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1.
利用小麦微卫星引物建立偃麦草Ee染色体组特异SSR标记   总被引:4,自引:0,他引:4  
选用40对小麦SSR引物对17份偃麦草(Thinopyrum sp.)、2份小麦(Triticum aestivum)材料进行了PCR扩增分析,从中筛选到引物Xgwm325能在不同偃麦草材料中扩增出4条长度分别为1400、440、120和100bp的特异DNA片段,可以作为偃麦草种质的特异SSR标记。利用小麦-二倍体长穗偃麦草(Th.elongatum)异代换系和异附加系对引物Xgwm325进行了扩增鉴定,结果只有100bp左右的片段出现在长穗偃麦草所有E^e组染色体上,该片段可以作为E^e染色体组的特异SSR标记。  相似文献   

2.
为了打破小麦抗赤霉病育种种质资源狭窄的局限,丰富抗赤霉病遗传材料,本研究通过对中国春-长穗偃麦草二体异代换系DS7E/7A与扬麦16杂交后代进行辐射诱变,以产生小麦-长穗偃麦草易位系选择群体,并结合赤霉病抗性鉴定和长穗偃麦草染色体特异分子标记辅助选择,最终获得了5个小麦-长穗偃麦草抗赤霉病易位单株,创建了一条有效获得小麦-长穗偃麦草抗赤霉病易位系的技术路线。小麦-长穗偃麦草抗赤霉病易位系的成功创建不仅为小麦抗赤霉病育种提供了重要的中间材料,也将为小麦近缘种的抗赤霉病研究和利用提供理论与实践参考。  相似文献   

3.
摘要:以普通小麦(Triticum aestivum)中国春、长穗偃麦草?穴Thinopyrum elongatum ?雪及其双二倍体、二体异附加系、二体异代换系为材料,采用SDS-PAGE分析了种子高分子量麦谷蛋白亚基。长穗偃麦草高分子量麦谷蛋白基因在中国春背景中编码一条高分子量麦谷蛋白亚基,其迁移率与中国春1By8亚基相同,命名为1E8亚基,控制该亚基的基因位点Glu-E1位于长穗偃麦草E组染色体第一同源群的长臂上。用高分子量麦谷蛋白y亚基基因重复区域的特异引物进行扩增,长穗偃麦草1E8亚基编码基因(Glu-E1)扩增出1 300 bp的片段,而中国春1By8亚基编码基因(Glu-B1y)扩增出1 950 bp的片段。  相似文献   

4.
小麦品系CH5383是源于中间偃麦草(Thinopyrum intermedium)的兼抗多种小麦病害的新种质。为明确其抗性来源和外源DNA片段的渗入位置,采用基因组原位杂交(GISH)和荧光原位杂交(FISH)对CH5383进行分析,GISH分析未发现外源信号,FISH结果观察到CH5383的3BL染色体端部与对照小麦(Triticum aestivum)品种中国春相比有明显的重组信号,初步推断CH5383染色体3BL端部可能有中间偃麦草DNA片段插入。用135对PLUG(PCR-based landmark unique gene)标记对CH5383、中国春和多个近缘物种进行扩增分析,发现位于3B染色体长臂端部的引物TNAC1383,能在CH5383和中间偃麦草中扩增出大约1 300 bp的特异DNA产物。从而进一步证实,CH5383是一个小麦-中间偃麦草小片段渗入系,携带抗条锈病基因的外源中间偃麦草DNA渗入片段位于3B染色体端部0.81-1.00区段。CH5383可以作为优异的小麦抗病育种新种质加以利用。  相似文献   

5.
摘要:为建立簇毛麦特定染色体上的特异性ISSR标记,以二倍体簇毛麦、小麦中国春等为材料,对96条ISSR引物进行PCR筛选,发现引物UBC848可在二倍体簇毛麦中扩出一条长388bp的特异性片段(命名为pDv848/388),而中国春等小麦均未扩出该片段。进而利用UBC848对小麦近缘种偏凸山羊草等进行扩增,发现它们均未扩出该片段。用一套中国春-二倍体簇毛麦附加系CSDA1V~CSDA7V对目标片段进行染色体定位,将目标片段确定在二倍体簇毛麦5V染色体上。进一步用UBC848对多年生簇毛麦、小麦-簇毛麦双二倍体、部分双二倍体及其衍生后代进行扩增,发现它们均能扩出目标片段,这表明,在小麦染色体工程育种中, pDv848/388可作为簇毛麦5V染色体上的一个特异性ISSR标记,用于快速跟踪检测簇毛麦遗传物质向栽培小麦中的导入。  相似文献   

6.
为了建立小麦背景中粗穗披碱草(Elymus trachycaulus)和纤毛披碱草(Elymus ciliaris)染色体的跟踪鉴定方法,本研究利用寡聚核苷酸Oligo-pSc119.2-1和Oligo-pTa-535-1为探针的双色荧光原位杂交(FISH)和以(GAA)8为探针的单色FISH,分别对4个小麦-粗穗披碱草附加系和5个小麦-纤毛披碱草附加系染色体进行分析。结果发现,经与小麦FISH核型比对后,建立了可用于追踪小麦背景中粗穗披碱草1St、5Ht、6Ht和7Ht染色体的标准FISH核型;而纤毛披碱草Sc和Yc染色体FISH信号较弱。FISH检测发现,小麦-粗穗披碱草1St附加系自交自发变成了1St(1BS·3BL)代换系,且在其染色体组中检测到了一对T1BL·3BS易位染色体,同时发现5AS端部寡聚核苷酸Oligo-pSc119.2-1序列发生了删除。另外,在小麦-粗穗披碱草5Ht附加系中发现2B染色体短臂末端删除现象,形成了2B-del和2BS-4AS·4AL易位染色体,而另一条2B和4A为正常的完整染色体。这种染色体结构重排事件表明,部分小麦-远缘物种附加系细胞学并不稳定,因此,繁种前后应对材料进行单株细胞学鉴定。上述小麦-粗穗披碱草染色体结构变异体的获得,为研究染色体结构重排与基因转录表达和表型变化的关系提供了研究基础。  相似文献   

7.
本研究利用形态学和分子细胞遗传学研究方法对中国春-长穗偃麦草二体附加系与中国春-柱穗山羊草(2C)二体附加系的杂交后代F1、F2、F3进行研究,旨在探讨杀配子染色体诱导小麦-外源染色体畸变的频率,为小麦遗传育种提供材料基础。结果表明,杀配子染色体(2C)的丢失与恢复成二体,造成杂种F1、F2的减数分裂行为异常,杂种F3较前2代单价体数明显下降,相对紊乱系数明显低于F2。同时,杀配子染色体连续2次作用,使得附加系中ABDE染色体组受到严重影响,部分染色体产生缺失和易位,在后代传递中易于丢失,而外源E组染色体片段渗入到小麦染色体组中,使得染色体配对异常,产生非理论值的单价体数,导致后代育性下降或败育。本研究共检测F3植株448株,得到易位24株,缺失36株,小麦间易位19株,易位频率为5.36%,缺失频率为8.04%,染色体畸变总频率为13.39%。断裂位点的分布比率依次为:B组A组D组。本研究表明,杀配子染色体诱发染色体畸变频率高,且具有一定的方向性,是创造小麦遗传育种新材料的重要途径。  相似文献   

8.
根据已报道的其他植物高亲和钾离子转运蛋白(HKT1;4)基因的保守序列设计一对简并性引物,以长穗偃麦草幼根总RNA为模板,采用RT-PCR方法克隆出长穗偃麦草HKT1;4,基因命名为EeHKT1;4。结果表明:该基因片段长度为614 bp,编码204个氨基酸,所得序列与其他植物氨基酸序列同源性均在80%以上,特别是和一粒小麦TmHKT1;4氨基酸同源性高达92%。这为长穗偃麦草HKT1;4全长基因的克隆及其耐盐分子机制的研究奠定基础。  相似文献   

9.
为建立簇毛麦(Dasypyrum)特定染色体上的特异性ISSR标记,以二倍体簇毛麦(Dasypyrum villosum)和小麦中国春(Triticum aestivum)等为材料,对96条ISSR引物进行PCR筛选,发现引物UBC848可在二倍体簇毛麦中扩出一条长388bp的特异性片段,命名为pDv848/388(GenBank登录号:EF411201),而中国春等小麦均未扩出该片段。利用UBC848对小麦近缘种偏凸山羊草(Aegilops ventricosa)、荆州黑麦(Secale cereale cv.Jingzhou rye)和节节麦(Ae.tauschii)等进行扩增,发现它们均未扩出pDv848/388。进而用一套中国春-二倍体簇毛麦附加系CSDA1V~CSDA7V对pDv848/388进行染色体定位,将目标片段确定在二倍体簇毛麦5V染色体上。进一步用UBC848对多年生簇毛麦(D.breviaristatum)、小麦-簇毛麦双二倍体、部分双二倍体及其衍生后代进行扩增,发现它们均能扩出pDv848/388,表明pDv848/388可作为簇毛麦5V染色体上的一个特异性ISSR标记,用于快速跟踪检测簇毛麦遗传物质向栽培小麦中的导入。  相似文献   

10.
为了能在分子水平上有效鉴定具有粘果山羊草(Aegilops kotschyi)、偏凸山羊草(A.ventricosa)、普通小麦变种斯卑尔脱(Triticum spelta)细胞质雄性不育系及其保持系90-110和8222,提高其在杂交小麦(Triticum aestivum L.)研究与应用中的定向遗传改良,本研究对其线粒体DNA进行了扩增片段长度多态性(amplified fragment length polymorphism,AFLP)标记和序列特征性片段扩增区域(sequence characterized amplified region,SCAR)标记。通过AFLP标记方法,应用64对引物组合EcoRⅠ-NNN/MseⅠ-NNN对小麦同核异质雄性不育系和保持系进行扩增,共扩增出682条带,其中113条为多态性条带。引物E-AGG/M-CTA组合在粘果山羊草细胞质雄性不育系中扩增出一条大小约300 bp的特异性条带,对该特异条带进行回收、测序,利用Primer Premier 5.0软件重新设计SCAR引物,并对这3种类型同核异质小麦细胞质雄性不育系和保持系进行扩增,其中引物YW1在3种细胞质雄性不育系和保持系中都扩增出条带,而引物YW2仅在粘果山羊草细胞质雄性不育系扩增出一条198 bp的特异性片段,结果表明,已成功地将AFLP标记转化为操作简便、表现稳定的SCAR标记。此片段与小麦线粒体基因组有很高的同源性(同源性为99%),为烟酰胺腺嘌呤二核苷酸(NADH)脱氢酶基因(GenBank登录号:EU534409.1)上的序列,该酶是线粒体中氧化磷酸化的入口酶,与小麦细胞质雄性不育密切相关。本研究可以用于粘类小麦细胞质雄性不育系分子标记辅助育种,也为小麦细胞质种性鉴定提供了技术支撑和理论依据。  相似文献   

11.
Forty-seven samples of date palm (Phoenix dactylifera L.) collected from eight locations in Egypt were studied using four sets of amplified fragment length polymorphism (AFLP) markers with near infrared fluorescence labeled primers. These samples belonged to 21 named accessions and 9 of unknown pedigrees. A total of 350 bands were scored and 233 (66.6%) were polymorphic. Twenty-seven Egyptian accessions and ‘Medjool’and ‘Deglet Noor’accessions from California could beclassified into the major cluster. This major cluster may represent a major group of date palm germplasm in North Africa. There were four other clusters, each containing one or two accessions. The variety ‘Halawy’and one accession of unknown provenance were most likely from hybridization between two clusters. Six groups of accessions of which had the same names, revealed similar but not identical AFLP profiles suggesting these accessions might derive from seedlings rather thanthrough clonal offshoot propagation.  相似文献   

12.
Sweet potato (Ipomoea batatas L.) is the fifth most important crop in the developing countries after rice, wheat, maize and cassava. The amplified fragment length polymorphism (AFLP) method was used to study the genetic diversity and relationships of sweet potato accessions in the germplasm collection of Sokoine University of Agriculture, Morogoro and Sugarcane Research Institute, Kibaha, Tanzania. AFLP analysis of 97 sweet potato accessions using ten primer combinations gave a total of 202 clear polymorphic bands. Each one of the 97 sweet potato accessions could be distinguished based on these primer combinations. Estimates of genetic similarities were obtained by the Dice coefficient, and a final dendrogram was constructed with the un-weight pair-group method using arithmetic average. AFLP-based genetic similarity varied from 0.388 to 0.941, with a mean of 0.709. Cluster analysis using genetic similarity divided the accessions into two main groups suggesting that there are genetic relationships among the accessions. Principal Coordinate analysis confirmed the pattern of the cluster analysis. Analysis of molecular variance revealed greater variation within regions (96.19%) than among regions (3.81%). The results from the AFLP analysis revealed a relatively low genetic diversity among the germplasm accessions and the genetic distances between regions were low. A maximally diverse subset of 13 accessions capturing 97% of the molecular markers diversity was identified. We were able to detect duplicates accessions in the germplasm collection using the highly polymorphic markers obtained by AFLP, which were found to be an efficient tool to characterize the genetic diversity and relationships of sweet potato accessions in the germplasm collection in Tanzania.  相似文献   

13.
利用102对微卫星引物对5份黑麦(Secale)、4份普通小麦(Triticum aestivum)和1份分枝小黑麦(Triticale)进行SSR分析,引物Xgwm614能在分枝小黑麦中扩增出一个387bp的特异DNA片段(记为FZ387,GenBank登录号为EF179137),而黑麦未能扩增出。序列比对结果显示该片段与一粒小麦(T. monococcum)(AY485644)和栽培二粒小麦(T. turgidum)(AY494981)A基因组中Gypsy Ty3-LTR反转座子fatima的一部分分别有94%和95%同源性。根据序列同源性比对结果,在FZ387内部设计1对特异引物FaF和FaR。引物Xgwm614F和FaR能在含有A基因组的物种中扩增出约350bp的条带(记为A350),而其不含A基因组的物种都未扩增出该条带。利用小麦二体和端体代换系材料对其进行定位,结果显示该片段分布在所有A染色体的长臂和断臂上。此外,引物FaF和Xgwm614R能在含有A、B或AB基因组的物种中扩增出约350bp的条带(记为AB350),而不含AB基因组的材料未扩增出目标条带。利用这两对特异引物对小麦属近缘物种进行PCR扩增,发现只有中国春能够扩增出A350和AB350。序列比对结果和FZ387两侧SSR引物结合区的规律性变化表明该反转座子在进化上可能存在属间多样性和属内相似性。A350和AB350也可以分别作为分子标记检测A染色体和AB染色体。  相似文献   

14.
A total of 242 accessions of common wheat (Triticum aestivum L.) released in China since the 1940s were evaluated with AFLP (amplified fragment length polymorphism) technique. Genetic diversity was analyzed using five pairs of polymorphic primer combinations with 245 polymorphic bands. The highest genetic diversity was found in the accessions of the 1950s, and in the next place was that in the 1940s. The genetic diversity began to descend in the 1960s, and fell to the lowest in the 1970s. After that, the genetic diversity came back to some extent in the 1980s, however, it became much lower in the 1990s compared with that in the 1940–1950s. Landraces and introduced accessions from foreign countries showed greater genetic diversity in comparison to improved varieties. In addition, greater genetic diversity was observed in winter wheat. It was emphasized that great attention should be paid on further exploration of genetic diversity in wheat breeding program.  相似文献   

15.
The genetic diversity was studied of 115 Agropyron cristatum accessions from 17 countries. Tetraploids were the most common (74.8%), followed by diploid (16.3%) and hexaploid (6.9%). We observed a relation between geographic distribution and ploidy level. The tetraploids, the most widespread, were found from Europe through Russia to East Asia. The diploids appeared over the same general range, except in Turkey, Iran and Georgia where no diploid accessions were found. Hexaploid accessions mainly came from a region comprising the east of Turkey, the north of Iran and Georgia. A selection of 71 accessions, including all three ploidy levels, were analyzed by capillary electrophoresis using six wheat simple sequence repeat (SSR) markers. All markers presented high levels of polymorphism, generating 166 different alleles ranging in size between 84 and 256 bp. Based on polymorphic information content values obtained (0.579–0.968), all the SSRs were classified as informative markers (values?>?0.5). According to the dendrogram generated, all the A. cristatum accessions were distinctly classified. Diploid, tetraploid and hexaploid accessions are not clearly differentiated from each other on the basis of SSR markers. A field experiment was conducted to morphologically characterize 18 accessions including the three ploidy levels. Significant differences were found between the accessions in spike length, spike width and number of spikelets per spike. All the cytological, molecular, and morphological data demonstrate the high genetic diversity present in A. cristatum, making it a valuable resource for future breeding programs.  相似文献   

16.
We used amplified-fragment-length polymorphism (AFLP) markers to evaluate genetic variation in a set of bread wheat (Triticum aestivum L.) landraces and improved materials. Landraces collected from different geographic and agro-ecological zones in Pakistan in 1987, 1989 and 1991 were separated into two groups based on their geographic origins: northern (Himalaya) and south-western (Balochistan) Pakistan. Six AFLP primer combinations detected 453 AFLP markers in the 43 landrace accessions and four high-yield varieties (HYVs). Of these, 225 (49.67%) were rare (shared with < 5% of all accessions). Among these rare alleles, 23 (10.22%) were common in the Himalaya (shared with > 10% of accessions collected there) but were not found in Balochistan. We conclude that there is a higher probability of collecting rare alleles at overall, but which are in contrast locally common ones in the Himalayan region. Gene diversity was 0.17 in the Himalayan group and 0.15 in the Balochistan group. Considerable genetic variability was found in both groups. Accessions from different agro-ecological zones were indistinguishable by cluster analysis, indicating intensive seed trading within the country. Cluster analysis indicated that the landraces and the HYVs are genetically distinct; suggesting that genetic erosion of wheat landraces has been unlikely taken in place. This study provides an example of how analysis of existing materials and data, can serve as a basis for future collection planning and conservation policies.  相似文献   

17.
Opaque endosperm is the main phenotypic indicator for waxy rice, but other phenotypic and genotypic variation among waxy rice accessions has largely been ignored. Previous studies showed that wide diversity in starch physiochemical properties exists in both indica and japonica waxy rices, especially for starch gelatinization temperature (GT) which could be divided into a high- and a low-GT group. In the present study, amplified fragment length polymorphism (AFLP) and inter-simple sequence repeat (ISSR) molecular markers were employed to examine genetic diversity and relationships of 56 waxy rice accessions. A total of 358 AFLP fragments were amplified with five primer combinations, showing a high level of polymorphism (78.3%). A total of 190 ISSR bands were generated with a single primer and a primer pair, showing a very high level of polymorphism (92.2%). The genetic distance matrices obtained from the two sets of markers were significantly correlated (r = 0.731, P = 0.004). The dendrogram generated with combined AFLP and ISSR markers could clearly differentiate the indica and japonica groups. Newly released varieties and breeding lines within each subspecies tended to be clustered together, whereas landraces were more distantly placed in the dendrogram. Only one AFLP band was found specific to the indica type, while no specific bands were found for starch GT. The implications for the conservation and breeding of waxy rice are discussed.  相似文献   

18.
A set of wheat cv. Chinese Spring (CS)-Lophopyrum elongatum addition and substitution lines has been analyzed for its potential to offer new sources of stripe rust resistance for wheat. The adult plants of these lines, together with CS-L. elongatum amphiploid and CS, were inoculated by new physiological races CYR-30 and CYR-31 of stripe rust in China. The resistance investigation indicated that chromosome 7E1 of L. elongatum was responsible for the adult resistance in CS background. Moreover, the expression of the adults plant resistance derived from chromosome 7E1 in wheat background was dependent on the wheat genotype. In addition, the different rust resistance and glutenin composition were observed in 2 CS-L. elongatum amphiploids, indicating the genetic diversity existed in different origins of L. elongatum.  相似文献   

19.
A comprehensive characterization of crop germplasm is critical to the optimal improvement of the quality and productivity of crops. Genetic relationships and variability were evaluated among 63 durum wheat landraces from the Mediterranean basin using amplified fragment length polymorphisms (AFLPs) and microsatellites markers. The genetic diversity indices found were comparable to those of other crop species, with average polymorphism information content (PIC) values of 0.24 and 0.70 for AFLP and microsatellites, respectively. The mean number of alleles observed for the microsatellites loci was 9.15. Non-metric multi-dimensional scaling clustered the accessions according to their geographical origin with the landraces from the South shore of the Mediterranean Sea closely related. The results support two dispersal patterns of durum wheat in the Mediterranean basin, one through its north side and a second one through its south side.  相似文献   

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