Fruit antioxidant capacity and self-incompatibility genotype of Ukrainian sweet cherry (Prunus avium L.) cultivars highlight their breeding prospects

Breeding for higher levels of health-promoting components are among the priorities for many fruit crops. Such an approach requires breeding material displaying great variations in the trait to be improved. Commercial sweet cherry cultivars demonstrate only limited variability in fruit antioxidant parameters and hence this study was carried out to characterize some Ukrainian sweet cherry cultivars for their antioxidant and antiradical capacity and total phenolic and anthocyanin contents. Ukrainian cultivars had significantly higher ferric reducing antioxidant power and total phenolic contents compared with commercial cultivars. Levels of all four parameters varied considerably in the Ukrainian germplasm, with some cultivars having outstanding values. A total of eight different self-incompatibility genotypes were assigned to the 14 accessions tested. Assigning cultivars to the appropriate incompatibility groups allowed the design of parental combinations which may result in new genotypes with enhanced functional properties and other valuable fruit traits. Based on the S-genotypes, parental combinations were proposed to ensure 50 or 100 % self-compatible seedlings in the offspring population. The most perspective Ukrainian cultivars like ‘Dagestanka’ and ‘Kodrinskaya’ also had reasonable fruit weight (over 8 g), high flesh to pit ratio (approx. 18) and soluble solid contents (over 16 %). Our results demonstrated that adequate variation is available in Ukrainian germplasm for improving fruit health benefits in sweet cherry through directed hybridizations.     

Inheritance of tree and fruit characters in progenies from crosses of sweet cherry (Prunus avium L.) cultivars

Summary Approximately 1000 seedlings from 20 combinations crossed in 1979, 1980 and 1981 (Theiler-Hedtrich 1985a) were tested for several characters: fruit set (yield), fruit size, fruit colour, formation of abscission layer and bleeding after fruit removal from fruit stalks, bacterial canker resistance, flowering and harvesting time. From progeny of crosses with Stella as pollinator, 56% (Vittoria × Stella) and 46% (Schüttler × Stella) of the seedlings were self-compatible, of which 14 were high yielding with good fruit size and quality. From the data recorded it can be concluded:fruit set is a recessive character; only 5 to 20% of very good yielding seedlings were obtained in different progeny, even if the parental plants were both very good croppers.Fruit juice and skin colour was in most progenies ‘black’ even if they were from combinations with ‘white’ varieties, e.g., Merton Glory or Schüttler. Only from the combination Schiittler (‘white’) × Stella (‘black’), 50% of the seedlings were ‘white’; Stella therefore is heterozygous for the character of fruit juice and skin colour.Fruit size is evenly distributed in progeny with respect to the fruit size of their parent plants.Abscission layer formation and non-bleeding is a genetically complex character. In combinations where both parent plants formed fruits with complete abscission layers and which were not bleeding after fruit removal from the stalk, this character was inherited only to 50% (Vittoria × Schüttler) or 85% (Vittoria × Frühe von der Weid) in the progeny. For the genetical control of this character further studies are necessary.Bacterial canker susceptibility was evenly distributed in seedlings from all combinations even if the highly resistant cv. Vittoria was used as one parent plant, thereby not confirming the expected results of a higher proportion of resistant seedlings from combinations with Vittoria.Flowering and harvest time of the seedlings from different combinations was within the range of the parent plants. Only in the combination of Vittoria × Stella (mid-to late-ripening season) one seedling out of 99 was found to form ripe fruits two weeks earlier than the parental plants. From the seedlings tested 40 have been chosen for further evaluation or genetical studies.     

Developmental changes in cell wall polysaccharides from sweet cherry (Prunus avium L.) cultivars with contrasting firmness

Firmness is a major quality attribute of fresh cherries, and is also a main factor affecting susceptibility to bruising and postharvest rots. In order to identify the factors determining the textural differences between genotypes, we evaluated the solubilization, depolymerization and monosaccharide composition of pectin and hemicelluloses from two cultivars with contrasting firmness (‘Sweetheart’, firm and ‘Newstar’, soft) at four different developmental stages. Firm ‘Sweetheart’ cherries had higher contents of cell wall material than soft ‘Newstar’ fruit. Moderate depolymerization of hemicellulose and tightly bound pectins was detected irrespective of cultivar firmness. The general pattern and extent of uronic acid solubilization was quite similar in both cultivars. Rhamnogalacturonan I (RG-I) seemed to be preferentially solubilized in firm ‘Sweetheart’ fruit as opposed to tightly bound homogalacturonans (HG) in soft cherries. Pectic polymers with higher neutral sugar to uronic acids ratio were found from early development in soft ‘Newstar’ fruit. Overall, soft ‘Newstar’ fruit had reduced wall content and higher branching of tightly bound pectins than firm ‘Sweetheart’ fruit. These factors may be associated with the varietal differences in cherry firmness.     

Determination of self-incompatible genotypes in sweet cherry (Prunus avium L.) accessions and cultivars of the German Fruit Gene Bank and from private collections

Sweet cherries are self-incompatible because of a gametophytic self-incompatibility system. S alleles in the style and pollen determine the crossing relationships. Knowledge of the S allele constitution of cultivars is very important for cherry growers and breeders, and recently, molecular methods have been developed to distinguish the S alleles in sweet cherries. The S allele genotypes of 149 sweet cherry cultivars and clones, including 126 not previously genotyped, were determined by using PCR analysis. Thirteen different S alleles in 40 combinations were distinguished and nine new incompatibility groups were documented. Two new S alleles were identified in five local sweet cherry processing cultivars from southwestern Germany using the second intron primers. The sequence of these alleles was determined and compared to all known sequences available in the NCBI database. The sequences obtained showed high similarities to the alleles S ₁₉ and S ₂₂, previously described only in wild cherries, Prunus avium L.     

Fertility and pollen tube growth in polyploid melons (Cucumis melo L.)

The high sterility of tetraploid melon (Cucumis melo L.) plants is a serious inconvenient to obtain economically useful polyploid lines. In order to study the alterations on diploid pollen tube growth and their influence on sterility, in vitro and in vivo pollen tube growth from three commercial varieties and its tetraploids was observed. Polyploid pollen grains show a lower germination percentage in vitro and a slower germination rate than haploid pollen. Furthermore, diploid pollen grains producing two or three tubes and tube branchings were observed on diploid pollen; this seems to indicate a decrease in its functionality. In vivo and in vitro pollen behaviours are quite similar, but an interaction between the pollen and the pistillar tissues was detected in vivo. In the diploid × tetraploid cross, where fruits were not obtained, pollen tubes showed a slow growth rate at the base of the pistil. The possible effects of these alterations in sterility are discussed in terms of the factors that could affect fecundation. This revised version was published online in July 2006 with corrections to the Cover Date.     

Development of a molecular marker for self‐compatible S4′ haplotype in sweet cherry (Prunus avium L.) using high‐resolution melting

Sweet cherry (Prunus avium L.) has stylar gametophytic self‐incompatibility, which is controlled by the multi‐allelic S‐locus and encompasses the highly polymorphic genes for the S‐ribonuclease (S‐RNase) and S‐haplotype‐specific F‐box (SFB), which are female and male determinants, respectively. The self‐compatible mutant SFB4′ corresponds to an allele variant of SFB4 and presents a frameshift mutation. Even though male‐determinant molecular markers can discriminate between SFB4 and SFB4′ alleles, the methods required are laborious, time‐consuming and expensive, and not suitable for massive analysis and integration into breeding programmes. Our aim was to develop molecular markers for the evaluation of self‐compatibility alleles in sweet cherry, that could be used as a high‐throughput screening strategy to identify SFB4 and SFB4′ alleles, based on a marker for male determinacy. Our results were consistent using primers flanking the mutation responsible for the SFB4′ allele. We designed a specific molecular marker and confirmed it in sweet cherry commercial varieties. This new molecular marker is feasible for self‐compatibility alleles in the male determinant in sweet cherry‐assisted breeding programs.     

南方高海拔地区栽培甜樱桃质地特性研究

以南方高海拔地区栽培的甜樱桃品种"美早"和"萨米脱"为试材,利用质构仪质地多面分析法(TPA)和整果穿刺法研究其贮藏过程中质地特性的变化。结果表明,贮藏过程中两个品种樱桃果肉的硬度与弹性、咀嚼性,咀嚼性与弹性均呈极显著正相关,且随贮藏时间的延长,果肉硬度、弹性、咀嚼性和果皮强度下降,果皮韧性增强,而果肉凝聚性和回复性则先上升后下降。贮藏42d后",美早"甜樱桃果肉的硬度、咀嚼性、凝聚性和回复性显著高于"萨米脱"甜樱桃,同时保持较低的果皮韧性,说明其更耐贮藏。     

The crossing of common wheat (Triticum aestivum L.) with cultivated rye (Secale cereale L.). I. crossability,pollen grain germination and pollen tube growth

Summary The crossing of common wheat (Triticum aestivum L.) with rye (Secale cereale L.), and especially the action of the crossability genes of wheat, was studied using the readily crossable wheat cv Chinese Spring (genotype kr ₁ kr ₁ kr ₂ kr ₂ >), the poorly crossable wheat cv Hope (genotype Kr ₁ Kr ₁ Kr ₂ Kr ₂ ), as well as the disomic substitution line of chromosome 5B of Hope into Chinese Spring (CS/Hope 5B, genotype Kr ₁ Kr ₁ kr ₂ kr ₂ ). By comparing crossability and actual fertilization, the poor crossability with rye of both cv Hope and the CS/Hope 5B substitution line was shown to result from absence of fertilization. Studies of pollen grain germination and pollen tube growth showed that the dominant alleles of the crossability genes manifested themselves through retardation and eventually inhibition of pollen tube growth at the style base and in the ovary wall. In Hope the growth of all pollen tubes was inhibited, whereas in CS/Hope 5B rarely fertilization was achieved. The recessive alleles of the crossability genes do not seem to have an influence on the growth of rye pollen tubes in wheat pistils.     

Germination of pollen of weed rye (Secale segetale L.) on wheat (Triticum aestivum L.) stigmas and the growth of the pollen tubes

Summary Two gene pairs have been found regulating the crossability of wheat and rye. It is concluded from our work that the same genes regulate crossability between wheat and weed rye. The crossing barrier was not observed in the stigma, style or ovary wall because pollen tubes of weed rye were seen in these tissues irrespective of the wheat variety used as the female parent.Pollen grains germinated within 6 min after pollination. The tubes reached the region of the micropyle after about 40 min.     

Towards sweet cherry (<Emphasis Type="Italic">Prunus avium</Emphasis> L.) breeding: phenotyping evaluation of newly developed hybrids

An increasing demand for cherry production (Prunus avium L.) in Greece led to the development of new high quality sweet cherry cultivars. Self-incompatibility in cherry is one of the most challenging issues for the species’ cultivation and top breeding priority. Τhe present study focuses on the development of new hybrids with improved traits such as productivity, fruit size, organoleptic characteristics and self-compatibility. For this purpose, thirty different cultivars were crossed and produced hybrids that were evaluated according to 34 morpho-physiological characteristics. The results were analyzed using the XLSTAT (version 2014.1) software and a dendrogram was constructed using the agglomerative hierarchical clustering method. Optimal hybrid clustering was achieved when characteristics of great economic importance such as fruit shape and size, growth habit and days to blooming were included in the analysis. Based on the results, new sweet cherry hybrids with the special character of self-compatibility were developed. Our findings provide crucial new information for sweet cherry future breeding programs and cultivation.     

Assessment of sweet cherry (<Emphasis Type="Italic">Prunus avium</Emphasis> L.) genotypes for response to bacterial canker disease

Integration of alleles for bacterial canker resistance into new sweet cherry cultivars requires information on the sources of resistance in the germplasm. Five market-leading sweet cherry cultivars, ‘Rainier’, ‘Sweetheart’, ‘Bing’, ‘Regina’ and ‘Chelan’, advanced selections ‘AA’, ‘BB’, ‘CC’, ‘DD’, ‘EE’, ‘GG’, and ‘PMR-1’ used as breeding parents in the Washington State University’s Sweet Cherry Breeding Program were evaluated. Comparative genotypic disease severity was obtained with three methods of inoculation (leaf wounding with carborundum, cut wounds in leaf mid-rib and shoot tip) on whole plants. Additionally, genotypic data on susceptibility of detached leaves versus fruit and an assessment of the movement of Pseudomonas syringae pv. syringae (Pss) population in inoculated shoots were obtained. Genotype susceptibility was significantly (P ≤ 0.05) influenced by inoculation method, with shoot inoculation providing the best separation of resistance levels among genotypes. A low correlation (r = 0.26, P = 0.21) was observed between disease responses measured on detached leaf versus fruit, while a moderately high correlation (r = 0.50, P = 0.10) was found among bacterial populations in the tissues and in the degree of symptoms expressed. By all comparative methods, the advanced selections, as well as, ‘PMR-1’, were less susceptible than the market-leading cultivars. Also, movement of Pss from shoot tip inoculation points to the shoot base was not detected for advanced selections ‘AA’, ‘BB’, ‘DD’, and ‘EE’. This study reveals that the advanced selections could be potential sources of resistance alleles to bacterial canker. This is the first evaluation of the advanced selections for bacterial canker disease.     

RFLP variability in apricot (Prunus armeniaca L.)

The level of polymorphism of the restriction fragment length polymorphisms (RFLPs) detected by 33 almond genomic and cDNA probes was studied in a set of 52 European and North-American apricot cultivars. Eighteen of these probes were polymorphic and yielded a total of 48 scorable bands, allowing the identification of 45 different phenotypes. Most cultivars (43) had an individually distinguishable RFLP phenotype, and three of the five clusters with the same phenotype contained cultivars that were likely to be synonymous. The group of Spanish cultivars (25) had a lower level of polymorphism than the others, suggesting that bottlenecks may have occurred in the recent history of the apricot that have eroded its genetic variability.     

The effect of temperature on pollen production in carnations

Summary For breeding carnations the supply of good pollen may be a problem. For this reason the effect of temperature on pollen production in carnations was studied.In phytotron experiments a temperature of 23°C proved to be the optimum.     

Pollen germination,pollen tube growth and fertilization following self and interspecific pollination of Paspalum species

Summary Crossability between most Paspalum species is very low. This study was undertaken to identify the impediments to hybridization. Accessions of P. intermedium Munro. ex Morong, P. jurgensii Hackel and P. dilatatum Poir were self-pollinated and crossed with one anther. Paspalum intermedium is essentially self-sterile but P. jurgensii and P. dilatatum are highly self-fertile. Following pollination, pollen germination and tube growth were studied by observing the pollinated pistils with fluorescent microscopy. Examination of self-pollinated pistils revealed that the pollen germinated shortly after contacting the stigmas. Germination was over 80% for the P. intermedium and P. dilatatum accessions but only 57% for P. jurgensii. Pollen tubes grew to the micropyle within 45 minutes after pollination in P. dilatatum and 1 hour and 15 minutes in P. jurgensii. However, in the P. intermedium accessions most tubes did not grow beyond the stigma and very few penetrated the style and ovary. Apparently stylar-incompatibility is the reason for the low selfed seed set. In the cross-pollinations, pollen germinated shortly after pollination and germination ranged from 57 to 88% for the different crosses. In all crosses the pollen tubes grew to the micropyle within 30 minutes to 2 hours after pollination indicating that a cross-incompatibility system is not the cause for low crossability among these species. By examining embryo sacs from P. intermedium × P. dilatatum, its reciprocal and P. intermedium × P. urvillei crosses, it was determined that gametes failed to unite in some crosses and this is a major reason for low crossability.     

In vitro pollen germination and tube growth of tomato (Lycopersicon esculentum Mill.) and its relation with plant growth

Summary In vitro pollen germination at 10°C, 14°C and 22°C of four groups of two pure line tomato varieties was compared with their plant growth at 19°C D/10°C N under controlled environmental conditions. Generally, pollen germination was slow at 10°C but after 6 h the percentage of germination was similar to that at 22°C. Maximum germination was obtained at 14°C already after 4 h. The longest pollen tubes occurred at 22°C. The two varieties within each group differed significantly from each other for percentage of pollen germination. For one group, this difference was greater at 10°C than at 22°C. The varieties in two groups also differed significantly for pollen tube growth. These differences in pollen tube growth rate were greater at 22°C than at 10°C. There was no clear relationship between pollen germination, pollen tube growth and plant growth in any of the four pairs of varieties. The results are discussed with regard to the possibility of pollen selection at low temperature in order to improve the efficiency of breeding for growth at low temperature.     

The effect of pollen load and pollen grain competition on fertilization success and progeny performance inFagopyrum esculentum

Summary Flowers of cultivated buckwheat (Fagopyrum esculentum Moench) often receive natural pollen loads of fewer than 10 pollen grains. The cultivated varieties also have high genetic variability. These observations raise the question of whether seed production in buckwheat is often limited by pollen delivery, and whether small increases in pollen load could result in gametophytic selection through pollen grain competition. In greenhouse-grown buckwheat plants, embryo sac penetration by pollen tubes was universal with 10 or more pollen grains. However, seed production increased with pollen load up to 30 grains per flower. Larger pollen loads, which intensify selection among gametophytes, resulted in more vigorous progeny. Seedlings produced from high pollen load (15–20 pollen grains) were larger (40% by weight) than those from low pollen load (5 pollen grains). These results are evidence that pollen grain competition can occur in buckwheat with benefits for progeny performance.     

Effect of high temperature on pollen morphology,plant growth and seed yield in quinoa (Chenopodium quinoa Willd.)

Quinoa (Chenopodium quinoa Willd.) has gained considerable attention worldwide during the past decade due to its nutritional and health benefits. However, its susceptibility to high temperatures has been reported as a serious obstacle to its global production. The objective of this study was to evaluate quinoa growth and pollen morphology in response to high temperatures. Pollen morphology and viability, plant growth and seed set, and several physiological parameters were measured at anthesis in two genotypes of quinoa subjected to day/night temperatures of 22/16°C as a control treatment and 40/24°C as the heat stress treatment. Our results showed that heat stress reduced the pollen viability between 30% and 70%. Although no visible morphological differences were observed on the surface of the pollen between the heat‐stressed and non‐heat‐stressed treatments, the pollen wall (intine and extine) thickness increased due to heat stress. High temperature did not affect seed yield, seed size and leaf greenness. On the other hand, high temperature improved the rate of photosynthesis. We found that quinoa has a high plasticity in response to high temperature, though pollen viability and pollen wall structure were affected by high temperatures in anthesis stage. This study is also the first report of quinoa pollen being trinucleate.     

The effect of nitrate nutrition on iron chlorosis and leaf growth in sunflower (Helianthus annuus L.)

The objective of this work was to investigate the effects of nutrient solution pH, nitrogen form (NO₃, NH₄NO₃), bicarbonate and different Fe concentrations in the nutrient solution on the Fe concentration in roots and on the development of Fe deficiency symptoms in sunflower plants (Helianthus annuus L.). High pH in the nutrient solution induced by nitrate supply or by a pH-stat device led to increased Fe concentrations in roots and low leaf Fe concentrations associated with a significant decrease in leaf chlorophyll concentration manifested by yellow leaves. Plants of the nitrate fed treatments with 1 μM Fe in the nutrient solution were also characterized by reduced leaf growth and by the suppression of new leaf formation. The reduced leaf growth and the suppression of new leaves only occurred with nitrate and not with NH₄NO₃ in all treatments with 1 μM Fe in the nutrient solution. All symptoms were removed by a high Fe concentration in the nutrient solution (100 μM) at low external pH proving that suppression of leaf formation, reduced leaf growth and low chlorophyll concentration were caused by Fe deficiency. In the nitrate treatment with a low Fe supply (1 μM Fe) and pH 4 in the nutrient solution leaf chlorophyll concentrations similar to the controls were found. In comparison to control plants (NH₄NO₃, 1 μM Fe), leaf growth was still significantly reduced, and new leaf formation was suppressed. The chlorophyll concentration and CO₂ assimilation rate did not differ from those of the control plants. These results show that Fe deficiency is also characterized by small green leaves and the suppression of leaf formation. At the onset of leaf development, leaf growth and new leaf formation may respond more sensitively to poor Fe efficiency than chlorophyll concentration. In experiments with NO₃ plus HCO₃, simulating soil solution conditions prevailing in calcareous soils, the Fe efficiency of the youngest leaves was poor, showing retarded leaf growth and low chlorophyll concentration.     

Incompatible pollen tube growth and protein composition in styles of Japanese pear following high temperature treatments

Summary Immersing the excised-prepollinated styles of self-incompatible Japanese pear, Pyrus serotina Rehd., into distilled water at 45°C for 1.5–2.5 min promoted the growth of self-pollen tubes; both the number of styles penetrating the pollen tubes through styles to cut ends and of pollen tubes protruding from cut ends were increased. The subsequent growth of the protruded pollen tubes on agar media was also enhanced by the treatments. Treatments at 40°C did not promote the growth of self-pollen tubes and those at 50°C led the styles to necrosis. In the stylar proteins analyzed by isoelectricfocusing polyacrylamide gel electrophoresis, several changes in banding pattern were found following the heat treatment; one of the bands was a glycoprotein that increases in quantity in developing styles as self-incompatibility of the style becomes stronger (Hiratsuka et al., 1986). This protein band was stained more weakly by Coomassie Brilliant Blue-G250 dye. Antigenic substances in the styles examined by immunodiffusion seemed not to be denatured by high temperature treatments.Abbreviations IEF-PAGE isoelectricfocusing polyacrylamide gel electrophoresis - CBB Coomassie Brilliant Blue     

杏ISSR反应体系的建立

为建立杏适宜的ISSR反应体系,以红玉杏为试材,探讨影响ISSR扩增的主要因素包括模板浓度、MgCl2浓度、dNTPs浓度、引物浓度及循环次数。通过筛选及优化,确定杏ISSR的适宜扩增条件。结果表明,在25 μl PCR反应体系中,各组分的适宜浓度配比为：模板1 μl（40 ng）、dNTPS 2.5 μl（2.5 mmol/L）、Taq DNA聚合酶0.5 μl（2.5 U）、引物1 μl（20 μmmol/L）、10×PCR Buffer 2.5 μl、MgCl2 2.5 μl（2.5 mmol/L）、ddH2O 15 μl,反应循环次数40次。同时利用建立的ISSR反应体系对3种普通杏品种进行扩增,证明该体系完全适合此标记对杏不同品种的遗传分析,从而为下一步杏资源遗传多样性的研究提供理论依据和参考价值。