Helia Pub Date : 2019-03-27 DOI: 10.1515/HELIA-2017-0024

V. Vasko, V. Kyrychenko

{"title":"Induced Mutagenesis for the Creation of New Starting Material in Sunflower Breeding","authors":"V. Vasko, V. Kyrychenko","doi":"10.1515/HELIA-2017-0024","DOIUrl":"https://doi.org/10.1515/HELIA-2017-0024","url":null,"abstract":"Abstract The article colligates data of studies on the variability of quantitative and qualitative traits in mutant sunflower M1-M3 generations affected by dimethyl sulfate (DMS) (0.01, 0.05 %) and gamma rays (120; 150 Gy), frequencies and range of mutations in M2 and their inheritance in mutant families, chromosome aberrations in meiosis, as well as on the breeding and genetic value of induced mutants and possibilities of their use in breeding. The methodical peculiarities of the mutational breeding of the cross-pollinating crop were defined, and new mutants with changed features were created. Investigating new homozygous self-pollinated sunflower lines, we observed a more negative mutagenic impact of gamma irradiation (120 and 150 Gy) on the germinability of M1 sunflower seeds in the field compared with the DMS effect (0.01 and 0.05 %). The field germinability of DMS-treated seeds was 83–87 % vs. 11–15 % of gamma-irradiated ones. The mutagenic effect of gamma rays (120 and 150 Gy) on M1 meiosis was shown to be stronger than that of DMS (0.01 and 0.05 %). The percentage of cells with alterations varied within 15.79–18.78 % (120 Gy) and 20.38–25.26 % (150 Gy) compared to 0–0.16 % in the control. The effect of gamma rays on the frequency of morphoses in M1 was stronger, in particular, after exposure to 120 Gy or 150 Gy of gamma irradiation, the number of plants with alterations was 43 %, whereas after DMS treatment (0.01 and 0.05 %) this parameter averaged 27–28 %. We determined the inheritance of mutations of quantitative and qualitative traits, which are important for breeding, in mutant M2 families and selected mutant families with inherited altered traits that can be considered as mutations. Among the best mutations, there are morphological mutants with marker traits, mutants with increased content of oil in seeds, increased 1000-seed weight, increased contents of behenic, linoleic and palmitoleic acids as well as with absolute resistance to downy mildew.","PeriodicalId":39086,"journal":{"name":"Helia","volume":"42 1","pages":"17 - 36"},"PeriodicalIF":0.0,"publicationDate":"2019-03-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1515/HELIA-2017-0024","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43125715","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 3

Genetic and Phenotypic Diversity of the Sunflower Collection of the Pustovoit All-Russia Research Institute of Oil Crops (VNIIMK) 俄罗斯石油作物研究所向日葵品种的遗传和表型多样性

Helia Pub Date : 2019-03-09 DOI: 10.1515/HELIA-2018-0021

S. Goryunova, D. Goryunov, A. Chernova, E. Martynova, A. E. Dmitriev, S. Boldyrev, A. Ayupova, P. Mazin, E.A. Gurchenko, A. Pavlova, D. Petrova, Y. Chebanova, L. Gorlova, S. Garkusha, Z. Mukhina, E. Savenko, Y. Demurin

{"title":"Genetic and Phenotypic Diversity of the Sunflower Collection of the Pustovoit All-Russia Research Institute of Oil Crops (VNIIMK)","authors":"S. Goryunova, D. Goryunov, A. Chernova, E. Martynova, A. E. Dmitriev, S. Boldyrev, A. Ayupova, P. Mazin, E.A. Gurchenko, A. Pavlova, D. Petrova, Y. Chebanova, L. Gorlova, S. Garkusha, Z. Mukhina, E. Savenko, Y. Demurin","doi":"10.1515/HELIA-2018-0021","DOIUrl":"https://doi.org/10.1515/HELIA-2018-0021","url":null,"abstract":"Abstract Publicly supported collections of cultivated germplasm are one of the key sources of new genes for crop improvement. VNIIMK is the leading organization in oil and essential oil crop breeding and seed growing in the Russian Federation with more than a century-long history. Sunflower varieties created by V.S. Pustovoit at VNIIMK became the basis for the development of the modern sunflower varieties worldwide. In the present study, 186 sunflower lines from the VNIIMK collection were characterized based on their genotype and general morphological and phenological economically-important traits. Additionally, for 99 sunflower lines fatty acid content, seed oil content, seed husk content, 100-seed weight, and seed number in the head were determined. Sequencing of RAD-libraries and the subsequent analysis have identified 65,553 variants including SNPs and indels. LD analysis revealed substantial variability across the genome. The longest LD blocks (>5,000 Kb) were found in the linkage groups 1, 5, and 17. The analysis revealed significant genetic and phenotypic diversity of the VNIIMK sunflower collection. Novel significant associations with linolenic acid content in the seeds were found on LGs 8, 9, and 17.","PeriodicalId":39086,"journal":{"name":"Helia","volume":"42 1","pages":"45 - 60"},"PeriodicalIF":0.0,"publicationDate":"2019-03-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1515/HELIA-2018-0021","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49073962","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 3

Sunflower Resistance to Race G of Broomrape (Orobanche Cumana Wallr.) In the Russian Federation: the Development of the Lines and the Study of Inheritance 向日葵对锦葵G种的抗性在俄罗斯联邦:路线的发展和继承的研究

Helia Pub Date : 2019-03-08 DOI: 10.1515/helia-2019-0003

S. Guchetl, T. Antonova, N. Araslanova, T. Tchelyustnikova

{"title":"Sunflower Resistance to Race G of Broomrape (Orobanche Cumana Wallr.) In the Russian Federation: the Development of the Lines and the Study of Inheritance","authors":"S. Guchetl, T. Antonova, N. Araslanova, T. Tchelyustnikova","doi":"10.1515/helia-2019-0003","DOIUrl":"https://doi.org/10.1515/helia-2019-0003","url":null,"abstract":"Abstract The objective of this work is the development of sunflower lines that are not affected by race G of broomrape, as well as the determination of genetic control of resistance. The testing of the resistance of VNIIMK’s collection accessions of a cultivated sunflower of various origins was carried out on an artificial background made from the seeds of race G of broomrape. 6 lines resistant to race G were developed by the method of inbreeding on the basis of the obtained unaffected forms. The genetic control of resistance of one of them was studied. The resistance was inherited monogeneuosly, with incomplete dominance. There was established the presence of a reciprocal effect and the dependence of the resistance characteristic on the genotype of a susceptible parent under crossbreeding with certain lines. The other 5 sunflower lines are in the process of a hybridological analysis in order to determine the genetic control of their resistance. The combination of different genes of resistance to the same race of broomrape in one sunflower genotype could contribute to the long-term resistance of the crop to the parasite. The results of the presented study are of high importance for breeders, since the gene that we studied provides a new source of resistance to race G, thereby ensuring the protection of sunflower from the spread of new Orobanche pathotypes.","PeriodicalId":39086,"journal":{"name":"Helia","volume":"42 1","pages":"161 - 171"},"PeriodicalIF":0.0,"publicationDate":"2019-03-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1515/helia-2019-0003","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45356609","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 2

Polygenic Inheritance of Bracts Number in Sunflower 向日葵苞片数的多基因遗传

Helia Pub Date : 2019-03-06 DOI: 10.1515/helia-2019-0004

A. Soroka, V. Lyakh

{"title":"Polygenic Inheritance of Bracts Number in Sunflower","authors":"A. Soroka, V. Lyakh","doi":"10.1515/helia-2019-0004","DOIUrl":"https://doi.org/10.1515/helia-2019-0004","url":null,"abstract":"Abstract Two inbreds of mutant origin, differing in the number of bracts, were crossed to obtain the F1 hybrid. One mutant line had 24.5 ± 1.01 bracts, while the other, 78.6 ± 1.69 bracts. The F1 hybrid had an average value between parents, which practically did not differ from the mean value in the F2 population. The variability of the trait under study in the F2 population was continuous, varying from 20 to 84 bracts. This indicated the probable participation of several genes with an additive effect in the control of the number of bracts. Assuming that the differences between the parental lines are due to two pairs of genes, the F2 plant population, grown in 2016, was divided into 5 classes. In that population the observed classes ratio turned out to be close to the theoretically expected ratio of 1 : 4 : 6 : 4 : 1. Over the next two years F2 populations were tested in a similar way. In all the cases, the calculated chi-square value did not exceed the critical value for 4 degrees of freedom and 5 % significance level. This gave reason to talk about the two-loci control of a such quantitative trait as the number of bracts. The participation of two non-allelic genes in the control of this trait is also proved by matching the actual number of plants in the parental classes to the theoretically expected number of plants. Thus, the number of bracts depends on the number of dominant alleles of two different genes in the genotype.","PeriodicalId":39086,"journal":{"name":"Helia","volume":"42 1","pages":"221 - 228"},"PeriodicalIF":0.0,"publicationDate":"2019-03-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1515/helia-2019-0004","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42697287","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Inheritance of Top Branching in Sunflower (Helianthus Annuus L.) Collection Samples 向日葵顶端分枝的遗传收集样品

Helia Pub Date : 2019-02-28 DOI: 10.1515/helia-2019-0001

K. Vedmedeva

{"title":"Inheritance of Top Branching in Sunflower (Helianthus Annuus L.) Collection Samples","authors":"K. Vedmedeva","doi":"10.1515/helia-2019-0001","DOIUrl":"https://doi.org/10.1515/helia-2019-0001","url":null,"abstract":"Abstract Aim of our research was to study the genetic diversity and establish the inheritance of top branching trait in the collection of 34 sunflower lines of the Institute of Oilseed Crops of the NAAS. Experiments were carried out in 2005–2016 according to classical cultivation methods, using manual castration, crossings, forced self-pollinating, isolation and visual assessment of the first and second generation of obtained descendants. The statistical reliability of the obtained ratio was confirmed by calculating the Pearson’s chi-squared test. Presence of two loci determining the inheritance of the branching trait in sunflower was established. In one locus, recessive alleles are responsible for manifestation of the branching trait. In the second locus, dominant alleles are responsible for the manifestation of the branching trait. In 23 lines of sunflower, it was established that a recessive homozygote for one gene causes phenotypical top and full branching. In 8 lines of the collection, full branching trait is due to the dominant allele of the gene. In the lines InK235, APS49, the presence of two genes was established, the dominant alleles of which determine full branching trait. In the APS56 line, full branching is controlled by the dominant alleles of three genes.","PeriodicalId":39086,"journal":{"name":"Helia","volume":"42 1","pages":"203 - 212"},"PeriodicalIF":0.0,"publicationDate":"2019-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1515/helia-2019-0001","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43424307","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 1

Correlation and Path Coefficient Analysis for Morphological and Biochemical Parameters in Sunflower (Helianthus Annuus L.) 向日葵形态生化参数的相关及通径系数分析

Helia Pub Date : 2019-02-19 DOI: 10.1515/HELIA-2018-0011

A. Tahir, M. Iqbal, R. Saif, Masood Qadir, R. Sultana

{"title":"Correlation and Path Coefficient Analysis for Morphological and Biochemical Parameters in Sunflower (Helianthus Annuus L.)","authors":"A. Tahir, M. Iqbal, R. Saif, Masood Qadir, R. Sultana","doi":"10.1515/HELIA-2018-0011","DOIUrl":"https://doi.org/10.1515/HELIA-2018-0011","url":null,"abstract":"Abstract The aim of this study was to evaluate the best performing lines in sunflower on the basis of phenotypic and genotypic correlation so that we can find out the which trait directly or indirectly effect the yield and quality of the sunflower because being an breeder our main aim is yield and quality and the lines which are performing best can be further used in the breeding programs. Sunflower is a valuable oil producing crop because it contains good quality oil composition and can be grown twice in a year. There is scarcity of oil in our country so that there is requisite to heighten the yield of sunflower in order to exploit its share in oilseed sector. The study was conducted at the research field of Rajawala farm, University of Agriculture, Faisalabad during year 2015–16 to study the correlation among yield related traits, oil and protein content in Sunflower (Helianthus annuus L.). 20 sunflower lines were sown in randomized complete block design with three replications. Data was assessed at maturity for plant height, leaves per plant, leaf area, head diameter, internode length, 100-achene weight, achene yield per plant, oil contents and protein contents. Genotypic and phenotypic correlation was estimated among these traits. The recorded data was subjected to statistical analysis of variance, correlation and path coefficient analysis. The line G-16 showed best performance for leaf area, head diameter and achene yield per plant. Line G-20 was good in 100 achene weight. The above mentioned lines could be used in future breeding programs for effective improvement in yield of sunflower. This data was helpful to select superior lines and these lines may also be used in further hybridization program to get better yield, oil and protein contents.","PeriodicalId":39086,"journal":{"name":"Helia","volume":"42 1","pages":"61 - 72"},"PeriodicalIF":0.0,"publicationDate":"2019-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1515/HELIA-2018-0011","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43460916","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 3

Developing a Selection Criterion Using Correlation and Path Coefficient Analysis in Sunflower (Helianthus Annuus L.) 利用相关和通径分析建立向日葵品种的选择标准

Helia Pub Date : 2019-02-12 DOI: 10.1515/helia-2017-0031

A. Riaz, M. H. Nadeem Tahir, M. Rizwan, S. Fiaz, S. Chachar, Khuram Razzaq, B. Riaz, Hafiza Sadia

{"title":"Developing a Selection Criterion Using Correlation and Path Coefficient Analysis in Sunflower (Helianthus Annuus L.)","authors":"A. Riaz, M. H. Nadeem Tahir, M. Rizwan, S. Fiaz, S. Chachar, Khuram Razzaq, B. Riaz, Hafiza Sadia","doi":"10.1515/helia-2017-0031","DOIUrl":"https://doi.org/10.1515/helia-2017-0031","url":null,"abstract":"Abstract In breeding programs, it is imperative to recognize the genetic variations and levels of association between traits that directly or indirectly affect the yield. Achene yield and related traits in forty-nine sunflower genotypes were studied for genetic variability, correlations and path coefficient analysis. The phenotypic coefficient of variation (PCV) were higher than the genotypic coefficient of variation (GCV) for the all the studied traits in sunflower. The most active association of achene yield was found with achene/head, 100-achene weight and plant height. However, oil contents had a negative association with achene filling percentage, 100-achene weight, number of leaves and stem diameter. The maximum direct influence on achene yield were exerted by head diameter followed by achene filling percentage, 100 achene weight, achene/head and number of leaves. In contrast, plant height, days to maturity and stem diameter had the negative direct impact on yield. The genetic correlations were higher than phenotypic correlations which suggest that environmental influences were very low. Our results showed that achene/head, 100 achene weight, plant height and head diameter are important plant traits which should be considered while planning any breeding program for higher achene yield in sunflower.","PeriodicalId":39086,"journal":{"name":"Helia","volume":"42 1","pages":"85 - 99"},"PeriodicalIF":0.0,"publicationDate":"2019-02-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1515/helia-2017-0031","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45870590","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 9

Economic Efficiency and Bioenergetic Assessment of Predecessors and Fertilizer Systems in the Sunflower Cultivation 向日葵栽培中前人和肥料系统的经济效益和生物能源评价

Helia Pub Date : 2019-02-12 DOI: 10.1515/helia-2017-0007

A. Kvashin, N. Neshchadim, S. Gontcharov, Ksenija N. Gorpinchenko

引用次数: 2

Treatment by Trait Biplot Analysis of Organic Manure and Nano-Fertilizers on Sunflower Production 有机肥料和纳米肥料对向日葵生产的性状双相分析

Helia Pub Date : 2018-12-01 DOI: 10.1515/helia-2017-0017

N. Sabaghnia, S. Yousefzadeh, M. Janmohammadi

{"title":"Treatment by Trait Biplot Analysis of Organic Manure and Nano-Fertilizers on Sunflower Production","authors":"N. Sabaghnia, S. Yousefzadeh, M. Janmohammadi","doi":"10.1515/helia-2017-0017","DOIUrl":"https://doi.org/10.1515/helia-2017-0017","url":null,"abstract":"Abstract This study was performed to study the effects of farmyard manure (zero (M1), 20 (M2) t ha−1) and nano-fertilizers [control (n1), Mn (n2), Fe (n3), Zn (n4)] on sunflower. Traits days to 50% flowering, days to maturity, leaf number, leaf area, plant height, achene yield and harvest index were measured.Treatment by trait (TT) analysis using biplot analysis was used to determine which treatment was best and for what trait which were generated using the standardized values of the traits means. Results showed that the first two principal components (PC1 and PC2) were used to create a two-dimensional TT biplot that accounted percentages of 94% of sums of squares of interaction. The most important vertex treatment in polygon of TT biplot was M2n4 (application of 20 tonnes ha−1 manure and Zn nano-micronutrient) which indicated high performance in leaf number, leaf area, plant height, achene yield and harvest index. The identification of ideal treatment, the treatment that is most favorable treatment among all treatments, showed that the M2n4 might be used in selecting superior traits and it can be considered as the candidate treatment. Finally, nano-fertilizer could increase crop yields and improve the efficiency of manure application. The results of this investigation showed that application of nanoparticles may alleviates the adverse environmental factors and improve the sunflower performance and the integrated application of organic manure and nano-micronutrients is more effective.","PeriodicalId":39086,"journal":{"name":"Helia","volume":"41 1","pages":"241 - 251"},"PeriodicalIF":0.0,"publicationDate":"2018-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1515/helia-2017-0017","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45108927","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 2

Characterization of Non-Target-Site Mechanisms in Imidazolinone-Resistant Sunflower by RNA-seq RNA-seq表征抗咪唑啉酮向日葵的非靶位点机制

Helia Pub Date : 2018-12-01 DOI: 10.1515/helia-2018-0012

M. Gil, T. Vega, S. Felitti, L. Picardi, S. Balzergue, G. Nestares

{"title":"Characterization of Non-Target-Site Mechanisms in Imidazolinone-Resistant Sunflower by RNA-seq","authors":"M. Gil, T. Vega, S. Felitti, L. Picardi, S. Balzergue, G. Nestares","doi":"10.1515/helia-2018-0012","DOIUrl":"https://doi.org/10.1515/helia-2018-0012","url":null,"abstract":"Abstract Imisun sunflowers (Helianthus annuus L.) are imidazolinone-resistant cultivars in which the two mechanisms of herbicide resistance coexist: (i) mutation in herbicide target-site (target-site resistance) and (ii) non-target-site resistance (NTSR). In Imisun technology, NTSR could be related to herbicide metabolism and might occur as a result of a constitutive up-regulation of resistance genes, or it can appear only after herbicide treatment. The objective of this study was to characterize NTSR in Imisun sunflower in response to imazethapyr using RNA-Seq and to determine whether these mechanisms are constitutive or herbicide-induced. Cypsels were germinated in plastic pots, watered by capillarity and growth in chamber under controlled conditions. Seven-day-old plants were treated with 0 (control) and 1 μM imazethapyr for 12 h. After leaf RNA purification, stranded, paired-end cDNA libraries were constructed. Sequencing was performed with Illumina HiSeq2000. Local mapping, with and without multihits, was carried out over the reference transcriptome HaT13l and differential expression was analysed. Sixty one and 47 contigs (according to mapping strategy) related to xenobiotic metabolism were found: cytochromes P450s, ABC transporters, glycosyltransferases, UDPglucuronosyl/glucosyltransferases and glutathione S-transferases. None of these contigs showed differential expression between control and imazethapyr-treated plants. Seventeen interesting contigs were verified by qRT-PCR. These results suggest that constitutive NTSR mechanisms may account for imidazolinone resistance in Imisun sunflower.","PeriodicalId":39086,"journal":{"name":"Helia","volume":"41 1","pages":"267 - 278"},"PeriodicalIF":0.0,"publicationDate":"2018-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1515/helia-2018-0012","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42064737","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 6

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