It was helpful for the wheat improvement to evaluate the genetic resources of Triticum turgidum L. ssp. turgidum landraces. In this study, 68 turgidum landraces accessions, belonging to four geographic populations in ...It was helpful for the wheat improvement to evaluate the genetic resources of Triticum turgidum L. ssp. turgidum landraces. In this study, 68 turgidum landraces accessions, belonging to four geographic populations in China, were investigated by using EST-SSR markers. A total of 63 alleles were detected on 22 EST-SSR loci, and the number of alleles on each locus ranged from 1 to 5, with an average of 2.9. The results of the analysis of molecular variance （AMOVA） indicated that 92.5% of the total variations was attributed to the genetic variations within population, whereas only 7.5% variations among populations. Although the four populations had similar genetic diversity parameters, Sichuan population was yet distinguished from other populations when comparing the population samples in pairs. Significant correlations were detected by the statistic analysis among six genetic diversity parameters among each other. The selection difference between heterozygosty and homozygosty was also observed among different EST-SSR locus. The genetic similarity （GS） ranged from 0.18 to 0.98, with the mean of 0.72, and all accessions could be clustered into 7 groups. The dendrogram suggested that the genetic relationships among turgidum accessions evaluated by EST-SSR markers were unrelated to their geographic distributions. These results implied that turgidum landraces from China had the unique characters of genetic diversity.展开更多
To further develop EST-SSR marker in rubber tree, we assembled the sequences downloaded from NCBI and Malaysia EST databases of rubber tree. By analyzing the assembled 3 733 unigenes, we identified 566 potential SSR s...To further develop EST-SSR marker in rubber tree, we assembled the sequences downloaded from NCBI and Malaysia EST databases of rubber tree. By analyzing the assembled 3 733 unigenes, we identified 566 potential SSR sites in this study. That is to say, there was one EST-SSR in every 3.96 kb. The di-nu-cleotide repeat was the most abundant type, fol owed by tri-, hexa-, tetra- and pen-ta-nucleotide repeat. The most common number of repeat units was 5, fol owed by more than 12, 6 and 7. Of 51 SSR motifs identified in this study, di-, tri-, tetra-, penta- and hexa-nucleotide repeats were 6, 26, 5, 3 and 11 types, respectively. The GA/CT di-nucleotide repeat was the most abundant motif, fol owed by TC/AG, AT/TA, CTT/GAA, TTC/AAG and TCT/AGA. In total, 158 new EST-SSRs were developed and amplified with the DNA of RRIM600 as a template. The results showed that the PCR products of 99 EST-SSRs generated clear amplifying bands. The EST-SSR markers developed in this study further enrich the number of molecular marker in rubber tree, and they wil be widely applied in DNA fingerprinting, genetic diversity, marker-assisted selection and genetic mapping, etc.展开更多
[Objective] This research aimed to develop walnut EST-SSR markers and design corresponding primers.[Method] 5213 EST sequences of walnut（Juglans regia Linn.） public online in NCBI were used for character analysis wi...[Objective] This research aimed to develop walnut EST-SSR markers and design corresponding primers.[Method] 5213 EST sequences of walnut（Juglans regia Linn.） public online in NCBI were used for character analysis with bioinformatics methods,and primers were designed for the selected EST sequences by using Primer 3.0 software.[Result] 207 SSRs were obtained from the EST sequences,including 188 non-redundant sequences,the detection rate was 3.97% with an average distribution distance of 21.12 kb.Totally 92 types of repeat motifs were involved,which were mainly composed of dinucleotide and trinucleotide,accounting for 31.40% and 35.27% of the total number of repeat motifs,respectively.30 pairs of primers were initially selected from the 50 randomly-selected SSR primers by PCR amplification.[Conclusion] This research would lay foundations for the development of EST-SSR molecular markers in walnut and design of the targeted EST-SSR primers by mining and analyzing the SSR sites in walnut EST sequences.展开更多
Developing expressed sequence tag-derived SSR （EST-SSR） markers is imperative in genetic research. In this paper, we reported 37 EST-SSR markers which were developed from 286 unigenes obtained from soybean cDNA libr...Developing expressed sequence tag-derived SSR （EST-SSR） markers is imperative in genetic research. In this paper, we reported 37 EST-SSR markers which were developed from 286 unigenes obtained from soybean cDNA library. Among the 286 markers designed for the 4 accessions of Glycine max and 6 of its wild progenitor （G. soja） within the subgenus Soja, 209 markers amplified DNA fragments, taking 73.1% and 37 markers appeared to be polymorphic, which was 12.9% of the total. The 37 loci detected a total of 142 alleles, while the PIC values varied from 0.194 to 0.794. Both the number of alleles per locus and PIC value were significantly related to the SSR motif. Six EST-SSR loci may be fixed for different alleles between G. max and G. soja since they were particularly polymorphic among the 6 G. soja accessions. A neighbor-joining tree placed the G. max accessions together as a group within the G. soja, though the average genetic distance among G. soja accessions was much higher. These new EST-SSRs markers will be useful for genetic diversity analysis, genetic mapping construction and gene discovery in Soja subgenus.展开更多
Faba bean （Vicia faba L.）, one of the most important legumes in the world, evolved different types of cultivars due to its partial cross-pollination. The development of simple sequence repeat （SSR） markers from ex...Faba bean （Vicia faba L.）, one of the most important legumes in the world, evolved different types of cultivars due to its partial cross-pollination. The development of simple sequence repeat （SSR） markers from expressed sequence tags （EST） provided a useful tool for investigation of its genetic diversity. The purpose of the present study was to investigate the genetic diversity of faba bean from China and Europe using EST-SSR markers. 5 031 faba bean ESTs from the NCBI database were downloaded and assembled into 1 148 unigenes. A total of 107 microsatellites in 96 unigenes were identified, indicating that merely 8.36% of sequences contained SSRs. The most abundant SSR within faba bean was tri-nucleotide repeat motif, and among all the tri-nucleotide repeats, the motif AAG/CTT was the most abundant type. Based on these results, 11 EST-SSR markers were used to assess the genetic diversity of 29 faba bean cultivars from China and Europe with two to three alleles per locus. The polymorphism information content value ranged from 0.0644 to 0.4278 with an average of 0.2919. Principal coordinate analysis （PCA） and phylogenetic clustering based on these 11 EST-SSR markers distinguished these cultivars into different groups. The results indicated that faba bean in China had a narrow genetic basis, and the additional sources of genetic cultivars/accessions should be introduced to enhance the genetic variability. The results of this study proved that the EST-SSR marker is very effective in evaluation of faba bean germplasm.展开更多
Fifteen expressed sequence tag (EST)-derived simple sequence repeats (EST-SSRs) were used to investigate genetic diversity in 139 plants obtained from seeds of 35 watermelon accessions collected from all the geographi...Fifteen expressed sequence tag (EST)-derived simple sequence repeats (EST-SSRs) were used to investigate genetic diversity in 139 plants obtained from seeds of 35 watermelon accessions collected from all the geographical provinces of Zimbabwe. In addition, 15 plants representing three commercial varieties developed in the United States (USA) were analyzed for comparison. A total of 65 alleles were detected among all the watermelon accessions. For the 13 polymorphic EST-SSR loci, number of alleles per locus varied from 2 to 13, with an average of 5 alleles per locus. Values for the polymorphic information content increased as the number of alleles increased, and varied from 0.15 to 0.77 with an average of 0.54 suggesting sufficient discriminatory power. Both cluster analysis and principal coordinate analysis (PCA) produced two major clusters;one with the 22 cow-melon accessions and the other with the 16 sweet watermelon accessions. Within the sweet watermelon group, two distinct sub-clusters formed, one of which contained only two of the commercial varieties from USA. Partitioning of genetic variation in the Zimbabwean material using analysis of molecular variation (AMOVA) revealed that 64% of the total variation resides between the two major forms, i.e. sweet watermelons and cow-melons, 28% between accessions within forms and 8% within accessions. The EST-SSR markers revealed a somewhat higher diversity in sweet watermelon accessions compared to that of cow-melons. This finding is contrary to previous reports using other markers (genomic SSR loci or RAPD) and/or a plant material that is likely to have experienced more stringent selection procedures compared to the landraces analyzed in our study.展开更多
In this study,64 498 ESTs of Melampsoraspp.was assembled into 1 998 contigs,and 604SSR loci were detected on these contigs,with 736.6 bp containing one SSR on average.Among these SSRs,trinucleotide repeats were the mo...In this study,64 498 ESTs of Melampsoraspp.was assembled into 1 998 contigs,and 604SSR loci were detected on these contigs,with 736.6 bp containing one SSR on average.Among these SSRs,trinucleotide repeats were the most abundant repeats（44.70%）.As for the composition of microsatellites, AC and AT repeats were the richest motif in dinucleotide repeats,and AGT and AAG repeats were the most frequent motifs in trinucleotide repeats,whereas（AAAN） n and（AAAAN） n repeats were dominant in tetranucleotide and pentanucleotide repeats,respectively.All the dominant repeat motifs of different types of SSRs were rich in A and T.In EST sequences of Melampsoraspp.genome,microsatellites longer than 20 bp accounted for about 15.07%.It was noticeable that microsatellites were highly rich in the expressed sequences of Melampsoraspp.genome,which implied that SSRs played a significant role in triggering the gene mutation in Melampsoraspp.genome.A total of 455 SSR primers were designed according to the detected microsatellites using Primer 5.0 and Oligo 6. 0,and 30 primer pairs were randomly selected for amplification test.Among these primer pairs,27 primer pairs succeed in amplification,with a successful rate of 90%.Eight primer pairs generated polymorphic fingerprints in Melampsoraspp.collected from different poplar genotypes,accounting for 26.7% of the total primer pairs.The EST-SSR markers developed fromMelampsoraspp.EST sequences provided important marker resources for studying Melampsoraspp.from the aspects of pathogen identification and survey of genetic variation.展开更多
In this study, 27 pairs of EST-SSR primers were employed to analyze the genetic diversity and genetic relationship of 100 wild tea plant germplasm re- sources and 22 cultivars, according to the results, a total of 88 ...In this study, 27 pairs of EST-SSR primers were employed to analyze the genetic diversity and genetic relationship of 100 wild tea plant germplasm re- sources and 22 cultivars, according to the results, a total of 88 polymorphic bands were amplified with 27 pairs of primers; the variation of effective alleles accounted for 69.01% ; a total of 183 genotypes were detected, with a variation range of 4 -11 ; averagely 6.78 genotypes were amplified with each primer pair; Shannon index （I） of 27 primer pairs ranged from 0.32 to I. 35, with an average of 0.88 ; the observed heterozygosity （0.52） was basically consistent with the expected het- erozygosity （0.52） ; the average polymorphism heterozygosity was 0.48, which was very close to 0.50 ; the average Nei＇s index was 0.51, which was higher than 0. 50 ; the average polymorphism information content （PIC） was 0.52, which was higher than 0.50, indicating high genetic diversity among wild tea germplasm resources in Yuunan Province. According to the clustering results, based on geographical origins and genetic backgrounds, 122 materials were clustered into 14 categories. Dendrogram based on Nei＇s genetic distance revealed complex genetic relationships among wild tea germplasm resources in Yunnan Province. This study provided certain reference for subsequent preservation, development and research of wild tea germplasm resources in China.展开更多
The present study was conducted to develop EST-SSR markers using the cDNA library from rice plant. Total RNA extracted from the leaves of brown plant hopper resistance gene originated from a rice cultivar “Cheongche...The present study was conducted to develop EST-SSR markers using the cDNA library from rice plant. Total RNA extracted from the leaves of brown plant hopper resistance gene originated from a rice cultivar “Cheongcheong” and sensitive rice cultivar “Nakdong” were used to synthesize a cDNA library. As a result of analyzing the cDNA library, the 17 EST-SSR primer sets were developed. This study enables to provide effective marker assisted selection (MAS) methods on the selection of white-backed planthopper resistance gene originated from a rice plant more simply, quickly and precisely. Furthermore, using this marker’s advantage of deriving from cDNA, it is possible to identify the white-backed planthopper resistance gene. In addition, this study introduces a technique for construction of a cDNA library safely without using radioactivity.展开更多
This commentary critically appraises the study by Li et al which pioneered the exploration of the triglyceride-glucose(TyG)index as a prognostic marker in hepatitis B virus-related advanced hepatocellular carcinoma pa...This commentary critically appraises the study by Li et al which pioneered the exploration of the triglyceride-glucose(TyG)index as a prognostic marker in hepatitis B virus-related advanced hepatocellular carcinoma patients undergoing combined camrelizumab and lenvatinib therapy.While we acknowledge the study’s clinical relevance in proposing an easily accessible metabolic biomarker,we delve into the mechanistic plausibility linking insulin resistance to immunotherapy response and angiogenic inhibition.We further critically examine the methodological limitations,including the retrospective design,the populationspecific TyG cut-off value,and unaddressed metabolic confounders.We highlight the imperative for future research to validate its utility across diverse etiologies and treatment settings,and to unravel the underlying immunometabolic pathways.展开更多
Notopterygium incisum C. C. Ting ex H. T. Chang(Apiaceae) is an endangered perennial herb in China. The lack of transcriptomic and genomic resources for N. incisum greatly hinders studies of its population genetics an...Notopterygium incisum C. C. Ting ex H. T. Chang(Apiaceae) is an endangered perennial herb in China. The lack of transcriptomic and genomic resources for N. incisum greatly hinders studies of its population genetics and conservation. In this study, we employed RNA-seq technology to characterize transcriptomes for the flowers, leaves, and stems of this endangered herb. A total of 56 million clean reads were assembled into 120,716 unigenes with an N50 length of 850 bp. Among these unigenes, 70,245(58.19%) were successfully annotated and 65,965(54.64%) were identified as coding sequences based on their similarities with sequences in public databases. We identified 21 unigenes that had significant relationships with cold tolerance in N. incisum according to gene ontology(GO) annotation analysis. In addition, 13,149 simple sequence repeats(SSRs) and 85,681 single nucleotide polymorphisms were detected as potential molecular genetic markers. Ninety-six primer pairs of SSRs were randomly selected to validate their amplification efficiency and polymorphism. Nineteen SSR loci exhibited polymorphism in three natural populations of N. incisum. These results provide valuable resources to facilitate future functional genomics and conservation genetics studies of N. incisum.展开更多
果肉颜色是猕猴桃重要的品质性状,不同果肉颜色的猕猴桃在色素成分、糖和维生素C含量等方面存在很大差异。为了探究猕猴桃果肉颜色形成的分子机制,本研究以红肉猕猴桃‘红实2号’和黄肉猕猴桃‘金实1号’为研究对象,采用RNA-seq技术,通...果肉颜色是猕猴桃重要的品质性状,不同果肉颜色的猕猴桃在色素成分、糖和维生素C含量等方面存在很大差异。为了探究猕猴桃果肉颜色形成的分子机制,本研究以红肉猕猴桃‘红实2号’和黄肉猕猴桃‘金实1号’为研究对象,采用RNA-seq技术,通过分析果实不同时期不同部位的转录组表达差异。结果表明,编码花青素合成的Achn385311(3GGT)基因可能是控制‘红实2号’内果皮呈红色的主效基因,‘金实1号’后期果肉呈深黄色可能与Achn158981(GT7)、Achn150731、Achn068721(PAO)、Achn282201(PAO)和Achn176251(UGT71A16)等基因的表达调控有关。针对差异表达的230个猕猴桃果肉颜色相关基因,利用Primer Premier 5.0设计出727对EST-SSR引物,以7份不同果肉颜色的猕猴桃基因组DNA为模板,随机选择112对引物进行有效性验证,筛选出了具有清晰的扩增产物的引物78对,有效扩增率为69.64%,58对具有多态性,多态性频率为74.36%。本研究结果为猕猴桃果实呈色机理研究及分子标记辅助育种提供了理论基础。展开更多
In recent years, Jatropha curcas L. has gained popularity as a potential biodiesel plant. The varying oil content, reported between accessions belonging to different agroclimatic zones, has necessitated the assessment...In recent years, Jatropha curcas L. has gained popularity as a potential biodiesel plant. The varying oil content, reported between accessions belonging to different agroclimatic zones, has necessitated the assessment of the existing genetic variability to generate reliable molecular markers for selection of high oil yielding variety. EST derived SSR markers are more useful than genomic markers as they represent the transcriptome, thus, directly linked to functional genes. The present report describes the in silico mining of the microsatellites (SSRs) using J. curcas ESTs from various tissues viz. embryo, root, leaf and seed available in the public domain of NCBI. A total of 13,513 ESTs were downloaded. From these ESTs, 7552 unigenes were obtained and 395 SSRs were generated from 377 SSR-ESTs. These EST-SSRs can be used as potential microsatellite markers for diversity analysis, MAS etc. Since the Jatropha genes carrying SSRs have been identified in this study, thus, EST-SSRs directly linked to genes will be useful for developing trait linked markers.展开更多
We analyzed the genetic differences of 16 poplar clones between genomic-SSR and EST-SSR markers. The statistical results show that the average number of alleles detected by genomic-SSR was 4.1, Shannon＇s index 1.0646...We analyzed the genetic differences of 16 poplar clones between genomic-SSR and EST-SSR markers. The statistical results show that the average number of alleles detected by genomic-SSR was 4.1, Shannon＇s index 1.0646, observed heterozygos- ity 0.4427 and expected heterozygosity 0.5523, while for the EST-SSR, the average number of alleles was 2.8, Shannon＇s index 0.6985, observed heterozygosity 0.2330 and expected heterozygosity 0.4684. Cluster analysis indicated that the EST-SSR capacity of genotypic identification was more precise than that of genomic-SSR. These resuks reveal that EST-SSR and genomic-SSR have statistically significant genetic differences in polymorphism detection and genotypic identification. These differences could provide a theoretical basis for the rational use of SSR markers in species diversity and other related research.展开更多
基金the National High Technology Research and Development Program of China (863 Program,2006AA10Z179 and 2006AA10Z1F8)the National Excellent Doctoral Dissertation of China (200357 and 200458)the Key Technologies R&D Program of China (2006BAD01A02-23 and 2006BAD13B02)
文摘It was helpful for the wheat improvement to evaluate the genetic resources of Triticum turgidum L. ssp. turgidum landraces. In this study, 68 turgidum landraces accessions, belonging to four geographic populations in China, were investigated by using EST-SSR markers. A total of 63 alleles were detected on 22 EST-SSR loci, and the number of alleles on each locus ranged from 1 to 5, with an average of 2.9. The results of the analysis of molecular variance （AMOVA） indicated that 92.5% of the total variations was attributed to the genetic variations within population, whereas only 7.5% variations among populations. Although the four populations had similar genetic diversity parameters, Sichuan population was yet distinguished from other populations when comparing the population samples in pairs. Significant correlations were detected by the statistic analysis among six genetic diversity parameters among each other. The selection difference between heterozygosty and homozygosty was also observed among different EST-SSR locus. The genetic similarity （GS） ranged from 0.18 to 0.98, with the mean of 0.72, and all accessions could be clustered into 7 groups. The dendrogram suggested that the genetic relationships among turgidum accessions evaluated by EST-SSR markers were unrelated to their geographic distributions. These results implied that turgidum landraces from China had the unique characters of genetic diversity.
基金Supported by the Fundamental Scientific Research Funds for Chinese Academy of Tropical Agricultural Sciences(1630022013028)National Natural Science Foundation of China(30960310,31200514,31270651)~~
文摘To further develop EST-SSR marker in rubber tree, we assembled the sequences downloaded from NCBI and Malaysia EST databases of rubber tree. By analyzing the assembled 3 733 unigenes, we identified 566 potential SSR sites in this study. That is to say, there was one EST-SSR in every 3.96 kb. The di-nu-cleotide repeat was the most abundant type, fol owed by tri-, hexa-, tetra- and pen-ta-nucleotide repeat. The most common number of repeat units was 5, fol owed by more than 12, 6 and 7. Of 51 SSR motifs identified in this study, di-, tri-, tetra-, penta- and hexa-nucleotide repeats were 6, 26, 5, 3 and 11 types, respectively. The GA/CT di-nucleotide repeat was the most abundant motif, fol owed by TC/AG, AT/TA, CTT/GAA, TTC/AAG and TCT/AGA. In total, 158 new EST-SSRs were developed and amplified with the DNA of RRIM600 as a template. The results showed that the PCR products of 99 EST-SSRs generated clear amplifying bands. The EST-SSR markers developed in this study further enrich the number of molecular marker in rubber tree, and they wil be widely applied in DNA fingerprinting, genetic diversity, marker-assisted selection and genetic mapping, etc.
基金Supported by Natural Science Foundation of Zhejiang Province(Y307469)Talent Start-up Fund Project of Zhejiang Agriculture and Forestry University~~
文摘[Objective] This research aimed to develop walnut EST-SSR markers and design corresponding primers.[Method] 5213 EST sequences of walnut（Juglans regia Linn.） public online in NCBI were used for character analysis with bioinformatics methods,and primers were designed for the selected EST sequences by using Primer 3.0 software.[Result] 207 SSRs were obtained from the EST sequences,including 188 non-redundant sequences,the detection rate was 3.97% with an average distribution distance of 21.12 kb.Totally 92 types of repeat motifs were involved,which were mainly composed of dinucleotide and trinucleotide,accounting for 31.40% and 35.27% of the total number of repeat motifs,respectively.30 pairs of primers were initially selected from the 50 randomly-selected SSR primers by PCR amplification.[Conclusion] This research would lay foundations for the development of EST-SSR molecular markers in walnut and design of the targeted EST-SSR primers by mining and analyzing the SSR sites in walnut EST sequences.
基金supported by the National High-Tech R&D Program of China(863Program,2006AA10A110 and 2006AA10Z164)National Basic Research Program of China(973 Program,2010CB125900 and 2004CB117203)the Academy and Institute Foundation for Basic Scientific Research in Institute of Crop Science,Chinese Academy of Agricultural Sciences
文摘Developing expressed sequence tag-derived SSR （EST-SSR） markers is imperative in genetic research. In this paper, we reported 37 EST-SSR markers which were developed from 286 unigenes obtained from soybean cDNA library. Among the 286 markers designed for the 4 accessions of Glycine max and 6 of its wild progenitor （G. soja） within the subgenus Soja, 209 markers amplified DNA fragments, taking 73.1% and 37 markers appeared to be polymorphic, which was 12.9% of the total. The 37 loci detected a total of 142 alleles, while the PIC values varied from 0.194 to 0.794. Both the number of alleles per locus and PIC value were significantly related to the SSR motif. Six EST-SSR loci may be fixed for different alleles between G. max and G. soja since they were particularly polymorphic among the 6 G. soja accessions. A neighbor-joining tree placed the G. max accessions together as a group within the G. soja, though the average genetic distance among G. soja accessions was much higher. These new EST-SSRs markers will be useful for genetic diversity analysis, genetic mapping construction and gene discovery in Soja subgenus.
基金supported by the Zhejiang Provincial Science and Technology Program, China (2007C32013)the Natural Science Foundation of Zhejiang Province, China (Y3090660)
文摘Faba bean （Vicia faba L.）, one of the most important legumes in the world, evolved different types of cultivars due to its partial cross-pollination. The development of simple sequence repeat （SSR） markers from expressed sequence tags （EST） provided a useful tool for investigation of its genetic diversity. The purpose of the present study was to investigate the genetic diversity of faba bean from China and Europe using EST-SSR markers. 5 031 faba bean ESTs from the NCBI database were downloaded and assembled into 1 148 unigenes. A total of 107 microsatellites in 96 unigenes were identified, indicating that merely 8.36% of sequences contained SSRs. The most abundant SSR within faba bean was tri-nucleotide repeat motif, and among all the tri-nucleotide repeats, the motif AAG/CTT was the most abundant type. Based on these results, 11 EST-SSR markers were used to assess the genetic diversity of 29 faba bean cultivars from China and Europe with two to three alleles per locus. The polymorphism information content value ranged from 0.0644 to 0.4278 with an average of 0.2919. Principal coordinate analysis （PCA） and phylogenetic clustering based on these 11 EST-SSR markers distinguished these cultivars into different groups. The results indicated that faba bean in China had a narrow genetic basis, and the additional sources of genetic cultivars/accessions should be introduced to enhance the genetic variability. The results of this study proved that the EST-SSR marker is very effective in evaluation of faba bean germplasm.
文摘Fifteen expressed sequence tag (EST)-derived simple sequence repeats (EST-SSRs) were used to investigate genetic diversity in 139 plants obtained from seeds of 35 watermelon accessions collected from all the geographical provinces of Zimbabwe. In addition, 15 plants representing three commercial varieties developed in the United States (USA) were analyzed for comparison. A total of 65 alleles were detected among all the watermelon accessions. For the 13 polymorphic EST-SSR loci, number of alleles per locus varied from 2 to 13, with an average of 5 alleles per locus. Values for the polymorphic information content increased as the number of alleles increased, and varied from 0.15 to 0.77 with an average of 0.54 suggesting sufficient discriminatory power. Both cluster analysis and principal coordinate analysis (PCA) produced two major clusters;one with the 22 cow-melon accessions and the other with the 16 sweet watermelon accessions. Within the sweet watermelon group, two distinct sub-clusters formed, one of which contained only two of the commercial varieties from USA. Partitioning of genetic variation in the Zimbabwean material using analysis of molecular variation (AMOVA) revealed that 64% of the total variation resides between the two major forms, i.e. sweet watermelons and cow-melons, 28% between accessions within forms and 8% within accessions. The EST-SSR markers revealed a somewhat higher diversity in sweet watermelon accessions compared to that of cow-melons. This finding is contrary to previous reports using other markers (genomic SSR loci or RAPD) and/or a plant material that is likely to have experienced more stringent selection procedures compared to the landraces analyzed in our study.
基金Supported by the Key Forestry Public Welfare Project(201304102)the Key Project for Universities of Jiangsu Province(10KJA180018)+1 种基金enabled by the Program for Innovative Research Team in Universities of Jiangsu Province and the Educational Department of Chinathe Priority Academic Program Development of Jiangsu Higher Education Institutions
文摘In this study,64 498 ESTs of Melampsoraspp.was assembled into 1 998 contigs,and 604SSR loci were detected on these contigs,with 736.6 bp containing one SSR on average.Among these SSRs,trinucleotide repeats were the most abundant repeats（44.70%）.As for the composition of microsatellites, AC and AT repeats were the richest motif in dinucleotide repeats,and AGT and AAG repeats were the most frequent motifs in trinucleotide repeats,whereas（AAAN） n and（AAAAN） n repeats were dominant in tetranucleotide and pentanucleotide repeats,respectively.All the dominant repeat motifs of different types of SSRs were rich in A and T.In EST sequences of Melampsoraspp.genome,microsatellites longer than 20 bp accounted for about 15.07%.It was noticeable that microsatellites were highly rich in the expressed sequences of Melampsoraspp.genome,which implied that SSRs played a significant role in triggering the gene mutation in Melampsoraspp.genome.A total of 455 SSR primers were designed according to the detected microsatellites using Primer 5.0 and Oligo 6. 0,and 30 primer pairs were randomly selected for amplification test.Among these primer pairs,27 primer pairs succeed in amplification,with a successful rate of 90%.Eight primer pairs generated polymorphic fingerprints in Melampsoraspp.collected from different poplar genotypes,accounting for 26.7% of the total primer pairs.The EST-SSR markers developed fromMelampsoraspp.EST sequences provided important marker resources for studying Melampsoraspp.from the aspects of pathogen identification and survey of genetic variation.
基金Supported by National Natural Science Foundation of China(31160175,31440034)Project for Protection and Utilization of Crop Germplasm Resources,Ministry of Agriculture(NB2012-2130135)+2 种基金Program of Technological Innovation Talents of Yunnan Province(2011CI068)Project for Construction of National Tea Modern Industrial Technology System of China(NYCYTX-23)Special Fund of Yunnan Academy of Agricultural Sciences(YAAS2012ZY002)
文摘In this study, 27 pairs of EST-SSR primers were employed to analyze the genetic diversity and genetic relationship of 100 wild tea plant germplasm re- sources and 22 cultivars, according to the results, a total of 88 polymorphic bands were amplified with 27 pairs of primers; the variation of effective alleles accounted for 69.01% ; a total of 183 genotypes were detected, with a variation range of 4 -11 ; averagely 6.78 genotypes were amplified with each primer pair; Shannon index （I） of 27 primer pairs ranged from 0.32 to I. 35, with an average of 0.88 ; the observed heterozygosity （0.52） was basically consistent with the expected het- erozygosity （0.52） ; the average polymorphism heterozygosity was 0.48, which was very close to 0.50 ; the average Nei＇s index was 0.51, which was higher than 0. 50 ; the average polymorphism information content （PIC） was 0.52, which was higher than 0.50, indicating high genetic diversity among wild tea germplasm resources in Yuunan Province. According to the clustering results, based on geographical origins and genetic backgrounds, 122 materials were clustered into 14 categories. Dendrogram based on Nei＇s genetic distance revealed complex genetic relationships among wild tea germplasm resources in Yunnan Province. This study provided certain reference for subsequent preservation, development and research of wild tea germplasm resources in China.
文摘The present study was conducted to develop EST-SSR markers using the cDNA library from rice plant. Total RNA extracted from the leaves of brown plant hopper resistance gene originated from a rice cultivar “Cheongcheong” and sensitive rice cultivar “Nakdong” were used to synthesize a cDNA library. As a result of analyzing the cDNA library, the 17 EST-SSR primer sets were developed. This study enables to provide effective marker assisted selection (MAS) methods on the selection of white-backed planthopper resistance gene originated from a rice plant more simply, quickly and precisely. Furthermore, using this marker’s advantage of deriving from cDNA, it is possible to identify the white-backed planthopper resistance gene. In addition, this study introduces a technique for construction of a cDNA library safely without using radioactivity.
文摘This commentary critically appraises the study by Li et al which pioneered the exploration of the triglyceride-glucose(TyG)index as a prognostic marker in hepatitis B virus-related advanced hepatocellular carcinoma patients undergoing combined camrelizumab and lenvatinib therapy.While we acknowledge the study’s clinical relevance in proposing an easily accessible metabolic biomarker,we delve into the mechanistic plausibility linking insulin resistance to immunotherapy response and angiogenic inhibition.We further critically examine the methodological limitations,including the retrospective design,the populationspecific TyG cut-off value,and unaddressed metabolic confounders.We highlight the imperative for future research to validate its utility across diverse etiologies and treatment settings,and to unravel the underlying immunometabolic pathways.
基金supported by the National Natural Science Foundation of China (31470400)Shaanxi Provincial Key Laboratory Project of Department of Education (grant no. 17JS135)+2 种基金the Shaanxi Provincial Education Department Serves Local Special Projects (grant no. 2018JC032)the programme for the Key Research and Development Plan in Shaanxi province (grant no. 2018ZDXM-SF-014)Public health specialty in the Department of Traditional Chinese Medicine (grants no. 2011-76, 201207002, 201213, 2013-135, 201407002, 2014-76, 2015-78, 2016-44, 2017-66)
文摘Notopterygium incisum C. C. Ting ex H. T. Chang(Apiaceae) is an endangered perennial herb in China. The lack of transcriptomic and genomic resources for N. incisum greatly hinders studies of its population genetics and conservation. In this study, we employed RNA-seq technology to characterize transcriptomes for the flowers, leaves, and stems of this endangered herb. A total of 56 million clean reads were assembled into 120,716 unigenes with an N50 length of 850 bp. Among these unigenes, 70,245(58.19%) were successfully annotated and 65,965(54.64%) were identified as coding sequences based on their similarities with sequences in public databases. We identified 21 unigenes that had significant relationships with cold tolerance in N. incisum according to gene ontology(GO) annotation analysis. In addition, 13,149 simple sequence repeats(SSRs) and 85,681 single nucleotide polymorphisms were detected as potential molecular genetic markers. Ninety-six primer pairs of SSRs were randomly selected to validate their amplification efficiency and polymorphism. Nineteen SSR loci exhibited polymorphism in three natural populations of N. incisum. These results provide valuable resources to facilitate future functional genomics and conservation genetics studies of N. incisum.
文摘果肉颜色是猕猴桃重要的品质性状,不同果肉颜色的猕猴桃在色素成分、糖和维生素C含量等方面存在很大差异。为了探究猕猴桃果肉颜色形成的分子机制,本研究以红肉猕猴桃‘红实2号’和黄肉猕猴桃‘金实1号’为研究对象,采用RNA-seq技术,通过分析果实不同时期不同部位的转录组表达差异。结果表明,编码花青素合成的Achn385311(3GGT)基因可能是控制‘红实2号’内果皮呈红色的主效基因,‘金实1号’后期果肉呈深黄色可能与Achn158981(GT7)、Achn150731、Achn068721(PAO)、Achn282201(PAO)和Achn176251(UGT71A16)等基因的表达调控有关。针对差异表达的230个猕猴桃果肉颜色相关基因,利用Primer Premier 5.0设计出727对EST-SSR引物,以7份不同果肉颜色的猕猴桃基因组DNA为模板,随机选择112对引物进行有效性验证,筛选出了具有清晰的扩增产物的引物78对,有效扩增率为69.64%,58对具有多态性,多态性频率为74.36%。本研究结果为猕猴桃果实呈色机理研究及分子标记辅助育种提供了理论基础。
文摘In recent years, Jatropha curcas L. has gained popularity as a potential biodiesel plant. The varying oil content, reported between accessions belonging to different agroclimatic zones, has necessitated the assessment of the existing genetic variability to generate reliable molecular markers for selection of high oil yielding variety. EST derived SSR markers are more useful than genomic markers as they represent the transcriptome, thus, directly linked to functional genes. The present report describes the in silico mining of the microsatellites (SSRs) using J. curcas ESTs from various tissues viz. embryo, root, leaf and seed available in the public domain of NCBI. A total of 13,513 ESTs were downloaded. From these ESTs, 7552 unigenes were obtained and 395 SSRs were generated from 377 SSR-ESTs. These EST-SSRs can be used as potential microsatellite markers for diversity analysis, MAS etc. Since the Jatropha genes carrying SSRs have been identified in this study, thus, EST-SSRs directly linked to genes will be useful for developing trait linked markers.
基金support provided by the National Department Public Benefit Research Foundation(No.201004009)the National High Technology Research and Development Program of China(863Program,No.2009AA10Z107)
文摘We analyzed the genetic differences of 16 poplar clones between genomic-SSR and EST-SSR markers. The statistical results show that the average number of alleles detected by genomic-SSR was 4.1, Shannon＇s index 1.0646, observed heterozygos- ity 0.4427 and expected heterozygosity 0.5523, while for the EST-SSR, the average number of alleles was 2.8, Shannon＇s index 0.6985, observed heterozygosity 0.2330 and expected heterozygosity 0.4684. Cluster analysis indicated that the EST-SSR capacity of genotypic identification was more precise than that of genomic-SSR. These resuks reveal that EST-SSR and genomic-SSR have statistically significant genetic differences in polymorphism detection and genotypic identification. These differences could provide a theoretical basis for the rational use of SSR markers in species diversity and other related research.
