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1.
高粱基因组遗传图谱构建的研究进展   总被引:8,自引:0,他引:8  
高粱 (Sorghum bicolor L. Moench) 基因组分子遗传连锁图谱的构建始于20世纪90年代。覆盖高粱全基因组的高密度分子遗传连锁图谱已构建完成,正在进行高粱遗传图谱与物理图谱的整合,把遗传连锁群与相应的染色体对应起来, 并已初步确定了染色体着丝粒、长短臂、核仁组织者区域(NOR)等染色体重要结构在遗传连锁图上的位置。随着研究的深入,高粱全基因组完全测序和高粱功能基因组学的研究即将全面启动.  相似文献   

2.
半滑舌鳎(Cynoglossus semilaevis)属于雌配子异形染色体性别决定机制(ZW/ZZ),其W染色体包含的大量重复序列阻碍了W染色体序列的精确组装.目前只有物理图谱可以有效地解决由重复序列所造成的组装难题.本研究在已有物理图谱和遗传图谱的基础上,利用3D(three-dimensional)-BAC(bacterial artificial chromosome)池构建和PCR筛选策略进行了性别连锁标记的物理定位.首先将所选44个384孔板的BAC克隆进行接种和预培养,再接种到4个96孔深孔板中.将每两个384孔微孔板的BAC克隆分别汇集获得相应的板池、行池和列池,在提取DNA后,再将相应板池的部分DNA混合得到超级池.最终构建的3D-BAC池,共包括22个超级池和440个基池,覆盖半滑舌鳎基因组4.2倍,完成一个阳性超级池的基池筛选只需24个反应(20个BAC基池和4个对照).然后选用159个半滑舌鳎性别连锁标记对这些克隆池进行两步PCR筛选.在对超级池的筛选中,有142个标记获得阳性扩增.对这些阳性超级池进行基池筛选后,最终有100个标记得到准确定位.定位到半滑舌鳎物理图谱的84个重叠群(contigs)和20个单个克隆(singletons)上,共计1 760个克隆,包含有21 927个共享条带,物理长度为42.4Mb,覆盖半滑舌鳎基因组约5.3%;确定一个定位关系(contig与标记之间的)平均使用了2.2个克隆.在这些标记中,有17个同时定位到了两个或两个以上的contigs上,有11个标记同时定位到contigs和singletons上.另外有19个contigs定位了2个或2个以上标记.这些性别连锁标记的物理定位将有助于半滑舌鳎性染色体基因组序列的精确组装.  相似文献   

3.
董娜  张新  王清连  李成奇  刘阳阳 《核农学报》2013,(10):1431-1440
短季棉育种是缓解我国当前粮棉争地矛盾、实现粮棉双丰收的有效途径。研究短季棉早熟及相关性状的遗传,寻找与其相关基因紧密连锁的分子标记对于短季棉育种具有重要意义。以百棉2号×TM-1组合构建的F2为作图群体,利用SSR标记构建遗传连锁图,采用复合区间作图法对短季棉早熟及相关性状在F2群体中进行QTL定位,构建了一张含152个SSR标记、40个连锁群、总长1010.3cM、覆盖棉花基因组的22.6%、标记间平均距离为5.91cM的短季棉分子标记遗传连锁图谱,得到的40个连锁群已定位到相应的21条染色体上,为短季棉分子遗传图谱进一步饱和奠定了良好的基础;检测出22个控制早熟及相关性状的QTL,其中检测到的主效QTL可用于短季棉标记辅助选择育种。  相似文献   

4.
阐述了近年来鲍分子遗传学方面的研究进展,总结了核型分析、等位酶、微卫星和小卫星、随机扩增多态性DNA、限制性片断长度多态性、线粒体DNA、表达序列标签研究和基因序列等技术在鲍种群遗传多样性、遗传分化、遗传结构及种质鉴定等方面的应用。并指出今后应加强鲍蛋白质组学、功能基因组学、遗传连锁图谱、数量性状基因座和标记辅助选择等方面研究。  相似文献   

5.
为了实现分子标记或基因辅助育种在牙鲆养殖中成功运用,并快速提高牙鲆产量,使用微卫星标记首次构建了国内第一张牙鲆遗传连锁图谱。采用基因组测序的方法,筛选出大量微卫星序列,从中随机挑选1 000条序列设计合成引物,利用2009年建立的第10号家系为作图群体,使用JoinMap4.0软件,构建了牙鲆(Paralichthys olivaceus)SSR标记遗传连锁图谱。雌雄图谱分别由24个连锁群组成,其中雌性图谱标记212个,总长度1 320.4 cM,覆盖率为77.7%;雄性图谱标记198个,总长度1 361 cM,覆盖率为76.1%。每个连锁群长度变动在9.3~116.1 cM之间,连锁群上的标记数在3~21个之间。各连锁群上的SSR标记并不是均匀分布的,其中1、3和8号连锁群存在标记密集区。该图谱能够进行初步的QTL定位分析和基因定位相关研究,为开展牙鲆基因和QTL的精细定位及分子标记辅助育种(MAS)等提供更有效的依据。  相似文献   

6.
聚合酶链式反应-单链构象多态性(polymerase chain reaction-single strand conformation polymorphism,PCR-SSCP)作为一种能检测基因组间DNA碱基微小差异的技术手段,具有操作简便、快捷且灵敏度高等优点.比较基因组学近年来已经成为研究生物基因组的最主要手段之一;大量基因组序列的产生为通过比较基因组学开发新标记和定位目标基因提供了大量的DNA序列资源.本研究以李氏超短纤维突变体(Ligon lintless-1,Li1)为材料,定位Li1基因,针对PCR扩增产物片段较大的样品(>400bp)进行PCR-SSCP技术的优化,用己发表的拟南芥(Arabidopsis thaliana)和棉花(Gossypium spp.)D基因组DNA序列为参考,探索PCR-SSCP技术与比较基因组学结合开发新标记应用于棉花基因定位的可能性.结果表明,电压为150V,胶浓度为10%,电泳温度为4℃时电泳结果最好;上样缓冲液与PCR扩增目的片段比例为5∶1为最佳比例.利用上述优化的PCR-SSCP技术将根据棉花与拟南芥同线性区段开发的分子标记构建了一个由28个标记组成的长度为149.6 cM的遗传图,该区域包含4个功能上与Li1位点密切相关的基因.此外,利用D基因组序列开发的标记构建了一个由17个SSCP标记和Li1位点组成,跨越40.3cM的遗传图谱,距其最近的两端标记W058和P095分别为0.6和0.3 cM.本研究为棉花Li1基因的精细定位及其候选基因的克隆提供了理论基础.  相似文献   

7.
采用磁珠富集法从AFLP片段中筛选微卫星标记,并对5个福建地方鸭品种的遗传多样性进行检测。基因组DNA用EcoRⅠ/MseⅠ内切酶酶切的同时与接头连接,酶切连接产物与用生物素标记的探针杂交,应用磁珠捕获100~2000bp含有微卫星序列的DNA片段并通过pGEM-T载体转化到大肠杆菌DH5a感受态细胞中,构建富集微卫星序列的基因组文库。随机挑选46个阳性克隆,测序获得14条含有微卫星的DNA序列并递交到GenBank(登录号:FJ599499~FJ599512)。设计合成14对微卫星引物,通过PCR优化从中选择5对引物用于5个福建地方鸭品种的遗传多样性分析。结果显示,5对微卫星引物共检测到31个等位基因,各微卫星基因座的有效等位基因数为1.969 7~2.834 4,多态信息含量和杂合度的平均值分别为0.5133和0.7480,遗传多样性丰富,说明磁珠富集法适合用于鸭微卫星标记的分离与筛选,筛选得到的5个微卫星位点可作为有效的遗传标记用于福建省地方鸭品种遗传多样性的研究。  相似文献   

8.
猪基因组AFLP指纹图谱的构建   总被引:7,自引:0,他引:7  
摘要:对猪(Sus scrofa)基因组AFLP指纹图谱构建的各项条件进行了优化,包括双酶切消化、接头连接、预扩增和选择性扩增、凝胶电泳银染技术及荧光标记检测法等,建立了稳定的猪基因组DNA的AFLP指纹图谱构建方法,采用E32/T32引物组合在45个猪种基因组混合DNA中检测到28个多态标记。  相似文献   

9.
橡胶是我国的重要的战略物资,深入研究橡胶树基因组具有重要意义。本实验以巴西橡胶树(Hevea brasiliensis)热研7-33-97幼叶为材料,采用酶解去壁低渗法制片,玻璃针显微分离法成功分离出橡胶树单染色体。单染色体经蛋白酶消化、Sau3A核酸内切酶酶切和两轮LA-PCR(接头连接PCR)扩增。得到250~2000bp之间的DNA片段,通过Southernblot对单染色体产物进行验证,结果表明扩增片段与巴西橡胶树基因组DNA之间有同源性。从而说明单染色体DNA已被成功的扩增。采用荧光原位杂交技术(fluorescence insitu hybridization,FISH)验证分离出的是第9号染色体,构建了橡胶树第9号染色体微克隆文库。克隆片段长度在300~1200bp,平均为650bp左右,文库包含48000个克隆,空载率为1%。本研究从巴西橡胶树单染色体入手,运用单染色体微分离、微克隆技术,构建了巴西橡胶树单染色微克隆文库,为橡胶树基因组研究提供了新的思路与方法,为更好地开展橡胶树分子辅助育种提供技术方法。  相似文献   

10.
叶片面积影响光合作用效率,是农作物产量的重要构成性状之一。野生黄瓜(Cucumis sativus var.hardwickii)的叶片较小,经过人工驯化后的栽培黄瓜(Cucumis sativus var.sativus)的叶片面积扩大了2~3倍。前人研究已经将控制黄瓜叶面积的主效基因之一ll(little leaf)定位在第6号染色体上。本研究以黄瓜小叶品系XF-24(P1)、大叶品系DF-32(P2)杂交产生的205个单株的F2群体为研究材料,用SAS软件对成熟期调查的各单株相同节位的叶面积进行正态性检验,结果服从正态分布,符合数量性状的遗传特征。为了有效地加快研究进程,在双亲测序情况下,采用插入缺失位点(insertion and deletion,InDel)标记进行基因定位。研究结合双亲全基因组测序数据,生物信息学分析得出双亲序列之间的InDel位点,用Primer Premier 5.0软件在所有染色体上均匀设计88对InDel标记引物,扩增采用分离群体分组混合分析法(bulked segregant analysis,BSA)组建大叶、小叶基因池,池间有多态的引物再进一步扩增F2群体DNA,筛选到7个与黄瓜叶面积基因ll2连锁的分子标记,位于黄瓜第7号染色体上,分别是InDel-1、InDel-2、InDel-3、InDel-4、InDel-5、InDel-6、InDel-7。建立遗传连锁图谱并进行区间作图寻找QTL位点,结果显示,遗传连锁图谱包含以上7个InDel标记,连锁区间为22.1 cM,包括1个控制黄瓜叶片大小的主效QTL位点ll2(little leaf 2),位于标记InDel-2与InDel-4之间,这两个标记之间物理距离为1.24 Mb。与前人的研究结果相比,定位区间更小且是7号染色体上首次发现的黄瓜叶面积QTL位点。截止目前,在黄瓜6号、7号染色体共发现了2个黄瓜叶面积主效QTL位点,分别是ll和ll2,表明黄瓜叶面积遗传机制复杂。叶面积主效QTL ll2的遗传定位,对于控制黄瓜叶片面积的遗传机制和分子机理研究以及分子标记辅助育种具有重要的意义。  相似文献   

11.
TriticumThinopyrum amphiploids arose from the need to obtain forage grasses highly resistant to pest, drought, soil salinity and frost and they can be used as efficient bridges to transfer desired genes from wheatgrass species to wheat. One of them is trigopiro SH16 INTA, it was introduced in Argentina in 1947 but its genomic composition was unknown. The aim of this work was to determine the chromosome number and genomic and chromosome composition of trigopiro SH16 INTA in order to use it in breeding programs. The simultaneous use of the in situ hybridization technique with different probes (genomic DNA of Th. ponticum (Podp.) Barkworth et D.R. Dewey, pSc119.2 and pAs1) allowed us to conclude that the chromosome number of trigopiro SH16 is 2n = 42 and the genome composition would be: 14 chromosomes of the B genome, the 2D and 4D chromosome pairs of wheat, 14 chromosomes of the J genome of Thinopyrum and the remaining chromosomes probably belong to the A genome of wheat.  相似文献   

12.
为了探索一种高效可靠的芸薹属植物核型分析方法,本文以甘蓝和白菜为实验材料,分别提取甘蓝基因组DNA用地高辛标记为探针,提取白菜基因组DNA作为封阻剂,对甘蓝有丝分裂中期染色体进行原位杂交,每对同源染色体上显示出了特定的原位杂交信号,将甘蓝的9对染色体进行了精细的分型。为区分像甘蓝这样的小型染色体提高了一条可行且精确的方法,并为研究甘蓝自交不亲和性信号传导相关基因在染色体上的定位奠定重要基础。  相似文献   

13.
The tricepiro “Don René INTA” is an artificial hybrid with 2n?=?42 chromosomes, including 14 rye (RR) and 28 wheat (AABB) chromosomes, with introgression of Thinopyrum ponticum (Podp.) Barkw. et Dewey on chromosome 6A. The aim of this work was to study the spatial distribution of the genomes and chromosomes of rye and wheat in metaphase cells from the root tip of this hybrid. The rye chromosomes were recognized by genome in situ hybridization using total genomic DNA as a probe of rye and wheat DNA as the blocking agent. Two points were determined in each cell: one representing the genomic mean distance of rye chromosomes (GRMD) and the other the genomic mean distance of wheat chromosomes (GWMD). The distance between the rye chromosomes and GRMD and GWMD showed no statistically significant difference, thus indicating that there would be no differential spatial domains for both genomes. The fluorescence in situ hybridization analysis of rye chromosomes using the pSc119.2 probe suggested that all chromosome pairs present somatic association in mitotic metaphase.  相似文献   

14.
高等植物基因组中,大部分序列为非表达序列,基因序列所占的比例很小,了解基因在基因组中的分布是研究基因组结构的一个重要方面。在美国能源部资助下,一个毛果杨无性系的基因组测序已经完成并对公众发布。杨树全基因组序列的完成,为我们了解林木基因组中基因的分布提供了一个特例。在本文中,我们利用泊松分析对杨树基因组中基因在各个染色体上的密度进行了检测,结果表明杨树基因组中各条染色体的基因含量存在显著差异。杨树全基因组测序项目揭示现代杨树基因组起源于一次古全基因组复制事件(称为杨柳科基因组复制),所以杨树基因组不同染色体间存在很大的同源复制片段。但是我们的研究显示,杨树基因组中大多数高度同源的染色体上基因的密度与染色体间的同源性没有明显关系,这说明杨柳科全基因组复制事件后,各个高度同源染色体上的基因发生了流失,且基因流失的速率是不一样的。同时本文还对近九万条毛果杨EST序列进行了比对分析,结果显示这些EST序列覆盖的基因仅占杨树基因组中基因总数的16.8%左右。EST测序虽然是发现基因的一个重要手段,但小规模EST测序对基因的覆盖度很低,所以小规模EST测序的应用价值是有限的。  相似文献   

15.
Reliable identification of hotspot areas with high numbers of threatened plant species has a central role in conservation planning. We investigated the potentiality of identifying the distribution, richness and hotspots of threatened plant species at a 25 ha resolution using eight state-of-the-art modelling techniques (GLM, GAM, MARS, ANN, CTA, GBM, MDA and RF) in a taiga landscape in north-eastern Finland. First, the individual species models developed based on occurrence records of 28 species in the 1677 grid squares and derived from different statistical techniques were extrapolated to the whole study area of 41 750 km2. Second, the projected presence/absence maps were then combined to create species richness maps, and the top 5% of grid cells ranked by species richness were classified as hotspots. Finally, we created an overall summary map by combining the individual hotspot maps from all eight modelling techniques and identified areas where the individual hotspots maps overlapped most. There were distinguishing differences in projections of the geographic patterns of species richness and hotspots between the modelling techniques. Most of the modelling techniques predicted several hotspot locations sporadically around the study area. However, the overall summary map showed the highest predictive performance based on Kappa statistics, indicating that the locations where the hotspot maps from the eight models coincided most harboured highest observed species richness. Moreover, the summary map filtered out the patchy structures of individual hotspot maps. The results show that the choice of modelling technique may affect the accuracy and prediction of hotspot patterns. Such differences may hamper the development of useful biodiversity model applications for conservation planning, and thus it is beneficial if the conservation decision-making can be based on sets of alternative maps and overlaying of predictions from multiple models.  相似文献   

16.
The chromosome morphology and the variation in the N-band distribution along the chromosomes were studied in 13 accessions of the wild wheat Aegilops geniculata Roth (2n=28, genome formula UUM°M°). The karyotype consisted of four metacentric, five submetacentric, three subtelocentric, and two satellited pairs of chromosomes. The N-banding technique stained differentially all 14 chromosome pairs. The most prominent bands were observed near the centromeres and in the intercalary regions of both arms. Telomeric bands were found in only seven chromosomes. All chromosomes produced a specific banding pattern, permitting their identification. Polymorphism for presence or absence of particular bands was observed among the different accessions. The variable bands were located predominantly at the terminal and subterminal chromosome regions, and were also unevenly distributed over chromosomes. This polymorphism resulted in up to eight distinct banding pattern variants for each of the Ae. geniculata chromosomes. Each accession in this study showed a unique overall karyotype, sharing 0 to 8 chromosomes of identical banding pattern with the other accessions. A generalized idiogram of Aegilops geniculata is presented.  相似文献   

17.
Genomic restructuring was detected in newly synthesized tritordeum by molecular and cytogenetic tools. Genomic stability is expected for advanced tritordeum lines (HchHchAABB; 2n = 42) with multiple generations of self-fertilization. This study intends to confirm or decline this hypothesis by characterizing three advanced tritordeum lines and their parental species using cytogenetics, inter-simple sequence repeat (ISSR) and retrotransposon-based markers. Mitotic chromosomes of each tritordeum line were hybridized with six synthetic oligonucleotide probes using non-denaturing fluorescence in situ hybridization. Polymorphic hybridization patterns and structural rearrangements involving SSR regions were detected. The same chromosome spreads were re-hybridized with genomic DNA of Hordeum chilense Roem. et Schult. and the 45S ribosomal DNA (rDNA) sequence pTa71. These FISH experiments allowed for parental genome discrimination, identification of nucleolar chromosomes, and detection of structural rearrangements, mostly involving rDNA loci. The chromosomes bearing SSR hybridization signals and/or chromosomes involved in structural rearrangements were identified. ISSR, retrotransposon-microsatellite amplified polymorphism, inter-retrotransposon amplified polymorphism and inter-primer binding site markers evidenced genomic reshuffling in all tritordeum lines relative to their parents. Line HT28 was considered the most genetically stable. This work demonstrated that cytogenetic and molecular monitoring of tritordeum is needed, even after several self-fertilization generations, to guarantee the selection of the most stable lines for improvement and sustainable agriculture.  相似文献   

18.
将外源λDNA导入受体普通小麦品系 81 45 2 7,获得一雄性不育变异体。对该不育变异体和受体花粉母细胞减数分裂过程进行观察。结果发现 ,不育变异体花粉母细胞染色体异常比率达到 1 8% ,而受体花粉母细胞染色体异常比例为 0 8% ,二者存在明显差异。染色体异常类型主要有单价体、染色体落后、染色体断片、染色体桥、微核及二分体、四分体异常等。染色体异常可能是外源DNA导入受体后 ,整合进受体染色体中的DNA片段影响其正常的遗传过程而造成的。花粉母细胞减数分裂过程染色体异常可以引起部分花粉败育 ,但不是该变异体败育的主要原因  相似文献   

19.
"航香糯"是经空间诱变育成的籼型香糯稻品种。与原种南丰糯相比,航香糯的产量和稻瘟病抗性有明显提高,粒形变细长,并带有香味。为深入了解空间诱变的机理以及初步鉴定与性状变异相关的区间,本研究用SSR标记对航香糯与南丰糯进行等位基因变异分析。结果表明,在12条染色体上选取的156个标记位点中检测到45个变异位点,变异频率为2...  相似文献   

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