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1.
抗芜菁花叶病毒转基因甘蓝型油菜的研究   总被引:31,自引:0,他引:31  
以子叶柄为材料,建立了甘蓝型油菜(BrassicanapusL.)双低品种的再生体系。通过子叶柄与农杆菌(AgrobacteriumtumefaciensLBA4404)共培养,将表达载体pBTu中芜菁花叶病毒外壳蛋白(TuMV-CP)基因以整合方式导入甘蓝型油菜,然后用卡那霉素进行筛选,获得了油菜再生植株。经PCR特异性扩增、点杂交和Southern印迹分析,证明再生植株基因组DNA中整合了TuMV-CP基因。攻毒实验表明,有TuMV-CP基因插入的工程油菜对TuMV均有不同程度的抗性。  相似文献   

2.
用RFLP标记分析甘蓝型油菜的遗传多样性   总被引:38,自引:2,他引:36  
孟金陵  钱秀珍 《遗传学报》1996,23(4):293-306
以甘蓝型油菜的28个基因组探针和两种限制性内切酶对包括46个中国品种、9个欧洲品种在内的59个甘蓝型油菜品种(系)的RFLP标记进行了分析。在放射自显影胶片上,共检测到410条具多态性的分子杂交带,表明甘蓝型油菜中存在着极为丰富的遗传变异。聚类分析结果表明,在相似性为45%的水平上,可把中国甘蓝型油菜划分为6组:胜利油菜组、跃进油菜组、中油821组、远缘种质组、优质油菜组和变异不详组。欧洲冬油菜与以上6组存在着较显著的遗传距离。主成分分析的结果与上述分组较为一致。以上结果表明,对于扩大中国甘蓝型油菜的遗传基础,欧洲冬油菜无疑是一个重要的种质资源。另一方面,用典型的中国甘蓝型油菜与欧洲冬油菜配制的杂交种,较易产生强大的杂种优势。从对已进行了染色体定位的61条放射自显影带的分析看,无论是上述分组内,还是分组间,RFLP的相对差异均主要表现在A基因组中。讨论了致使A基因组遗传变异较大的可能因素。  相似文献   

3.
我国芥菜型油菜品种遗传多样性初探   总被引:8,自引:0,他引:8  
用RAPD技术对包括春、冬芥菜型采及国外品种在内的36个芥菜型油菜品种的遗传多样性进行了分析。在扩增得到的128条DNA带中,多态性DNA片段达88.28%。分析表明:春 油菜间遗传差异较大,国内冬性芥菜型油菜地方种多样性水平较高,25份冬性品种分属Ⅰ、Ⅱ和Ⅲ类群,而春性芥菜型 采地方种均归于Ⅳ类;印度的RLM198与四川的珙县金黄油菜、澳大利亚品种与我国春油菜品种亲缘关系密切。  相似文献   

4.
抗白粉病小麦——中间偃麦草异附加系的细胞学和RAPD鉴定   总被引:21,自引:5,他引:16  
利用细胞学和RAPD技术,对从小麦与中间偃麦草杂种后代中选育的抗白粉病异附加系DAL66进行了鉴定。结果证明DAL66根尖细胞染色体数为44,花粉母细胞减数第一分裂中期(PMC MI)杂色体模型为2n=22Ⅱ。对DAL66及其双亲进行RAPD分析,从40个随机引物中筛选出1个特异引物(OPE-02)能够稳定地扩增出特异带型。  相似文献   

5.
RAPD应用于蕈菌研究中的条件优化探讨   总被引:4,自引:0,他引:4  
在进行鹅膏菌属蕈菌遗传多样性的RAPD分析时,对RAPD分析过程中的DNA提取方法、DNA纯度、模板DNA和dNTP浓度、循环次数、DNA不同来源等影响因素进行了大量的实验探索,结果表明:DNA不同提取方法具有相同的扩增产物,DNA模板中的RNA对扩增产物无影响,模板浓度在一个相当大的范围内(50~400μg)不影响扩增结果.dNTP浓度达0.75mmol/L时无带谱出现,引物浓度达1μmol/L时出现非特异性带谱,从菌丝体和子实体中提取的DNA可获得一致的扩增产物.扩增循环40~45周期条件下扩增效果较好,本实验证明了RAPD产物具有很好的重复性,建立了适合蕈菌RAPD分析的PCR程序及条件,为RAPD应用于蕈菌的遗传研究打下了良好的基础.  相似文献   

6.
RAPD技术与桂花品种分类和鉴定   总被引:11,自引:1,他引:10       下载免费PDF全文
朱诚  刘非燕   《广西植物》1999,19(2):190-192
简介了随机扩增多态DNA(RAPD)技术的原理,及对桂花品种分类鉴定的必要性,并建议开展RAPD技术在桂花品种分类鉴定的应用研究。  相似文献   

7.
植物RAPD标记的可靠性研究   总被引:71,自引:0,他引:71  
随着RAPD技术应用范围越来越广,RAPD的优缺点也不断反映出来。相左实验结果的出现,对RAPD标记可靠性提出了置疑。本文从重复性、序列同源性和系统学价值三个方面,就RAPD标记的可靠性研究的现状进行了综述。并就怎样提高RAPD标记的可靠性进行了探讨  相似文献   

8.
甘蓝型油菜与蓝花子远缘杂交及双二倍体的合成研究   总被引:16,自引:5,他引:11  
徐利远  罗鹏  兰泽蘧 《遗传学报》1996,23(2):124-130
甘蓝型油菜品种奥罗×蓝花子杂种F-1,平均配对构型为12.1Ⅰ+6.53Ⅱ+0.41Ⅲ+0.18Ⅳ+0.18Ⅴ,A、C染色体组与R染色体间存在配对,它们之间具有一定的同源性。在甘蓝型油菜与蓝花子的杂种F-1代中,存在一种染色体不配对的减数分裂类型。这一类型中有少量可形成平衡的不减数配子。提供了油菜与蓝花子远缘杂种回交结实的细胞学根据。在MS+0.2mg/LNAA+3mg/LBA+1g/L秋水仙碱+30g/L蔗糖+8g/L琼脂的培养基中,接种甘蓝型油菜奥罗与蓝花子的杂种F-1进行加倍处理,经快速繁殖后,获得大量的染色体数为2n=56的双二倍体幼苗。上述双二倍体自交结实,在减数分裂中绝大多数细胞形成28个二价体,个别形成26个二价体和1个四价体。上述技术在油菜与蓝花子远缘杂交中首次解决了用常规方法不易获取远种杂种稳定双二倍体的难题。甘蓝型油菜品种Altex×蓝花子杂种F-1代,长时间的快速繁殖后,出现了染色体丢失和加倍,有形成19条染色体的配子回复到甘蓝型油菜染色体组成的趋势。在油菜远缘杂种中发现了类似于球茎大麦远缘杂种中染色体丢失的现象。  相似文献   

9.
白鲢和镛鱼的扩增多态DNA分析   总被引:12,自引:0,他引:12  
根据鱼类外周血细胞都有核的特点,采用从冷冻和低渗双重处理分离的细胞核的取基因组DNA。以此法获得的白鲢和镛鱼的基因组DNA为模板,和Operon公司生产的OPN和OPM两个组共40个随机引物,对这两种鱼进行了随机扩增多态DNA(RAPD)分析;确定了对这两种鱼基因组相关区域可进行随机PCR扩增的有效引物,特别是哪些可产生种群内或群体的RAPD遗传标记即可产生个体特异性和群体特异性RAPD带谱的引物  相似文献   

10.
油菜分子标记图谱构建及抗菌核病性状的QTL定位   总被引:30,自引:2,他引:28  
以油菜品种085(抗菌核病)与湘油13号杂交得到的F2代作为作图群体,以RAPD标记来构建油菜连锁图谱。通过对300个10-mer随机引物的筛选,共获得200个多态性RAPD位点。经Mapmaker/EXP3.0软件处理,构建了1张含193个标记位点、19个连锁群、覆盖长度为1324cM的连锁图谱。在此基础上利用Mapmaker/QTL 1.1将抗菌核病基因定位在第四、八和十四3个连锁群上,即:S  相似文献   

11.
Zhao YT  Wang M  Fu SX  Yang WC  Qi CK  Wang XJ 《Plant physiology》2012,158(2):813-823
MicroRNAs (miRNAs) and small interfering RNAs are important regulators of plant development and seed formation, yet their population and abundance in the oil crop Brassica napus are still not well understood, especially at different developmental stages and among cultivars with varied seed oil contents. Here, we systematically analyzed the small RNA expression profiles of Brassica napus seeds at early embryonic developmental stages in high-oil-content and low-oil-content B. napus cultivars, both cultured in two environments. A total of 50 conserved miRNAs and 9 new miRNAs were identified, together with some new miRNA targets. Expression analysis revealed some miRNAs with varied expression levels in different seed oil content cultivars or at different embryonic developmental stages. A large number of 23-nucleotide small RNAs with specific nucleotide composition preferences were also identified, which may present new classes of functional small RNAs.  相似文献   

12.
A G Sharpe  D J Lydiate 《Génome》2003,46(3):461-468
Recent oilseed rape breeding has produced low glucosinolate cultivars that yield proteinaceous meal suitable for animal feed. The low glucosinolate character was introduced into modern cultivars from Brassica napus 'Bronowski', a cultivar that is agronomically inferior in most other respects. Residual segments of 'Bronowski' genotype in modern cultivars probably cause reduced yield, poorer winter hardiness, and lower oil content. The quantity and distribution of the 'Bronowski' genotype in the modern oilseed rape cultivar Brassica napus 'Tapidor' was investigated using a segregating population derived from a cross between 'Tapidor' and its high glucosinolate progenitor. This population was analyzed with 65 informative Brassica RFLP probes and a genetic linkage map, based on the segregation at 77 polymorphic loci, was constructed. The mapping identified 15 residual segments of donor genotype in 'Tapidor', which together occupy approximately 29% of the B. napus genome. Mapping the loci that control variation for the accumulation of total seed glucosinolates in the segregating population has identified three loci that together explain >90% of the variation for this character. All of these loci are in donor segments of the 'Tapidor' genome. This result shows the extent to which conventional breeding programmes have difficulty in eliminating residual segments of donor genotype from elite material.  相似文献   

13.
The genetic control of self-incompatibility in Brassica napus was investigated using crosses between resynthesized lines of B. napus and cultivars of oilseed rape. These crosses introduced eight C-genome S alleles from Brassica oleracea (S16, S22, S23, S25, S29, S35, S60, and S63) and one A-genome S allele from Brassica rapa (SRM29) into winter oilseed rape. The inheritance of S alleles was monitored using genetic markers and S phenotypes were determined in the F1, F2, first backcross (B1), and testcross (T1) generations. Two different F1 hybrids were used to develop populations of doubled haploid lines that were subjected to genetic mapping and scored for S phenotype. These investigations identified a latent S allele in at least two oilseed rape cultivars and indicated that the S phenotype of these latent alleles was masked by a suppressor system common to oilseed rape. These latent S alleles may be widespread in oilseed rape varieties and are possibly associated with the highly conserved C-genome S locus of these crop types. Segregation for S phenotype in subpopulations uniform for S genotype suggests the existence of suppressor loci that influenced the expression of the S phenotype. These suppressor loci were not linked to the S loci and possessed suppressing alleles in oilseed rape and non-suppressing alleles in the diploid parents of resynthesized B. napus lines.  相似文献   

14.
单核苷酸多态性及其在油菜分子标记辅助育种中的应用   总被引:3,自引:0,他引:3  
油菜是一种重要的油料作物,油菜的基因组功能研究已引起了各国政府和科研工作者的高度重视。SNP是目前为止分布最广泛,存在数量最多且标记密度最高的一种遗传多态性标记。对油菜等作物的基因组功能研究提供了丰富的信息资源。综述了近来单核苷酸多态性的研究进展以及其在油菜的分子标记辅助品质育种中的可能用途。  相似文献   

15.
杂交育种中,亲本选配是育种成败的关键。本研究以重庆市油菜工程技术研究中心提供的180份甘蓝型黄子油菜亲本种质为材料,应用分布于不同连锁群的60对SSR标记进行了分析,共检测出308个标记位点,每对引物在不同亲本材料之间的等位基因数在1~11个之间,平均位点为5.1个。其中多态性位点207个,多态率达67.2%。对SSR扩增结果进行UPGMA分析,在遗传距离0.566处,180个品种(系)分为3个类群,聚类结果与种质来源比较一致,本研究为甘蓝型油菜黄子杂交育种和优势组合的选配提供了理论依据。  相似文献   

16.
An efficient protocol for Agrobacterium tumefaciens-mediated transformation of six commercial Brassica napus winter cultivars is described. Two B. napus spring cultivars were analysed for comparison. Five strains of A. tumefaciens with different combinations of nopaline and octopine chromosomal backgrounds and virulence plasmids were used for cocultivation. Selection of putative regenerated transgenic plants was performed on kanamycin- or hygromycin-containing media. The scores of transgenic plants were calculated on the basis of GUS (-glucuronidase) activity, detected by the histochemical X-Gluc test. Target tissue derived from the cut surface of cotyledon petioles resulted in successful transformation with all the winter cultivars tested. Target tissue from hypocotyl segments resulted in a successful transformation with only one winter cultivar. The transformation rates for B. napus winter cultivars in this study were higher than in previous reports. Southern blot analysis revealed that integration of marker genes occurred in single and in multiple copies and at multiple loci in the genome. The transgenic plants all grew normally and developed fertile flowers after a vernalization period. After self-pollination, Southern blot analysis of selected GUS active F1 plants revealed that introduced marker genes were stably inherited to the next generation. These data demonstrate that morphologically normal, fertile transgenic plants of B. napus winter cultivars can be achieved with both nopaline- and octopine-derived A. tumefaciens strains. This protocol should have a broad application in improvement of Brassica napus winter cultivars by introduction of foreign genes  相似文献   

17.
Phyllotreta cruciferae is an important insect pest of spring-planted Brassica crops, especially during the seedling stage. To determine the effect of early season P. cruciferae infestation on seed yield, 10 genotypes from each of two canola species (Brassica napus L. and Brassica rapa L.) and two mustard species (Brassica juncea L. and Sinapis alba L.) were grown in 2 yr under three different P. cruciferae treatments: (1) no insecticide control; (2) foliar applications of endosulfan; and (3) carbofuran with seed at planting plus foliar application of carbaryl. Averaged over 10 genotypes, B. rapa showed most visible P. cruciferae injury and showed greatest yield reduction without insecticide application. Mustard species (S. alba and B. juncea) showed least visible injury and higher yield without insecticide compared with canola species (B. napus and B. rapa). Indeed, average seed yield of S. alba without insecticide was higher than either B. napus or B. rapa with most effective P. cruciferae control. Significant variation occurred within each species. A number of lines from B. napus, B. juncea, anid S. alba showed less feeding injury and yield reduction as a result of P. cruciferae infestation compared with other lines from the same species examined, thus having potential genetic background for developing resistant cultivars.  相似文献   

18.
甘蓝型油菜若干农艺性状与单株产量的关系分析   总被引:12,自引:0,他引:12  
以38个甘蓝型油菜品种为材料,对单株产量、有效分枝部位高度、主轴花序长度等11个农艺性状进行了方差分析、通径分析和聚类分析.结果显示:所有调查性状在品种间存在显著的差异;主花序长度、主序有效角果数与单株产量都有显著的遗传相关性,两者都通过有效分枝部位高度对单株产量产生较大的正向间接作用;有效分枝部位高度对单株产量的影响也较显著,高产品种具有相对较高的有效分枝部位高度;总角果数对单株产量的直接遗传通径系数虽小,但它具有最大的直接表型通径系数,它通过主序有效角果数和有效分枝数对单株产量产生了两个较大的间接作用;要获得3130kg/hm~2的产量,重点应具备果多、枝多的特征,总角果数不少于657个,有效分枝数不少于18个.  相似文献   

19.
真空渗透法转化油菜及转化种子的筛选   总被引:4,自引:0,他引:4  
王道杰  杨翠玲  陆鸣 《植物学报》2009,44(2):216-222
真空渗透遗传转化法首先在拟南芥(Arabidopsis thaliana)中获得成功, 是一种简便、快速且无需经过组织培养阶段即可获得大量转化植株的基因转化方法。该研究以不同遗传背景的3个甘蓝型油菜(Brassica napus)品种(系)陕3B、L45和Mg23为材料, 对真空渗透遗传转化方法中真空渗透时间和Silwet L-77浓度与遗传转化效果的关系进行了比较, 同时对转化种子的筛选方法进行了优化。结果表明, 卡那霉素(Km)对油菜种子的萌发影响不显著, 但对其生长发育有明显的抑制作用。不同油菜品种对卡那霉素的敏感性不同, 各自的致死浓度也不一样。在0%-0.05%的浓度范围内, 随着Silwet L-77浓度的增加, 在相同的真空渗透时间内, 3个油菜品种的转化率逐渐升高。当Silwet L-77浓度为0.05%时, 10分钟的真空渗透时间可获得最高转化效率, 此时陕3B、L45和Mg23的转化率分别达到1.97%、2.09%和2.30%。  相似文献   

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