外源赤霉素诱导‘藤稔’葡萄果实差异表达基因的cDNA-RAPD分析     

王西成  宋长年  张彦苹  冷翔鹏  孙欣  房经贵 《果树学报》2012,(5):733-739

【目的】为鉴定葡萄果实赤霉素诱导响应基因,【方法】应用cDNA-RAPD技术,以鲜食葡萄品种‘藤稔’为研究对象,对赤霉素处理后果实发育过程中基因表达进行了mRNA指纹分析。【结果】通过16条随机引物的筛选,共分离得到61个差异表达的转录衍生片段(TDF),其中增强表达或赤霉素诱导下特异性表达TDF42条,抑制表达19条。对其中18个TDF进行了克隆、测序和序列分析发现,17个TDF与NCBI已有序列同源,其中10个为已知功能基因,另有1个TDF无同源序列。对选取的6个TDF进行半定量RT-PCR验证,结果表明3个基因片段在处理条件下均特异表达或上调表达,另外3个基因片段在处理条件下均未见表达或下调表达,这与cDNA-RAPD表达谱结果相一致。【结论】利用cDNA-RAPD技术成功分离了部分受赤霉素诱导的差异表达基因。  相似文献   

苹果MdGLRs家族基因生物信息学鉴定和表达分析   总被引：2，自引：1，他引：1  

罗华  胡大刚  张连忠  郝玉金 《园艺学报》2012,39(3):425-435

 利用生物信息学策略对苹果MdGLRs家族基因编码蛋白的氨基酸序列的基本特征、二级结构、跨膜区域、亲疏水性、基因亚细胞定位及保守结构域进行了预测和分析,与拟南芥AtGLRs家族的20个基因进行了氨基酸序列比对和进化树分析,并对这些基因在不同营养生长器官中进行了表达分析。结果表明,苹果MdGLRs家族包含32个基因,分为3个亚族,大部分基因编码800个氨基酸以上,多含有3个跨膜区域,二级结构主要以α–螺旋、无规则卷曲、折叠延伸链为主;亚细胞定位预测发现,MdGLRs蛋白主要定位在内质网和质膜上;MdGLRs蛋白的保守结构域是S1-M1-M2-M3-S2-M4;大多数基因在根、茎、叶中普遍都有表达,在叶中的表达量最高。  相似文献   

Additional evidence for oligogenic inheritance of durable host resistance to coffee berry disease (<Emphasis Type="Italic">Colletotrichum kahawae</Emphasis>) in arabica coffee (<Emphasis Type="Italic">Coffea arabica</Emphasis> L.)     

H. A. M. van der Vossen  D. J. Walyaro 《Euphytica》2009,165(1):105-111

Breeding for host resistance to coffee berry disease (CBD) in arabica coffee (Coffea arabica) was initiated some 35–40 years ago in Kenya, Ethiopia and Tanzania in response to severe CBD epidemics. The release of CBD resistant cultivars to the coffee growers has been in progress since 1985. The resistance of cultivars like Ruiru 11 (Kenya) and Ababuna (and other cvs in Ethiopia) appears to be of a durable nature, since confirmed cases of a breakdown of host resistance under field conditions have not been reported over the past 20 years. Host resistance to the hemibiotrophic fungus Colletotrichum kahawae is of a quantitative nature, but nevertheless can be practically complete in some genotypes of arabica coffee. There is still no consensus on the genetics of CBD resistance, some claiming convincing evidence for oligogenes (1–3 major genes) and others for polygenes determining CBD resistance. Results from genetic studies with germplasm from the centre of genetic diversity for C. arabica in Ethiopia are presented here. These together with the recent identification of molecular markers associated with and the mapping of one major gene, provides additional evidence for oligogenic inheritance of CBD resistance. The development of cultivars combining yield and quality with durable host resistance to CBD has contributed greatly to increased sustainability of arabica coffee production in Africa. It has also considerable relevance to arabica coffee in Latin America and Asia, where CBD is still a quarantine disease but with a risk of becoming endemic one day, just as has happened earlier with coffee leaf rust (Hemileia vastatrix).  相似文献   

Validation of molecular markers linked to fertility restorer gene(s) for WA-CMS lines of rice     

N. K. Sheeba  B. C. Viraktamath  S. Sivaramakrishnan  M. G. Gangashetti  Pawan Khera  R. M. Sundaram 《Euphytica》2009,167(2):217-227

The present study was carried out with the objective to validate the molecular markers, which have been previously reported to be linked to fertility restorer (Rf) gene(s) for WA-CMS lines of rice. Two mapping populations involving fertility restorer lines for WA-cytoplasm, viz., (i) an F₂ population derived from the cross IR58025A/KMR3R consisting of 347 plants and (ii) a BC₁F₁ population derived from the cross IR62829A/IR10198R//IR62829A consisting of 130 plants were analyzed. Nine SSR and three CAPS markers reported to be linked to Rf genes along with two previously unreported SSR markers were analyzed in the mapping populations. In both the populations studied, the trait of fertility restoration was observed to be under digenic control. Eight SSR markers (RM6100, RM228, RM171, RM216, RM474, RM311, MRG4456 and pRf1&2) showed polymorphism between the parents of the F₂ population, while the SSR markers RM6100 and RM474 showed polymorphism between the parents of both the F₂ and BC₁F₁ populations. Only one CAPS marker, RG146FL/RL was polymorphic between the parents of the BC₁F₁ population. RM6100 was observed to be closely segregating with fertility restoration in both the mapping populations and was located at a distance of ~1.2 cM. The largest phenotypic variation was accounted for the region located between RM311 and RM6100. Using the marker-trait segregation data derived from analysis of both the mapping populations, a local linkage map of the genomic region around Rf-4, a major fertility restoration locus on Chromosome 10 was constructed, and RM6100 was observed to be very close to the gene at a distance of 1.2 cM. The accuracy of the marker RM6100 in predicting fertility restoration was validated in 21 restorers and 18 maintainers. RM6100 amplified the Rf-4 linked allele in a majority of the restorers with a selection accuracy of 94.87%. Through the present study, we have established the usefulness of the marker RM6100 in marker-assisted selection for fertility restoration in segregating populations and identification of restorers while screening rice germplasm for their fertility restoration ability.  相似文献   

Molecular mapping of genic male-sterile genes ms₁₅, ms₅ and ms₆ in tetraploid cotton     

D. Chen    Y. Ding    W. Guo    T. Zhang 《Plant Breeding》2009,128(2):193-198

Two genic male sterile (GMS) lines, Lang-A conditioned by ms ₁₅ and Zhongkang-A conditioned by ms ₅ ms ₆ duplicate recessive genes in Gossypium hirsutum L., were chosen to map GMS genes. These two lines were crossed with Gossypium barbadense cv. 'Hai7124' to produce segregating populations. The ms ₁₅ gene was mapped on chromosome 12, and was flanked by two simple sequence repeat (SSR) markers, NAU2176 and NAU1278, with a genetic distance of 0.8 and 1.9 cM respectively. The ms ₅ and ms ₆ genes were mapped to one pair of homoeologous chromosomes, ms ₅ on chromosome 12 flanked by three SSR markers, NAU3561, NAU2176 and NAU2096, with genetic distances of 1.4, 1.8 and 1.8 cM, respectively, and ms ₆ on chromosome 26 flanked by two SSR markers, BNL1227 and NAU460, with a genetic distance of 1.4 and 1.7 cM respectively. These tightly linked markers with the ms ₁₅ , ms ₅ and ms ₆ genes can be used in the marker-assisted selection among segregating populations in a breeding programme, and provide the foundation for gene isolation by map-based cloning for these three genes.  相似文献   

狐貉源犬瘟热病毒融合蛋白基因的同源性分析   总被引：3，自引：0，他引：3  

张莉  华育平  曾祥伟  刘澜澜 《东北林业大学学报》2008,36(3):70-72,77

2004—2006年,从黑龙江省和内蒙等地发病饲养狐、貉中分离获得6株犬瘟热病毒(CDV),分别命名为MS01、SC01、ZD01、GN01、HL01、NM01分离株。根据Genbank上发表的的CDV F基因序列设计了1对特异性引物,应用RT-PCR方法分别对6个分离株F基因进行克隆、测序,并推导其氨基酸序列,绘制出F基因的遗传进化树。结果表明:6个CDV分离株F基因的开放阅读框架(ORF)均为1 989 bp,编码663个氨基酸,蛋白结构中的13个Cys残基位点和4个潜在的糖基化位点均高度保守,其裂解位点的氨基酸序列为220R-R-Q-R-224R。遗传进化分析表明:6个分离株与CDV代表强毒株A75/17属于同一谱系,均为强毒株。其中,HL01株与海豹瘟热病毒2型(PDV-2)亲缘关系较近,与其它5个分离株关系相对较远。  相似文献   

玉米果穗秃尖性状遗传分析   总被引：5，自引：0，他引：5  

孟昭东  张发军  丁照华  孙琦  汪黎明  郭庆法  王洪刚 《中国农业科学》2008,41(1):280-285

 【目的】采用秃尖与不秃尖两种结实类型的育种材料分析玉米果穗秃尖性状的遗传特点。【方法】试验以不秃尖类型自交系lx01-3和秃尖类型自交系wx04-1及其F1、F2、BC1、BC2群体为试验材料,采用低密度种植以促进植株个体发育,分析各世代群体的果穗秃尖表现。【结果】F1代植株的果穗均为秃尖类型。果穗秃尖与不秃尖类型的分离比例在F2群体中为12.78﹕1,在BC1群体中为2.75﹕1。经卡方测验,符合2对基因间存在重叠作用的遗传特点。SPSS分析表明,F2群体中秃尖类型果穗的秃尖长度性状分布曲线呈明显的偏态分布。【结论】（1）结实类型是由2对存在重叠作用的基因所控制的质量性状。秃尖类型对不秃尖类型为显性;（2）秃尖长度属于存在主效基因作用的数量性状。  相似文献   

cDNA微阵列鉴定差异表达基因的可靠性分析     

王冰林  柳俊  李媛媛  谢从华 《华北农学报》2008,23(3)

应用马铃薯晚疫病水平抗性相关的1 009条表达序列标签(ESTs)所制备的cDNA微阵列和荧光信号Cy3,Cy5杂交系统,通过同一样品RNA的自身比较试验及与不同样品RNA的差异分析试验,对该技术的重复性和可靠性进行了验证分析。结果表明,在自身比较试验中,芯片杂交的假阳性率仅为0.79%,相关系数高达0.981 1;在差异分析试验中,同向标记、正反向标记、同一芯片内2个重复杂交结果的相关系数分别为0.966 0,0.889 5,0.980 2。研究结果证实,cDNA芯片技术具有高度的可靠性,可以鉴定出显著差异表达基因,并构建高质量的基因表达数据库,通过3次生物学重复和2次技术重复试验(采用正、反向标记)即可克服绝大部分试验误差。  相似文献   

转基因林木中安全标记基因的研究进展   总被引：1，自引：0，他引：1  

续晨  王伟东  查琳  诸葛强 《分子植物育种》2008,6(5)

近年来,林木转基因研究进展迅速。但是,由于基因工程技术中使用选择标记基因可能对生态、环境、非靶标生物等带来危害,标记基因的安全性问题已越来越受到人们的关注。据报道在基因工程研究特别是植物转基因研究中使用安全标记基因遗传转化系统方面已作了大量尝试。本文主要综述林木植物转基因研究中应用安全标记基因遗传转化系统的研究现状,重点介绍了正选择标记的使用,可重组系统去除标记基因,利用位点特异性重组去除叶绿体DNA中标记基因的叶绿体遗传转化表达载体系统,及未来基于高转化率和大量快捷PCR筛选的完全无选择标记基因的遗传转化体系等研究进展与应用前景。  相似文献   

利用KASP标记检测青海和西藏小麦品种中光周期基因分布     

权有娟  袁飞敏  刘德梅  李想  陈志国 《麦类作物学报》2019,(10):1165-1172

为了解青海和西藏小麦品种中光周期基因的分布情况,采用KASP标记对青海和西藏249份小麦品种光周期基因 Ppd-D1、 Ppd-B1和 Ppd-A1等位变异组成进行检测。结果显示,在 Ppd-B1位点上,237个品种携带光周期不敏感型等位变异 Ppd-B1a(95.18%),12个品种携带光周期敏感型等位变异 Ppd-B1b(4.82%);在 Ppd-A1位点上,233个品种携带光周期不敏感型等位变异 Ppd-A1a(93.57%),16个品种携带光周期敏感型等位变异 Ppd-A1b(6.43%);在 Ppd-D1位点上,221个品种携带光周期不敏感型等位变异 Ppd-D1a(88.76%),28个品种携带光周期敏感型等位变异 Ppd-D1b(11.24%)。光周期不敏感型等位变异 Ppd-B1a和 Ppd-A1a分别在青海和西藏小麦品种光周期反应中占主导地位。西藏和青海小麦品种中共存在6种等位变异组合类型,其中青海小麦品种中存在5种等位变异组合类型,不存在 Ppd-D1a/Ppd-B1b/Ppd-A1a类型,西藏农家品种中存在4种等位变异组合类型,不存在含光周期敏感型等位变异 Ppd-A1b的类型。等位变异组合 Ppd-D1a/Ppd-B1a/Ppd-A1a在青海和西藏小麦品种中分布最广。  相似文献