Analysis on Genetic Diversity of 40 Flowering Cherry Cultivars and Construction of Molecular ID Based on SSR Markers

Studying on the genetic diversity and genetic relationship of flowering cherry cultivars is extremely important for germplasm conservation, cultivar identification and breeding. Flowering cherry is widely cultivated as an important woody ornamental plant in worldwide, especially Japan, China. However, owning to the morphological similarity, many cultivars are distinguished hardly in non-flowering season. Here, we evaluated the genetic diversity and genetic relationship of 40 flowering cherry cultivars, which are mainly cultivated in China. We selected 13 polymorphicprimers to amplify to allele fragments with fluorescent-labeled capillary electrophoresis technology. The population structure analysis results show that these cultivars could be divided into 4 subpopulations. At the population level, Na and Ne were 6.062, 4.326, respectively. Ho and He were 0.458 and 0.670, respectively. The Shannon’s information index (I) was 1.417. The Pop3, which originated from P. serrulata, had the highest Ho, He, and I among the 4 subpopulations. AMOVA showed that only 4% of genetic variation came from populations, the 39% variation came from individuals and 57% (p < 0.05) came from intra-individuals. 5 polymorphic SSR primers were selected to construct molecular ID code system of these cultivars. This analysis on the genetic diversity and relationship of the 40 flowering cherry cultivars will help to insight into the genetic background, relationship of these flowering cherry cultivars and promote to identify similar cultivars.

Genome of tetraploid sour cherry (Prunus cerasus L.) ‘Montmorency’ identifies three distinct ancestral Prunus genomes

Sour cherry(Prunus cerasus L.)is a valuable fruit crop in the Rosaceae family and a hybrid between progenitors closely related to extant Prunus fruticosa(ground cherry)and Prunus avium(sweet cherry).Here we report a chromosome-scale genome assembly for sour cherry cultivar Montmorency,the predominant cultivar grown in the USA.We also generated a draft assembly of P.fruticosa to use alongside a published P.avium sequence for syntelog-based subgenome assignments for‘Montmorency’and provide compelling evidence P.fruticosa is also an allotetraploid.Using hierarchal k-mer clustering and phylogenomics,we show‘Montmorency’is trigenomic,containing two distinct subgenomes inherited from a P.fruticosa-like ancestor(A and A’)and two copies of the same subgenome inherited from a P.avium-like ancestor(BB).The genome composition of‘Montmorency’is AA’BB and little-to-no recombination has occurred between progenitor subgenomes(A/A’and B).In Prunus,two known classes of genes are important to breeding strategies:the self-incompatibility loci(S-alleles),which determine compatible crosses,successful fertilization,and fruit set,and the Dormancy Associated MADS-box genes(DAMs),which strongly affect dormancy transitions and flowering time.The S-alleles and DAMs in‘Montmorency’and P.fruticosa were manually annotated and support subgenome assignments.Lastly,the hybridization event‘Montmorency’is descended from was estimated to have occurred less than 1.61 million years ago,making sour cherry a relatively recent allotetraploid.The‘Montmorency’genome highlights the evolutionary complexity of the genus Prunus and will inform future breeding strategies for sour cherry,comparative genomics in the Rosaceae,and questions regarding neopolyploidy.

Molecular characterization of the SAUR gene family in sweet cherry and functional analysis of PavSAUR55 in the process of abscission

Small auxin up RNA(SAUR) is a large gene family that is widely distributed among land plants. In this study, a comprehensive analysis of the SAUR family was performed in sweet cherry, and the potential biological functions of PavSAUR55 were identified using the method of genetic transformation. The sweet cherry genome encodes 86 SAUR members, the majority of which are intron-less. These genes appear to be divided into seven subfamilies through evolution. Gene duplication events indicate that fragment duplication and tandem duplication events occurred in the sweet cherry. Most of the members mainly underwent purification selection pressure during evolution. During fruit development, the expression levels of Pav SAUR16/45/56/63 were up-regulated, and conversely, those of Pav SAUR12/61were down-regulated. Due to the significantly differential expressions of PavSAUR13/16/55/61 during the fruitlet abscission process, they might be the candidate genes involved in the regulation of physiological fruit abscission in sweet cherry. Overexpression of PavSAUR55 in Arabidopsis produced earlier reproductive growth, root elongation, and delayed petal abscission. In addition, this gene did not cause any change in the germination time of seeds and was able to increase the number of lateral roots under abscisic acid(ABA) treatment. The identified SAURs of sweet cherry play a crucial role in fruitlet abscission and will facilitate future insights into the mechanism underlying the heavy fruitlet abscission that can occur in this fruit crop.

Impact of black cherry on pedunculate oak vitality in mixed forests:Balancing benefits and concerns

The vitality of European forests continues to decline due to new pests and diseases,climate-change related disturbances and high loads of atmospheric nitrogen deposition.Deteriorating soil health is a major factor underpinning the low vitality of West-European forests.Selecting tree species with soil ameliorative traits is proposed as an avenue to counteract soil acidification and improve overall forest vitality.Here we evaluate the impact of black cherry(Prunus serotina Ehrh.),a known rich litter species,on the vitality of neighboring pedunculate oak(Quercus robur L.)in ten mixed forests on sand in Germany,Belgium and the Netherlands.We found that black cherry admixture increases foliar N and P to a surplus whereas it causes deficiencies in foliar Mg,thereby resulting in an overall negative effect on oak foliar nutrient concentrations.Contrary,defoliation of oak leaves by herbivory decreases with the proximity of black cherry.Using structural equation modelling(SEM),we tested the hypothesized‘improved soil health’pathway.Our analyses showed that black cherry admixture leads to lower accumulation in the humus layer,resulting in higher soil base saturation which has a positive effect on foliar Ca yet a negative effect on total chlorophyll.Moreover,the SEM illustrated that herbivory of oak leaves decreases when black cherry is admixed,both via dilution and improved soil health.Indirect effects of black cherry on oak vitality via“improved soil health”in our SEM are however small in comparison to direct relations.Hence,our study showed that the combined positive and negative impacts of black cherry on oak vitality are limited,which tempers the potential benefits of using the rich litter species to counteract oak decline via improved soil health–yet,the concern of black cherry as an invasive alien species negatively affecting the vitality of mature pedunculate oak trees may also be exaggerated.

Study on Control Techniques of Virus Diseases in Facility Cherry

The diseases of sweet cherry caused by viruses and viroids have occurred in the main cherry producing areas of China and are increasing year by year, which has become an important factor affecting the yield and quality of sweet cherry. For this reason, this paper elaborated the prevention and countermeasures of virus diseases from six aspects, including soil treatment, cultivation of disease-free seedlings, prevention of disease and pest damage, reasonable pruning, water management, and treatment of virus diseases, in order to provide technical guidance for the disease resistance, high quality and high yield of facility cherry, as well as information reference for the further improvement of the technical system of virus disease prevention and cultivation of facility cherry.

Effects of Controlled Atmospheres with High_O_2 or High_CO_2 Concentrations on Postharvest Physiology and Storability of "Napoleon" Sweet Cherry

Sweet cherries ( Prunus avium L. cv. Napoleon) were stored in controlled atmospheres (CA) of high O(2) (70% O(2) + 0% CO(2)) or high CO(2) (5% O(2) + 10% CO(2)), in modified atmosphere package (MAP, (13% - 18%) O(2) + (2% -4%) CO(2)) and in air (control) at 1 degreesC to investigate the effects of different O(2) and CO(2) concentrations on physiological properties, quality and storability of the fruits during storage. The results indicated that compared with other treatments, CA with high O(2) concentration decreased fruit decay and ethanol content, but increased the accumulation of malondialdehyde (MDA) and stimulated browning. CA with high CO(2) concentration inhibited polyphenol oxidase (PPO) activity, reduced MDA content, maintained vitamin C content and firmness, decreased fruit decay and browning. Soluble solids contents (SSC) were not significantly affected by different atmosphere treatments. 'Napoleon' fruits stored in 5% O(2) + 10% CO(2) for as long as 80 d were of good quality, but only 40, 20 and 30 d stored in MAP, 70% O(2) + 0% CO(2) and air, re-spectively.

Marketing Strategies for Chinese Specialty Agricultural Products under the Internet Environment : Taking Tianshui Qinzhou Big Cherry Industry as an Example

Combined with the current situation of marketing of Chinzhou big cherries in Tianshui,we find out the problems in the development of network marketing of Chinzhou big cherries and put forward corresponding countermeasures and suggestions to improve its marketing level and solve the problem of imbalance between supply and demand of Qinzhou big cherries.

Effects of Probiotics on Growth Performance and Blood Biochemical Indexes of Cherry Valley Ducks

In order to investigate the effects of probiotics on growth performance, blood biochemical indexes and economic benefits of Cherry Valley ducks, different probiotics were added into the feed of Cherry Valley ducks. Total 240 1-d-old Cherry Valley ducks were selected, and they were randomly divided into 4 groups, including 1 control group and 3 treatment groups. There were 3 replicates for each group and there were 20 ducks for each replicate. The ducks in the control group were fed the basal diet, and the ducks in the 3 treatment groups were fed the basal diet added with Lactobacillus, Bacillus natto and florfenicol, respectively. The test lasted for 42 d. The results showed that the addition of Lactobacillus and Bacillus natto could improve the average daily weight gain and final average weight （P〈0.05） of Cherry Valley ducks, and reduce the glutamic-pyruvic transaminase ac- tivity, alkaline phosphatase activity and serum urinary nitrogen content in duck blood （P〈0.05） and reduce the creatinine content and total cholesterol content in Cherry Valley ducks （P〉0.05）; at the same time, adding probiotics to feed could also in- crease the grass economic benefits of Cherry Valley ducks. In overall, the effects of adding probiotics were better than those of adding antibiotic.

Effect of Weak Light on the Peroxidation of Membrane_Lipid of Cherry Leaves

The effect of weak light on the peroxidation of membrane_lipid of one_year_old cherries ( Prunus pseudocerasus L. 'Laiyang') was studied by whole_tree shading. The results showed that the net photosynthetic rate of cherry leaves under weak light was remarkably lower; the activity peroxidase (POD) increased when light intensity decreased; the activity of catalase (CAT) showed an opposite trend, and it was positively correlated with light intensity; the activity of superoxide dismutase (SOD) increased under 366 μmol·m -2 ·s -1 and 533.8 μmol·m -2 ·s -1 light intensity, but decreased under 228.8 μmol·m -2 ·s -1 and 83.9 μmol·m -2 ·s -1 light intensity. A remarkable increase of malondialdehyde (MDA), a product of membrane_lipid peroxidation, was also observed in cherry leaves when treated with weak light, indicating more serious peroxidation in the membrane.

Effects of Different Preservative Treatments on Physiological Metabolism and Preservation of Sweet Cherry

[Objective] This study aimed to investigate the effects of different preservative treatments on physiological metabolism and preservation of sweet cherry. [Method] Sweet cherry （Prunus avium var. Summit） was soaked into benziothiazolinone （1 000 ppm）, lysozyme （500 ppm）, lysozyme （500 ppm） ＋ NPS polysaccharide （5 000 ppm） and water for 5 min, respectively. Non-treated sweet cherry was set as control. All the sweet cherries were then put into 3 mm thick PE bags and preserved at （-0.5±0.5） ℃. [Result] The results showed that the malate dehydrogenase （MDH） activity of benziothiazolinone treatment researched a significant peak on the 14 th d, while the MDH activity of Lysozyme （500 ppm）, Lysozyme （500 ppm） ＋ NPS polysaccharide （5 000 ppm） and water treatments began to increase on the 20 th d; the polyphenol oxidase （PPO） activity in various treatments showed a decreasing trend during the experiment, which researched a significant peak on the 14 th d, while that in water treatment was decreased consistently; on the 21 st d, the PPO activity in each treatment increased slowly; the titratable acid （TA） content in preservative treatments was higher than that in both water treatment and control; the soluble solid （SSC） content showed no significant difference between various treatments and between experimental treatment and control; on the 40 th d, the healthy fruit rate in preservative treatments was significantly higher than that in water treatment and control. [Conclusion] Benziothiazolinone, lysozyme and other preservatives show good effects on preservation of sweet cherry; lysozyme treatment can decrease the activity of malate dehydrogenase, maintain the relatively high content of organic acid and significantly improve the healthy fruit rate within a certain period of time.

Application of Landfill Composed of Biogas Residue and Slurry in Cherry Production

In order to explore the effects of landfill composed of biogas residue and slurry on the improvement of soil nutrients and cherry fruit yield, three treatments： control （no landfill）, landfilling with biogas residue and water （BR）, landfilling with biogas residue and slurry （BS） were conducted in a cherry orchard. The results showed that compared with the control, soil water content around the landfills with a radius of 60 cm within 30 d was increased in BR and BS treatments. The poment- age of short shoots and the total number of shoots of cherry trees were also signif- icantly increased by BR and BS treatments; the cherry fruit yield per tree and the single-fruit weight in BS treatment were increased by 21.76% and 28.89%, respec- tively. In addition, BS treatment obviously improved the contents of soil organic matter, soil available nitrogen, s0il available phosphorus, soil available potassium and other nutrients. The positive effects of BR treatment on the improvement of soil nutrients and cherry fruit yield were lower than those of BS treatment, indicating that the combined use of biogas residue and biogas slurry as landfill can improve the soil water and fertilizer status in orchards, and thus can be promoted in the cultivation of fruit trees.

Cluster Analysis of Characteristics of Different Sweet Cherry Varieties

In order to compare the characteristics of different varieties of sweet cherry and to formulate corresponding pruning scheme, hierarchical cluster analysis was conducted for the 14 sweet cherry varieties that were mainly planted in Shanxi Province. The results showed that the 14 varieties of sweet cherry could be divided into two types, Hongmanao and Rainier. Fruit setting rate, branching rate, medium fruit shoot proportion, spur proportion and yield per plant were significantly different between these two types of sweet cherry. The key points of pruning management, to improve the yield of Rainier type, were to increase the fruit setting rate and spur proportion, and to control properly the long and medium fruit shoot proportion.

Study on Chilling Requirement of Different Fruit Branch of a Sweet Cherry Cultivar ‘Hongmanao'

With ＇Hongmanao＇, a sweet cherry cultivar from central Shanxi Province, as the experimental material, the chilling requirements of different types of fruit branches （including mixed fruit branch, long fruit branch, medium fruit branch, short fruit branch and blossom fruit branch） were studied by the 0-7.2 ℃ model in a solar greenhouse during 2012-2013. In the greenhouse, the fruit branches of ＇Hongmanao＇ were cultured in water in a thermostatic environment. The results showed that the chilling requirement of leaf buds of ＇Hongmanao＇ was lower than that of its flower buds; and the chilling requirements of the five types of fruit branches ranked as blossom fruit branch＇s〈short fruit branch＇s〈medium fruit branch＇s〈long fruit branch＇s〈mixed fruit branch＇s,

Mapping of Quantitative Trait Loci （QTLs） Controlling Cherry and Green Bean Characters in the Robusta Coffee （Coffea canephora Pierre）

In coffee breeding practice, two quantitative traits, namely cherry and green bean characters are the important phenotypic selection index. The synchronous of cherry maturation, size, weight and shape of green bean are desirable traits for the future breeding. In order to increase the breeding efficiency, a set of quantitative trait loci （QTLs） analysis controlling these traits was carried out. The QTL analysis was performanced in cross pollinated population of Coffea canephora using single nucleotide polymorphisms （SNPs） and simple sequence repeats （SSRs） markers. Totally 12 QTLs determined desirable cherry and green bean traits were identified on 9 linkage groups （LG）, where these QTLs mapped forming clusters in 11 different chromosomal regions. The desirable traits which can be detected by their QTLs are synchronous of cherry maturation （LG B and LG G）, cherry size （LG A, LG F and LG G）, bean shape （LG B, LG D and LG J）, bean weight （LG H）, bean size （LG A and LG E） and cherry-bean size （LG K） The gene action of these QTLs was dominance or an interaction of alleles （epistasis between alleles at the studied locus） effect. The yielded linkage map and the detected QTL can provide a tool for further genetic analysis of two traits and be potential for maker-assisted selection in C. canephora breeding.

Regeneration of Leaves in vitro in Cherry Rootstock Colt

Plants were regenerated from leaves of cherry rootstock Colt by two methods,the frequencies were 48 3% and 21 3%.Leaves were cultured on MS medium supplemented with NAA 1 0mg/L,KT 3 0mg/L and ZT 0 25mg/L.After 5～7 days leaves dedifferentiated,and formed callus.About 25 days later,callus grew into greenish or pink compact ones,the induction frequency was 100%.The color and structural feature of callus depended on the medium,culture condition and physiology phase of leaves.To induce callus,NAA was the main factor.Leaves rooted in medium only with NAA.In order to inhibit rooting and make callus grow fast,KT and ZT were added to the medium.Leaves cultured without light could quickly form callus during the initial stage of dedifferentiation.Young leaves dedifferentiated more easily than old leaves.After had been cultured about 50 days,callus were transferred to MS medium supplemented with NAA 0 2mg/L,IAA 0 5mg/L,6 BA 0 5mg/L,KT 1 0 mg/L and GA 0 5mg/L,and redifferentiated at a frequency of 21 3%.The callus of Colt leaves were difficult to redifferentiate.GA was important in the redifferentiation of callus.The result showed that callus could redifferentiate in all the mediums with GA,and could not redifferentiate without GA.The ability of redifferentiation of big callus was higher than that of small ones.And the physiology character of callus was also important.High regeneration frequency was obtained from greenish or pink compact callus.The frequency of regeneration of petioles was higher than that of leaves.Leaf petioles' regeneration need high concentration of cytokinin(more than 5 0mg/L).Leaf petioles were cultured on MS medium supplement with 6 BA 6 0mg/L,NAA 1 0mg/L and GA 0 5mg/L,and regenerated at a frequency of 48 3%.

Interaction Between Cryptococcus laurentii,Monilinia fructicola,and Sweet Cherry Fruit at Different Temperatures

The present study was performed mainly to investigate the antagonist-pathogen-host interaction in wounds of the sweet cherry fruits. The antagonistic yeast Cryptococcus laurentii could significantly reduce the brown rot of the sweet cherry fruit caused by Monilinia fructicola at 25 and 1 ℃. The populations of yeast increased faster in the presence of the pathogen initially, but then decreased rapidly. In the fruits inoculated with M. fructicola alone or combined with C. laurentii, an induction of lipid peroxidation as well as activities of the antioxidant enzymes, such as, superoxide dismutases （SOD）, catalase （CAT）, and peroxidase （POD）, was observed. The isoenzyme pattern of polypheno/oxidase （PPO） changed greatly after the symptoms appeared, with new PPO isoforms being induced. By contrast, the induction of lipid peroxidation and activities of SOD, CAT, and POD were low, although no significant changes were found in the PPO isoenzyms in the fruits inoculated with antagonist C. laurentii alone. The inhibition of brown rot during the antagonist- pathogen-host interaction in wounds of the sweet cherry fruits was mainly on account of the stimulated growth of C. laurentii as well as the induction of antioxidant enzymes of the fruits by M. fructicola.

Changes of Aroma Components in Hongdeng Sweet Cherry During Fruit Development

To study the changes of aroma components in sweet cherry during fruit development, the aroma components in sweet cherry fruit from the green stage, the color stage, the commercial stage, and the ripe stage were collected using head-space solid phase microextraction （HS-SPME）, and were analyzed using a gas chromatograph-mass spectrophotometer （GC- MS）. A total of 37 compounds were identified from the sample extracts. Aldehydes, alcohols, and esters were the major constituents. The aroma components behaved differently during the fruit developmental period. C6 aldehydes and aromatic aldehydes were the main aldehydes in the sweet cherry fruit. The contents of C6 aldehydes increased quickly to 84.16% in the color stage, then decreased as ripening proceeded, and then, the contents decreased to 59.20 and 55.58% at the commercial stage and the ripe stage, respectively. The aromatic aldehydes （benzaldehyde） increased as ripening proceeded, and the maximum was found at the ripe stage. Alcohols of sweet cherry fruit included C6 alcohols and aromatic alcohols. The content of （E）-2-hexen-l-ol increased as ripening proceeded. The maximum was found at the commercial stage; alcohol was only found at the ripe stage. Ester components included ethyl acetate, butanoic acid ethyl ester, hexanoic acid ethyl ester, which increased as the fruit ripened. Hexanal, （E）-2-hexenal, benzaldehyde, （E）-2-hexen-l-ol, ethyl acetate, and hexanoic acid ethyl ester were the characteristic aroma components of sweet cherry fruit. These aroma components started to form drastically at the color stage, and the main aroma was formed at the commercial stage, which then turned bad at the ripe stage because of the presence of alcohol. So the optimal harvest time of sweet cherry was at the commercial stage.

Impact of chilling accumulation and hydrogen cyanamide on floral organ development of sweet cherry in a warm region

The microscopic investigation of the floral development of sweet cherry（Prunus avium L. cv. Hongdeng） from a warm winter climate（Shanghai） and cold winter climate（Tai＇an, Shandong Province, China） was conducted to explore the reason of low fruit set. The effect of hydrogen cyanamide（HCN） on floral development under warm winter conditions was also investigated. Trees grown in Shanghai with insufficient chilling accumulation exhibited little difference in the progression of microspore development compared to trees in Tai＇an that accumulated adequate chilling, but showed substantial delays in ovule and embryo sac development. The growth of nucelli did not proceed beyond the macrospore mother cell and macrospore stages with abortion rates of 13, 15 and 45% by 6, 3 and 0 d before full bloom, respectively. These abnormalities in the ovule and embryo sac in the Shanghai-grown trees were eliminated by HCN application. These results suggest that chilling regulates the development of female floral organs in winter dormancy; therefore, insufficient chilling accumulation, causing abnormality of the female floral organs, restricts the cultivation of sweet cherry in warm winter regions. Interestingly, HCN application, which decreased the chilling requirements for Hongdeng, may be a potential strategy for sweet cherry cultivation in warm winter regions.

Flavonoid content and radical scavenging activity in fruits of Chinese dwarf cherry(Cerasus humilis) genotypes

The Chinese dwarf cherry（Cerasus humilis（Bge.） Sok.） is a small shrub with edible fruits. It is native to northern and western China. This species was included as a medicinal plant in the ‘‘Chinese Pharmacopeia’’ and has emerged as an economically important crop for fresh fruit consumption, processing into juice and wine and nutraceutical products as well. To gain a better understanding of flavonoid biosynthesis and help develop value added products and better cultivars with greater health benefits, we analyzed total flavonoid content（TFC）, composition, and radical scavenging activities in fruit extracts of 16 Chinese dwarf cherry genotypes. Fruit peel TFC ranged from 33.5 to72.8 mg/g REáFW（RE： rutin equivalent, FW： fresh weight）while fruit flesh TFC ranged from 4.3 to 16.9 mg/g REáFW.An HPLC analysis revealed that fruit extracts contained 14 flavonoids with considerable variation in their profiles across genotypes. The most abundant flavonoids in most genotypes were proanthocyanidin B1（PA-B1）, proanthocyanidin B2（PA-B2）, phloretin 20-O-glucoside（PG）, and phloretin 20,40-O-diglucoside（PDG）. Principal component analysis showed that PG, PA-B1, and PA-B2 had large,positive factor loading values in the first principal component for each genotype. Increased scavenging activity of2,2-diphenyl-1-picrylhydrazyl（DPPH） radicals was apparent in genotypes ‘Nongda 4’, ‘Nongda 3’, ‘Nongda 6’,‘Wenfenli’, and ’10-32’, suggesting promising applications in the production of nutraceutical products. In summary, our results will aid in breeding, fruit processing, and developing medicinal uses of the Chinese dwarf cherry.

Complete Genome Sequence Analysis of Duck Circovirus Strains from Cherry Valley Duck

To investigate molecular epidemiology of DuCV in Cherry Valley ducks in China, the complete genomes of six DuCV strains, which were detected from Cherry Valley ducks in China between 2007 and 2008, were sequenced. Sequence and phylogenetic analysis were carried out to compare these six strains with another 27 DuCV strains from Mulard duck, Muscovy duck, Pekin ducks and Mule duck. The analysis showed that the six DuCV strains exhibited typical genetic features of the family of DuCV, such as a stem-loop structure, three major open reading frames (Rep, Cap and ORF3), four intergenic repeats and the conserved motifs for rolling circle replication and for the dNTP binding domain located in the Rep protein. Phylogenetic analysis of the nucleotide sequences of the complete genome and Cap gene of these strains together with those that have been previously published demonstrated two distinct DuCV genotypes. The DuCV strains with complete genomes containing 1988 and 1989 nucleotides clustered in genotype A, whereas the strains with complete genomes containing 1991, 1992, 1995 and 1996 nucleotides lay in genotype B. The six DuCV strains from Cherry Valley ducks were divided into the two groups. The results of the study provides some insight into the variation of DuCVs in Cherry Valley ducks.