Waraporn Heedchim, Chakriya Niha, S. Yenchon, S. Te-chato
{"title":"Development of somatic embryogenesis of mangrove fan palm (Licuala spinosa Thunb.) from culturing immature zygotic embryo","authors":"Waraporn Heedchim, Chakriya Niha, S. Yenchon, S. Te-chato","doi":"10.1017/s1479262123000540","DOIUrl":"https://doi.org/10.1017/s1479262123000540","url":null,"abstract":"\u0000 Mangrove fan palm is an endemic plant that grows well and has for a long period been related to traditional ceremonies in southern Thailand. It is an endangered plant with a slow growth and is difficult to propagate in mass production. Tissue culture technique can be used to produce a large number of plants within a short time and to preserve in vitro germplasm resulting in the plant not becoming extinct from natural habitats. Thus, the objectives of this research were to study effects of auxins and embryo preparation on somatic embryogenesis. The sterile immature zygotic embryo was cultured on MS medium with different types and concentrations of auxins. The results showed that 1 mg/l dicamba gave the best results in callus induction at 100% and callus diameter at 4.67 mm after 8 weeks of culture. For callus proliferation, callus chopped at 100 times and transferred to 0.1 mg/l dicamba containing oil palm culture medium gave the highest callus diameter at 10.33 mm after 4 weeks of culture and somatic embryos were also produced. Upon transferring somatic embryos to MS medium with 0.2 M sorbitol, a large number of various developmental stages of secondary somatic embryos were induced within 4 weeks. Therefore, the finding of this study could be highlighted that dicamba and chopping callus have an important role to induce callus and somatic embryo. This is the first report achieved on in vitro propagation of this plant and it will be beneficial for genetic conservation and biotechnological applications in the future.","PeriodicalId":20252,"journal":{"name":"Plant Genetic Resources: Characterization and Utilization","volume":null,"pages":null},"PeriodicalIF":1.1,"publicationDate":"2023-08-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"74896695","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Genetic variation in Bambara ground nuts as revealed by agro-morphological and DArTseq markers and selection for improved yield performance","authors":"Michael M. Chipeta, D. Gimode","doi":"10.1017/s1479262123000503","DOIUrl":"https://doi.org/10.1017/s1479262123000503","url":null,"abstract":"\u0000 Bambara groundnut (Vigna subterranea (L.) Verdc) has been neglected in terms of variety selection and development which has resulted in farmers growing a mixture of landraces that are not genetically characterized and are low yielding. With the need to set up a breeding programme in Malawi, it was necessary to thoroughly understand the genetic diversity (GD) present in the available germplasm. The objectives of the study were to assess Bambara genotypes GD using agro-morphological traits and SNP markers, and to identify and select high yielding Bambara genotypes. Field trials were conducted for two seasons at Bunda College. Later, genotypes were genotyped using DartSeqLD SNP markers. All data were analysed using R Package. Significant genetic variations (P < 0.001) were observed for most traits including grain yield, which suggests that genetic variability exists in Bambara groundnuts which can be exploited in breeding programmes aimed at developing high performing varieties. Based on grain yield, the study identified 18 top performing genotypes across the evaluation seasons which will be tested under farmers’ fields’ conditions. DArTseqLD grouped the genotypes into three clusters. It was noted that majority of the genotypes from the same origin clustered together. High genetic distances were observed between genotypes from Southern African and West African regions and this has important implications in parental selection for the genetic improvement of Bambara. Our results provide valuable insights about the extent of genetic variability and how parental lines can be selected for improved genetic gain in Bambara groundnuts.","PeriodicalId":20252,"journal":{"name":"Plant Genetic Resources: Characterization and Utilization","volume":null,"pages":null},"PeriodicalIF":1.1,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"77416526","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Samira Montahae Dargah, M. Rezaei, M. Ghanbari Jahromi, S. Kalateh Jari, Shahin Jahangirzadeh Khiavi
{"title":"Genetic resources diversity of tea (Camellia sinensis (L.) Kuntze) in the southern region of the Caspian Sea","authors":"Samira Montahae Dargah, M. Rezaei, M. Ghanbari Jahromi, S. Kalateh Jari, Shahin Jahangirzadeh Khiavi","doi":"10.1017/s147926212300045x","DOIUrl":"https://doi.org/10.1017/s147926212300045x","url":null,"abstract":"\u0000 In the fields of agricultural, food and medical research, the potential impacts of tea on human health are of special interest because tea (Camellia sinensis (L.) Kuntze) is continuously consumed by many people in the world. The evaluation of the diversity of genotypes found in tea germplasm can aid in the improvement of the breeding programme. In this study, the genetic diversity of 30 tea genotypes from two commercial sites of tea production in Iran was investigated by using morphological and 12 ISSR markers. Morphological analysis showed that the diversity between samples of tea was limited, and the narrow matching range was calculated. In cluster analysis at level 0.63, samples were divided into four groups. The application of 12 ISSR primers produced 91 polymorphic bands. PIC test showed a range of 0.41–0.48. Based on the ISSR data, the matching range was obtained in the range of 0.24–0.93. In cluster analysis, samples at level 0.58 were divided into five groups. According to the results, it can be understood that these series of traits and primers can very well recognize genetic differences. Using these markers, genetic diversity was observed among tea genotypes, but this diversity was not such as to be able to separate genotypes of various regions from each other. The results showed that the tea genotypes in Iran had high genetic diversity. As a consequence, the findings of our study will help the development of tea germplasm conservation strategies and their sustainable use in breeding programmes.","PeriodicalId":20252,"journal":{"name":"Plant Genetic Resources: Characterization and Utilization","volume":null,"pages":null},"PeriodicalIF":1.1,"publicationDate":"2023-07-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87927223","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Hyun Min Kim, M. Roh, Yong Kwon Yoo, K. Yoo, Y. Joung
{"title":"Leaf morphology, genetic analysis and low temperature requirement for flowering of Verbascum blattaria","authors":"Hyun Min Kim, M. Roh, Yong Kwon Yoo, K. Yoo, Y. Joung","doi":"10.1017/s1479262123000497","DOIUrl":"https://doi.org/10.1017/s1479262123000497","url":null,"abstract":"\u0000 Verbascum blattaria L., commonly known as moth mullein, naturalized in the USA that produces white or yellow flowers could be considered as a potential ornamental plant. However, genetic characterization using molecular markers and leaf morphology, colourimetric analysis and flowering of V. blattaria influenced by low temperature treatments was not investigated to evaluate as a potential horticultural and landscape plant use. The basal leaves developed during the rosette-growth stage were oblanceolate with an obtuse leaf apex and incisions at the margin. Leaves produced on the stem during the reproductive development were ovate or lanceolate with an obtuse or acute leaf apex. Regardless of the colour of the petiole and leaf blade during the rosette-growth stage, there were no differences in the sequences of nuclear ribosomal ITS and chloroplast interspacer. All plants produced creamy white flowers with a purplish base corolla. All leaves formed during the vegetative and reproductive development were glabrous. Numerous stalked glandular trichomes were observed in the sepal, pedicel and bracts. Seeds started to germinate in 10 days at 25°C and reached the plateau in 30 days after sowing. The earliest flowering occurred in 131 days when the plants received 20 days of low temperature treatment (CD) (20 CD), producing 76 flowers, the highest number compared to the number of flowers produced by plants that received 0, 40 and 60 CD. Plants that received 20 CD exhibited early flowering, probably because of the early transition from vegetative growth to reproductive development, as judged by the short stem to the first flower.","PeriodicalId":20252,"journal":{"name":"Plant Genetic Resources: Characterization and Utilization","volume":null,"pages":null},"PeriodicalIF":1.1,"publicationDate":"2023-07-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75925394","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Apostolos N. Konstantopoulos, Sofia Pozoukidou, M. Irakli, I. Tsialtas
{"title":"Testa and hilum colour associations with seed traits of a Greek field pea landrace","authors":"Apostolos N. Konstantopoulos, Sofia Pozoukidou, M. Irakli, I. Tsialtas","doi":"10.1017/s1479262123000527","DOIUrl":"https://doi.org/10.1017/s1479262123000527","url":null,"abstract":"Pea landraces may hold genetic variation that can be exploitable in breeding/selecting new cultivars. In a small-seeded pea landrace, four phenotypes were distinguished according to testa (green, non-spotted and green, spotted) and hilum colour (white, black). The four phenotypes were tested for two growing seasons in the field for pod (seeds/pod) and seed traits (1000-seed weight, toughness, total phenols and tannins, testa colour, protein and carbon concentration, C/N ratio, bruchid infection). Significant differences were found for testa colour parameters, phenolic, tannin and carbon concentration and bruchid tolerance. The larger-seeded, spotted peas had darker testa and more phenols, while white-hilumed peas had lighter testa and more tannins. The spotted, black-hilumed phenotype, with the highest carbon concentration and C/N ratio was the most tolerant to bruchids. However, grouping the phenotypes, neither spotted nor black-hilumed ones showed to be more tolerant compared with their counterparts. Concluding, our results showed that phenotyping variation in seeds of a pea landrace revealed variation in seed traits, which could be exploitable. Since testa and hilum colour were associated with specific seed traits, they could, alone or in combination, be used as biomarkers of seed quality traits in pea. Testing of larger number of phenotypes is needed to solidify our findings.","PeriodicalId":20252,"journal":{"name":"Plant Genetic Resources: Characterization and Utilization","volume":null,"pages":null},"PeriodicalIF":1.1,"publicationDate":"2023-07-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"74557806","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
K. Chaturvedi, U. Kumara, Anuradha Sane, Pritee Singh, Prakash Kumar, P. Tripathi
{"title":"Exploring the genetic diversity of Aegle marmelos (L.) Correa populations in India","authors":"K. Chaturvedi, U. Kumara, Anuradha Sane, Pritee Singh, Prakash Kumar, P. Tripathi","doi":"10.1017/s1479262123000485","DOIUrl":"https://doi.org/10.1017/s1479262123000485","url":null,"abstract":"\u0000 Bael is an important sub-tropical fruit crop in family Rutaceae that is widely distributed throughout South-East Asia. For local communities, the nutritious composition of its fruits and leaves offers tremendous economic and social possibilities to exploit. However, its underutilized status, as well as man-made threats to its natural habitat, make it imperative to implement concrete strategies for its cultivation and conservation. To fully grasp the ability of this adaptable fruit tree for human health and environmental well-being, it is necessary to characterize the genetic diversity. The goal of this study was to use morphological (13 quantitative traits), biochemical (9 attributes) and molecular (10 SRAP primers) characterization to evaluate 24 bael genotypes from two agroecological zones of India. Fruit and pulp weight ranged from 79.0– to 1478.8 g and 15.0– to 894.3 g with mean values of 448.67 and 233.3 g, respectively. Traits such as fruit, pulp, and seed weight (g), fruit length (cm) and width (cm), number of fruits per tree, number of seeds per fruit, shell weight (g) and shell thickness (mm) recorded highly significant differences. High phenol (11.65–24.38 mg GAE/g fw) and flavonoid (12.32–74.63 mg CE/g fw) content was observed in fruit pulp indicating significant antioxidant potential of this fruit. Several morphological and biochemical characters were found to have significant positive correlations. Principal component analysis revealed that first five components contributed 96.76% to total variation. Hierarchical cluster analysis separated the populations into two distinct clusters, while analysis of molecular variance (AMOVA) using SRAP markers revealed that 70% of the total marker variation was due to interpopulation variance, while 30% was attributed to intrapopulation.","PeriodicalId":20252,"journal":{"name":"Plant Genetic Resources: Characterization and Utilization","volume":null,"pages":null},"PeriodicalIF":1.1,"publicationDate":"2023-07-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75025321","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Sabhyata Sabhyata, Arun Gupta, D. Aggarwal, R. Tiwari, G. Singh, G. Singh
{"title":"Diversity in Indian wheat (T. aestivum L.) germplasm for various agro-morphological traits","authors":"Sabhyata Sabhyata, Arun Gupta, D. Aggarwal, R. Tiwari, G. Singh, G. Singh","doi":"10.1017/s1479262123000461","DOIUrl":"https://doi.org/10.1017/s1479262123000461","url":null,"abstract":"\u0000 A better understanding of diversity in landraces is essential for planning crosses for the development of trait specific varieties with better adaptability and stability. In the present study, 120 wheat genotypes comprised of landraces, genetic stocks, released varieties and improved genotypes were evaluated in randomized block design for 14 agro-morphological traits. The clustering method and principal component analysis (PCA) programme of Statistical Package for Agricultural Research (SPAR1) was used for grouping the genotypes. These 120 genotypes were grouped into nine clusters based on agro-morphological traits. These nine clusters differed significantly on the basis of mean values for 14 agro-morphological traits. PCA showed that the two principal components (PC1 to PC2) exhibited about 49% of the total variability. Scatter plot was constructed by plotting scores of PC1 and PC2. Based on mean values obtained over two years, the diverse superior genotypes were identified for utilization in hybridization programme. From the present study, we conclude that cluster analysis grouped the landraces with greater agro-morphological similarity into one group rather than geographical isolation indicating that geographical origin may not be the only factor causing diversity. Further, released varieties exhibited superiority for grain yield, while many landraces had higher values for number of tillers in a meter, biomass and thousand-grain weight (TGW). Thus, for the improvement in TGW in released varieties, the hybridization between superior landraces for TGW from cluster ‘E’ and released wheat varieties from cluster ‘C’ could give desirable segregates.","PeriodicalId":20252,"journal":{"name":"Plant Genetic Resources: Characterization and Utilization","volume":null,"pages":null},"PeriodicalIF":1.1,"publicationDate":"2023-07-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"82230522","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Tomoko Nozoye, Y. Gorafi, Naoki Ube, Fan Wang, H. Nakanishi, A. Ishihara, Takayoshi Ishii, H. Tsujimoto
{"title":"Diversity in the genome of Aegilops tauschii, a wild wheat relative, to generate Fe-biofortified and Fe-deficiency-tolerant wheat","authors":"Tomoko Nozoye, Y. Gorafi, Naoki Ube, Fan Wang, H. Nakanishi, A. Ishihara, Takayoshi Ishii, H. Tsujimoto","doi":"10.1017/s1479262123000424","DOIUrl":"https://doi.org/10.1017/s1479262123000424","url":null,"abstract":"\u0000 Iron (Fe) is an essential element for all organisms. Fe deficiency can limit plant production and cause anaemia in humans. The improvement of Fe homoeostasis could resolve both problems. Fe homoeostasis in Aegilops tauschii, the D genome donor of bread wheat, is not fully understood. Here, we analysed physiological traits in 42 accessions of Ae. tauschii associated with Fe homoeostasis, i.e. mugineic acid family phytosiderophores (MAs), phenylamides, SPAD values and metal concentrations. All traits showed diversity, suggesting the presence of candidate genes in the Ae. tauschii accessions, which could improve Fe homoeostasis in bread wheat. All accessions mainly produced and secreted mainly 2′-deoxymugineic acid among MAs, but eight of them secreted unknown products from their roots under Fe deficiency. It was revealed that 15 kinds of phenylamides and 2 kinds of bread wheat phytoalexins were produced in Fe-deficient roots of Ae. tauschii. Multivariate and principal component analyses showed that chlorophyll content was correlated with shoot Fe concentration. Genome-wide association study analysis associated several genomic markers with the variations in each trait analysed. Our results suggest that Ae. tauschii has alleles that could improve Fe homoeostasis to generate Fe-deficiency-tolerant or Fe-biofortified bread wheat.","PeriodicalId":20252,"journal":{"name":"Plant Genetic Resources: Characterization and Utilization","volume":null,"pages":null},"PeriodicalIF":1.1,"publicationDate":"2023-07-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75841829","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
T. Preethy, M. Murugan, Nimisha Mathews, Astile Kuriakose
{"title":"Insights into the genetic diversity of Indian cardamom [Elettaria cardamomum (L.) Maton]: for a future research perspective","authors":"T. Preethy, M. Murugan, Nimisha Mathews, Astile Kuriakose","doi":"10.1017/s1479262123000369","DOIUrl":"https://doi.org/10.1017/s1479262123000369","url":null,"abstract":"\u0000 One of the most essential prerequisites in plant breeding is to have a maximum collection of germplasm materials with all sorts of variability. As a part of the programme under the All India Coordinated Research Projects on Spices, totally 196 germplasm accessions of small cardamom are being maintained as field gene bank repository at the Cardamom Research Station, Pampadumpara. Tropical evergreen forests of the Indian Western Ghats and Sri Lankan Central Highlands are recognized as the home of cardamom. The area and production of cardamom are maximum in Kerala followed by Karnataka and Tamil Nadu. Variations among the germplasm collections in morphological and biometrical characters as well as for yield have been studied and reported in this paper. Accessions with distinct morphological marker characters, such as compound panicle types, terminal panicle bearing, narrow leaf types, pink pseudostem types, dark green bold capsules with high-yield potential and biotic-stress tolerant types, are being evaluated and conserved in this repository. The assessment of genetic diversity is an essential prerequisite for undertaking crop breeding activities to evolve suitable area and region-specific variety. Sixty-seven cardamom accessions were studied for genetic diversity by evaluating 14 unique characters for 3 years (2016–2018). Almost all accessions have shown significant variability for most of the biometric and biotic stress characters. Results have indicated a greater magnitude of genetic diversity of small cardamom present among accessions representing whole evergreen tropical forest of the Western Ghats.","PeriodicalId":20252,"journal":{"name":"Plant Genetic Resources: Characterization and Utilization","volume":null,"pages":null},"PeriodicalIF":1.1,"publicationDate":"2023-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89202287","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
A. Kronberga, I. Nakurte, L. Kaļāne, Alekss Vecvanags, D. Jakovels, J. Filipovs, I. Mežaka
{"title":"Cultivation potential of Taraxacum campylodes G.E.Haglund wild populations: morphological and biochemical variation","authors":"A. Kronberga, I. Nakurte, L. Kaļāne, Alekss Vecvanags, D. Jakovels, J. Filipovs, I. Mežaka","doi":"10.1017/s1479262123000436","DOIUrl":"https://doi.org/10.1017/s1479262123000436","url":null,"abstract":"\u0000 The common dandelion (Taraxacum campylodes G.E.Haglund) is known for its medicinal properties and as a non-toxic, highly nutritious edible plant. Even though the herb is mostly collected in the wild, the food and pharmaceutical industries prefer cultivated plant material. Cultivation offers the possibility to achieve predictable yields, ensure the botanical identity of the plants and products can be certified according to organic or biodynamic standards; therefore the interest in domestication is growing. Due to the scarcity of research on dandelion cultivation in agricultural systems, the current study focused on determining the quantitative and qualitative characteristics of 13 wild populations from Latvia in an organic farming system. Populations collected at Gluda, Priekuli, Mundigciems, Marupe as well as variety TA008 had the highest dry root weight (>100 g per plant) and exceeded the root weight of variety Nouvelle more than two-fold. The highest concentration of chlorogenic acid (CHA) (0.12%) and chicoric acid (CCA) (0.15%) was found in the wild population collected in Turaida. The wild populations from Marupe and Mundigciems had a high average root weight as well as a relatively high CHA and CCA content, making them suitable for commercial growing. The present study examined the feasibility of cultivating wild dandelion populations in an organic farming system. The results revealed a high degree of variation within and between populations; therefore, dandelion wild populations can be used for domestication, breeding and selection of accessions most suitable for the pharmaceutical or food industries.","PeriodicalId":20252,"journal":{"name":"Plant Genetic Resources: Characterization and Utilization","volume":null,"pages":null},"PeriodicalIF":1.1,"publicationDate":"2023-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85819581","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}