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1.
与棱果沙棘性别相关的RAPD标记   总被引:4,自引:0,他引:4  
应用RAPD技术筛选与棱果沙棘性别相关的分子标记,对棱果沙棘雌雄株的基因组DNA进行混合分组分析(BSA),在194条随机引物中有50条引物能够在雌雄DNA反应池间形成多态性条带,应用这50条引物分别对棱果沙棘雌雄个体(雌雄个体各选取5个)进行RAPD分析,其中引物S10扩增得到1个约为1030 bp的与雌性相关的RAPD标记。该标记的获得进一步表明棱果沙棘雌雄株间存在基因水平的差异,为棱果沙棘的性别研究提供分子依据。进一步利用该雌性特异位点设计出更加稳定的SCAR标记,可望用于棱果沙棘的早期性别的准确鉴别。  相似文献   

2.
谭清苏铁性别相关的RAPD标记研究   总被引:1,自引:0,他引:1       下载免费PDF全文
以谭清苏铁(Cycas tanqingii D.Y.Wang)雌雄植株半年生羽叶为材料,用优化的CTAB法分别提取其全基因组DNA,进行RAPD单因子梯度实验和正交实验以优化扩增条件。应用160个RAPD随机引物检测基因组DNA,雌雄植株均扩增出1450多条带,其中引物S0465扩增出与谭清苏铁雌株高度相关的RAPD标记,其大小约为500bp,该标记与雄株没有关联。  相似文献   

3.
高产王浆西蜂DNA分子中的相关基因标志筛选及其鉴定   总被引:5,自引:0,他引:5  
为了探讨西蜂与高产王浆相关特异基因标记 ,用 12种随机引物 (P1~P12 )对产王浆量不同的4品系西蜂的基因组DNA进行了RAPD PCR分析 ,分别获得了产王浆量高、低不同西蜂的DNA多态性图谱 ,并从P2 引物的DNA多态性图谱中筛选出一差异DNA片段P2 316bp .将P2 316bp差异DNA片段用地高辛标记制备成探针 ,进行Southern杂交鉴定 .实验显示 ,探针与高产王浆西蜂基因组DNA的扩增产物出现了阳性杂交信号 ,而与低产王浆西蜂基因组DNA的扩增产物未出现阳性杂交信号 .结果表明 ,该差异性基因片段P2 316bp是西蜂高产王浆优良性状相关的遗传标记 ,序列为 30 5个核苷酸 .  相似文献   

4.
用随机扩增多态性DNA产物做探针产生鸡的DNA指纹图   总被引:2,自引:0,他引:2  
我们用12个随机扩增多态性DNA(RAPD)引物对来自不同品系的4只鸡进行了RAPD分析,在扩增出的共99条带中,表现多态性的带为38条,占总带数的38%.回收了4个表现个体特异性的RAPD产物,当用鸡的基因组总DNA探针与它们杂交时,其中3个表现阳性,说明RAPD方法扩增出的高变异产物含有重复序列.用含重复序列的个体特异性RAPD产物作探针,与无关个体鸡基因组DNA的HaeⅢ酶切产物进行DNA印迹,获得了变异性较高的DNA指纹图谱.因此,高变异的RAPD产物可以有效地用作DNA指纹探针.  相似文献   

5.
葎草雄性连锁的RAPD标记的克隆与SCAR标记的建立   总被引:1,自引:0,他引:1  
应用随机扩增的DNA多态性(RAPD)技术可以快速简便获得雌雄异株植物雌雄株间的基因组差异信息,本文采用该技术对律草雌雄混合基因池进行了扩增,在选用的100条10bD的随机引物中,引物S1519和S2142分别扩增出长度为1207bp和762bp的雄性连锁标记,对其进行回收克隆和测序后发现这两个片段富含AT序列,AT含量分别为64%、54.7%,经核苷酸数据库BLAST检索未发现其同源序列。根据测序结果设计的SCAR引物可以将这两个雄性连锁的RAPD标记转化为稳定性和特异性更好的SCAR标记。  相似文献   

6.
随机扩增杂交微卫星(Random amplified hybridization microsatellites,RAHM)是一种复合RAPD扩增和寡核苷酸扫描的方法。该方法能够从RAPD产物凝胶上获得更多的信息,具有分析方法快速,高敏感性,能检测到高水平的多态性等优点。RAHM方法通过对RAPD扩增的DNA片段进行微卫星杂交来替代限制性内切酶对基因组DNA的消化,有助于揭示微卫星基因组克隆,进行微卫星引物的筛选。本文采用随机扩增杂交微卫星方法来检测小型哺乳动物大仓鼠(Techerskia triton)种群的遗传多态性,结果表明RAHM方法能够检测到大仓鼠种群中较高的多态性以及种群间的差异,这些条带模式可能代表真核基因组中另一种多态性标记的来源,可用于检测小型哺乳动物种群的遗传多态性。  相似文献   

7.
雌雄异株葡萄的性别鉴定研究   总被引:1,自引:0,他引:1  
通过扫描电镜观察花粉的形态;分析叶片超氧化物歧化酶同工酶(SOD);RAPD分析基因组DNA多态性,来鉴别葡萄的性别。结果表明:葡萄雌雄株的花粉形态存在着较大的差异;在本实验条件下葡萄雌雄株叶片SOD同工酶谱带一致;一些引物的RAPD产物间存在着多态性,在葡萄雌雄株的基因组DNA序列中存在着碱基排列顺序的差异。实验指出,花粉形态和基因组DNA的多态性分析是进行性别鉴定的有效方法。  相似文献   

8.
利用RAPD技术寻找银杏(Ginkgo biloba L.)中与性别相关的分子标记.筛选了1 200个10 bp的随机引物,产生了8 372个RAPD条带.只有S1478产生一条大小为682 bp、雄性特异的分子标记,该分子标记被命名为S1478-682,出现在所有雄性植株中,而所有雌性植株都不具有该分子标记.通过在北京和沈阳种植的银杏植株的RAPD推广验证,说明该分子标记可以用来检测银杏植株的性别.  相似文献   

9.
银杏性别相关分子标记   总被引:12,自引:0,他引:12  
利用RAPD技术寻找银杏(Ginkgo biloba L.)中与性别相关的分子标记。筛选了1200个10bp的随机引物,产生了8372个RAPD条带。只有S1478产生一条大小为682bp、雄性特异的分子标记,该分子标记被命名为S1478—682,出现在所有雄性植株中,而所有雌性植株都不具有该分子标记。通过在北京和沈阳种植的银杏植株的RAPD推广验证,说明该分子标记可以用来检测银杏植株的性别。  相似文献   

10.
河南农业大学牧医工程学院杨霞、陈陆、许兰菊等五位科学工作者以鸡鲍氏志贺菌,鸡白痢沙门菌和痢疾志贺菌为对象,分别提取基因组DNA,利用6条随机引物以随机扩增多态性DNA(RAPD)技术对其基因组DNA进行了分析,其结果:4条随机引物能较好地在这3种菌中检测到多态性分子标记,共扩增出了59个DNA片段,其中3个菌株共有的谱带7条,而显示多态性的片段有52条,占88.1%。  相似文献   

11.
 A 400-bp RAPD marker generated by a primer of random decamer sequence has been found associated with the male sex phenotype in 14 dioecious cultivars and accessions of hemp (Cannabis sativa L.). The primer OPA8 generates a set of bands, most of which polymorphic among all the individual plants tested, and 1 of which, named OPA8400, present in all male plants and absent in female plants. A screening of 167 plants belonging to different genotypes for the association of the OPA8400 marker with the sex phenotype revealed that only in 3 cases was the 400-bp band was present in plants phenotypically female; on the contrary, in male plants the band was never missing, while in monoecious plants it was never present. Despite this sex-specific association, the sequences corresponding to OPA8400 were present in both staminate and carpellate plants, as revealed by Southern blotting and hybridization with the cloned RAPD band. The RAPD marker was sequenced, and specific primers were constructed. These primers generated, on the same genotypes used for RAPD analysis, a SCAR marker 390 bp in length and male-specific. This SCAR is suitable for a precise, early and rapid identification of male plants during breeding programs of dioecious and monoecious hemp. Received: 16 January 1998 / Accepted: 30 April 1998  相似文献   

12.
Genetics of control mechanisms that underlies sex differentiation in date palm is not known. Sex of the plants becomes known only at the time of first flowering, which takes around 5 years. In comparison, molecular diagnosis (if available/feasible) promises quick and reliable identification of sex types very early when plantlets are growing in seedbeds. To develop such an assay, genomic DNA from 45 individual plants (25 female and 20 male) belonging to different varieties of date palm was subjected to PCR amplification using 100 random amplified polymorphic DNA (RAPD) and 104 intersimple sequence repeat (ISSR) primers. Initially, two bulk genomic DNA samples (each made by pooling DNA from ten male and female plants, separately) were used. A primer showing sex-specific band in bulked samples was further used for amplification of the genomic DNA of the individual samples of that bulk. Only one RAPD primer, OPA-02, amplified a fragment of ~1.0 kb in all the individual samples of male genotypes, whereas this fragment was absent in all the female genotypes. This male-specific fragment was cloned and sequenced (GenBank accession no. JN123357), and a sequence-characterized amplified region (SCAR) primer pair was designed that amplified a 406-bp fragment in both female and male genotypes and a unique fragment of 354 bp in only male genotypes. The SCAR marker was further validated using 25 female and ten male date palm plants belonging to different varieties collected from different locations.  相似文献   

13.
The sex chromosomes of the silkworm, Bombyx mori, are designated ZW for the female and ZZ for the male. We previously characterized a female-specific randomly amplified polymorphic DNA (RAPD) marker, designated Female-218, from the translocation-bearing W chromosomes. These W chromosomes contain a region of the second chromosome, which carries visible larval markers of the p loci. We used strain TWPB in which female larvae have black skin due to the p(B) gene (T(W;2)p(B), +p/+p) while male larvae have whitish skin (+p/+p). To determine whether the Female-218 RAPD marker is derived from the "W region" or a "second chromosome fragment", we induced a detachment of the translocated W chromosome, T(W;2)p(B), by treating the eggs with hot water at an early developmental stage. After hot water treatment, we obtained 27 white female larvae out of 4850 female larvae. The Female-218 RAPD marker was not amplified in 26 out of 27 white female larvae, and was amplified from one white female larva. Moreover, we obtained 11 black male larvae out of 5377 male larvae. Eight out of 11 black male larvae became adult moths, and the Female-218 RAPD marker was amplified from all eight male moths. Examination of the genetic relationship between the Female-218 RAPD marker and the second chromosome fragment of the translocated W chromosome strongly indicates that the Female-218 RAPD marker is amplified from the region of second chromosome fragment of the T(W;2)p(B) chromosome.  相似文献   

14.
Identification of sex in hop (Humulus lupulus) using molecular markers.   总被引:1,自引:0,他引:1  
A Polley  M W Ganal  E Seigner 《Génome》1997,40(3):357-361
The rapid identification of sex in the dioecious hop (Humulus lupulus) is important for the breeding of this cultivated plant because only unfertilized flowers of the female plants are used as an ingredient in the production of beer. It is thought that a sex-chromosome mechanism controls the development of male or female plants. We have compared pools of male and female plants derived from a hop cross to identify molecular markers associated with the Y or male-specific chromosome. Of 900 functional RAPD primers, 32 revealed fragments specific for male plants that were absent in female plants of this cross. Subsequently, the 32 positive primers were tested on unrelated male and female plants. Three of these 32 primers were specific for the Y chromosome in all lines. The Y-specific product derived from one of these primers (OPJ9) was of low copy in hybridization experiments and predominantly present in male plants. Primers developed from the DNA sequence of this product provide a marker for rapid sex identification in crosses of hop by means of PCR.  相似文献   

15.
The random amplified polymorphic DNA (RAPD) technique was used to determine the sex of a dioecious species, Carica papaya L., with three sex types, male, female and hermaphrodite. A 450 bp marker fragment, named PSDM(Papaya Sex Determination Marker), exists in all male and hermaphrodite plants but not in the female plants so far analyzed. The DNA sequence of PSDM exhibited no significant similarity to previously reported sequences. A sequence-characterized amplified region (SCAR) marker, SCARps, was developed from PSDM to determine the sex of papaya. Southern hybridization, using PSDM as a probe, showed that PSDM exists in the male and hermaphrodite genomes, but not in the female genome. This result strongly suggests that PSDM is located on the chromosome region that is specific to the male and the hermaphrodite. SCARps is a suitable marker for the precise and rapid diagnosis of sex in papaya. Received: 1 February 2001 / Accepted: 22 May 2001  相似文献   

16.
High-yielding dwarf clones of Hevea brasiliensis are tolerant to wind damage and therefore useful for high-density planting. The identification of molecular markers for the dwarf character is very important for isolating true-to-type high-yielding dwarf hybrid lines in the early stage of plant breeding programs. We have identified a dwarf genome-specific random amplified polymorphic DNA (RAPD) marker in rubber tree. A total of 115 random oligonucleotide 10-mer primers were used to amplify genomic DNA by PCR, of which 19 primers produced clear and detectable bands. The primer OPB-12 generated a 1.4-kb DNA marker from both natural and controlled F1 hybrid progenies (dwarf stature) derived from a cross between a dwarf parent and a normal cultivated clone as well as from the dwarf parent; it was absent in other parent (RRII 118). To validate this DNA marker, we analyzed 22 F1 hybrids (13 with a dwarf stature and nine with a normal stature); the dwarf genome-specific 1.4-kb RAPD marker was present in all dwarf-stature hybrids and absent in all normal-stature hybrids. This DNA marker was cloned and characterized. DNA marker locus specificity was further confirmed by Southern blot hybridization. Our results indicate that Southern blot hybridization of RAPD using probes made from cloned DNA fragments allows a more accurate analysis of the RAPD pattern based on the presence/absence of specific DNA markers than dye-stained gels or Southern blot analysis of RAPD blots using probes made from purified PCR products. Detection of RAPD markers in the hybrid progenies indicates that RAPD is a powerful tool for identifying inherited genome segments following different hybridization methods in perennial tree crops.  相似文献   

17.
目的:探讨利用种子贮藏蛋白SDS—PAGE电泳、酯酶同工酶超薄等电聚焦电泳和RAPD分子标记鉴定五个两系杂交水稻组合F_1种子纯度的可行性。方法和结果:利用SDS—PAGE电泳技术未能找到所试五个组合各自的父本特征蛋白质带;利用酯酶同工酶超薄等电聚焦电泳技术和RAPD分子标记可找到所试五个组合的父母本特征酶带和RAPD标记,但酯酶同工酶的多态性同时也受种子萌发时间的影响。酯酶同工酶超薄等电聚焦电泳和RAPD分子标记可用于所试五个两系杂交水稻组合F_1种子纯度的鉴定。  相似文献   

18.
In order to develop the specific genetic marker for Korean native cattle (Hanwoo), randomly amplified polymorphic DNA (RAPD) analysis of 6 different cattle breeds was attempted by using 38 decamer primers. In comparison of RAPD patterns, two distinctive DNA bands specific for Hanwoo were detected. One was 296 bp of DNA fragment found to be specific only for female Hanwoo when primer GTCCACACGG was employed. In individual analysis of this RAPD marker was observed only in female individuals with the possibility of 85.3%. The other was 521 bp of RAPD marker amplified using TCGGCGATAG and AGCCAGCGAA primers, which showed 83.0% of genetic frequency in 85 male and 68 female individuals tested. Nucleotide sequencing of these genetic markers revealed that 296 bp marker has a short microsatellite-like sequence, ACCACCACAC, and a tandem repeat sequence of microsatellite GAAAAATG in the determined sequence. Two distinctive tandem repeats of microsatellite sequences, AAC and GAAGA, were also appeared in 521 bp DNA marker. In BLAST search, any gene having high homology with these markers was not found  相似文献   

19.
The anti-thrombotic effects of specific paf-acether antagonist BN 52021 were compared to the effects of Ginkgo Biloba extracts A, B, (A + B), and C. Local superfusion of BN 52021 over an experimentally injured arterial segment embolizes an existent paf-acether induced platelet thrombus. When applied before paf-acether, BN 52021 prevents local thromboformation in this model. Applied intravenously, BN 52021 reduces local thromboformation in a significant way. As compared to this BN 52021 standard, only Ginkgo Biloba B and the (A + B)-mixture present major thromboreductive activity.  相似文献   

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