为更好地了解葡萄胚与果实发育之间的关系.本研究以大粒六月紫、早玛瑙等6个早、中、晚熟葡萄品种为试材,通过观察果实干鲜重、果型指数以及胚畸形率等性状对果实与胚发育动态进行了分析.结果表明,葡萄果实鲜重、干重变化总体呈“快-慢-快”的“双S”型曲线.浆果先纵向生长,后横向生长,果型指数先升高后逐渐降低.胚珠畸形率早熟品种大粒六月紫最高达88.6％;中熟品种最高达50.9％;晚熟品种最高仅12.5％.胚发育变化时间上早于果实发育变化,葡萄胚(种子)败育变化分为不完全败育和完全败育两种.

miRNA prediction algorithms often fail to predict the accurate location of the mature miRNA in a precursor sequence with nucleotide-level precision. miRNAs-rapid amplification of cDNA ends (miR-RACE) is an efficient method to determine the precise sequences of computationally predicted microRNAs (miRNAs). miR-RACE includes the following steps: miRNA-enriched library preparation, two specific 5'- and 3'-miRNA RACE (miR-RACE) PCR reactions, and sequence-directed cloning. The most challenging step is the two gene-specific primers designed for the two RACE reactions. The miR-RACE protocol is rapid and can be executed and completed in 2-3 days.

MicroRNAs (miRNAs) play critical regulatory roles mainly through cleaving their target mRNAs or repressing gene translation during plant development. Grapevines are among the most economically important fruit crops with available whole genome sequences. Studies on grapevine miRNAs (Vv-miRNAs) are also widely available. However, studies on the regulation mode of Vv-miRNAs on their target mRNAs during grapevine development have not been studied well, especially at the transcriptome-wide level. Here, six small RNA and mRNA libraries from various grapevine tissues were constructed for Illumina and Degradome sequencing. Subsequently, we systematically analyzed the spatiotemporal variations in the regulation of the target genes of regulation of Vv-miRNAs. In total, 242 known and 132 novel Vv-miRNAs and 193 target mRNAs were identified, including 103 target mRNAs for known and 90 target mRNAs for novel miRNAs, were validated in one or more of the tissues examined. More than 50 % of novel miRNAs were expressed exclusively in the flowers and berries, where they cleaved their target genes in a tissue-specific manner, especially, the breadth of their cleavage sites in flower tissues. Moreover, six novel miRNAs in berries responded to exogenous gibberellin and/or ethylene under a quantitative real time RT-PCR analysis, which confirmed their regulatory functions during berry development. Up to 93.6 % of the known miRNAs were highly conserved in various tissues, where their expression levels exhibited dynamic variations during grapevine development. Significantly, some Vv-miRNA families had one key member that acted as the main regulator of their target genes during grapevine development.

In order to deeply understand the status of the usage of DNA fingerprinting in the plant cultivars identification, more importantly, to promote DNA fingerprinting techniques’application in plant cultivars identification, this paper summarized the DNA fingerprinting techniques developed currently that used to identify plant cultivars and pointed out the advantages and disadvantages of the existing DNA fingerprinting identification techniques. The analysis measures applying to the DNA fingerprinting in cultivars identification were also reviewed. Because of the lack of the ideal measures, the techniques limited the transformation between DNA fingerprinting and plant identification information. Therefore, DNA markers have not really effectively applied in production practice. As a result, it was very important to propose a manual cultivar identification diagram (MCID) method based on the DNA fingerprinting. This article also focuses on the implementation of MCID.

Fruit taste depends on the contents and types of soluble sugars and organic acids. The large-scale identification and comparison of genes involved in sugar biosynthesis in apple, grape, and sweet orange based on the genome and expressed sequence tag information have not been reported. In this study, we found that sorbitol-6-phosphate dehydrogenase (S6PDH) was one of the most important genes in sorbitol biosynthesis in apple leaf, and was decomposed by NAD(+) -dependent sorbitol dehydrogenase (NAd(+)SDH) in apple fruit. The high expression level of soluble acidic invertase (AIV) gene possibly caused the high soluble sugar content (i.e., glucose and fructose) in grape. The expression level of sucrose synthase (SUS) in grape and sweet orange was also significantly higher than that in apple. The results show that SUS was involved in the resynthesis of fructose and UDP-glucose into sucrose in grape and sweet orange. By contrast, sucrose was transported into a vacuole and decomposed into glucose and fructose by AIV. Thus, ADP-glucose pyrophosphorylase (AGP), granule-bound starch synthase (GBSS), isoamylase I (ISA1), and isoamylase 2 (ISA2) could be the key genes in starch biosynthesis, whereas a-amylase (AMY3), isoamylse 3 (ISA3), and pullulanase (PUL) could be the key genes in starch degradation in apple. This study provides new insight into the isolation and comparison of candidate genes involved in the sugar biosynthetic pathway among multiple plants. (C) 2013 Elsevier B.V. All rights reserved.

In plant and animal species FK506-binding protein (FKBP) family genes are important conserved genes and it is defined as the receptors of FK506 and rapamycin, where they work as PPIase and protein folding chaperones. FKBP have been isolated from Arabidopsis thaliana, Oryza sativa, and Zea mays. In grape, twenty-three genes containing the FK506-binding domain (FKBP_C) were first time identified by HMMER and blast research, they were classified into three groups and 17 out of the 23 genes were located on 11 chromosomes (Chr1, 3, 5, 7, 8, 14, 15, 16, 17, 18, and 19). The predicted gene expression pattern and semi-quantitative RT-PCR results revealed that five VvFKBPs were expressed in all tissues, while seven VvFKBPs were expressed only in some of the tissues, and the remaining VvFKBPs were not expressed in leaf, stem, inflorescences, flowers, and a mixture of fruit tissues (small, medium and big-sized fruits). Most of the VvFKBPs in grapevine 'Summer Black' were similar to those predicted one in 'Pinot Noir' except for VvFKBP16-4 and VvFKBPa. VvFKBP12, FaFKBP12 and PpFKBP12 were cloned from 'Summer Black', 'Sweet Charlie' and 'Xiahui 6'. Protein structure analysis confirmed that homologous genes have some differences during the process of protein structure construction. In this study, we characterized and verified 23 FKBP family genes in grapevine (Vitis vinifera L.) as well as their sub-cellular and chromosome location. The successful cloning of CDS regions and protein structural analysis of VvFKBP12, FaFKBP12, and PpFKBP12 can provide useful information for further study.

Fruit skin coloration is a unique phase in the life cycle of fruiting plants and is mainly attributed to anthocyanin pigments. Anthocyanins are the largest and most diverse group of plant pigments derived from the phenyl propanoid pathway. They are water-soluble phenolic compounds that form part of a large and common group of plant flavonoids. Coloration encompasses several physiological and biochemical changes that happen through differential expression of various developmentally regulated genes. Due to research importance and economic value, Arabidopsis thaliana (chromosome no. = 5) and Vitis vinifera (chromosome no. = 19) have been used for investigations of the structural genes involved in anthocyanin biosynthesis. Thus for this review, V. vinifera is used as a model crop. In anthocyanin biosynthesis, a wide range of constructive genes including phenylalanine ammonia lyase, chalcone synthase and anthocyanidin synthase that are regulated by MYB transcription factors are involved. These genes are coordinately expressed and their levels of expression are positively related to the anthocyanin concentrations. Expression or suppression of the constructive genes contributes to a variety of changes that make fruits visually attractive and edible. Transgenic approaches also have discovered a strong relationship between phenyl propanoid/flavonoid gene expressions for fruit skin coloration. In this study, various developments that have taken place in the last decade with respect to identifying and altering the function of color-related genes are described.

Background With the completion of genome sequencing projects for more than 30 plant species, large volumes of genome sequences have been produced and stored in online databases. Advancements in sequencing technologies have reduced the cost and time of whole genome sequencing enabling more and more plants to be subjected to genome sequencing. Despite this, genome sequence qualities of multiple plants have not been evaluated. Methodology/Principal Finding Integrity and accuracy were calculated to evaluate the genome sequence quality of 32 plants. The integrity of a genome sequence is presented by the ratio of chromosome size and genome size (or between scaffold size and genome size), which ranged from 55.31% to nearly 100%. The accuracy of genome sequence was presented by the ratio between matched EST and selected ESTs where 52.93% ∼ 98.28% and 89.02% ∼ 98.85% of the randomly selected clean ESTs could be mapped to chromosome and scaffold sequences, respectively. According to the integrity, accuracy and other analysis of each plant species, thirteen plant species were divided into four levels. Arabidopsis thaliana, Oryza sativa and Zea mays had the highest quality, followed by Brachypodium distachyon, Populus trichocarpa, Vitis vinifera and Glycine max, Sorghum bicolor, Solanum lycopersicum and Fragaria vesca, and Lotus japonicus, Medicago truncatula and Malus × domestica in that order. Assembling the scaffold sequences into chromosome sequences should be the primary task for the remaining nineteen species. Low GC content and repeat DNA influences genome sequence assembly. Conclusion The quality of plant genome sequences was found to be lower than envisaged and thus the rapid development of genome sequencing projects as well as research on bioinformatics tools and the algorithms of genome sequence assembly should provide increased processing and correction of genome sequences that have already been published.

This experiment aimed to study the distribution of single sequence repeats(SSRs) in the expressed sequence tags(EST) of apple,develop new EST-derived SSR markers,and validate the application of EST-SSR markers in apple genetic diversity analysis.A total of 63 708 EST sequences of apple was obtained from the National Centre for Biotechnology Information(NCBI) and were screened for SSR motifs using MISA software.Forty-nine pairs of primer were designed using the Primer 5.0 Plus software and the PCR products by these primers were detected by PAGE.A summary on the characteristics of PCR products was done,and some of which were recovered for sequencing.The results displayed 6 153 SSRs identified from 5 423 apple EST sequences out of the entire apple collection of 63 708 ESTs.Dinucleotide,trinucleotide and hexanucleotide repeats were the dominant SSR types with proportion of 50.38%,14.85% and 15.47%,respectively.Electrophoresis results showed that among the 49 pairs of EST-SSR primers,33 could amplify distinct bands and gave expected products,and among the 33 pairs,25 were polymorphic.The dendrogram using SSR markers from apple ESTs was consistent with Fuji apple family relationship.

波尔多液是在果树上应用时间最长、应用范围最广、药效最为稳定的杀菌剂。为更好、更科学地将波尔多液应用于果树生产,在总结前人研究的基础上,结合正在开展的有关研究,针对波尔多液的作用机理、配制与施用、药害以及相关microRNA398对植物铜离子胁迫的影响进行了简要介绍。

A bioinformatics strategy is applied to clone a full length cDNA of TIR1 and Pt-F-box gene from citurs by blasting search of EST database with homologous gene cDNA of Arabidopsis thaliana and identified.According to the cDNA of above sequence,the 5′-end and 3′-end sequence were obtained from cDNA library of opening flower of Poncirus trifoliata(L.) Raf.using two gene specific primers by 5′RACE and 3′RACE methods,respectively.The full length cDNA of TIR1 and F-box gene from Poncirus trifoliata was spliced based on 5′-end and 3′-end sequence.The complete cDNA,designated as Pt-TIR1 and Pt-F-box,were 2 048,1 695 bp,respectively.These sequences were deposited in GenBank database,accession numbers FJ502240 and FJ502241,with an open reading frame encoding 569 and 468 amino acids,respectively.T Bioinformatics analysis showed that the cDNA of Pt-TIR1 and Pt-F-box have the recognition sites of microRNA393 and microRNA394.Pt-TIR1 and Pt-F-box with other plant F-boxes have the same amino acid sequences that are highly conserved as the designed F-box domains.Recombinant plasmid 35S-GW-FJ502240/FJ502241-GFP was introduced into onion epidermal cells by the particle bombardment method with a PDS1000/He.Transformed cells were incubated for 24 h at 22℃ in the dark and green fluorescence was monitored under a laser scanning confocal microscope.Subcellular localization results showed that the Pt-TIR1 and Pt-F-box were localized in the nucleus.

【Objective】 It is a fundamental work to identify grape germplasm materials and cultivars for exploration and collecting of germplasm resources as well as plant variety protection.A new approach(manual cultivar identification diagram,MCID) by using RAPD marker was adopted to distinguish 72 cultivars of grape germplasm from different countries and areas.【Method】 In this study,based on the optimization of RAPD through increasing the length of the RAPD primers and strict screening PCR annealing temperature,6 effective primers were screened from 35 RAPD arbitrary 11-mer primers,and in which 6 effective primers had clear amplification products and amplified polymorphic bands,and furthermore to conduct PCR amplification.Using this strategy of MCID,all the cultivars were gradually and completely separated by a combination of the 6 primers.【Result】A manual cultivar identification diagram(MCID) of the 72 grape cultivars was constructed using polymorphic bands from the DNA fingerprints.The MCID method could separate all the cultivars from each other,with the polymorphic bands employed for identifying the cultivars and the corresponding primers being marked in the correct position on the MCID.【Conclusion】So far,the MCID method is the best and most effective one to realize the identification of fruit cultivars by using DNA molecular markers.The MCID of 72 grape cultivars has high operability and practicability in identifying varieties using DNA markers,which can be of much help in grape cultivars identification for protection of cultivar rights and for early identification of seedlings in the nursery industry,thus providing a theoretical basis for germplasm evaluation and cultivar identification.

The color of grape berry is determined by the quantity of anthocyanins,and the biosynthesis of these anthocyanins is controlled both by structural genes and regulatory genes.Grape berry coloring is a complex system in that different grape species and even different cultivars of same grape cultivar group are different in coloring mechanism.In recent years,with the discovery of Myb-related genes,an important regulatory gene family,the anthocyanin biosynthesis was much better understood.

In order to verify the application effect of gene in-silico expression analysis platform for grape(Vitis vinifera L.) on expression prediction and rapid retrieval of specific sequences of fruit development genes,the prediction result on expression of four genes including VvANR,VvCHI,VvCHS2 and VvDFR in different tissues of V.vinifera was verified by means of mass ESTs on NCBI and technology of RT-PCR and semi-quantitative PCR.And the brief introduction of the platform function on rapid retrieval of specific sequences was carried out,too.The prediction results show that numbers of non-redundant ESTs of VvANR,VvCHI,VvCHS2 and VvDFR in different tissues of V.vinifera are all much more with sequences of 33,36,55 and 46,respectively.The expression amount of four genes has a certain difference,in which,the expression amount of VvANR is relatively high in inflorescence and bud,that of VvCHI relatively high in inflorescence,fruit and bud,that of VvCHS2 relatively high in fruit,bud,inflorescence and flower,while that of VvDFR relatively high in inflorescence,bud,flower,fruit and root.The results of semi-quantitative PCR and RT-PCR show that VvANR mainly expresses in inflorescence,flower and small-sized fruit,VvCHI predominantly in small-sized fruit,flower,shoot and inflorescence,VvCHS2 primarily in shoot,inflorescence,flower and small-sized fruit,and the order of VvDFR expression from high to low is flower,inflorescence,shoot,leaf,small-sized fruit,medium-sized fruit,big-sized fruit.The verification result indicates that the platform prediction result is basically in accordance with the experiment result in recognizing tissues with high expression,but it is not quite effective in predicting tissues with low expression.The information of specific cDNA libraries in specific tissues or status can be easily and quickly retrieved through five steps with this platform.

MicroRNAs (miRNAs) are an extensive class of newly identified small RNAs that regulate gene expression at post-transcription level by mRNA cleavage or translation. In our study, we used qRT-PCR and found that Vv-miR164 is expression in grapevine leaves, stems, tendrils, inflorescences, flowers and fruits. In addition, two potential target genes for Vv-miR164 were also found and verified by PPM-RACE and RLM-RACE. The results not only maps the cleavage site of the target mRNA but allowed for detection the expression pattern of cleaved fragments that can indicate the regulatory function of this miRNA on its target genes. These target genes were explored by qRT-PCR where some exhibited different expression patterns from their corresponding miRNA, indicating the cleavage mode of the miRNA on its target genes. The efficient and powerful approach used in this study can help in further understanding of how miRNAs cleaved their target mRNAs. Results from this study prove the importance of Vv-miR164 in regulating development and growth of grapes, and adds to the existing knowledge of small RNA-mediated regulation in grapes.

Fruit crops are important cash crop with high nutrition value.The scientific nomenclature of fruit cultivars not only was of importance in the unification and the specification of fruit cultivars' name,but also benefits the researches in pomology areas and the development of fruit industries.However,the nomenclature of the fruit crops was affected by region,culture and some other factors.In order to understand the situation of the nomenclature of fruit cultivars in China,and to provide more comprehensive referential data for the nomenclature in the future,we analyzed the nomenclature modes of the cultivars of apple,peach,litchi,chestnut,hazelnut,mei,hawthorn,longan,loquat,kixifruit,jujube,persimmon,citrus,lie,apricot,pear,cherry,strawberry,grapevine and banana in total of 20 important and representative fruit crops selected.

NAC gene family is a specific kind of important transcriptional factors to plants,which are involved extensively in plant growth,development and organogenesis and stress responses.Now,grape has been focused on molecular biology research areas,of which the informationmining and analysis of grape genome is very significant for the progress of grape functional genomics.In this article,by utilizing the bioinformatics method,we analyzed the phylogenetic of 143 NAC family protein sequence and NAC genome localization firstly,and then predicted and analyzed their amino acid composition,physical and chemical characteristics,as well as secondary and tertiary structures,meanwhile,we still assayed the relation of NAC gene family between grapevine and Arabidopsis thaliana.The result showed that these 143 amino acid sequences could be clustered into sixty groups together with 94 NAC sequences of Arabidopsis thaliana,which illuminated that NAC genes had highly conservative between grapevine and Arabidopsis thaliana.Further genetic mapping of NAC genome localization found that they could be distributed on 17 chromosomes.The research still found that the number of amino acid and hydrophobic of amino acid sequences in different subfamilies presented some differences,however,the results of secondary structure predicted that the main composition of 143 amino acid sequences was random curl and the tertiary structure of all the sequences were quite similar.The results of this investigation could definitely provide a significant foundation for further research on the function analysis of grapevine NAC gene family.

葡萄是世界性的重要果树,具有很高的经济效益。提高葡萄的育种水平和培育新品种是葡萄产业持续发展的重要基础,而葡萄基因组学与分子生物学的开展对葡萄的遗传育种具有重大的推动作用。葡萄遗传与基因组学也成为世界许多国家的重要研究内容。本文根据第10届国际葡萄育种与基因组学会议递交论文情况对国际葡萄遗传与基因组学研究现状进行究领域简要分析。