首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到18条相似文献,搜索用时 171 毫秒
1.
由致病疫霉菌(Phytophthora infestans)引起的晩疫病是世界范围内最具毁灭性的马铃薯(Solanum tuberosum)病害。以含有晚疫病抗性基因R11的材料MaR11和不含已知抗性基因的品种Katahdin为亲本进行有性杂交,对获得的F1分离群体的83个基因型进行了晚疫病菌株接种鉴定和遗传分析。结果表明,R11为主效单基因,在MaR11中以单式形式(R11r11r11r11)存在。应用比较作图和分离群体分组分析(BSA),开发了6个与R11连锁的分子标记,并将R11定位于11号染色体长臂末端。R11距C2_At5g59960标记最近,约为2.4 cM。通过遗传图谱比较表明,R11较晚疫病抗性基因R3a和R10更靠近染色体端粒区。本研究所获得的遗传图谱为进一步构建R11高密度遗传图谱提供了基础。  相似文献   

2.
采用Dynal磁珠富集法构建了香椿微卫星富集文库,通过测序结果对文库的特性进行了分析。实验使用改良CTAB法提取香椿基因组DNA,用(AC)8、(AG)8和(ATG)123种带有生物素标记的探针与香椿基因组DNA的酶切片段进行杂交,将磁珠捕获的含有微卫星序列的DNA片段插入p MD 18-T载体,并转入感受态细胞Trans 5α构建克隆,经筛选后得到含356个克隆的香椿基因组微卫星富集文库。从富集文库中挑选插入片段长度为400~800 bp的128个克隆进行测序,其中含有SSR的序列77条,得率达60.16%,其中完美型占82.69%,非完美型8.65%,混合型8.65%。上述结果为SSR位点的进一步开发奠定了基础。  相似文献   

3.
棉花抗黄萎病相关基因筛选与亚克隆文库构建   总被引:3,自引:0,他引:3  
以黄萎病菌诱导下差异表达的棉花抗病相关基因片段PR8为探针,利用杂交方法,对优质、抗病海岛棉品种Pima90-53 BAC文库进行筛选。从30 336个BAC克隆中筛选到含有PR8基因片段的4个阳性克隆,分别为127K13,128D14,169J3和178C5。用Sau3AI对其中的169J3克隆进行酶切,回收2~4 kb的DNA片段并连接到载体pUC118BamHI-BAP上,构建了含有该基因片段的亚克隆文库,共含有4 224个克隆。经电泳检测,插入片段在1~3 kb,平均为2 kb。该亚克隆文库的构建为克隆PR8基因奠定了基础。  相似文献   

4.
磁珠富集法筛选马尾松微卫星标记   总被引:2,自引:0,他引:2  
将马尾松核基因组DNA用Sau3A Ⅰ酶切后,电泳回收300~1 000 bp片段.在回收的片段上连接接头PCR后与用生物素标记的微卫星探针(AC)15、(AG)15杂交运用磁珠富集含有微卫星序列的DNA片段.将获得的序列通过PCR扩增后,连接pGEM-T载体,转化入感受态大肠杆菌,得到微卫星序列文库.然后用PCR法直接对文库进行扩增,获得58个阳性克隆,经测序分析,获微卫星序列33个,并成功设计出马尾松微卫星引物19对.  相似文献   

5.
应用Dynal磁珠-生物素标记的微卫星探针(AC)8,(AG)8和(ATG)12与地黄基因组DNA酶切片段杂交,捕获含有微卫星序列的DNA片段,连接到pMD 18-T栽体上,转入感受态细胞Trans 5 α,构建地黄富集微卫星文库.利用M13F和M13R载体序列引物筛选文库,对插入片段长度为400~ 800 bp的克隆进行测序.共获得96条序列,48条(50%)含有微卫星位点,其中完美型占66%,非完美型22%,混合型12%.微卫星重复基元中,二核苷酸(AG)n和三核苷酸(CAT)n最为常见.  相似文献   

6.
为了研究马铃薯内源BAG基因在晚疫病抗性建立中的作用,利用拟南芥和水稻的BAG基因保守序列,在马铃薯基因库中比对获得马铃薯内源StBAG3的基因序列。以StBAG3基因序列为模板设计引物,通过RT-PCR克隆得到了StBAG3基因cDNA序列。序列分析的结果表明,该基因开放阅读框为1026bp,编码341个氨基酸。蛋白二级结构预测结果表明,StBAG3具有UBQ和BAG2个保守功能域。StBAG3基因的表达谱分析结果显示,不同马铃薯品种中StBAG3基因表达水平存在差异,其中在冀张薯12号中表达量最高,而夏坡蒂中表达量最低。该基因在马铃薯不同组织中表达量测定结果表明,马铃薯茎中的表达水平最高,老叶中表达水平最低。接种晚疫菌后,StBAG3能够被显著上调并在接种48h后达到最高值,预示着StBAG3基因在马铃薯晚疫病抗性建立过程中具有一定的调控作用。  相似文献   

7.
NAC转录因子是特异存在于植物中具有多种生物功能的新型转录因子,其家族成员N端含有保守氨基酸序列,C端为高度变异的转录激活区。本项研究以南瓜叶片为材料,根据甘蓝型油菜NAC1、番茄NAC、辣椒NAC保守结构域设计一对简并引物,采用RT-PCR方法扩增得出长度约为440 bp大小的DNA片段,将其克隆至pMDl9-T载体上,而后对重组克隆进行测序,用BLAST和DNAMAN软件对核酸及氨基酸序列进行分析,结果表明所获得的南瓜NAC基因片段由442个碱基组成,编码147个氨基酸,命名为CmNAC。该基因片段具有其他植物NAC基因中存在的保守区,并且属于NAC家族中ATAF1/2亚家族。  相似文献   

8.
广谱抗性基因的挖掘是马铃薯高抗晚疫病品种选育的基础。本研究以288份国际马铃薯中心筛选的晚疫病抗性群体为试验材料,经过连续2年田间调查,计算AUDPC和sAUDPC值,评估群体晚疫病抗性;利用SLAF-seq方法进行群体简化基因组测序,通过对晚疫病抗性表型数据的全基因组关联分析,挖掘晚疫病抗性相关的遗传位点和候选基因,为晚疫病抗性品种选育和抗病机理研究提供一定的理论和材料基础。结果表明,晚疫病抗性在288份材料间存在着广泛的遗传差异;基于5种分析模型,共鉴定到82个与晚疫病抗性显著关联的位点;在关联区间关联到54个已知或可能与晚疫病抗性相关的基因。其中,23个基因为抗性基因,包括晚疫病抗性基因R1同源基因、Sw-5同源基因(R8)和Rpi-vnt1以及编码多效性耐药蛋白基因;5个基因编码MAPK蛋白和WRKY转录因子;1个基因参与茉莉酸途径;3个基因与水杨酸途径相关;6个基因是病程相关的基因;3个基因参与苯基丙酸类合成途径;其他与晚疫病抗性相关的基因,如HMGR基因(2个)、细胞色素P450(21个)。  相似文献   

9.
用磁珠富集法分离亚麻基因组微卫星分子标记   总被引:5,自引:0,他引:5  
应用Dynal磁珠-生物素标记的微卫星探针(CT)15与亚麻基因组DNA酶切片段杂交,捕获300~1 500 bp含有微卫星序列的DNA片段,连接到pMD18-T载体中,构建富集微卫星序列的小片段插入文库。利用接头引物和根据微卫星核心序列设计的引物VRV(CT)15使用PCR方法直接对文库筛选,从422个转化子中获得了104个阳性克隆,对其进行测序分析,获得了97个微卫星序列,微卫星序列的富集效率达到22.99%,PCR扩增筛选效率93.27%。对97个微卫星序列进行比对分析,其中51个重复序列的两端序列高度相似,据其设计的特异引物对阳性克隆进行2次筛选,能淘汰相似度高的同类序列,提高筛选亚麻微卫星标记的效率。  相似文献   

10.
以本实验室分离的Bt菌株B-DLL质粒DNA为模板,利用cry8类基因特异性引物JJX5和JJX3扩增出3.5kb的片段,将该片段插入克隆载体pMD 18-T中,筛选获得一个含新基因的克隆pMD-cry8new.序列测定表明,该基因编码区为3 525 bp,编码的蛋白质由1 174个氨基酸残基组成,理论分子量为133....  相似文献   

11.
12.
C. M. Lu  W. Y. Yang  B. R. Lu 《Euphytica》2005,143(1-2):75-83
The devastating late blight pathogen Phytophthora infestans infects foliage as well as tubers of potato. To date, resistance breeding has often focused on foliage blight resistance, but tuber blight resistance is becoming more and more important in cultivated potatoes. In this study, a reliable tuber assay for resistance assessment was developed and foliage and tuber blight resistance (R) was compared in four mapping populations. In the RH4X-103 population, tuber blight resistance inherited independently from foliage blight resistance. Three specific R genes against P. infestans were segregating. The Rpi-abpt and R3a genes function as foliage-specific R genes, whereas the R1 gene acts on both foliage and tuber. In the segregating populations SHRH and RH94-076, tuber and foliage blight resistance correlated significantly, which suggests that resistance in foliage and tuber is conferred by the same gene (could be R3b) and quantitative trait loci (QTL), respectively. In the CE population neither tuber nor foliage resistance was observed.  相似文献   

13.
马铃薯是世界上重要的粮食作物之一,晚疫病则是当今危害马铃薯生产最为严重的病害。重点介绍了四种常用的分子标记技术RFLP、RAPD、AFLP和SSR,以及国内外利用这些标记技术在马铃薯晚疫病研究中的应用。分析了马铃薯晚疫病菌的遗传多样性、菌株抗药性、有性杂交后代的遗传分离以及抗病种质资源的遗传多样性,介绍了遗传图谱的构建、抗性基因及与晚疫病抗性相关的QTL定位、体细胞杂种及回交后代的晚疫病抗性检测。这些对今后中国学者开展相关领域的研究具有重要的借鉴意义。  相似文献   

14.
The objectives of this study were to evaluate the use of potato (Solanum tuberosum L.) late blight (Phytophthora infestans (Mont.) de Bary) resistant parents in cultivar development and identify superior clones possessing moderate to high late blight resistance combined with acceptable maturity and tuber quality. Ninety-five crosses were made between eight unadapted parents with reported late blight resistance (B0718-3, Bertita, Bzura, Greta, Libertas, Stobrawa, Tollocan and Zarevo) and susceptible parents (cultivars or advanced breeding clones) adapted to North American growing conditions. A total of 408 field selected clones were assessed for late blight resistance in the greenhouse and in the field using a mixture of US8 P. infestans isolates (A2 mating type, metalaxyl resistant) that overcame all known R-genes except R8 and R9. Clones with ≤ 10% infected foliar area in the greenhouse test or ≤ 0.30 RAUDPC (relative area under the disease progress curve) value in the field in 1998 were re-tested in 1999. A total of 118 (29% of 408) putative late blight resistant clones were selected. The eight late blight resistant parents differed in both the ability to transmit late blight resistance and in the level of resistance transmitted to the progeny. The Tollocan and B0718-3 families (half-sib progeny) had the greatest degree of resistance and frequency of resistant clones. Scott-Knott cluster analysis ranked 79 clones (67% of 118) in the high and moderate late blight resistant groups. Among these 79 clones, 19 clones had vine maturity equal to or earlier than mid-season combined with acceptable tuber quality. Further selection in 2000 resulted in eight advanced selected clones (six from Tollocan and two from B0718-3 families) with the same level of resistance as the parent combined with vine maturity and tuber quality equivalent to Atlantic, a standard cultivar for chip processing in North America. The results indicate that this breeding approach can be used to select parents for late blight resistance breeding and to identify superior clones with high levels of late blight resistance and marketable vine maturity and tuber quality. This revised version was published online in July 2006 with corrections to the Cover Date.  相似文献   

15.
分子标记辅助培育双抗稻瘟病和白叶枯病杂交稻恢复系   总被引:10,自引:3,他引:7  
本研究利用常规回交育种结合分子标记辅助选择技术,将来自C750的抗白叶枯病基因Xa23和抗稻瘟病基因pi9聚合到感病杂交稻恢复系闽恢3139中.最后在回交后代中获得了4个双抗稻瘟病和白叶枯病改良系ZR21-sk1、ZR21-sk2、ZR21-sk3和ZR21-sk4,且其遗传背景恢复率达96%以上.用来自福建省具有代表性的24个稻瘟病菌株和白叶枯病广致病型菌系P6对改良株系进行人工接菌鉴定,结果表明,聚合了Xa23和pi9基因的改良系同时抗稻瘟病和白叶枯病,且抗性与抗谱与供体亲本C750相似.农艺性状分析显示,改良株系所配组合基本保持闽恢3139的农艺性状和配合力,可直接应用于生产或作抗性育种亲本.  相似文献   

16.
Identification of resistance gene analogs in cotton (Gossypium hirsutum L.)   总被引:4,自引:0,他引:4  
Sequence analyses of numerous plant disease resistance genes have revealed the presence of conserved motifs common to this class of genes, namely a nucleotide binding site (NBS) and leucine rich repeat region. In this study, thirty-three resistance gene analogs (RGAs) were cloned and sequenced from cotton (Gossypium hirsutum L.) following PCR with degenerate primers designed from the conserved NBS motif of plant resistance (R) genes. Phylogenetic analysis of the predicted amino acid sequences grouped the RGAs into four distinct classes from which several subgroups were delineated based on nucleic acid sequences. Gene database searches with the consensus protein sequences of each of the four classes and respective subgroups of cotton RGAs revealed their conserved NBS domains and homology to RGAs and known resistance genes from a variety of plant genera. Given the complete lack of knowledge regarding molecular organization of R genes in cotton, the cloned RGAs described here may be useful as probes to map, characterize, and manipulate R genes of the cotton genome. This revised version was published online in July 2006 with corrections to the Cover Date.  相似文献   

17.
云南抗白叶枯病稻种的RGA初析   总被引:1,自引:1,他引:1  
根据水稻抗白叶枯病Xa21基因的富含亮氨酸重复区域(LRR)和番茄抗细菌性斑点病(Pseudomonas syringae pv. tomato)的Pto基因编码蛋白质激酶的DNA序列,设计2对引物用于扩增抗水稻白叶枯病品种中的抗病基因同源序列。经聚丙烯酰胺凝胶电泳和聚类分析,结果表明供试抗病品种间具有丰富的RGA多态性,用同一引物测定的属于同一簇  相似文献   

18.
The R10 late blight differential of potato, 3681ad1, exhibits good field resistance. Progeny from the cross between 3681ad1 and the susceptible cultivar ‘Katahdin’ were assessed for late blight resistance to three Phytophthora infestans isolates, using a detached leaf assay. Progeny differed in response to the three isolates. Resistance to isolates IPO‐0 and 99018 was controlled by quantitative trait loci (QTL), whereas resistance to isolate 89148‐9 was inherited as a dominant R gene, designated as R10 in this study. Statistical analysis revealed that one of the resistance QTLs to isolates IPO‐0 and 99018 is linked to the R10 gene, which maps to chromosome 11 in a region where a complex late blight resistance locus has been reported previously. A high‐resolution map of R10 was constructed using a large segregating population, and the gene was delimited to a genetic interval of 0.26 cM. The clustering of the qualitative gene R10 with resistance QTLs could explain the field resistance observed with 3681ad1.  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号