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小麦黄矮病是由大麦黄矮病毒(Barley yellow dwarf virus, BYDV)引起的小麦重要病毒病。分离于小麦–中间偃麦草易位系的蛋白激酶编码基因TiDPK1, 是一个抗小麦黄矮病相关基因。本文报道利用酵母双杂交技术和双分子荧光互补技术对TiDPK1与BYDV外壳蛋白(coat protein, CP)互作的研究结果。酵母双杂交分析结果表明, TiDPK1能够与BYDV-GAV、-PAV株系的CP互作, 双分子荧光互补分析结果进一步表明, TiDPK1可与BYDV的CP互作、产生双分子荧光互补信号, 说明TiDPK1确可与BYDV CP相互作用, 该结果对了解TiDPK1在小麦抗BYDV反应机制具有一定意义。 相似文献
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利用BSMV-VIGS技术快速分析小麦TNBL1基因的抗黄矮病功能 总被引:1,自引:0,他引:1
小麦黄矮病是由蚜虫介导的大麦黄矮病毒(Barley yellow dwarf virus, BYDV)侵染引起的小麦重要病害之一。利用cDNA-AFLP分析,筛选出在抗黄矮病小麦易位系YW642中特异表达的长度为292 bp的 cDNA片段,以此片段为启始序列,利用RACE和RT-PCR技术克隆出该基因的全长cDNA序列,推导该基因编码1个NBS-LRR蛋白,将其命名为TNBL1。本研究利用大麦条纹花叶病毒(BSMV)诱导的基因沉默(virus-inducing gene silencing, VIGS)技术,快速分析TNBL1是否参与小麦抗黄矮病反应。通过PCR添加酶切位点、定向酶切与连接,将TNBL1特异的292bp片段反向整合到BSMV-γ链的多克隆位点上,获得重组载体BSMV-γ: TNBL1as,体外转录BSMV-VIGS载体的3个组分(BSMV-TNBL1as、BSMV-α和BSMV-β),等量混合、摩擦接种到抗黄矮病的小麦易位系YW642幼苗叶片上,使YW642中TNBL1基因沉默,然后接种BYDV病原进行黄矮病抗性鉴定。结果表明,TNBL1基因沉默后的YW642对BYDV敏感、显现感病症状,其体内BYDV含量较未发生基因沉默的YW642中的明显增加,证明TNBL1基因是正向调控小麦抗BYDV反应的1个重要基因。 相似文献
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抗黄矮病小麦生物技术育种研究进展 总被引:3,自引:0,他引:3
由蚜虫介导的大麦黄矮病毒引起的小麦黄矮病,难以预测,一旦发病尚无药可治,有“小麦癌症”之称,近年有扩展和危害加重的趋势。综述了国内外在小麦近缘抗病基因发掘与利用、基于BYDV自身基因和蚜虫传毒蛋白基因进行生物技术、基因工程育种方面的进展,介绍了利用生物技术培育抗黄矮病小麦新种质与品种的情况,植物抗黄矮病基因遗传与定位的研究进展,提出了今后研究与发展方向。 ; 相似文献
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研究分子标记鉴定大麦抗黄矮病基因Yd2的有效性,可为Yd2基因在大麦抗病育种中的广泛应用提供快速有效的分子辅助选择工具。利用与Yd2基因紧密连锁的YLM、CAPS-Ylp和ASPCR-Ylp标记同时检测52份国内外大麦品种(系)与4份大麦F1杂种的Yd2基因型,同时结合生物学抗性检测的表型分析其有效性。通过对Yd2基因型已知的20份大麦品种(系)及4个F1杂种的Yd2基因型分析,表明YLM、CAPS-Ylp与ASPCR-Ylp标记可以有效判断大麦Yd2基因型。进一步用这3个标记检测32份Yd2基因型未知的大麦的基因型,鉴定出基因型为Yd2-/Yd2-的品种(系) 27份,基因型为Yd2+/Yd2+的品种(系) 5份。在回交育种的分子辅助选择实例中,从BC2F2世代中选出了16个基因型为Yd2+/Yd2+的单株。3个分子标记结合应用能够快速有效地鉴定大麦Yd2基因型,可用于Yd2基因回交育种中的大规模分子标记辅助选择。 相似文献
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小麦抗病基因类似序列BRG1的分离与功能分析 总被引:1,自引:0,他引:1
利用cDNA-AFLP技术筛选到1个在抗黄矮病的小麦-中间偃麦草易位系YW642中特异表达的小麦抗病基因类似序列(BYDV resistance-related gene1, BRG1)的基因片段。利用cDNA末端快速扩增技术(RACE)和RT-PCR技术,从YW642中分离出BRG1基因全长cDNA序列,获得了一个通读的抗病同源基因cDNA序列,编码由645个氨基酸组成的蛋白质,包含1个NB-ARC保守结构域和3个LRR结构域,该蛋白属于nucleotide-site binding, leucine-rich repeats (NBS-LRR)家族。荧光定量或半定量RT-PCR表达分析表明,BRG1在抗病小麦易位系YW642叶片中优势表达,受BYDV的诱导,BYDV接种后48 h表达量最高,BRG1在感病小麦亲本中8601中表达量始终较低,随BYDV接种时间延长呈轻微的下调趋势;而且外源激素水杨酸(SA)与茉莉酸(JA)处理可上调该基因在YW642中的表达。利用病毒介导的基因沉默技术分析了BRG1基因的功能,结果表明该基因沉默的抗病小麦易位系YW642中BYDV相对含量增加,但未造成抗病性表型显著改变,说明该基因参与抗黄矮病反应,但不是小麦抗黄矮病重要基因。 相似文献
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大麦黄矮病毒引起的小麦黄矮病是我国小麦生产上重要的病毒病。BYDV GAV株系是我国小麦黄矮病的主要病原,目前有关BYDV GAV编码蛋白质P1、P2和CP的功能研究鲜有报道,因此,研究P1、P2和CP的功能,可为深入研究BYDV GAV的致病机制奠定基础。通过Mega 7.0构建系统进化树,对BYDV GAV株系编码蛋白质P1、P2和CP的核苷酸序列进行同源进化分析;通过构建P1、P2和CP的YFP表达载体,转化GV3101农杆菌后浸润本氏烟,激光共聚焦显微镜观察确定3个蛋白质的亚细胞定位;构建BYDV GAV 5个编码蛋白质的双分子荧光互补载体,转化GV3101农杆菌后分别与P1、P2和CP共浸润本氏烟叶片,通过激光共聚焦显微镜观察确定这3个蛋白质与其他病毒蛋白质的体内互作情况;通过构建P1、P2和CP的马铃薯X病毒(PVX)异源表达载体,转化GV3101农杆菌后接种本氏烟,接种后5 d观察症状形成,并采集系统叶进行病毒积累量检测,研究3个蛋白质对该病毒致病性的影响。结果表明,BYDVs的4种分离物中,PAV与GAV在核苷酸水平上的亲缘关系最近。BYDV GAV编码的P1、P2和C... 相似文献
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青海省大麦黄矮病毒的种类鉴定及基于CP基因的分子进化研究 总被引:2,自引:1,他引:1
由大麦黄矮病毒(Barley Yellow Dwarf Viruses,BYDVs)引起黄矮病是世界范围的主要的经济危害严重的禾谷类作物病毒病。BYDVs可侵染大麦、小麦、青稞等多种禾谷类作物,2010年该病害在青海省东部麦区中度流行。为了明确青海麦区大麦黄矮病毒株系种类,应用酶联免疫吸附法和核酸斑点杂交方法对采集到的112个麦类黄矮病标样进行检测。结果显示,GAV为当地大麦黄矮病毒的流行株系。测定了12个青海GAV分离物的外壳蛋白基因序列;核苷酸和氨基酸序列间比对分析表明:青海GAV分离物与中国各地GAV分离物外壳蛋白基因相似性非常高,变异很小。掌握青海东部麦区大麦黄矮病毒的株系分布及分子变异情况,对麦类作物的抗病性育种工作提供有价值的参考,同时对指导该地区小麦黄矮病的防治有着非常重要的意义。 相似文献
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J. Ovesná J. Vacke L. Kucera J. Chrpová I. Nováková A. Jahoor V. ip 《Plant Breeding》2000,119(6):481-486
The inheritance of resistance to barley yellow dwarf virus (BYDV) was studied in the selected 24 spring and winter barley cultivars that showed a high or intermediate resistance level in 1994‐97 field infection tests. The polymerase chain reaction diagnostic markers YLM and Ylp were used to identify the resistance gene Yd2. The presence of the Yd2 gene was detected with both markers in all the resistant spring barley cultivars and lines from the CIMMYT/ICARDA BYDV nurseries. The results of field tests and genetic analyses in winter barley corresponded with marker analyses only when the Ylp marker was used. Genes non‐allelic with Yd2 were detected by genetic analyses and the Ylp marker in moderately resistant spring barley cultivars ‘Malvaz’, ‘Atribut’ and ‘Madras’, and in the winter barley cultivars ‘Perry’ and ‘Sigra’. Significant levels of resistance to BYDV were obtained by combining the resistance gene Yd2 with genes detected in moderately resistant cultivars. The utilization of analysed resistance sources in barley breeding is discussed. 相似文献
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A breeding programme was developed to obtain barley yellow dwarf virus (BYDV)-resistant winter genotypes using the Yd2 gene. The aim was to incorporate the Yd2 allele into the new high-yielding genotypes to release cultivars that allow barley cultivation in areas where BYDV is endemic. The resistant lines were developed using pedigree selection. An ICARDA resistant line (83RCBB130) carrying the Yd2 gene was crossed with three susceptible, high-yielding winter varieties and their F1 lines were either selfed or backcrossed to the matching susceptible parent. The best lines selected from subsequent selfing generations were evaluated in replicated trials in the presence or absence of BYDV, starting from F6 and BC1F5 to F8 and BC1F7 generations. Four genotypes with superior agronomic traits and BYDV resistance were selected. 相似文献
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应用GISH与STS标记鉴定小麦-中间偃麦草抗黄矮病端体系 总被引:3,自引:3,他引:0
由大麦黄矮病毒引起的小麦黄矮病毒病是一个严重病害,至今在小麦属内还没有发现抗源。中间偃麦草2Ai-2染色体携带一个高抗黄矮病基因,对该基因的染色体臂定位将为制定抗病基因向小麦转移策略,筛选、开发特定的、与抗性连锁的分子标记的研究提供重要信息。本文对由小麦-中间偃麦草二体附加系Z6衍生的3个抗黄矮病端体系进行鉴定,通过分析端体的遗传构成、筛选与端体共分离的STS标记以确定端体在遗传上的染色体臂归属,从而明确BYDV抗病基因的染色体位置。以拟鹅冠草基因组[Pseudoroegneria strigosa (M. Bieb.) Löve,St]DNA为探针,中国春基因组(Triticum aestivum L., ABD) DNA作封阻分别对抗病亲本Z6及抗病端体系N530的根尖体细胞染色体进行原位杂交,结果表明,N530体细胞中有2个端体显示出与Z6中外源染色体2Ai-2短臂相似, 而与长臂不同的杂交信号。以小麦第2同源群的5个RFLP探针的DNA序列为基础,设计了6对PCR引物,对小麦-中间偃麦草二体异附加系、二体代换系和端体系进行扩增,结果表明,基于短臂探针psr126,psr131序列设计的2对引物,可在含有2Ai-2染色体及端体的抗黄矮病材料中特异扩增,而基于长臂探针psr112序列设计的1对引物,可在含有2Ai-2染色体的抗黄矮病材料中特异扩增,但不能在端体系进行特异扩增,证明外源端体为2Ai-2染色体的短臂。本研究不仅将黄矮病抗性基因定位于2Ai-2染色体的短臂上, 而且由RFLP探针psr126、psr131和psr112序列转化的标记STS126 (sequence tagged site) STS131和STS112还可分别作为追踪2Ai-2染色体短臂和长臂的分子标记,用于抗病易位系辅助选择。 相似文献
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The enzyme-linked immunosorbent assay (ELISA) was used to determine the concentration of barley yellow dwarf virus (BYDV) in leaves of spring wheat Triticum aestivum L. cv. ‘Fukuhokomugi’, Leymus angustus (Trin.) Pilger (Altai wild rye) and their F1 hybrids inoculated at the 2- to 3-leaf stage. The BYDV isolate “Cloutier” (serotype PAV) was used. The results showed that L. angustus confers immunity or strong resistance to the virus. Significant multiplication of the virus occurred in ‘Fukuhokomugi’ which expressed an intermediate level of tolerance to the disease under field conditions. The F1 haploid hybrids of ‘Fukuhokomugi’×L. angustus expressed a level of resistance almost equal to that of their wild parent, with a low concentration of virus appearing 16 days after inoculation and no more afterwards. The discovery of BYDV resistance in L. angustus and the expression of this character in the F1 hybrids provide new opportunities for the enlargement of the gene pool for BYDV resistance in wheat. 相似文献
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Barley yellow dwarf disease (BYDD) is one of the main viral diseases of small grain cereals. This disease, reported on numerous plant species of the Poaceae family, is caused by a complex of viral species including the species Barley yellow dwarf virus‐PAV (BYDV‐PAV, family Luteoviridae, genus Luteovirus), frequently found in western Europe. Resistance sources towards BYDD are scarce. Indeed, breeding‐resistant genotypes is a long and expensive process. Thus, estimating the durability of the resistance genes before the achievement of selection would be an asset for breeders. One isolate of BYDV‐PAV has been serially passaged on two hosts, ‘Zhong ZH’ and ‘TC14’, carrying a gene for partial resistance. The resulting viral population showed an increase of the speed of development of the infection in controlled conditions. In this study, these viral populations were evaluated in a 3‐year field trial, including a susceptible host, ‘Rendezvous’, and a host carrying the resistance gene of ‘TC14’ in a ‘Rendezvous’ background, to assess the effect of serial passages in field conditions. Results indicate that isolates issued from serial passages on hosts carrying a gene for partial resistance induced increased damage in field conditions when compared with the initial isolate. Yield losses are mainly due to a decrease of the number of kernels per square metre. The interest on using partial resistance gene to control BYDD is discussed. 相似文献
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Marker-assisted backcross introgression of the Yd2 gene conferring resistance to barley yellow dwarf virus in barley 总被引:4,自引:0,他引:4
S. P. Jefferies B. J. King A. R. Barr P. Warner S. J. Logue P. Langridge 《Plant Breeding》2003,122(1):52-56
YLM, a codaominant polymerase chain reaction (PCR) marker linked to Yd2, could substantially improve the precision and efficiency of barley yellow dwarf virus (BYDV) resistance breeding. The aim of this study was to assess the effectiveness of YLM in a marker‐assisted introgression programme and to quantify associations between the presence of Yd2 and other agronomic and quality traits. The Yd2 gene was introgressed into a BYDV‐susceptible background through two cycles of marker‐assisted backcrossing. BC2 F2‐derived lines, either carrying or not carrying the YLM allele associated with resistance, were compared in the presence and absence of BYDV. The YLM marker was shown to be effective in the introgression of Yd2. Lines carrying the YLM allele associated with resistance produced significantly fewer leaf symptoms and showed a reduction in yield loss when infected with BYDV. There were no deleterious effects associated with the introgression of Yd2 on grain yield, grain size or malting quality. The implications of marker‐assisted selection for Yd2 on barley improvement are discussed. 相似文献
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K. K. Nkongolo 《Euphytica》1996,90(3):337-344
Summary The Barley Yellow Dwarf Virus disease (BYDV) and the Russian wheat aphid (RWA) Diuraphis noxia (Mordvilko) have caused significant losses to wheat and barley in several areas of the world. Important sources of resistance to both BYDV and RWA have been found in Triticale. Different generations of interspecific wheat x Triticale crosses were produced and the progenies were screened for BYDV and RWA tolerance. Plants with equal chromosome numbers showed different levels of fertility. A significant correlation was observed between pollen fertility and seed set in primary florets (r=0.57). In generaL, pollen fertility, seed set and the number of euploid plants (2n=42) increased from one generation to the next. The expression of BYDV tolerance varied from population to population. Additive effects were predominant in F1 and some backcross populations. A dominant effect of rye tolerance genes was also observed in few populations. A monogenic trait or a quantitative (polygenic) character would not agree with the observed segregation patterns. The heritability of this oligogenic tolerance was quite different between populations and in many populations the tolerance genes were only partially expressed. Some transgressive segregation for tolerance and sensitivity was demonstrated. The genes controlling tolerance to RWA in Triticale lines, Muskox 658 and Nord Kivu were not expressed in advanced lines resistant to BYDV. This indicates that tolerance genes for BYDV and RWA in these lines are located on different chromosomes. 相似文献
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抗黄矮病小麦种质的分子标记 总被引:10,自引:0,他引:10
应用基因组原位杂交技术分析了抗小麦黄矮病种质的遗传组成,研究表明小麦一中间但麦草部分双二倍体无芒中4(2n=56)具有40条小麦染色体、5对中间僵麦草染色体、3对小麦/中间僵麦草易位染色体,其中1对是罗伯逊氏易位染色体。结果表明无芒中4与远中5的遗传组成有明显差异,是两种不同类型的材料。抗黄矮病小麦种质F940418, T10 相似文献