) and sugar (sucrose, glucose, fructose, sorbitol) contents. QTLs were detected for all fruit components analysed, except for fruit colour. The QTLs for nearly all components were present on two linkage groups. For productivity, fresh weight, pH, quinic acid, sucrose and sorbitol content, all the detected QTLs displayed the same effect as the parental phenotypes. By contrast, for maturity date, titratable acidity, malic and citric acids and fructose, some QTLs displayed the same effect as the parental phenotypes while others displayed the opposite effect. The fraction of the total variation in each trait throughout the population explained by the QTLs was very high and reached more than 90% for some characters. For most of the characters analysed, epistasis was observed between QTLs. Received: 10 October 1997 / Accepted: 18 August 1998     

In vitro callus induction from adult tissues of peach (Prunus persica L. Batsch)

We describe an efficient protocol for callus induction from adult tissues of Prunus persica (L.) Batsch. Three different commercial peach genotypes, Early May®, Zise May®, and UFO-3®, plus three other genotypes from hybrid crosses performed in February 2006, PS108, PS208, and PS708, were used in the study. Thirteen explant treatments were tested using nine different plant parts. Murashige and Skoog and woody plant medium salts were assayed with several concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D), kinetin (KN), and thidiazuron, and two different photoperiods were tested, a 16-h photoperiod or continuous darkness. In terms of the quantitative response, two parameters were assessed: the number of d to callus induction and relative callus growth recorded after 30 d. Woody plant medium supplemented with 2,4-D and KN significantly increased the rates of callus induction in the majority of treatments. And no significant differences among the P. persica genotypes were found. The explants derived from stem and calyx produced up to 85 and 96% callus induction, respectively. The protocol described here could be used for efficient callus induction in a range of Prunus spp.     

Characterization of microsatellite markers in peach [Prunus persica (L.) Batsch]

Microsatellites have emerged as an important system of molecular markers. We evaluated the potential of microsatellites for use in genetic studies of peach [Prunus persica (L.) Batsch]. Microsatellite loci in peach were identified by screening a pUC8 genomic library, a λZAPII leaf cDNA library, as well as through database searches. Primer sequences for the microsatellite loci were tested from the related Rosaceae species apple (Malus×domestica) and sour cherry (Prunus cerasus L.). The genomic library was screened for CT, CA and AGG repeats, while the cDNA library was screened for (CT)_n- and (CA)_n-containing clones. Estimates of microsatellite frequencies were determined from the genomic library screening, and indicate that CT repeats occur every 100 kb, CA repeats every 420 kb, and AGG repeats every 700 kb in the peach genome. Microsatellite- containing clones were sequenced, and specific PCR primers were designed to amplify the microsatellite- containing regions from genomic DNA. The level of microsatellite polymorphism was evaluated among 28 scion peach cultivars which displayed one to four alleles per primer pair. Five microsatellites were found to segregate in intraspecific peach-mapping crosses. In addition, these microsatellite markers were tested for their utility in cross-species amplification for use in comparative mapping both within the Rosaceae, and with the un- related species Arabidopsis thaliana L. Received: 18 June 1999 / Accepted: 6 December 1999     

Genetic mapping of the evergrowing gene in peach [Prunus persica (L.) Batsch

In temperate locations, terminal apices on evergrowing (also called evergreen) peach trees keep growing in winter until killed by low temperatures, while the lateral buds go into dormancy. A recessive allele of a single gene (evergrowing or evg) controls this trait in peach. The amplified fragment length polymorphism (AFLP) technique and bulked segregant analysis were applied to construct a local genetic linkage map for the evg gene from the cross Empress op op dwarf x Evergrowing (P.I. 442380). This map, comprising nine AFLP markers and the evg locus, covers a total genetic distance of 79.3 cM. Four dominant AFLP markers (EAT/MCAC, ETT/MCCA2, EAT/MCTA, and ETT/MACC) were linked to the evg locus at distances of 1, 5.3, 6.7, and 11.7 cM, respectively. EAT/MCAC and EAT/MCTA were converted into polymorphic sequence-tagged sites. Microsatellite markers in the evg region were developed from peach bacterial artificial chromosome (BAC) clones that hybridized to the AFLP marker fragments. Using three microsatellite anchor markers (pchgms12, pchgms17, and pchgms19), the local genetic linkage map was integrated into one minor linkage group of a previously constructed peach rootstock genetic linkage map. Three AFLP markers from the rootstock genetic linkage map were found linked to the evg locus.     

New high-quality peach (Prunus persica L. Batsch) genome assembly to analyze the molecular evolutionary mechanism of volatile compounds in peach fruits

Peach (Prunus persica L. Batsch) is an economically important fruit crop worldwide. Although a high-quality peach genome has previously been published, Sanger sequencing was used for its assembly, which generated short contigs. Here, we report a chromosome-level genome assembly and sequence analysis of Chinese Cling, an important founder cultivar for peach breeding programs worldwide. The assembled genome contained 247.33 Mb with a contig N50 of 4.13 Mb and a scaffold N50 of 29.68 Mb, representing 99.8% of the estimated genome. Comparisons between this genome and the recently published one (Lovell peach) uncovered 685 407 single nucleotide polymorphisms, 162 655 insertions and deletions, and 16 248 structural variants. Gene family analysis highlighted the contraction of the gene families involved in flavone, flavonol, flavonoid, and monoterpenoid biosynthesis. Subsequently, the volatile compounds of 256 peach varieties were quantitated in mature fruits in 2015 and 2016 to perform a genome-wide association analysis. A comparison with the identified domestication genomic regions allowed us to identify 25 quantitative trait loci, associated with seven volatile compounds, in the domestication region, which is consistent with the differences in volatile compounds between wild and cultivated peaches. Finally, a gene encoding terpene synthase, located within a previously reported quantitative trait loci region, was identified to be associated with linalool synthesis. Such findings highlight the importance of this new assembly for the analysis of evolutionary mechanisms and gene identification in peach species. Furthermore, this high-quality peach genome provides valuable information for future fruit improvement.     

Relationship of changes in the fatty acid compositions and fruit softening in peach (Prunus persica L. Batsch)

Fatty acid compositions of peach (Prunus persica L. Batsch) mesocarp tissues from ‘Kawanakajima Hakuto’ and its firm-fleshed mutant ‘Shuangjiuhong’ were examined by gas chromatography during the developmental stages from 20 days before to 20 days after fruit ripening. Fruits were harvested at 4-day intervals from July to September. The predominant fatty acids were linoleic, palmitic and linolenic acids with 27.66–48.93 %, 23.59–31.65 %, and 12.08–28.35 % in ‘Shuangjiuhong’, and 32.64–42.79 %, 23.53–28.95 %, 16.14–39.15 % in ‘Kawanakajima Hakuto’, respectively. Saturated fatty acids (palmitic and stearic acids) remained relatively constant throughout the ripeness period. On the contrast, from 15 days before ripening, notable decline in oleic acid and increase of linoleic and linolenic acids were observed in both cultivars. In addition, from 10 days before ripening, much lower levels of oleic and linolenic acids and higher proportion of linoleic acid were observed in ‘Shuangjiuhong’ than those found in ‘Kawanakajima Hakuto’. And notably higher SFA level, lower levels of UFA and IUFA in the firm-fleshed peach were investigated during those stages. Correlation analysis showed that oleic acid and SFA had very significantly positive, whereas linolenic acid, UFA and IUFA had significantly negative correlation with fruit firmness. These results above suggest that lower levels of oleic and linolenic acids, UFA and IUFA, and higher linoleic acid and SFA content represent fruits with firmer flesh and help to retain the fruit texture.     

The effect of autumn N supply on the architecture of young peach (Prunus persica L.) trees

Irrigation and fertilisation were recently considered as useful tools to control tree shape, and reduce pruning costs. The role of the N reserves, which determined spring growth, was considered to be essential. We intended therefore to evaluate its effects on peach tree architecture. Four levels of N fertilisation were applied on 1-year-old trees, from the end of shoot growth to leaf fall. In subsequent spring, each bud fell into one of the ten classes of positions previously defined within the crown. Its development was followed weekly from burst to June. Fertilisation promoted growth until a threshold level, since no differences were evidenced between the three highest N treatments. Fall N did not affect burst but the further transformation of the buds into rosettes, proleptic or ramificated axes. Crown base was little affected. Fall N increased the number of proleptic axes on most median and upper positions. Axes lengthening and thickening were limited on the median positions, promoted at crown top. The variations concerned the mean internodes lengths, not the number of phytomers per axis. Sylleptic ramification was limited to the crown outer parts, and decreased with fall N. Treatment did neither affect the fruit dry weights, nor the ratio between the number of leaves and the number of fruits. Fruit number was proportioned to vegetative growth by blossoming and fruit set. We conclude that a moderate autumn fertilisation improved orchard productivity, but favoured vegetative growth in the crown outer parts. Additional pruning may therefore be required to control tree shape.     

A first insight into peach [Prunus persica (L.) Batsch] SNP variability

Three factors may have reduced the diversity at both individual gene and whole genome levels in cultivated peach: its self-compatible mating system, the narrow genetic basis of most commercial cultivars, and the recent strong selection towards agronomically interesting traits. Previous diversity analyses with markers such as simple sequence repeats (SSRs) have revealed low levels of genetic variability. Here, we sequenced 23 genome-wide distributed DNA fragments in 47 occidental peach varieties, also observing reduced variability levels. On average, there was one single nucleotide polymorphism (SNP) every 598 bp and one indel every 4,189 bp. As expected, variability was higher in non-coding than in coding regions (one SNP every 390 non-coding bp versus one in 1,850 bp in coding DNA). In general, SNPs were observed at relatively high frequency, mean minor allele frequency?=?0.225, meaning that a large proportion of the SNPs discovered by sequencing similar germplasm will be useful for other purposes, such as association mapping. The average heterozygosity of the varieties was 0.28, with a low correlation between SSR and SNP heterozygosity. The whole sequence of two candidate genes, a pectate lyase 1 candidate for fruit firmness (CGPAA2668) and a sucrose synthase 1 candidate for sugar content (CGPPB6189), in the 47 varieties revealed that they both may have suffered a process of balancing selection.     

Effects of carbohydrates and nitrogen on the development of anthocyanins of a red leaf peach (Prunus persica (L.) Batsch) in vitro

Heterozygous red leaf peach (Prunus persica (L.) Batsch) shoots were implanted on media with varying nitrogen and carbohydrate regimes to identify a combination which elicited maximum anthocyanin production in explants. A medium with relatively low nitrogen (5 mM NH₄₊ and 10 mM NO_3-) and high sucrose (234 mM) was most effective in stimulating anthocyanin production. Sucrose was more effective as a carbon source than glucose, fructose, or starch under given nitrogen levels. The major anthocyanin in red leaf peach was tentatively identified as cyanidin 3-glucoside based on PC and HPLC analysis.     

Prognosis of iron chlorosis in pear (Pyrus communis L.) and peach (Prunus persica L. Batsch) trees using bud, flower and leaf mineral concentrations

Background and Aims

The possibility of using tree materials in early phenological stages, such as dormant buds and flowers, for the prognosis of Fe deficiency occurring later in the year has been studied in peach and pear trees.

Methods

Thirty-two peach trees and thirty pear trees with different Fe chlorosis degrees were sampled in different commercial orchards. In peach, samples included flower buds, vegetative buds, bud wood, flowers and leaves at 60 and 120?days after full bloom (DAFB). In pear, samples included buds, bud wood, flowers and leaves at 60 and 120?days DAFB. Leaf chlorophyll was assessed (SPAD) at 60 and 120 DAFB. Sampling was repeated for 3–5?years depending on the materials. Mineral nutrients measured were N, P, K, Ca, Mg, Fe, Mn, Zn and Cu.

Results

The relationships between the nutrient concentrations in the different materials and leaf SPAD were assessed using four different statistical approaches: i) comparison of means depending on the chlorosis level, ii) correlation analysis, iii) principal component analysis, and iv) stepwise multiple regression. In all cases, significant associations between nutrients and SPAD were found. The best-fit multiple regression curves obtained for the multi-year data set provided good prediction in individual years.

Conclusions

Results found indicate that it is possible to carry out the prognosis of Fe chlorosis using early materials such as buds and flowers. The relationships obtained were different from those obtained in previous studies using a single orchard. The different methods of analysis used provided complementary data.     

Mapping quantitative trait loci associated with blush in peach [Prunus persica (L.) Batsch]

Blush is an important trait for marketing peaches. The red skin pigmentation develops through the flavonoid and anthocyanin pathways, and both genetic and environmental stimuli, and their interaction, control the regulation of these pathways. The molecular basis of blush development in peach is yet to be understood. An F₂ blush population (ZC²) derived from a cross between two peach cultivars with contrasting phenotypes for blush, “Zin Dai” (～30 % red) and “Crimson Lady” (～100 % red), was used for linkage map construction and quantitative trait loci (QTLs) mapping. The segregating population was phenotyped for blush for 4 years using a visual rating scale and quantified using a colorimeter (L*, a*, and b*) 1 year. The ZC² population was genotyped with the IPSC 9 K peach single-nucleotide polymorphism (SNP) array v1, and a high-density ZC² genetic linkage map was constructed. The map covers genetic a distance of ～452.51 cM with an average marker spacing of 2.38 cM/marker. Four QTLs were detected: one major QTL on LG3 (Blush.Pp.ZC-3.1) and three minor QTLs on LG 4 and 7 (Blush.Pp.ZC-4.1; Blush.Pp.ZC-4.2; Blush.Pp.ZC-7.1), indicating the presence of major and minor genes involved in blush development. Candidate genes involved in skin and flesh coloration of peach (PprMYB10), cherry (PavMYB10), and apple (MdMYB1/MdMYBA/MdMYB10) are located within the interval of the major QTL on LG3, suggesting the same genetic control for color development in the Rosaceae family. Marker-assisted selection (MAS) for blush is discussed.     

Molecular characterisation of sweet cherry (Prunus avium L.) genotypes using peach [Prunus persica (L.) Batsch] SSR sequences

A total of 76 sweet cherry genotypes were screened with 34 microsatellite primer pairs previously developed in peach. Amplification of SSR loci was obtained for 24 of the microsatellite primer pairs, and 14 of them produced polymorphic amplification patterns. On the basis of polymorphism and quality of amplification, a set of nine primer pairs and the resulting 27 informative alleles were used to identify 72 genotype profiles. Of these, 68 correspond to unique cultivar genotypes, and the remaining four correspond to three cultivars that could not be differentiated from the two original genotypes of which they are mutants, and two very closely related cultivars. The mean number of alleles per locus was 3.7 while the mean heterozygosity over the nine polymorphic loci averaged 0.49. The results demonstrate the usefulness of cross-species transferability of microsatellite sequences allowing the discrimination of different genotypes of a fruit tree species with sequences developed in other species of the same genus. UPGMA cluster analysis of the similarity data divided the ancient genotypes studied into two fairly well-defined groups that reflect their geographic origin, one with genotypes originating in southern Europe and the other with the genotypes from northern Europe and North America.     

Microsatellite DNA in peach (Prunus persica L. Batsch) and its use in fingerprinting and testing the genetic origin of cultivars.

We isolated and sequenced 26 microsatellites from two genomic libraries of peach cultivar 'Redhaven', enriched for AC/GT and AG/CT repeats, respectively. For 17 of these microsatellites, it was possible to demonstrate Mendelian inheritance. Microsatellite polymorphism was assayed in 50 peach and nectarine cultivars. Of the 1300 PCRs carried out, all but two produced amplified products of the expected size. All microsatellites were polymorphic, showing 2-8 alleles per locus. Heterozygosity ranged from 0.04-0.74 (mean 0.47); the discrimination power (PD) ranged from 0.04-0.84 (mean 0.60). Cultivar heterozygosity varied greatly, with one cultivar ('Independence') being homozygous at all loci. The set of microsatellites discriminated all cultivars investigated, except several sport mutations, i.e., 'Dixitime' vs. 'Springcrest', 'Compact Redhaven' vs. 'Redhaven', and two pairs of cultivars, 'Venus' vs. 'Orion' and 'Elegant Lady' vs. 'Rome Star', whose pedigrees are controversial. We were able to analyze the paternity of several cultivars. In most cases, the parenthood was confirmed. The comparison of three long-living 'Redhaven' accessions supplied by different repositories did not provide any evidence of somatic instability of microsatellites. Hence, microsatellites, ranked according to their information content, are recommended as markers of choice for peach fingerprinting and suggestions are provided for interpreting band profiles and the correct sizing of alleles.     

QTL mapping for brown rot (Monilinia fructigena) resistance in an intraspecific peach (Prunus persica L. Batsch) F1 progeny

Brown rot (BR) caused by Monilinia spp. leads to significant post-harvest losses in stone fruit production, especially peach. Previous genetic analyses in peach progenies suggested that BR resistance segregates as a quantitative trait. In order to uncover genomic regions associated with this trait and identify molecular markers for assisted selection (MAS) in peach, an F1 progeny from the cross “Contender” (C, resistant)?×?“Elegant Lady” (EL, susceptible) was chosen for quantitative trait loci (QTL) analysis. Over two phenotyping seasons, skin (SK) and flesh (FL) artificial infections were performed on fruits using a Monilinia fructigena isolate. For each treatment, infection frequency (if) and average rot diameter (rd) were scored. Significant seasonal and intertrait correlations were found. Maturity date (MD) was significantly correlated with disease impact. Sixty-three simple sequence repeats (SSRs) plus 26 single-nucleotide polymorphism (SNP) markers were used to genotype the C?×?EL population and to construct a linkage map. C?×?EL map included the eight Prunus linkage groups (LG), spanning 572.92 cM, with an average interval distance of 6.9 cM, covering 78.73 % of the peach genome (V1.0). Multiple QTL mapping analysis including MD trait as covariate uncovered three genomic regions associated with BR resistance in the two phenotyping seasons: one containing QTLs for SK resistance traits near M1a (LG C?×?EL-2, R ²?=?13.1–31.5 %) and EPPISF032 (LG C?×?EL-4, R ²?=?11–14 %) and the others containing QTLs for FL resistance, near markers SNP_IGA_320761 and SNP_IGA_321601 (LG3, R ²?=?3.0–11.0 %). These results suggest that in the C?×?EL F1 progeny, skin resistance to fungal penetration and flesh resistance to rot spread are distinguishable mechanisms constituting BR resistance trait, associated with different genomic regions. Discovered QTLs and their associated markers could assist selection of new cultivars with enhanced resistance to Monilinia spp. in fruit.