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【目的】开发一组能够区分中国主栽葡萄品种的竞争性等位基因特异性PCR（KASP）分子标记,为中国葡萄品种保护、品种登记和市场维权等提供技术支撑。【方法】基于前人筛选的60个葡萄高多态性SNP位点,转化为KASP标记。分别使用23份代表性品种和76份主要栽培品种对转化成功的KASP标记进行初筛、复筛、验证,获得一组高质量的KASP标记,并用于构建76份葡萄品种的DNA指纹图谱。【结果】60个SNP位点有3个不具备基因组特异性,6个无法设计KASP-PCR引物,最终51个SNP成功转化为KASP标记,转化率达到89.47%。利用LGC-SNPline平台对23份代表性品种进行基因分型,51个KASP标记全部成功分型。基于次要等位基因频率（MAF）大于0.25、多态性信息含量（PIC）大于0.35、缺失率小于0.2、杂合率小于0.6等参数,初步筛选出27个优质KASP标记。使用构建指纹图谱的76份品种复筛出22个高质量KASP标记。22个KASP标记的缺失率均小于0.12,PIC均大于0.3,杂合率在0.4—0.6的标记占77.27%,MAF大于0.3的标记占95.45%。此外,利用这22个标记对同一品种不同树体提取的23份代表性品种的DNA扩增检测,前后两次检测分型结果稳定一致,表明这22个标记具有较好的重复性和稳定性。进一步将基于22个标记获得的76份葡萄品种分型结果转化为二元编码数据,得到76份葡萄品种的指纹图谱。经邻接聚类分析和群体结果分析,可将76份葡萄品种分为3类,并能正确区分二倍体和多倍体。仅用10个标记（VIT_15_18567587、VIT_6_4258638、VIT_8_3320936、VIT_12_22228357、VIT_11_19390306、VIT_16_17950801、VIT_18_11138668、VIT_12_739916、VIT_16_13454358、VIT_16_21202286）就能区分70份品种,其中有54份品种达到品种鉴定的标准（差异位点数≥2）。【结论】从60个葡萄SNP中成功转化51个KASP标记,并筛选出22个高质量KASP标记,构建了76份葡萄品种的SNP指纹图谱,首次验证了KASP技术在我国葡萄品种鉴定中的可行性。 相似文献
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Single nucleotide polymorphisms （SNPs） are the most abundant sequence variations found in plant genomes and are widely used as molecular genetic markers in cultivar identification and genetic diversity studies. The objective of this study was to identify SNP markers useful for discrimination of citrus cultivars, since large numbers of expressed sequence tags （ESTs） of sweet orange are available from the National Center for Biotechnology Information （NCBI）. We now have the opportunity to discover SNP markers suitable for determining the haplotypes with which to distinguish very closely related cultivars and to assess genetic diversity within or between related species of citrus. SNPs and small insertions/deletions （Indels） from ESTs of sweet orange and satsuma were identified by the in silico SNP discovery strategy. 55 296 EST sequences of sweet orange and 2 575 of satsuma retrieved from the NCBI repository were mined for potential SNPs. Cleaved amplified polymorphic sequences （CAPS） and sequencing approaches were used to validate putative SNPs in a sample of 30 citrus accessions. A total of 3 348 putative SNPs were identified based on the abundance of sequences and haplotype cosegregation. Of these 3 348 SNPs, the transitions, transversions and Indels ratios were 47.9, 36.1 and 16.0%, respectively. The SNPs occurred on average at a frequency of 1 per 164 bp in the coding region of citrus. 14 SNPs were randomly selected and genotyped according to 30 citrus accessions including 23 accessions of sweet orange; 11 SNPs displayed polymorphism with an average polymorphism information content （PIC） of 0.20 among 30 citrus accessions. The genetic diversity present in sweet orange was low, so the 14 SNP markers failed to discriminate different cultivars of sweet orange, but they did succeed in distinguishing accessions of inter-species of citrus. In this study, SNPs were mined from EST sequences of sweet orange and satsuma, which displayed potential capability as molecular markers to discriminate inter-species accessions of citrus. It is anticipated that these putative SNPs could be applied in citrus genetics research and breeding. 相似文献
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为了高效快速地区分浙茄类型茄子品种,并为鉴定浙茄品种的特异性和真实性提供依据,利用46份茄子材料,对48个单核苷酸多态性(SNP)标记进行核心SNP标记筛选,并利用筛选到的核心SNP标记,构建了6个浙茄类型茄子品种的SNP指纹图谱。结果表明:48个SNP标记均可转化为KASP标记,对46份茄子材料进行SNP标记的KASP分型,根据分型结果和多态性信息含量,34个SNP标记可作为核心标记,利用这些核心标记构建了6个浙茄类型茄子品种的指纹图谱;通过指纹图谱可以快速高效区分6个浙茄类型茄子品种,为茄子品种鉴定和知识产权的保护提供了理论依据。 相似文献
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利用基于RAPD标记的MCID法快速鉴定72个葡萄品种 总被引：3，自引：1，他引：2
【目的】葡萄种质材料和品种的鉴定是葡萄种质资源研究和新品种保护的基础。本研究应用一种新的基于DNA分子标记的人工绘制品种鉴别示意图方法（manual cultivar identification diagram, MCID）对来源于不同国家和地区的72个葡萄品种进行鉴定。【方法】通过增加引物长度以及严格筛选PCR退火温度优化的RAPD技术体系,从35个长度为11个碱基的引物中筛选出6个条带清晰且多态性高的引物,进一步对72个葡萄品种进行PCR扩增获得DNA指纹,然后利用MCID法成功将将72个葡萄品种区分开来。【结果】利用这6条引物扩增的多态性谱带构建了72个葡萄品种的MCID。该葡萄MCID很好地提供对其中某些品种进行鉴定所需要的引物以及需要参考的特征性谱带。【结论】到目前为止,MCID方法是实现利用DNA分子标记鉴定果树品种能力最好的有效途径。该研究所获得的MCID具有高操作性和实用参考性,对于葡萄种质资源研究、品种保护和品种纯度早期鉴定具有重要帮助。 相似文献
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Identification and validation of novel loci associated with wheat quality through a genome-wide association study 
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下载免费PDF全文 PU Zhi-en YE Xue-ling LI Yang SHI Bing-xin GUO Zhu DAI Shou-fen MA Jian LIU Ze-hou JIANG Yun-feng LI Wei JIANG Qian-tao CHEN Guo-yue WEI Yu-ming ZHENG You-liang 《农业科学学报》2022,21(11):3131-3147
Understanding the genetic basis of quality-related traits contributes to the improvement of grain protein concentration (GPC), grain starch concentration (GSC), and wet gluten concentration (WGC) in wheat. In this study, a genome-wide association study (GWAS) based on a mixed linear model (MLM) was performed on 236 wheat accessions, including 160 cultivars and 76 landraces, using a 55K single nucleotide polymorphism (SNP) array in multiple environments. A total of 12 stable QTL/SNPs that control different quality traits in this populations in at least two environments under stripe rust stress were identified. Among these 12, three, seven and two QTLs associated with GPC, GSC and WGC were characterized, respectively, and they were located on chromosomes (chr) 1B, 1D, 2A, 2B, 2D, 3B, 3D, 5D, and 7D with the phenotypic variation explained (PVE) ranging from 4.2 to 10.7%. Compared with the previously reported QTLs/genes, five QTLs (QGsc.sicau-1BL, QGsc.sicau-1DS, QGsc.sicau-2DL.1, QGsc.sicau-2DL.2, and QWgc.sicau-5DL) were potentially novel. KASP markers for the SNPs AX-108770574 and AX-108791420 on chr5D associated with wet gluten concentration were successfully developed. The phenotypes of the cultivars containing the A-allele in AX-108770574 and the T-allele in AX-108791420 were extremely significantly (P<0.01) higher than those of the landraces containing the G- or C-allele with respect to the wet gluten concentration in each of the environments. The KASP markers developed and validated in this study could be utilized in molecular breeding aimed at improving the quality of wheat. 相似文献
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Bing-ru CHEN Chun-yu WANG Ping WANG Zhen-xing ZHU Ning XU Gui-shan SHI Miao YU Nai WANG Ji-hong LI Jia-ming HOU Shu-jie LI Yu-fei ZHOU Shi-jie GAO Xiao-chun LU Rui-dong HUANG 《农业科学学报》2019,18(11):2446-2456
Starch is the most important component in endosperm of sorghum grain. Usually, two types of starch are present: amylose (AM) and amylopectin (AP). The levels of AM and AP contents play a significant role in the appearance, structure, and quality of sorghum grains and in marketing applications. In the present study, a panel of 634 sorghum (Sorghum bicolor (L.) Moench) accessions were evaluated for starch, AM, and AP contents of grain, which included a mini core collection of 242 accessions from the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT) in India, and 252 landraces and 140 cultivars from China. The average starch content was 67.64% and the average AM and AP contents were 20.19 and 79.81%, respectively. We developed a total of 260 000 high-confidence single nucleotide polymorphism (SNP) markers in the panel of 634 accessions of S. bicolor using specific locus amplified fragment sequencing (SLAF-seq). We performed genome-wide association studies (GWAS) of starch, AM, and AM/AP of grain and SNP markers based on a mixed linear model (MLM). In total, 70 significant association signals were detected for starch, AM, and AM/AP ratio of grain with P<4.452×10–7, of which 10 SNPs were identified with significant starch, 51 SNPs were associated with AM, and nine SNPs were associated with the AM/AP ratio. The Gene Ontology (GO) analysis identified 12 candidate genes at five QTLs associated with starch metabolism within the 200-kb intervals, located on chromosomes 1, 5, 6, and 9. Of these genes, Sobic.006G036500.1 encodes peptidyl-prolyl cis-trans-isomerase CYP38 responsible for hexose monophosphate shunt (HMS) and Sobic.009G071800 encodes 6-phospho-fructokinase (PFK), which is involved in the embden-meyerhof pathway (EMP). Kompetitive allele specific PCR (KASP) markers were developed to validate the GWAS results. The C allele is correlated with a high starch content, while the T allele is linked with a low level of starch content, and provides reliable haplotypes for MAS in sorghum quality improvement. 相似文献
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24份葡萄种质亲缘关系的ISSR分析 总被引：2，自引：0，他引：2
利用ISSR标记对24份葡萄材料进行了基因组多态性分析,从50条引物中筛选出6条扩增稳定且多态性丰富的引物用于葡萄的ISSR扩增。共扩增出59条条带,其中多态性条带53条,多态性百分率为90%。根据ISSR扩增结果,利用NTSYSpc2.10e软件进行Jaccard相似性系数分析,24份葡萄材料的遗传相似系数为0.42～0.88,平均遗传相似系数为0.6495。在遗传相似系数为0.55处,24份葡萄材料明显分为2大类群。第1类包含10个欧美杂种、7个欧亚种、1个华欧杂种,第2类包含3个美洲杂交种、1个河岸葡萄、1个冬葡萄、1个东亚葡萄。由此可见,美洲杂交种与欧美杂种、欧亚种葡萄的亲缘关系较远,欧美杂种与欧亚种葡萄之间亲缘关系较近。此外,引物BC820与引物BC847分别在我国自主选育砧木品种华佳8号与抗砧3号中扩增出一条特异条带,可为利用ISSR标记鉴定葡萄品种或品系提供参考依据。 相似文献
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BoCAL基因是控制花椰菜花球发生的关键基因之一。为了提高对花椰菜BoCAL基因型鉴定效率,根据该基因第5个外显子中第16个碱基（Ex5+16）G-T的功能性单核苷酸多态性（SNP）突变,开发了竞争性等位基因特异性PCR（KASP）标记(BoCAL-KASP1)。利用该标记对甘蓝类7个变种的蔬菜作物与野生种共计86份材料进行了BoCAL等位基因的分型,发现共有36份材料的基因型是T:T,包括28份花椰菜、6份青花菜与2份引进的地方种材料;43份材料的基因型是G:G,包括2份花椰菜、9份青花菜、6份结球甘蓝、5份芥蓝、3份苤蓝、7份羽衣甘蓝、2份抱子甘蓝、4份引进的地方种与5份甘蓝野生种材料;7份材料显示杂合基因型T:G,包括2份青花菜、2份羽衣甘蓝和3份引进的地方种材料。这86份材料的BoCAL基因型KASP鉴定结果和酶切扩增多态性标记(BoCAL-CAPS1)的鉴定结果完全一致。以上结果说明BoCAL-KASP1标记可以高效准确地鉴定BoCAL基因目标位点的基因型,从而为花椰菜花球发育研究和分子育种提供可靠的功能性标记。 相似文献
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葡萄黑痘病是危害葡萄生产的重要真菌病害之一,进行葡萄黑痘病抗性鉴定筛选抗病种质具有重要的意义。采用田间自然鉴定的方法,分析48份葡萄材料(北美种群内种间杂种、中国野葡萄、欧美杂种及欧亚种)对黑痘病的抗性差异。结果表明:不同葡萄材料对黑痘病抗性有着很大的差异。北美种群2个砧木品种、中国野葡萄华东葡萄湖南-1和山葡萄通化3号、欧美杂种均较欧亚种抗病,5BB、SO4、湖南-1、通化3号、京亚、8611、夏黑、翠峰和巨峰等表现为高度抗病;欧亚种对黑痘病的抗性表现为抗病和感病2种类型,其中紫地球、红提、玫瑰香等表现感病。研究结果为葡萄生产栽培和抗黑痘病育种提供理论依据。 相似文献
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葡萄属植物种质资源遗传多样性的RAPD分析 总被引：5，自引：1，他引：4
以起源于中国的野葡萄12个种23个株系、欧美杂种6个品种、河岸葡萄3份、15个欧洲葡萄品种共47份材料为试材,采用RAPD技术对葡萄属植物种质资源遗传多样性进行研究。从520个随机引物中获得16个多态性好的引物用于RAPD分析,共获得DNA条带258个,其中多态性条带186个,DNA条带大小介于100~3 500 bp之间。应用STATISTICA获得了47份葡萄材料树谱图,供试材料可分为6类。欧洲葡萄、欧美葡萄杂种、河岸葡萄与中国野葡萄亲缘关系较远。在中国野葡萄中,山葡萄与其他种亲缘关系较远,刺葡萄次之。 相似文献
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Fingerprinting 146 Chinese chestnut(Castanea mollissima Blume) accessions and selecting a core collection using SSR markers 下载免费PDF全文
下载免费PDF全文 Xing-hua NIE Ze-hua WANG Ning-wei LIU Li SONG Bo-qian YAN Yu XING Qing ZHANG Ke-feng FANG Yong-lian ZHAO Xin CHEN Guang-peng WANG Ling QIN Qing-qin CAO 《农业科学学报》2021,20(5):1277-1286
Chinese chestnut is an important nut tree around the world. Although the types of Chinese chestnut resources are abundant, resource utilization and protection of chestnut accessions are still very limited. Here, we fingerprinted and determined the genetic relationships and core collections of Chinese chestnuts using 18 fluorescently labeled SSR markers generated from 146 chestnut accessions. Our analyses showed that these markers from the tested accessions are highly polymorphic, with an average allele number(N_a) and polymorphic information content(PIC) of 8.100 and 0.622 per locus, respectively. Using these strongly distinguishing markers, we successfully constructed unique fingerprints for 146 chestnut accessions and selected seven of the SSR markers as core markers to rapidly distinguish different accessions. Our exploration of the genetic relationships among the five cultivar groups indicated that Chinese chestnut accessions are divided into three regional type groups: group I(North China(NC) and Northwest China(NWC) cultivar groups), group II(middle and lower reaches of the Yangtze River(MLY) cultivar group) and group III(Southeast China(SEC) and Southwest China(SWC) cultivar groups). Finally, we selected 45 core collection members which represent the most genetic diversity of Chinese chestnut accessions. This study provides valuable information for identifying chestnut accessions and understanding the phylogenetic relationships among cultivar groups, which can serve as the basis for efficient breeding in the future. 相似文献
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利用SNP标记构建茶树品种资源分子身份证 总被引：3，自引：0，他引：3
【目的】建立茶树品种的SNP分子标记数据库,结合茶树品种基本信息,将SNP位点组成的DNA指纹图谱构建28位数字组成的茶树品种资源分子身份证,便于茶树品种资源的保护与精准管理,避免“同名异物、同物异名”的现象。【方法】通过挖掘茶树的表达序列标签,获得大量的高质量表达序列标签,将其进行装配后,开发候选位点,将候选位点与茶树全基因组进行BLAST,得到其在全基因组染色体上的位置与具体关联基因。以铁观音、福鼎大白茶、龙井43、云抗10号等103份国内外不同类型的茶树品种资源为供试材料,提取基因组DNA,利用预扩增技术和微流体芯片法对供试茶树品种资源进行SNP基因分型,获得SNP位点数据及候选SNP位点的信息指数、观测杂合度、期望杂合度等信息,将多态性从高到低进行排序,进行SNP位点组合筛选,得到最优SNP位点组合后,结合茶树品种基本信息构建茶树品种资源分子身份证。【结果】从茶树的表达序列标签数据库中挖掘出1 786个候选SNP位点。根据序列保守性,筛选出96个SNP标记位点,与最新茶树基因组比对发现候选位点较均匀地分布于茶树全基因组的15条染色体上;对茶树品种资源的候选SNP位点的多态性信息进行分析,剔除10个不具多态性的位点,剩余86个位点的信息指数平均值为0.517,观测杂合度平均值为0.370,期望杂合度平均值为0.346,固定指数平均值为-0.036,次等位基因频率平均值为0.269。从86个SNP位点中筛选出24个多态性高的SNP位点,组成DNA指纹图谱,可区分出全部参试茶树品种资源。对24个SNP位点组成的DNA指纹图谱并结合茶树品种资源基本信息进行数字编码,最终形成由28位数字组成的茶树品种资源分子身份证。【结论】依据SNP标记的多态性信息,筛选SNP位点,精准区分全部供试茶树品种,并将24个SNP位点所构建的茶树品种资源DNA指纹图谱及品种资源的基本属性信息编码成特定的数字串,使每份茶树品种资源具有唯一的分子身份证,并生成相应的条形码和二维码,可快速被扫码设备识别。 相似文献
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【目的】近年来,油茶（Camellia oleifera）产业发展迅速,已成为中国四大油料之一。油茶良种不断涌现,但品质参差不齐,“同名异物、同物异名”等现象时有发生。建立油茶品种资源的单核苷酸多态性（single-nucleotide polymorphism,SNP）分子标记数据库,筛选重要SNP位点,开发油茶品种资源DNA指纹图谱,构建油茶品种资源的分子身份证,为品种鉴别、品种追溯等提供分子水平鉴别技术支撑。【方法】以221份普通油茶品种资源为材料,提取未成熟种子RNA,进行转录组测序。以二倍体南荣油茶基因组为参考,识别供试油茶品种资源的SNP位点并基因分型,利用SNP数据分析油茶群体及亚群的遗传多样性,分析SNP位点的观测杂合度、期望杂合度、多态信息含量（PIC）等信息,筛选核心SNP位点并采用Sanger测序验证,得到最优SNP位点组合后,结合品种资源基本信息构建油茶品种资源分子身份证。【结果】从油茶转录组中共检测到1 849 953个高质量SNP位点。群体遗传多样性分析发现,油茶群体观测杂合度为0.2966,期望杂合度为0.2462,固定指数为-0.2048,PIC为0.2... 相似文献
15.
Xiao-dan XU Jing FENG Jie-ru FAN Zhi-yong LIU Qiang LI Yi-lin ZHOU Zhan-hong MA 《农业科学学报》2018,17(1):37-45
Powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), is one of the most damaging diseases to wheat in the world. The cultivation of resistant varieties of wheat is essential for controlling the powdery mildew epidemic. Wheat landraces are important resources of resistance to many diseases. Mapping powdery mildew resistance genes from wheat landraces will promote the development of new varieties with disease resistance. The Chinese wheat landrace Baiyouyantiao possesses characteristic of disease resistance to powdery mildew. To identify the resistance gene in this landrace, Baiyouyantiao was crossed with the susceptible cultivar Jingshuang 16 and seedlings of parents and F1, BC1, F2, and F2:3 were tested with Bgt isolate E09. The genetic results showed that the resistance of Baiyouyantiao to E09 was controlled by a single recessive gene, tentatively designated PmBYYT. An Illumina wheat 90K single-nucleotide polymorphism (SNP) array was applied to screen polymorphisms between F2-resistant and F2-susceptible DNA bulks for identifying the chromosomal location of PmBYYT. A high percentage of polymorphic SNPs between the resistant and susceptible DNA bulks was found on chromosome 7B, indicating that PmBYYT may be located on this chromosome. A genetic linkage map of PmBYYT consisting of two simple sequence repeat markers and eight SNP markers was developed. The two flanking markers were SNP markers W7BL-8 and W7BL-15, with genetic distances of 3 and 2.9 cM, respectively. The results of this study demonstrated the rapid characterization of a wheat disease resistance gene and SNP marker development using the 90K SNP assay. The flanking markers of gene PmBYYT will benefit marker-assisted selection (MAS) and map-based cloning in breeding wheat cultivars with powdery mildew resistance. 相似文献
16.
【目的】 开发一系列高通量、低成本的桃重要性状竞争性等位基因特异PCR（Kompetitive Allele Specific PCR,KASP）标记,包括果皮有毛/无毛、果形扁平/圆形、果肉硬质/非硬质、DBF（Dominant Blood Flesh）红肉/非红肉、抗/感蚜等5对性状,加速桃优良品种培育,缩短桃育种年限。【方法】 本研究在控制这些性状的候选基因及位点附近300 kb内,利用已有桃种质资源的基因组序列比对,开发KASP标记,对已知表型的桃种质材料进行基因分型验证,最终获得与目标性状紧密连锁的KASP标记。【结果】 利用开发的5个性状的KASP分子标记对桃杂交分离群体和自然群体进行基因型检测,结果表明,标记鉴定结果与已知表型完全一致,准确率为100%。其中,杂交群体中果皮有毛/无毛性状分离比例为30﹕30,果形扁平/圆形分离比例为31﹕29,果实硬质/非硬质分离比例为27﹕26,抗蚜/感蚜分离比例为49﹕46,均符合孟德尔遗传1﹕1的分离定律。【结论】 开发的KASP标记可高效检测桃果实外观、抗性、肉质等重要性状相关基因的等位变异,在基因型鉴定、亲本选配和杂种后代的分子标记辅助选择中有很好的应用前景。 相似文献
17.
【目的】周8425B是中国小麦重要骨干亲本之一,小偃81是李振声院士选育的高产、优质、多穗型品种。千粒重是影响小麦产量的重要因素,发掘周8425B和小偃81的千粒重及相关性状的QTL,并分析不同生态区小麦品种所含QTL的单倍型与千粒重的关系,发掘优异单倍型,为不同生态区提高小麦产量及分子辅助育种提供基因型参考。【方法】以周8425B×小偃81衍生的重组自交系群体（F8）为研究对象,分别于2015和2016年在陕西杨凌（早晚播）进行田间种植,收获后对籽粒相关性状进行测量。利用90K芯片标记构建的高密度遗传图谱对3个环境下的千粒重、籽粒长、籽粒宽和籽粒厚进行QTL定位。同时,针对稳定的主效QTL开发相应的KASP分子标记,并以479份国内外小麦种质组成的自然群体为材料进行分子检测,结合自然群体的千粒重等性状进行关联分析;此外,从479份小麦中挑选出106份含有660K芯片基因型数据的当前黄淮麦区推广的小麦品种,以主效QTL置信区间的差异SNP为基础,进行目标QTL定位区间的单倍型分析,从而判断黄淮麦区中陕西、河南和山东种质材料中的优势类群。【结果】QTL定位结果显示,3个环境下共在8条染色体上检测到22个QTL,表型变异解释率（PVE）范围为4.77%—19.95%,12个位点为主效QTL（PVE>10%）,其中Qkgw.nwafu-6B可能为新QTL。4A、6A、6B、7D染色体上的QTL在多个环境中被检测到,其中,4A和7D染色体处QTL与已报道的相关位点位置相同或接近。6A染色体上的QTL区间包含已知千粒重基因TaGW2-6A,根据TaGW2-6A的分子功能标记检测结果,周8425B和小偃81同时含有TaGW2-6A,此外,基于单倍型分析结果,二者存在于不同的类群中,因此,该位点不同于TaGW2-6A,也可能为新的QTL。单倍型分析结果显示,Qkgw.nwafu-6A总共分为5种单倍型,在不同产区占比超过20%的为6A_h1,其在3个地点千粒重数据均高于其他单倍型;Qkgw.nwafu-6B总共分为8种单倍型,在不同产区占比超过20%的为6B_h6,在河南两点千粒重数据较高,推测含有这两类单倍型的材料为优势类群。此外,针对Qkgw.nwafu-6B开发出共分离的KASP标记,并在479份材料组成的自然群体显著性检测中证明该位点与千粒重表型显著相关。【结论】Qkgw.nwafu-6A和Qkgw.nwafu-6B可能为新的与千粒重相关的主效QTL位点,6A_h1和6B_h6为优势单倍型,开发了一个与Qkgw.nwafu-6B共分离的分子标记KASP_IWA349,可用于分子标记辅助育种。 相似文献
18.
绿豆抗豆象基因PCR标记的构建与应用 总被引：5，自引：1，他引：5
采用PCR分子标记技术，对16个绿豆品种（系）进行了遗传分析。在选用的56个随机引物中，发现抗豆象品种与感豆象品种间有一定差异。根据聚类分析结果将它们分成抗豆象野生种（TC1966）、抗豆象栽培种（V2709）、抗豆象杂交后代（VC3890A2/TC1966-23）和混合类型4个大组。以绿豆抗豆象和感豆象品种及抗豆象品种×感豆象品种组合的F2群体为试验材料，利用BSA法，对抗（感）豆象品种池和一个组合F2的抗（感）豆象池进行了鉴定，获得一个共显性标记。经F2分析，在抗豆象个体中扩增出约1.79 kb的特异片段或2个特异片段（1.79 kb/1.03 kb）；在感豆象个体中仅扩增出约1.03 kb的特异片段。初步认为此标记与TC1966的抗豆象基因位点紧密联锁，可用于绿豆抗豆象种质鉴定和遗传育种的分子标记辅助选择。 相似文献
19.
Li-ning WANG Wei GAO Qiong-ying WANG Ji-bin QU Jin-xia ZHANG Chen-yang HUANG 《农业科学学报》2019,18(3):580-589
Agaricus bisporus is one of the most widely cultivated mushrooms in the world. Commercial cultivars are usually phenotypically alike and easy to be copied by isolating tissue cultures. This brings great challenges to distinguish different cultivars and to protect new varieties. Thus, techniques for the accurate identification of cultivars are essentially required. In this study, we accurately identified 11 commercial cultivars of A. bisporus released in China by using microsatellite (SSR, simple sequence repeat) markers. SSR markers were developed by mining the genome sequence. A total of 3 134 SSRs were identified, of which 1 490 SSRs were distributed in gene models, and 1 644 in the intergenic regions. A total of 17 polymorphic primer pairs were developed and SSR fingerprints were constructed for all the commercial cultivars. These SSR markers generated a total of 73 alleles, with an average of 4.29 per locus. Specifically, the primer combination of AB_SSR_2341 and AB_SSR_2590 could distinguish all the 11 commercial cultivars. The similarity coefficients of the 11 commercial cultivars were between 0.56 and 0.95 indicating that some of them were close related. Our results provide an efficient technique for the identification of A. bisporus cultivars in China, which can also facilitate the marker-assisted breeding in the future. 相似文献
20.
【目的】 类胡萝卜素裂解双脱氧酶基因（Carotenoid Cleavage Dioxygenases 4,CCD4）控制桃果肉颜色（白/黄）,CCD4存在3种等位基因。本研究利用Indel、SSR荧光标记毛细管电泳及SNP鉴定等基因分型技术分析我国主要桃黄白肉品种（系）中CCD4等位基因的差异,为主要黄/白肉品种（系）的基因型鉴定、亲本选配和选择相应的分子标记对不同来源子代的果肉颜色进行鉴定奠定基础。【方法】利用已经报道的桃不同果肉颜色中CCD4等位基因3种突变类型,合成不同引物进行PCR扩增,LTR反转录转座子插入突变经1%的琼脂糖凝胶电泳检测,CT单元重复的PCR产物在ABI3730XL测序仪上进行SSR荧光标记毛细管电泳检测,SNP标记经Sanger测序后利用ContigExpress软件分析CCD4等位基因的碱基替换（A→T）。综合以上结果,统计每份材料中CCD4等位基因的突变类型与果肉颜色的一致性。【结果】通过对不同来源的122份桃品种（系）材料进行基因型分析,发现CCD4发生LTR反转录转座子插入突变材料的基因型共有31份,占总材料的25.4%,其中纯合插入突变材料的片段扩增长度为729 bp,共有8份,占总突变的25.8%;CCD4发生微卫星重复序列突变材料存在2 bp的插入,扩增片段长度为179 bp,该类型共有68份,占总材料的55.7%,其中纯合插入材料25份,占总突变的36.8%;CCD4发生A→T碱基替换突变的材料较少,仅有1份,占总材料的0.82%,实际应用中可以不考虑该种类型。CT和LTR插入的两种突变类型的黄肉品种（系）有7份,占总材料的5.7%。研究结果表明,LTR反转录转座子插入突变和微卫星序列重复突变是黄肉桃中CCD4等位基因的主要突变类型。其中CCD4发生一种纯合突变或两种杂合突变桃品种（系）为黄肉类型,分子标记鉴定结果与调查的122份桃品种（系）黄白肉表型性状完全一致,准确率为100%。【结论】采用分子标记明确了122个桃品种（系）黄/白肉性状的基因型,为不同亲本组合子代表型鉴定的标记类型选择提供了技术支撑,为建立桃种质材料黄/白性状的分子辅助育种体系和黄肉桃的选育奠定了基础。 相似文献