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The identification of self-incompatibility genotype (S-genotype) will be useful for selection of pollinizers and design of crossing in cultivar improvement of sand pear. This paper reported the identification of self-incompatibility genotypes of seven Chinese and two Japanese sand pear cultivars using PCR-RFLP analysis and S-RNase sequencing. The Sgenotypes of these cultivars were determined as follows: Huali 1 S1S3, Shounan S1S3, Xizilti S1S4, Qingxiang S3S7, Sanhua S2S7, Huangmi (Imamuranatsu) S1S6, Huali 2 S3S4, Baozhuli S7S33, Cangxixueli S5S15. S-RNase alleles (S1 to S9) in sand pear could be identified effectively by PCR-RFLP analysis.  相似文献   

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Porcine skeletal muscle genes play a major role in determining muscle growth and meat quality. Construction of a full-length cDNA library is an effective way to understand the expression of functional genes in muscle tissues. In addition, novel genes for further research could be identified in the library. In this study, we constructed a full-length cDNA library from porcine muscle tissue. The estimated average size of the cDNA inserts was 1 076 bp, and the cDNA fullness ratio was 86.2%. A total of 1 058 unique sequences with 342 contigs (32.3%) and 716 singleton (67.7%) expressed sequence tags (EST) were obtained by clustering and assembling. Meanwhile, 826 (78.1%) ESTs were categorized as known genes, and 232 (21.9%) ESTs were categorized as unknown genes. 65 novel porcine genes that exhibit no identity in the TIGR gene index of Sus scrofa and 124 full-length sequences with unknown functions were deposited in the dbEST division of GenBank (accession numbers: EU650784-EU650788, GE843306, GH228978-GH229100). The abundantly expressed genes in porcine muscle tissue were related to muscle fiber development, energy metabolism and protein synthesis. Gene ontology analysis showed that sequences expressed in porcine muscle tissue contained a high percentage of binding activity, catalytic activity, structural molecule activity and motor activity, which involved mainly in metabolic, cellular and developmental process, distributed mainly in intracellular region. The sequence data generated in this study would provide valuable information for identifying porcine genes expressed in muscle tissue and help to advance the study on the structure and function of genes in pigs.  相似文献   

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Porcine skeletal muscle genes play a major role in determining muscle growth and meat quality.Construction of a full-length cDNA library is an effective way to understand the expression of functional genes in muscle tissues.In addition,novel genes for further research could be identified in the library.In this study,we constructed a full-length cDNA library from porcine muscle tissue.The estimated average size of the cDNA inserts was 1 076 bp,and the cDNA fullness ratio was 86.2%.A total of 1 058 unique sequences with 342 contigs(32.3%)and 716 singleton(67.7%)expressed sequence tags(EST)were obtained by clustering and assembling.Meanwhile,826(78.1%)ESTs were categorized as known genes,and 232(21.9%)ESTs were categorized as unknown genes.65 novel porcine genes that exhibit no identity in the TIGR gene index of Sus scrofa and124 full-length sequences with unknown functions were deposited in the dbEST division of GenBank(accession numbers:EU650784-EU650788,GE843306,GH228978-GH229100).The abundantly expressed genes in porcine muscle tissue were related to muscle fiber development,energy metabolism and protein synthesis.Gene ontology analysis showed that sequences expressed in porcine muscle tissue contained a high percentage of binding activity,catalytic activity,structural molecule activity and motor activity,which involved mainly in metabolic,cellular and developmental process,distributed mainly in intracellular region.The sequence data generated in this study would provide valuable information for identifying porcine genes expressed in muscle tissue and help to advance the study on the structure and function of genes in pigs.  相似文献   

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TSARG7 is a novel member of the acyltransferase family since its sequence possesses the highly conserved phosphate acyltransferase (PlsC) domain existing in all acyltransferase-like proteins. The porcine TSARG7 had been identified by cloning in silico but had not been confirmed experimentally. The full-length mRNA of porcine TSARG7 gene was sequenced and two splice variants were discovered. The full-length cDNA of TSARG7 variant 1 was 2 513 bp and variant 2 was 2 634 bp. The putative porcine TSARG7 proteins, which were located in the cytoplasm, encoded 458 and 456 amino acids, respectively. Real-time PCR analysis showed that TSARG7 gene was expressed in various tissues, but at different levels. The expression levels of this gene were higher in the skeletal muscle, heart, and testis than that in other tissues, suggesting that the TSARG7 gene played a role in procine skeletal muscle, heart, and testis functions.  相似文献   

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The effect ofAphelenchoides besseyi on 27 cultivars of rice (23japonica and 4 indica) was assessed in the field for two seasons during 2010 and 2011. The vigorous pathogenic nematodes culturing on Botrytis cinerea were used for this experiment. Inoculation was carried out at the tilling stage; the growth parameters and nematode population were recorded at the end of growth of rice plants. The results showed that the cultivars differed in their response to infection. Most of cultivars were lack of the characteristic symptom of white tip, which was seen less frequently than the other two symptoms, namely small grains and erect panicles; moreover, the expression of symptoms was probably hereditary. The infection lowered the values of all the measured biological parameters, namely length of the stem and of the panicle, the number of filled grains per panicle, and 100-grain weight, in all the cultivars. The final nematode population indicated that the threshold of economic damage had also been exceeded in 10 cultivars, and none of them was immune. Three japonica cultivars proved most vulnerable whereas Tetep, an indica type, showed a level of resistance potentially useful in controlling A. besseyi.  相似文献   

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[Objective] The study aimed to identify Alternaria Nees. from some areas of China at molecular level by analyzing the rDNA ITS sequence.[Method] The DNA sequences coding for the 5.8S rDNA and the flanking internal transcribed spacers (ITS1 and ITS2) were amplified by PCR with universal primers ITS4 and ITS5 and subsequently sequenced for 34 Alternaria isolates from different areas of China.[Result] Sequences analysis showed that 5.8S rDNA was 159 bp and no variation in tested 34 isolates. There had variables sites in ITS. The isolates that had same sequences as A.tenuissima or A.alternata all put up eurytopicity to area and host. The variables sites of the isolates showed the diversity of Alternaria in the hosts of Oleaceae, Rosaceae and Solanaceae. At the same time that ITS could not clearly separated the isolates was indicated. The results indicated that the phylogenetic relationship were not closely related to the geographical origin and hosts of these isolates.[Conclusion] The sequence analysis of ITS region could provide theory basis for the identification of Alternaria Nees..  相似文献   

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[Objective] The aim was to study the molecular identification and cultivar fingerprints of Prunus persica (L.) Batsch germplasms.[Method] Sixty peach genotypes,representing China common local cultivars and European samples were screened by microsatellites (simple sequence repeats,SSRs) and Inter-Simple Sequence Repeat (ISSR) markers.[Result] 26 reproducible bands were amplified by Nine SSR primers,and 24 of which were polymorphic; 236 bands were amplified by 30 ISSR primers,and 113 of which were polymorphic.31 genotypes were discriminated with 1-3 distinct polymorphic bands generated from the primers ISSR and SSR.Seven cultivar-specific ISSR fragments and two SSR unique alleles obtained from this study were available to be converted into Sequence Characterized Amplified Region (SCAR) markers.The genetic similarity coefficient (GS) estimated from these molecular data averaged were 0.939 (ranged from 0.856 to 0.983) for ISSR and 0.646 (ranged from 0.240 to 1.000) for SSR,respectively.The combined grouping association indicated that most local Chinese peach cultivars and exotic accessions were clustered together.This could be related to the mode of introduction and maintenance of the peach cultivars involving limited foundation germplasm,exchange of cultivars between plantations,and periodic development of new recombinant cultivars following sexual reproduction.[Conclusion] The results obtained in this work would help to improve the conservation,molecular identification and management of peach germplasm in breeding.  相似文献   

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    通过逆转录PCR(RT-PCR及快速扩增cDNA末端(RACE)技术从中国白梨品种'雪花'梨中克隆到S16-RNase基因的全长cDNA序列(GenBank接受号为DQ991388)该cDNA克隆总长1101 bp,包含1个完整的开放阅读框,编码228个氨基酸S16-RNase表现出梨S-RNase基因的基本结构特征,即其具有5个保守区(C1,C2,C3,RC4及C5)和1个高变区在推导氨基酸水平上,S16-RNase与梨其他S-RNase基因的相似性为63%至74%,但与苹果S9-RNase的相似性高达95%通过多序列比对构建进化树,分析梨S16-RNase与蔷薇科植物其他S-RNase基因的遗传进化关系结果表明,S16-RNase与苹果亚科S-RNase基因形成一个亚科特异而非种特异的S-RNase基因类群;且不同S-RNase基因间存在属内种间遗传距离远于属间种问遗传距离现象  相似文献   

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11个中国杏品种S-RNase基因的检测与序列分析   总被引:2,自引:0,他引:2  
以11个未知基因型的中国杏品种为试材,根据李属植物S-RNase基因保守区设计2对引物组合检测各品种S-RNase基因,共获得22条等位扩增片段,电泳检测表明所有品种的扩增条带集中在300~1100bp的范围内,且表现出一定的长度多态性。序列分析进一步确定22个S-RNase基因为10个不同的等位基因,其中6个为首次发现,根据Gen-Bank中已登陆的杏S-RNase基因的顺序,分别命名为S19、S20、S23、S24、S25、S26,序列登陆号为:EF185300、EF185301、EU037262、EU037263、EU037264、EU037265。推导氨基酸序列的同源分析表明,杏的S-RNase与李属植物的S-RNase表现较高的同源性,为59.3%~100%;与苹果和梨的S-RNase同源性较低,为19.6%~31.6%。试验确定11个中国杏品种资源的自交不亲和基因型分别为:‘大果杏’S19/S20,‘张公园’S24/S25,‘二红’S9/S11,‘黄口外’S11/S26,‘植丸子’S11/S17,‘宇宙红’、‘大丰’S17/S23,‘超仁’、‘虹桥’S8/S11,‘冀光’、‘中华大杏梅’S8/S9;部分品种的田间杂交授粉座果与花粉管荧光显微镜观察证实了所鉴定基因型的可靠性。  相似文献   

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通过酵母双杂交的方法寻找苹果‘国光’花柱中与S-RNase互作的非S因子。以苹果‘国光’花柱为试材,构建了酵母cDNA文库,检测插入片段大小在300~2 000bp之间,符合库容要求。将S1-RNase成熟区cDNA序列S1-mat构建到pGBKT7载体上作为诱饵,筛选‘国光’花柱酵母cDNA文库。经文库筛选,获得一个大小为371bp的片段,与苹果全基因组序列比对后发现,该片段位于第9号染色体,其全长序列为552bp。NCBI BLAST比对及蛋白结构域分析显示其与拟南芥钙调素结合蛋白的同源性最高,且具有钙调素结合蛋白特有的磷酸二酯酶结构域。同时,酵母互作实验显示其与‘国光’花柱钙调素(CaM)有强烈互作,故认为此基因是苹果钙调素结合蛋白基因,命名为MdCaMBP。半定量RT-PCR结果显示其在‘国光’叶片及花的各组织中均有表达,与苹果花柱S1-、S2-、S9-RNase成熟多肽区均有互作且作用强烈。推测MdCaMBP可能作为一种S-RNase辅助因子参与了自交不亲和反应。  相似文献   

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利用已报道的李属S-RNase基因特异PCR的2对引物,分别对中国樱桃的5个品种进行了S-RNase基因的PCR扩增。用2对引物先优化PCR体系,然后比较扩增效果。结果表明:2对引物对中国樱桃品种的扩增效果不同。其中,引物EM-PC2consFD+EM-PC3consRD扩增效果最好;其次为PaConsⅠ-F+PaConsⅡ-R引物。因此,EM-PC2consFD+EM-PC3consRD及相应的PCR体系适于中国樱桃S-RNase基因的PCR扩增。本结果建立了中国樱桃S-RNase基因扩增的技术体系,为探讨中国樱桃自交亲和分子基础奠定了技术基础。  相似文献   

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    为鉴定中国梨S-RNase基因及其S基因型,根据日本梨S-RNase基因保守区设计引物,对13个中国梨品种进行基因组PCR特异扩增,并对PCR产物克隆、测序、序列分析.结果鉴定出13个S等位基因,其中11个S基因与GenBank中已知的S1, S7, S12, S15, S16, S18, S19, S22, S27, S29, S34-RNases相同,另外2个则为新的S-RNase基因,命名为S37-RNase和S38-RNase,GenBank接受号分别为DQ839238、DQ839239.13个S等位基因中,在推导氨基酸水平上,S18与S27相似性最低,为58%,S7与S27,S12与S19,S15与S37及S15与S38间相似性最高,为94%.经分析,S19为中国梨中出现频率最高的等位基因,对其DNA序列进行限制性酶切位点分析,建立了一种快速、经济鉴定S19的方法,该方法无需测序而仅使用特异的限制性内切酶AflⅡ对PCR产物进行酶切消化.根据分子鉴定结果,13个中国梨品种S基因型被确定为:'冰糖'(S16S19), '六棱' (S16S19), '锦香'(S34S37), '鹅酥' (S15S38), '蜜梨'(S19S29), '甜橙子'(S7S12), '大青皮'(S19S34), '秋白' (S19S34), '紫酥' (S19S34), '花长把'(S19S22), '灌阳雪梨'(S18S27), '早蜜'(S19S29),'青面'(S1S18).  相似文献   

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【目的】鉴定南疆扁桃品种自交不亲和S-RNase基因型,可为科学合理地配置授粉品种提供依据,有效提高南疆扁桃的产量。【方法】以南疆10个扁桃品种的叶片为试材,利用蔷薇科通用引物PaConsⅡ-F+PaConsⅡ-R、EM-PC2consFD+EM-PC3consRD对叶片基因组DNA进行S-RNase特异性扩增,将克隆测序结果在GenBank上进行同源性比对。【结果】显示10个扁桃品种均获得2条带,共20条带,包含16种不同的S基因核苷酸序列,其中4个为GenBank上登录的已知S-RNase基因序列,分别为S1(1 370 bp)、S12(2 479bp)、S14(740 bp)、S52(1 626 bp),12个为新的S-RNase基因序列,暂时分别命名为SA(780 bp)、SB(530 bp)、SC(1 261 bp)、SD(432 bp)、SE(1 266 bp)、SF(270 bp)、SG(1 263 bp)、SH(272 bp)、SI(690 bp)、SJ(1 523bp)、SK(755bp)、SL(1 486 bp)。【结论】鉴定出10个品种的S-RNase基因型。  相似文献   

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新疆杏品种自交不亲和S-RNase基因特异PCR扩增引物的筛选   总被引:1,自引:1,他引:0  
【目的】筛选鉴定新疆杏自交不亲和相关S-RNase基因的有效引物。【方法】以新疆30个杏品种为试材,选用已报道的蔷薇科李属通用的5对引物组合进行S-RNase基因特异PCR扩增,并依据扩增系数和扩增成功率两个指标对扩增效果进行评价。【结果】(1)5对引物组合对30个杏品种均扩出了条带,除了洛浦2号只扩出1条带之外,其余的29个品种都扩增出了两条带;(2)5对引物组合对新疆主栽杏的扩增效果差异较大,其中EM-PC2consFD+EM-PC3consRD的扩增成功率和扩增系数均为最高(86.67%,95.00%),扩增出两条带的品种有25个;其次,引物组合PaConsⅡ-F+PaConsⅡ-R的扩增成功率达66.67%,扩增系数达83.33%,扩出两条带的品种有20个。【结论】5对引物中EM-PC2consFD+EM-PC3consRD对新疆杏品种S-RNase基因的鉴定效果最好。  相似文献   

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【目的】建立可同时检测李属坏死环斑病毒(Prunus necrotic ringspot virus,PNRSV)、李矮缩病毒(Prune dwarf virus,PDV)、樱桃小果病毒2(Little cherry virus-2,LChV-2)的多重RT-PCR检测方法。【方法】以复合感染3种病毒的甜樱桃病株叶片为材料,采用CTAB法提取样本总RNA,选用随机六聚体引物对植物样本总RNA进行反转录,所得cDNA作为多重RT-PCR的扩增模板。根据GenBank中PNRSV、PDV、LChV-2基因组序列共设计6对特异引物,分别通过单一RT-PCR和多重RT-PCR筛选出可用于同时检测3种甜樱桃病毒的引物组合。对多重RT-PCR的退火温度及循环数进行优化,以筛选出各引物组合的最适扩增条件。分别以单一感染PNRSV、PDV、LChV-2、复合感染3种病毒、单一感染樱桃病毒A(Cherry virus A,CVA)及甜樱桃无毒苗为样本,对多重RT-PCR引物的特异性进行分析。选取复合感染3种病毒的甜樱桃总RNA的反转录产物为初始模板,按照梯度稀释法依次将模板稀释为2、22、23、24、25倍,在相同PCR反应体系及反应条件下分别对各引物组合的灵敏度进行分析。多重RT-PCR的扩增条带经凝胶回收试剂盒回收纯化后连接至pMD18-T vector,克隆测序,以验证多重RT-PCR检测的准确性。并应用该方法对山东泰安地区甜樱桃生产园中间隔栽培的中国樱桃进行检测。【结果】筛选到2个可以应用的引物组合,组合1“PNRSV-S1/A1、PDV-S2/A2、LChV2-S1/A1”可分别特异性地扩增733、467、337 bp的片段。组合2“PNRSV-S1/A1、PDV-S3/A3、LChV2-S1/A1”可分别扩增得到733、265、337 bp的片段。扩增产物大小与预期相符。多重RT-PCR反应条件优化结果显示,在退火温度52℃、35个循环条件下,2个引物组合的检测效果均较为理想。特异性分析结果显示,2个引物组合均能特异性检测其各自的靶病毒。灵敏度分析结果显示,2个引物组合在cDNA的23×稀释液中仍能特异性扩增,但扩增条带的强度稍有差异,其对植物总RNA的反转录产物的最低检测浓度为107.9 ng•μL-1。克隆测序及序列分析表明,2个引物组合对各自靶病毒的检测结果可靠。应用该方法对9个中国樱桃样本进行检测,结果显示,测试样品均至少感染了2种病毒,其中5个样品复合感染了3种病毒,2个样品同时感染PDV和LChV-2,2个样品同时感染PNRSV和LChV-2。【结论】应用建立的多重RT-PCR检测方法可稳定、准确、灵敏的同时检测单一或复合侵染的3种甜樱桃病毒。  相似文献   

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甜樱桃DFR基因内含子2和内含子3的多态性   总被引:1,自引:0,他引:1  
【目的】研究70个甜樱桃品种DFR基因多态性与果皮颜色的相关性。【方法】通过DNA序列分析,以不同果皮颜色的10个甜樱桃品种(Prunus avium L.)为材料,检测DFR基因的多态性。根据多态性出现的位点设计特异引物,通过PCR扩增检测70个甜樱桃品种DFR基因的多态性。【结果】获得甜樱桃DFR基因约1 kb的片段,测序结果用BLAST分析发现,其核苷酸序列与樱桃李(Prunus cerasifera)的核苷酸相似性为80 %,预测的氨基酸序列与已知的甜樱桃(P. avium)DFR氨基酸序列相似性达99 %。该片段由3个外显子和3个内含子组成,2个多态性位点分别在内含子2和内含子3上。在黄色、黄底红晕和红色果皮3个组的70个甜樱桃品种中发现3个单倍型,共5种单倍型组合。通过SAS 9.0软件分析发现,所检测到的DFR基因的等位基因频率在黄底红晕果皮品种组和红色品种果皮组中的分布无显著差异。【结论】本研究在甜樱桃DFR基因座上得到2个差异位点,分别在内含子2和内含子3内,其优势基因频率依次为:0.864和0.679;但未发现DFR基因内含子2和内含子3的多态性与果皮颜色之间存在直接关系。  相似文献   

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【目的】克隆李属甜樱桃自交不亲和性花粉S-决定子基因,为今后果树配子体自交不亲和性机理研究奠定理论基础。【方法】根据GenBank登录的16个S-locus F-box同源基因保守区设计兼并引物,利用RT-PCR、RACE等手段,从甜樱桃品种红灯花粉cDNA中克隆到两个编码376-氨基酸多肽的全长基因。【结果】GenBank Blast分析显示,克隆的两个基因中一个基因编码的蛋白产物与数据库甜樱桃自交不亲和性S3-单元型特异的PaSFB3(AB096857)编码的氨基酸序列完全一致。另一个基因编码一新的PaSFB同源序列,其推测的氨基酸序列N-端同SFB3一样具有明显的F-box基序,与PaSFB1~6的一致性为76%~82%。研究显示该基因在花粉组织中特异性表达,并表现出S9-单元型特异的连锁信号。【结论】新基因为甜樱桃自交不亲和性花粉S-决定子候选基因PaSFB家族中一新成员,命名为PaSFB9 (GenBank登录号:DQ422809),红灯自交不亲和基因型确定为S3S9。  相似文献   

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