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1.
抗白粉病小麦-中间偃麦草双体异附加系的鉴定 总被引：14，自引：2，他引：14
对从中间偃麦草与普通小麦品种烟农15杂交后代(BC3F6)中选育的双体异附加系山农Line15的形态学、白粉病抗性、细胞学、基因组荧光原位杂交(GISH)及RAPD进行鉴定分析.结果表明它的主要形态性状介于双亲之间;白粉病抗性鉴定结果表明山农Line15对白粉病高抗近免疫;根尖细胞染色体数目为2n=44,PMC MI染色体构型为2n=22Ⅱ;以中间偃麦草总基因组DNA为探针的基因组荧光原位杂交(GISH),结果表明山农Line15是在小麦的遗传背景中附加了2条中间偃麦草染色体,为小麦-中间偃麦草双体异附加系;遗传分析表明山农Line15的抗白粉病基因基本上可以确定来源于中间偃麦草的染色体;RAPD分析表明:在供试的120个随机引物中有1个引物S170(-5'-ACA ACG CGA G-3'-)能在山农Line15中稳定地扩增出特异带型,可以作为山农Line15所附加的中间偃麦草染色体的特异分子标记. 相似文献
2.
抗白粉病小麦-中间偃麦草染色体小片段易位系的选育与鉴定 总被引：1，自引：0，他引：1
对从普通小麦与中间偃麦草杂交后代中选育的一个抗白粉病新品系(AF-1)进行了形态学、细胞学和原位杂交(GISH)鉴定。结果表明:AF-1具有与小麦亲本相似的农艺性状,根尖细胞染色体数目为2n=42,花粉母细胞减数分裂中期(PMCMI)染色体构型为2n=21Ⅱ,且未见其他结构变异,细胞学上十分稳定。GISH结果表明,AF-1为中间偃麦草与普通小麦的一个小片段易位系,易位点位于一对染色体臂的中部偏着丝粒的位置。结合抗病调查结果,推断该片段上携带了一个来自偃麦草的抗白粉病基因。该片段可能是在杂种早期世代通过部分同源染色体配对或者单价染色体错分裂而形成的。 相似文献
3.
建立马铃薯晚疫病菌抗甲霜灵SCAR(sequenced characterized amplified region,序列特征扩增区域)标记,以马铃薯晚疫病菌对甲霜灵高抗菌株HD01-3和对甲霜灵高感菌株DK98-1为亲本,通过无性单游动孢子分离和有性杂交获得菌株HD01-3无性后代群体、菌株DK98-1无性后代群体以及F1代分离群体,以此为试验材料,利用BSA法(bulked segregant analysis,分离群体分组分析法)构建抗感基因池对后代菌系的甲霜灵抗性进行RAPD(random amplified polymorphic DNA,随机扩增多态性DNA)分析。从178条RAPD随机引物中找到一条特异性引物S2054,其可以扩增出一个与晚疫病菌对甲霜灵抗性相关的遗传标记,将该特异条带回收、克隆、测序,发现此标记大小为457bp,根据测序结果设计特异PCR引物,用于扩增抗感基因池,成功地将特异RAPD标记转化为SCAR标记。初步建立了马铃薯晚疫病菌抗甲霜灵SCAR标记,辅助监测晚疫病菌对甲霜灵的抗性。 相似文献
4.
本研究对以小麦-中间偃麦草异附加系L1和小麦-中间偃麦草部分双二倍体‘无芒中4’为抗源选育出的抗黄矮病小麦新品系进行分子检测和抗病性鉴定.通过应用RAPD、SSR、SCAR 3种分子标记OPF15、Xgwm37、SC-W37进行分子检测,并采用人工接种和大田自然感病的方式进行抗黄矮病鉴定,筛选到了‘93646’、‘2003-2’等高抗黄矮病的小麦新品系.分子检测抗黄矮病基因与田间抗病鉴定结果基本一致,应用的3种PCR标记都可以检测出抗病材料,但SCAR标记SC-W37特异性强、稳定性好,可在小麦抗黄矮病育种早代选择过程中发挥重要作用. 相似文献
5.
对从中间偃麦草与普通小麦品种烟农15杂交后代(BC3F6)中选育的双体异附加系山农Line15的形态学、白粉病抗性、细胞学、基因组荧光原位杂交(G ISH)及R A PD进行鉴定分析。结果表明,它的主要形态性状介于双亲之间;白粉病抗性鉴定结果表明,山农Line15对白粉病高抗近免疫;根尖细胞染色体数目为2n=44,PM CMⅠ染色体构型为2n=22Ⅱ;以中间偃麦草总基因组D N A为探针的基因组荧光原位杂交(G ISH),结果表明,山农Line15是在小麦的遗传背景中附加了2条中间偃麦草染色体,为小麦—中间偃麦草双体异附加系;遗传分析表明,山农Line15的抗白粉病基… 相似文献
6.
西瓜抗小西葫芦黄花叶病毒基因的连锁分子标记研究 总被引：10，自引：0，他引：10
小西葫芦黄花叶病毒中国株系(Zucchini yellow mosaic virus Chinese strain,ZYMV-CH)是危害我国西瓜的主要病毒。本实验以抗病毒病西瓜野生种质P.I.595203与感病的普通西瓜自交系98R为亲本,采用单粒传方式得到109个E代株系,分别对亲本、F1及109个F3代株系群体进行了苗期抗ZYMV-CH接种鉴定,通过F3代群体的抗感分离情况,推测得到F2代各单株的基因型,采用集团分离分析法(bulked segregant analysis,BSA)在F2代建立抗感基因池,以亲本、F1和抗感基因池为模板,对640条RAPD引物进行PCR扩增筛选,其中引物AK13在亲本、F1和抗感基因池之间扩增出一条多态性片段(644bp),在F2代群体上验证该多态性条带与ZYMV-CH的抗性基因呈现连锁关系,遗传连锁距离为8cM,定名为AK13-644,该连锁标记在ZYMV-CH抗性转育后代自交系上得到了验证。最终将此RAPD标记成功转化成SCAR标记SCAK13-644,该标记可以作为西瓜抗病毒病辅助选择的分子标记。 相似文献
7.
小麦抗叶锈病基因Lr38位于中间偃麦草 (Agropyron intermedium)第7组的一条染色体上,是抗性很强的基因,国内外至今尚未发现对 Lr38有毒性的菌株,是一个应用潜力很大的抗病基因。通过对目前已克隆的抗病基因结构分析,大部分抗病基因存在一些保守区域,利用保守域序列设计简并引物,PCR扩增抗病基因相似序列(Re- sistance gene analogs,RGA),来进行抗病基因克隆是一条很好的途径。本研究根据已克隆基因的蛋白激酶类保守序列设计简并引物RAF3和RAR4,对TcLr38及感病亲本Thatcher进行PCR扩增,旨在明确与小麦抗叶锈病基因Lr38相关的蛋白激酶,为克隆Lr38奠定基础。 相似文献
8.
小麦新抗源CH223抗条锈性的遗传分析及细胞学鉴定 总被引：1，自引：0，他引：1
用条中(CYR)30、31、32和33号对小麦抗病新品系CH223及其亲本进行抗性评价,分析其对条中32号的抗性遗传方式,并研究其细胞学特征。结果显示,CH223苗期和成株期对上述4个生理小种表现出免疫或近免疫的抗性水平,并具有与其抗性供体TAI7047及其野生亲本中间偃麦草相似的抗病反应型;抗×感的F1代均为免疫,反应型为0~0;型,且F2、F2:3、BC1代的抗、感分离比均符合1对显性基因控制的分离模式;CH223及其与小麦品种"中国春"等杂种F1的染色体数目均为2n=42,绝大多数的花粉母细胞具有2n=21Ⅱ的配对构型,并能与小麦染色体完好配对。说明CH223不含较大的外源染色体片段,是一个携带偃麦草抗条锈病基因的隐形异源渐渗系,对条中32号的成株抗性受1对显性核基因控制。 相似文献
9.
不同猕猴桃品种RAPD分析及其与抗溃疡病的关系 总被引：1，自引：0，他引：1
对不同猕猴桃品种的分子生物学试验表明：猕猴桃的DNA浓度在920 μg/mL符合RAPD分析的要求。通过60个随机引物的PCR扩增,报道了6个不同品种和类型猕猴桃种质资源的RAPD多态性,计算了它们之间的遗传距离,构建了聚类图,并讨论了其亲缘关系。聚类分析图反映出来源于安徽省主要猕猴桃产区的6个样品可以分为3组,其中抗病与感病的相对较为集中,由此可推断出现这种聚类的原因可能是由于它们基因组中有相同的DNA片段。抗病品系都有一条1 458 bp DNA片段,而感病品系均没有该带。故该片段可能与猕猴桃植株抗溃疡病相关。RAPD多态性从分子水平上反映出了猕猴桃种质资源不同品种及不同类型间复杂的遗传背景,为抗病育种的亲本选配提供了依据,也为合成猕猴桃抗溃疡病探针并用于检测猕猴桃抗溃疡病种质和分子标记辅助育种奠定了基础。 相似文献
10.
为了分析韩国栗疫病的抗病品种和感病品种的遗传变异和抗病性的筛选,利用抗病性的快速检测法和RAPD(random amplified polymorphic DNA)方法对13个栗树品种进行了抗病性检测和RAPD标记分析。抗病性的快速检测选出了5个抗病品种、5个感病品种和3个中度抗病(或中度感病)品种,并且这一结果与该品种的田间表现相一致。利用筛选的12个随机引物,扩增了100个多态性RAPD片段,但未发现与抗病性或感病性相关的特异RAPD片段。聚类分析结果表明,12个品种大致分为抗病、感病和中度抗病(或中度感病)等3个大组,并与抗病性的快速检测结果基本一致。抗病品种“MANSEKI”表现出了相对于12个品种较远的亲缘关系。 相似文献
11.
目前,番茄晚疫病(Phytophthora infestans)在世界各番茄主产区普遍发生,并造成严重的经济损失,已经成为番茄生产的主要障碍之一.防御番茄晚疫病最有效的方法是培育抗病品种.依靠常规方法耗时、耗力,同时由于人为鉴定标准的差异,会直接影响鉴定结果,延长育种周期,分子标记技术的发展为解决此问题提供了可能.分子标记可被用于在苗期进行大量的抗病鉴定,不受时间、地域的限制,从而加速育种工作进程[1].Qiu等[2]已找到1条与抗晚疫病Ph-3基因连锁的RAPD标记,本研究旨在将此RAPD标记转化为稳定的CAPS标记,为抗晚疫病分子辅助选择育种(MAS)奠定良好的基础. 相似文献
12.
Polymerase chain reaction (PCR) assays were used to detect phytoplasmas in foliage samples from Chinaberry ( Melia azedarach ) trees displaying symptoms of yellowing, little leaf and dieback in Bolivia. A ribosomal coding nuclear DNA (rDNA) product (1·8 kb) was amplified from one or more samples from seven of 17 affected trees by PCR employing phytoplasma-universal rRNA primer pair P1/P7. When P1/P7 products were reamplified using nested rRNA primer pair R16F2n/R16R2, phytoplasmas were detected in at least one sample from 13 of 17 trees with symptoms. Restriction fragment length polymorphism (RFLP) analysis of P1/P7 products indicated that trees CbY1 and CbY17 harboured Mexican periwinkle virescence (16SrXIII)-group and X-disease (16SrIII)-group phytoplasmas, respectively. Identification of two different phytoplasma types was supported by reamplification of P1/P7 products by nested PCR employing X-disease-group-specific rRNA primer pair R16mF2/WXint or stolbur-group-related primer pair fSTOL/rSTOL. These assays selectively amplified rDNA products of 1656 and 579 bp from nine and five trees with symptoms, respectively, of which two trees were coinfected with both phytoplasma types. Phylogenetic analysis of 16S rDNA sequences revealed Chinaberry yellows phytoplasma strain CbY17 to be most similar to the chayote witches'-broom (ChWBIII-Ch10) agent, a previously classified 16SrIII-J subgroup phytoplasma. Strain CbY1 resembled the Mexican periwinkle virescence phytoplasma, a 16SrXIII-group member. The latter strain varied from all known phytoplasmas composing group 16SrXIII. On this basis, strain CbY1 was assigned to a new subgroup, 16SrXIII-C. 相似文献
13.
KENTARO YASUDA AZUSA YANO YUICHIRO NAKAYAMA HIROFUMI YAMAGUCHI 《Weed Biology and Management》2002,2(1):11-17
Polymerase chain reaction (PCR) and polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) techniques were applied for establishing the reliable practice in identification of Echinochloa oryzicola Vasing. and E. crus-galli (L.) Beauv. (barnyardgrass). Total DNA was extracted from 18 accessions and 86 individuals of E. oryzicola , 33 accessions and 140 individuals of E. crus-galli var. crus-galli , 23 individuals of E. crus-galli var. praticola , and six individuals of E. crus-galli var. formosensis that were collected from Japan. A partial region of intergenetic spacer between trn T and trn L, and an intron of trn L were amplified separately using a trn-a and trn-b1 primer set, and a trn-c and trn-d primer set, respectively. All individuals of E. oryzicola showed the same fragment amplified by the trn-a and trn-b1 primer set. The fragment was 481 bp in length, and was undigested by Eco R I, whereas all individuals of E. crus-galli , including three botanical varieties, showed the same fragment with a 449-bp length. The fragment was digested by Eco R I into two fragments (178 and 271 bp). The fragment amplified by the trn-c and trn-d primer set in all individuals of E. oryzicola was digested by Alu I into two fragments (174 and 452 bp), but undigested by Dra I. In contrast, the fragment amplified by the trn-c and trn-d primer set in all individuals of E. crus-galli was digested by Dra I into two fragments (134 and 487 bp), but undigested by Alu I. There was no intraspecific variation in these regions; thus, these two species are easily identifiable by using our method. 相似文献
14.
Tomato spotted wilt virus was recorded for the first time in Jordan on tomato plants. Severe disease symptoms were observed in different tomato farms in the Jordan Valley. Using a specific primer pair a fragment of the capsid protein gene of the virus has been amplified by RT-PCR and IC-RT-PCR. The amplified PCR product was cloned and sequenced. Sequence analysis revealed that the Jordanian isolate of TSWV shared high nucleotide similarities with other isolates from different countries. The sequence of the capsid protein gene was deposited in GenBank under the accession number AY646682 . The response of different tomato breeding lines and hybrids, previously developed for resistance against Tomato yellow leaf curl virus (TYLCV) were tested for their reaction to TSWV infection. All tested lines and hybrids were susceptible to TSWV infection. This has been confirmed at the molecular level by using the SCAR 421 marker linked to the TSWV resistance gene Sw-5 . 相似文献
15.
用 ELISA 法鉴定了小麦近缘种赖草属(Leymus)、披碱草属(Elymus)、鹅冠草属(Roegneria)3个属的21个种,其中17个种抗 BYDV。21145份小麦品种中筛选到症状轻、病毒含量高的耐病品种忻县冬麦、江西早等29份。3604份大麦品种中筛选到症状轻、病毒含量低的抗病品种C13208、小麦近缘种(Agropyronintemedium)和普通小麦杂交的异源八倍体中4无芒,中5,远中7,陇远45、46,远中1001,忻4079以及附加系 L1。现已获得抗 BYDV 的以中4无芒、L1为亲本的杂交后代。 相似文献
16.
亚麻品系9801-1抗白粉病基因的RAPD标记 总被引：2，自引：0，他引：2
F2 populations were obtained from the cross between 9801-1 and DIANE.Bulked segregate and RAPD analyses were employed to identify molecules linked to the resistance to powdery mildew.OPP02 amplified about 792 bp polymorphic band in all individuals from 9801-1 and resistant bulk,but absent in all individuals from DIANE and susceptible bulk.By further analysis in F2 segregating population,the polymorphic band was found to be cosegregated with the resistant gene possibly.The fragment was sequenced, 相似文献
17.
Downy mildew is a destructive disease of spinach worldwide. There have been 10 races described since 1824, six of which have been identified in the past 10 years. Race identification is based on qualitative disease reactions on a set of diverse host differentials which include open-pollinated cultivars, contemporary hybrid cultivars, and older hybrid cultivars that are no longer produced. The development of a set of near-isogenic open-pollinated spinach lines (NILs), having different resistance loci in a susceptible and otherwise common genetic background, would facilitate identification of races of the downy mildew pathogen, provide a tool to better understand the genetics of resistance, and expedite the development of molecular markers linked to these disease resistance loci. To achieve this objective, the spinach cv. Viroflay, susceptible to race 6 of Peronospora farinosa f. sp. spinaciae, was used as the recurrent susceptible parent in crosses with the hybrid spinach cv. Lion, resistant to race 6. Resistant F(1) progeny were subsequently backcrossed to Viroflay four times with selection for race 6 resistance each time. Analysis of the segregation data showed that resistance was controlled by a single dominant gene, and the resistance locus was designated Pfs-1. By bulk segregant analysis, an amplified fragment length polymorphism (AFLP) marker (E-ACT/M-CTG) linked to Pfs-1 was identified and used to develop a co-dominant Sequence characterized amplified region (SCAR) marker. This SCAR marker, designated Dm-1, was closely linked ( approximately 1.7 cM) to the Pfs-1 locus and could discriminate among spinach genotypes that were homozygous resistant (Pfs-1Pfs-1), heterozygous resistant (Pfs-1pfs-1), or homozygous susceptible (pfs-1pfs-1) to race 6 within the original mapping population. Evaluation of a wide range of commercial spinach lines outside of the mapping population indicated that Dm-1 could effectively identify Pfs-1 resistant genotypes; the Dm-1 marker correctly predicted the disease resistance phenotype in 120 out of 123 lines tested. In addition, the NIL containing the Pfs-1 locus (Pfs-1Pfs-1) was resistant to multiple races of the downy mildew pathogen indicating Pfs-1 locus may contain a cluster of resistance genes. 相似文献
18.
Severe mosaic, yellowing and stunting symptoms were observed on petunia (Petunia hybrida L.) growing in pots at NBRI and in various gardens of Lucknow, India. The association of Cucumber mosaic virus (CMV) with the mosaic disease was detected based on positive bioassay on susceptible hosts, isometric cored virus particles of ~28?nm during electron microscopic observations in leaf dip preparations and positive amplification of expected size (~650?bp) during RT-PCR using coat protein gene specific primers. Further, the complete RNA 3 genomic fragment of virus isolate was amplified by RT-PCR using RNA 3 specific primers. The obtained amplicons of ~2.2 Kb were cloned and sequenced. The analysis of sequence data of RNA 3 revealed highest sequence identities (96%) with several CMV strains which belong to subgroup IB. The virus isolate also showed closest phylogenetic relationships with banana strain of CMV of subgroup IB (Acc. EF178298) reported from India. To the best of our knowledge, we report the first molecular characterization of CMV strain of subgroup IB causing severe mosaic disease on petunia in India. 相似文献
19.
云南泡桐丛枝病植原体核糖体蛋白基因片段序列分析 总被引：3，自引：0，他引：3
应用植原体核糖体蛋白基因通用引物对rpF1/rpR1,对采自云南省曲靖市的泡桐丛枝病植原体DNA (PaWB-QJ)进行PCR扩增,得到1.3 kb的特异片段,证明此病株中存在植原体。将此片段与pGEM-T Easy载体连接并转化大肠杆菌JM109感受态细胞,进行PCR鉴定、核糖体蛋白基因部分核苷酸序列测定及分析。结果表明,该株系(PaWB-QJ)核糖体蛋白基因片段长1 244 bp,包含rps19、rpl22和rps3基因。对PaWB-QJ株系的核糖体蛋白基因序列的同源性比较结果显示与16S rI-B亚组的翠菊黄化(Aster yellows,AY)、长春花黄化(Periwinkle yellows,PY)和泡桐丛枝德国株系(Paulownia witches'-broom,PaWB-German)的亲缘关系最近,达到99.0%以上,而与其它组中的株系明显低于97.0%,所以认为该植原体株系属于翠菊黄化组B亚组(16SrI-B)。 相似文献