The differentially expressed circRNA（DEcircRNA）of hazel ovule were screened at different developmental stages,and their target differentially expressed mRNA（DEmRNA）was further identified to clarify the regulatory effect of circRNAs on ovule development. Samples were collected from ovule formation stage（Ov1）,early growth stage（Ov2）,rapid growth stage（Ov3）and maturation stage（Ov4）of hazel. The DEcircRNA involved in ovule development was identified based on the available results of mRNA sequencing analysis at four developmental stages. A total of 7 122 circRNAs were identified,and 59.54% of them came from exon region. In pairwise comparisons of Ov1-vs-Ov2,Ov1-vs-Ov3,Ov1-vs-Ov4,Ov2-vs-Ov3,Ov2-vs-Ov4 and Ov3-vs-Ov4,123,162,187,85,105 and 21 DEcircRNAs were identified respectively. The expression pattern of DEcircRNAs was developmental stage specific. Correlation analysis showed that 199 target mRNAs of DEcircRNA were also differentially expressed. The expression changes of DEcircRNA and their responding DEmRNA were highly synchronized. The results of GO（Gene Ontology）classification indicated that DEcircRNAs were involved in the regulation of metabolism during ovule development. Four DEcircRNAs and their target differentially expressed mRNAs at four ovule development stages were randomly selected and subjected to qRT-PCR analysis,and their expression levels changes were consistent with the results of high-throughput sequencing of microRNAs.
cDNA library of strawberry fruit at different stages was constructed and the proteins interacting with strawberry mitogen activated kinase FvM4K1 were screened which revealed the molecular mechanism of FvM4K1 in regulating strawberry fruit development. The total RNA ofFragraria vesca ‘Hawaii 4'was extracted at different fruit development stages related to FvM4K1,and the strawberry cDNA library was established. Specifically,the library capacity was 9.6 × 10 6 CFU,the recombination rate was 96%,and the average length of inserted fragment was more than 1 000 bp. At the same time,the bait vector pGBKT7-FvM4K1 was constructed,and no self-activation activity was detected in yeast. Then 13 proteins interacting with FvM4K1 were screened from the library by co-transformation. The function of these candidate proteins mainly focused on floral development such as pollen development and pollen tube elongation,also affecting flowering time and so on. Meanwhile,the Gene Ontology（GO）pathway annotation of these proteins showed that they mainly participated in cell metabolism process and molecular processes with catalytic activity. Besides,the 13 proteins screened had different expression patterns during fruit development,and four of them were predicted to be closely related to strawberry fruit development.
A calcineurin B-like phosphatase subunit protein gene VaCBL6 from Vitis amurensis ‘Zuoshan-1'was cloned and functionally analyzed. VaCBL6is located on chromosome 4 and has an open reading frame of 777 bp,encoding 258 aa;protein domain analysis shows that the protein contains four EF-hands and one transmembrane domain. Phylogenetic analysis using amino acid sequence showed that high similarities between VaCBL6 and VvCBL9,VaCBL6 and VvCBL6 are 98.84% and 94.96%,respectively. VaCBL6 is located in the nucleus and cell membrane,and is highly expressed in roots and tendrils,and its expression is induced by low temperature,drought,NaCl and ABA. Analysis of transcriptional activation based on the yeast system displayed that VaCBL6 had no transcriptional activation activity. Experiments on the growth stage ofArabidopsis seedlings overexpressingVaCBL6with different treatments found that 100-125 μmol · L -1 NaCl and 0.25-1.25 μmol · L -1 ABA treatments inhibited the germination of VaCBL6 transgenic Arabidopsis seeds,and the transgenic Arabidopsis seedlings increased the sensitivity to NaCl and ABA. The results indicated that VaCBL6 plays an important role in plant stress response.
CRs is an recently discovered clubroot resistance gene in Chinese cabbage. In this study,through PCR amplication and sequencing of four CRs linked markers in eight resistant materials and eight susceptible materials,we found six sequence variations between resistant and susceptible materials. Among which,five variations（9 956 808,9 956 888,9 956 904,10 707 299 and 10 707 385 bp in chromosome A08）were novel,G/A SNP（Single Nucleotide Polymorphisms）variation in 10 539 495 bp of A08 is consistent with previous study. Five KASP（Kompetitive allele specific PCR）markers, Probe55K1,Probe55K2,Probe64K1,Probe64K2 and Probe60K1,were developed in terms of variations in 9 956 808,9 956 888,10 707 299,10 707 385 and 10 539 495 bp. It turned out that all the five KASP markers can successfully differentiate the resistance from the susceptible materials. Probe55K1 and Probe64K1 were further genotyped in 45 resistant and 48 susceptible individuals, which were part of a BC1P1 population. The results showed that the consistency rate between genotypes and phenotypes of the above 93 individuals was high to 94.62%. Thus,the KASP markers developed in this study can be efficently used for marker-assisted selection（MAS）of clubroot resistant gene in breeding.
For investigating the cytoplasmic effects of cytoplasmic male sterility（CMS）on major agronomic and quality characters of cauliflower hybrids,six cauliflower sterile lines（A lines）with different cytoplasm from rape and cabbage,etc,and their maintainer lines（B lines）were used to make hybrids of A lines × test lines（AF 1）and B lines × test lines（BF 1）. The effects were indicated by AF1-BF1 or （AF1-BF1）/BF1（relative effect）. The results showed that significant positive effects were observed on days of curd appearing,days of curd harvesting,and vitamin C content. Comparison of cytoplasmic effects of different CMS sources,the rape cytoplasm had significant positive effects on growth period and number of rosette leaves. The cabbage cytoplasm showed significant positive effects on curd weight,lengthways diameter of curd,vitamin C content and days of curd harvesting,but a significant negative effect on chlorophyll content and number of rosette leaves. The cauliflower cytoplasm presented significant positive effects on growth period and vitamin C content. Comparison of cytoplasmic effects of the six cauliflower CMS lines which showed that NB65A had the highest positive effect on curd weight,lengthways diameter of curd,transverse diameter of curd and soluble sugar content,XG108A had the highest positive effect on cotent of vitamin C and soluble protein. Both of them could be used as the female parent for breeding high yield or high quality combinations. These results indicated that the cytoplasmic of different CMS sources of cauliflower showed complex effects on major agronomic and quality characters,and the adverse effects on some characters could be alleviated or overcome by nucleus of a appropriate male parent.
The BoPID was found as one of the interactive protein of BoSPx through yeast two-hybrid screening assay in Brassica oleracea. Its characters such as coding sequence,spatiotemporal expression and interactive proteins were further analyzed,with the purpose of studying its possible role in interacting with the calcium-responsive protein BoSPx in the process of self-incompatibility. The results showed that BoPID encodes 439 amino acids and contains two exons and one intron. BoPID may have kinase activity based on the kinase phosphorylation prediction analysis of its sequence,its sequence is highly homologous with AtPIDat the chromosome level and has strong species specificity when clustering with cruciferous plants such as turnip,Arabidopsis and Brassica napusin the phylogeny and collinearity analysis. Additionally,its promoter area contains multiple response elements such as ABA,IAA,abscisic acid,jasmonic acid,salicylic acid and stress response. The spatiotemporal expression analysis indicated thatBoPID expressed higher in the stigma than that in other parts of floral organs,while it was up-regulated after 15 minutes of self-pollination and then down-regulated. In the analysis of subcellular localization,we found BoPID protein localized in the cell membrane and cytoplasm,indicating that it might be an amphibious protein and anchored on the membrane and synapses in the cytoplasm. We further found the proteins of BoCML12,BoCaM2 and BoPIN1 interact with BoPID in the assays of yeast two-hybrid and GST-Pull down. Based on these results,we reason that BoPID might play a role in regulating calcium response to auxin through interacting with BoSPx,BoCML12 and BoCaM2 in the process of self-incompatibility.
The regulatory effects of ethylene metabolism and energy status on the vase quality of cut Itoh peony‘Bartzella’flowers were studied by using nano silver and preservative solution treatment. Cut Itoh peony flowers with a developmental level of stageⅡwere employed as test materials and were pretreated with distilled water（the control）or 20 mg · L-1 nano silver（NS）for 2 h. The samples were then vased in preservative solution（PS）directly after distilled water treatment or after nano silver treatment （NS + PS）. The release rate of ethylene,the contents of 1-aminocyclopropane-1-carboxylic acid（ACC）,adenine nucleotides（ATP + ADP + AMP）,sugar,and the activities of enzymes related to ethylene and sugar metabolism during the vase insertion process were measured. The results demonstrated that nano-silver pretreatment（NS）inhibited the peak of ethylene release by 40.9%,and reduced ACC content and ACC synthetase（ACS）activity,while increased the activities of succinic dehydrogenase（SDH）and H+-ATPase,and contents of sugar and ATP,which maintained a high energy charge（EC）level,therefore,prolonged vase life by 40.7 h. The PS also inhibited the peak of ethylene release by 35.1%,and increased SDH activity and sugar and ATP contents,which maintained a high EC value,and increased the maximum flower diameter by 3.5 cm. Moreover,NS + PS group showed inhibition of ethylene release peak by 31.2%,and reduction of ACC content and ACS activity,while SDH,CCO,and H+-ATPase activities,and the contents of sugar and ATP were all increased,which also maintained high EC levels and increased the soluble protein content,and therefore,increased flower vase life by 46.4 h,and the maximum flower diameter by 3.6 cm. The results showed that inhibition of ethylene metabolism and enhancement of energy status could delay the senescence and improve the vase quality of Itoh peony‘Bartzella’cut flowers.
Total number of eight qualitative traits related to flower branch characteristics including stem height uniformity,stem strength,stem filling degree,flower branch angle,flower horizontal curvature,flower vertical distribution,flower horizontal convergence,flower branch hardness of 188 commercial varieties of cut chrysanthemums were observed and classified in this experiment. The traits were evaluated based on grey relational analysis. The results indicated that the weighted values of eight factors which were determined by grey relational analysis showed flower horizontal convergence,stem strength,flower branch hardness,flower branch curvature had the most important pact on the evaluation,and the values were 0.1524,0.1430,0.1312,0.1261,respectively. Five excellent varieties of ZQ2-83,Q6-12,Q3-8,Q1-31 and Q4-13 were screened via grey ralational analysis,their weighted correlations were all greater than 0.8000. Then the 188 varieties were divided into four grades by cluster analysis,there were excellent,good,medium,and poor varieties which accounted for 9.04%,26.06%,50.00% and 14.89%,respectively.
Using Oncidium flexuosum‘Honey Angle'and‘Million Dollar'cut flowers as test material and the reasons for the senescence ofO. flexuosum flowers caused by the anther cap shedding was investigated. The results revealed that the anther cap shedding of‘Honey Angle'and‘Million Dollar'resulting in petal dehydration and shrinkage earlier than the control for two days and three days,and began to release endogenous ethylene after 16 hours. The endogenous ethylene contents of‘Honey Angle'and ‘Million Dollar'increased 55.42 times and 8.57 time respectively compared to the control. After the fall of the anther cap,‘Honey Angle'and‘Million Dollar'reached their peak of endogenous ethylene contents on the 3rd and 6th day,the ethylene contents were 351.59 and 82.57 nL · g-1 · h -1,respectively. Scanning electron microscope showed that there was no block flat structure on the junction between the anther cap and stamen of‘Honey Angel',whilst on the‘Million Dollar'matching block flat structure was present which may reduce abscission of the anther cap. Moreover,exogenous ethylene treatment accelerates petal senescence of‘Honey Angle'and‘Million Dollar'. Petal dehydration and shrinkage appear on the 3rd and 4th days after treatment in both varieties,respectively. Furthermore,the water content of both varieties significantly decreased by 12.57% and 20.48%,respectively,while the malonaldehyde contents significantly increased. The petal water content of‘Honey Angle'without exogenous ethylene treatment for one day is about 34.61%-43.39% of‘Million Dollar'. The results indicate that petal water content is one of the main factors of the senescence ofO. flexuosum cut flower.
HOS1（High expression of osmotically responsive genes 1）encodes a protein with E3 ubiquitin ligase activity,and is a key negative regulator of cold response signal transduction pathway of model plant Arabidopsis,but its molecular mechanisms involved in the low temperature stress response in perennial Vitis vinifera are unclear. The VvHOS1 gene was cloned from the leaves of Vitis vinifera ‘Chardonnay'. The full length of VvHOS1 was 2 931 bp,coding 976 aa,and the presence of a RING-type zinc finger domain was located at 74-106 aa（zf-C3H4_3）. Phylogenetic tree analysis showed that VvHOS1 protein in Vitis vinifera is most closely related to tomato,tobacco,and tea tree. Furthermore, the VvHOS1 antigen epitope rich fragment（1-320 aa）was constructed into the prokaryotic expression vector pET28a-SUMO at the BamHⅠandHind Ⅲ restriction sites. The pET28a- VvHOS11-320was found to be highly expressed inEscherichia coliRosetta（DE3）with isopropyl-β-D-thiogalactoside（IPTG）induction. The fusion proteins were mainly in the form of inclusion body proteins. The purified recombinant protein was used as antigen to immunize white rabbits,and the resulting anti-VvHOS1 polyclonal antibody was obtained and detected by ELISA. The potency was 1︰51 200 with a strong Western specificity. The antiserum specifically detects VvHOS1 protein in grapevine leaves after cold stress（4 ℃）,and Western blot analysis indicated that the expression of VvHOS1 was significantly up-regulated at 6,36 and 48 h.
Transcriptome comparison found that preferentially expressed genes（PEGs）were mainly involved in metabolism in the yellow-fleshed ripened‘Jinyan'kiwifruit,while they were generally related to development in red-fleshed‘Hongyang'. Meanwhile,PEGs from the peripheral part consisting of the peel and the outer layer of kiwifruits flesh and the remained core flesh part were related to their own hallmark function namely photosynthesis and nutrient reservoir,respectively. Kiwifruit possessed relatively abundant MYB genes of R2R3 type,and nearly all types of MYB genes were expressed in fruits.MYB-A72 transcript could only be found in‘Hongyang',and it possessed extremely high level of expression in the core flesh part,and additionally it was located within the clade including MYB genes for anthocyanin biosynthesis. Expression pattern clustering analysis revealed thatbHLH-B36 and WD-A126 might participate in forming the MYB-bHLH-WD40 regulatory complex,and that CHI2,FLS1,and CYP98A might be the targets under its regulation.
To survey the effect of calcium treatment on the microstructure,wax composition of the fruit skin and aroma emission of pear cultivar‘Nanguoli',the fruits at 15 d before commercial maturity date were treated by 0.04 kg · L-1 calcium chloride. As a result,the significant differences of microstructure,cell arrangement and wax composition were observed between the fruit with calcium treatment and control（sprayed water）at commercial maturity and five days during post-maturity. In detail,the abscission of wax layer and the intercellular spaces of epidermal cells were larger in the fruit with calcium treatment compared to the control. However,the intactness of lenticels in the fruit with calcium treatment were better than that in the control fruit. The wax composition of fruit skin of‘Nanguoli'pear comprised by fatty acids,esters,fatty alcohol,aldehydes,alkanes,phenols and alkenes. Of these compounds,fatty acids,Fatty alcohol,alkanes and aldehydes had higher contents than the other compounds. Noteworthy,the contents of esters,fatty acids and aldehydes were increased in the fruits with calcium treatment compared to the control. The total contents of volatile aroma compounds in calcium-treated fruit were 3 959.75 and 6 676.80 ng · g-1 at commercial ripening stage and five days during post-maturity,respectively,which were 20.92% and 24.16% higher than those of the control. These results suggest that calcium treatment promotes aroma emission by affecting the microstructure and wax composition of the fruit skin of‘Nanguoli'pear.
In this study,both tomato spotted wilt orthotospovirus（TSWV）universal primers and TSWV synthetic primers were used to detect the virus disease samples collected from parts of Chifeng,Inner Mongolia China,and the amplified sequences were analyzed. Then,the nucleotide sequence containing N gene amplified by using specific primers was sequenced for phylogenetic analysis. The results showed that TSWV was detected in all 18 tomato fruit samples. The phylogenetic analysis of sequences including 914 nt fragment containingN gene showed that TSWV tomato isolate from Inner Mongolia was more closely to isolates from Yunnan and Jiangsu. After sap inoculation,pepper showed the symptoms of chlorosis in leaf and dwarfing. TSWV was detected by RT-PCR on the symptomatic leaves of pepper. This is the first time to prove that TSWV is harmful to tomato by molecular methods in Inner Mongolia.
The mechanism of enzymatic browning and the current states of 11 kinds of physical anti-browning technology of fresh-cut fruit and vegetables at home and abroad are systematically discussed. The browning mechanism in deep,and the synergistic effect of various treatment methods were prospected,which could provide reference for the research and application of fresh-cut fruit and vegetables browning prevention.
Melatonin is a plant growth regulator and bio-stimulator,which can regulate plant growth and development,and improve plant stress tolerance. In recent years,the roles of melatonin in fruit development regulation and postharvest preservation have been hot researches. This paper summarizes the content and distribution of melatonin in different fruit. The physiological functions and mechanisms of melatonin in regulating fruit growth and development,promoting postharvest fruit ripening and delaying fruit senescence are emphasized. It points out the shortcomings in the present studies and puts forward suggestion for the future research direction of melatonin,and provides reference for melatonin to be widely used in the regulation of fruit development and preservation.
The authors conducted a systematic review of the main promoters which are used for the transgenic research of ornamental plants in the past 30 years,and contrastively analyzed the effects of three types of promoters（constitutive,tissue-specific and inducible promoters）on the plant genetic breeding. The problems on the research and application of promoters amongst ornamental plants were further discussed. Taken together,this systematic review will be helpful for improving the effectiveness and specificity of gene expression in the genetic engineering studies of ornamental plants.
The plum cultivar‘Fulu'was selected from the hybrids of‘Suili 3'（female parent）and ‘Dali'（male parent）. The fruit is heart-shaped and freestone,with average fruit weight of 57 g and maximum fruit weight of 110.5 g. It tastes sour and sweet and the soluble solids content is 12.5%-15.8%. The maturation period of‘Fulu'is in early August in Jilin. The appearance is good and most fruit surface are covered by purplish red. The fresh quality is good.
‘Meilong 1'is a new red flesh pitaya cultivar selected from the hybrid offsprings of Costa Rican native species（Hylocereus costaricensis）and white flesh species of Vietnam pitaya‘Baiyulong'（Hylocereus undatus）. The fruit has long oval shape,bright green squam and red peel. The average single fruit weight is 525 g;the total soluble solids content is 20.1%;edible rate is 76%. Its flesh is delicate,sweet,slightly fragrant. The fruit development period is 30-50 d,and the last fruit batches matures at the end of December in Nanning. The plant has strong flowering ability,good self-flowering compatibility,high and stable yield,and is suitable for cultivation in south Guangxi and the similar climatic regions.
Sporulated monokaryons prepared from Auricularia heimuer,strains‘916'and‘Xinke',were intercrossed using symmetric hybridization and asymmetric hybridization methods.‘Lier 3'was directive breeded among 89 hybrid offspring strains through multi-year and multi-sites trails. The results showed that the resistance and yield of‘Lier 3'were better than that of the parent strain‘Xinke',meanwhile,the number of the reverse side wrinkles of fruiting body was less than that of parent strain‘916'. The new cultivar has excellent agronomic characters such as early maturity,high yield in autumn-winter period,good quality and strong resistance.
‘Zhejiao 8',a new early maturing double-harvest water bamboo cultivar developed by systematic selection from‘Suozijiao'mutants after 10 years,has compact plant type and medium growth vigor. The harvest period of‘Zhejiao 8'is 5-6 days earlier than that of‘Suozijiao'in fall. Gross harvested weight of‘Zhejiao 8'in fall is 107.8 g per each,net weight is 70.2 g per each,and the average yield is 17 910 kg · hm-2. The harvest period of‘Zhejiao 8'is 8-10 days earlier than that of ‘Suozijiao'in summer. Gross harvested weight of‘Zhejiao 8'in summer is 123.7 g per each,the net weight is 85.1 g per each,and the average yield is 34 170 kg · hm-2. The succulent stem has 3-5 joints and the bud is white. It has glossy surface,fine and tender quality. The cultivar was suitable for planted in Zhejiang Province.