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1.
棉花gDNA体细胞染色体FISH技术 总被引：17，自引：6，他引：11
介绍了棉花基因组ＤＮＡ（ｇｅｎｏｍｅＤＮＡ,简为ｇＤＮＡ）体细胞染色体荧光原位杂交〔ＦＩＳＨ〕的技术流程,并着重分析和讨论了影响试验结果的关键因素,包括染色体和探针的变性条件、染色体的蛋白酶Ｋ处理技巧等。试验中作为靶ＤＮＡ的体细胞染色体采用棉属异源四倍体种海岛棉;探针和封阻均采用ｇＤＮＡ,材料是棉属二倍体种Ａ染色体组（Ａｇｅｎｏｍｅ）的棉种（亚洲棉和草棉）和Ｄ染色体组（Ｄｇｅｎｏｍｅ）的棉种（瑟伯氏棉、雷蒙德氏棉、戴维逊氏棉等）,分别交互使用。试验结果比较理想,获得良好的ＦＩＳＨ片子,而且重复性好。 相似文献
2.
【目的】荧光原位杂交技术可以实现DNA序列直观准确的染色体定位,是基因组深入研究的重要技术之一。染色体特异探针的获得是该技术应用的关键。本研究旨在建立棉花寡核苷酸荧光原位杂交技术。【方法】利用已经公布棉花基因组序列数据,采用生物信息学方法获得染色体特异的寡核苷酸库,随后用乳化聚合酶链式反应方法标记成荧光探针,在棉花有丝分裂中期染色体上进行原位杂交。【结果】建立了一套棉花寡核苷酸荧光原位杂交技术体系。【结论】该体系可用于棉花单染色体识别鉴定。 相似文献
3.
《棉花学报》2017,(1)
【目的】研究二倍体野生棉与四倍体栽培棉间的遗传亲缘关系,进一步探索各棉种间的起源与进化。【方法】以5个二倍体基因组的代表种B1(异常棉)、C1(斯特提棉)、E2(索马里棉)、F1(长萼棉)以及G1(比克氏棉)的基因组DNA(gDNA)为探针,以2个四倍体栽培种(陆地棉中棉所16、海岛棉新海7号)有丝分裂中期染色体为靶DNA,进行了基因组原位杂交(Genomic in situ hybridization,GISH)分析。【结果】以B1、E2和F1gDNA为探针时,杂交信号主要分布在2个栽培种较长的13对A亚组染色体上;各产生3对较强的GISH-NOR信号,其中1对分布在较长的A亚组上,2对分布在较短的D亚组上,其GISH-NOR信号强度与分布情况与以D基因组棉种为探针时相似。说明二倍体B、E、F基因组与四倍体棉A亚基因组具有较高的同源性,亲缘关系更近。这一点与它们的地理分布情况相符;而它们基因组中的45S rDNA重复序列与二倍体D基因组的45S rDNA重复序列同源性较高。C1和G1中以gDNA为探针时,杂交信号分布在2个栽培种全部26对染色体上,无法区分开A或D亚组染色体,都有3对较强的GISH-NOR信号。这一现象与D基因组拟似棉(D6)gDNA为探针的GISH相似,表明二倍体C和G基因组与四倍体棉的A和D亚基因组均具有较高的同源性,或者C和G基因组同时含有A基因组(或其他非洲棉基因组)和D基因组成分,进一步证实了其基因组成分的杂合性;而它们基因组中的45S rDNA重复序列同属D基因组类型。【结论】这些发现可为棉花杂交育种和棉属起源与演化研究提供有用信息。 相似文献
4.
[目的]研究二倍体野生棉与四倍体栽培棉间的遗传亲缘关系,进一步探索各棉种间的起源与进化.[方法]以5个二倍体基因组的代表种B1(异常棉)、C1(斯特提棉)、E2(索马里棉)、F1(长萼棉)以及G1(比克氏棉)的基因组DNA(gDNA)为探针,以2个四倍体栽培种(陆地棉中棉所16、海岛棉新海7号)有丝分裂中期染色体为靶DNA,进行了基因组原位杂交(Genomic in situ hybridization,GISH)分析.[结果]以B1、E2和F1gDNA为探针时,杂交信号主要分布在2个栽培种较长的13对A亚组染色体上;各产生3对较强的GISH-NOR信号,其中1对分布在较长的A亚组上,2对分布在较短的D亚组上,其GISH-NOR信号强度与分布情况与以D基因组棉种为探针时相似.说明二倍体B、E、F基因组与四倍体棉A亚基因组具有较高的同源性,亲缘关系更近.这一点与它们的地理分布情况相符;而它们基因组中的45S rDNA重复序列与二倍体D基因组的45SrDNA重复序列同源性较高.C1和G1中以gDNA为探针时,杂交信号分布在2个栽培种全部26对染色体上,无法区分开A或D亚组染色体,都有3对较强的GISH-NOR信号.这一现象与D基因组拟似棉(D6) gDNA为探针的GISH相似,表明二倍体C和G基因组与四倍体棉的A和D亚基因组均具有较高的同源性,或者C和G基因组同时含有A基因组(或其他非洲棉基因组)和D基因组成分,进一步证实了其基因组成分的杂合性;而它们基因组中的45S rDNA重复序列同属D基因组类型.[结论]这些发现可为棉花杂交育种和棉属起源与演化研究提供有用信息. 相似文献
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棉花染色体序号的研究 总被引：3，自引：0，他引：3
回顾棉花染色体研究历史,讨论与染色体序号研究有关的技术方法及其在棉花上的应用,并阐明陆地棉染色体物理序号的来源。目前,公认的陆地棉染色体物理序号是以可鉴别的与特定染色体相关的细胞遗传学特征被发现的先后顺序命名的。这与以核型分析为依据的染色体物理序号还未达成一致。棉花BAC-FISH技术和粗线期染色体制片技术的成熟,为棉花染色体核型分析提供了可靠的技术平台。对棉花染色体序号重新整理,使目前两个陆地棉染色体序号命名体系达到统一,获得可实际操作的物理序号;同时,对海岛棉、亚洲棉、草棉等棉花的染色体序号进行规范,使染色体物理序号和遗传序号相一致,这些将为棉花基因组测序、棉花基因组的起源与变异和染色体细胞形态学研究等奠定良好的基础。 相似文献
6.
细菌人工染色体(Bacterial artificial chromosome, BAC）文库是开展基因组测序、基因图位克隆、分子标记、物理作图等研究的重要基因组资源。本文在构建了二倍体野生棉阿非利加棉（Gossypium herbaceum var. africanum）BAC文库的基础上,就棉花细菌人工染色体基因组文库构建过程中高分子量基因组DNA的提取、部分酶切片段选择、DNA的回收、连接转化以及BAC文库的保存等过程中一些细节和注意事项进行了比较详细的分析比较,希望能为棉花BAC文库的构建提供一些可供借鉴的经验。 相似文献
7.
以1套13个陆地棉A亚组染色体特异的BAC克隆为探针, 与海岛棉Pima90-53、红星草棉和阿非利加棉染色体进行荧光原位杂交。这些探针均在染色体上产生了特异信号,从而鉴别了这3个棉种基因组的单染色体。因此,这套BAC克隆也可以作为这3个基因组染色体的细胞遗传学标记。在此基础上,对3个棉种的A（亚）组染色体物理序号进行了命名。根据这些分子标记和连锁群的关系,染色体物理序号和遗传连锁图谱间可实现相互转化。这将有助于研究棉属不同棉种间的起源演化关系,以及新的遗传标记的染色体物理定位和构建染色体物理图谱等。 相似文献
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9.
甘蓝自交不亲和基因MLPK与SSP的FISH定位 总被引：4，自引：1，他引：3
利用FISH技术, 对自交不亲和基因MLPK与SSP在甘蓝有丝分裂前中期染色体、减数分裂早粗线期染色体以及伸长DNA纤维等3种分辨率水平的靶DNA载体上进行物理定位。结果表明, 在有丝分裂前中期, MLPK探针信号位于一对近中着丝粒同源染色体的短臂中部, 距着丝粒的百分距离约为53.41±3.16;SSP探针信号位于一对具有随体的近端着丝粒同源染色体的长臂端部, 距着丝粒的百分距离约为78.36±4.26。综合3种载体上的FISH结果表明, MLPK与SSP在甘蓝染色体组中可能都只有一个同源序列座位, 具有在单倍体基因组中的单拷贝性。重复FISH杂交表明, MLPK与5S rDNA位于同一对染色体。依据Armstrong的核型分析标准, 初步判断MLPK与SSP分别位于甘蓝的2号和7号染色体, 与S位点不存在连锁关系。另从比较基因组学角度对定位结果进行了讨论。 相似文献
10.
百棉系列棉花自交系品种（系）SSR指纹图谱构建 总被引：8，自引：3，他引：5
利用20对SSR核心引物构建了百棉系列棉花自交系品种（系）的DNA指纹图谱。20对引物分属棉花15条染色体,共检测到116个等位基因,平均5.8个;PIC值和MI值分别平均为0.718和4.384。引物组合NAU1028/NAU5104、NAU4903/NAU3110/NAU1043、NAU0905/NAU1255、NAU3839/NAU4024、NAU5104/NA U4024、NAU2121/NAU5104和NAU1043/NAU3254/NAU4024可分别将百棉1号、百棉2号、百棉13号、百棉5号、百棉011、百棉19号和百棉985与其他所有材料区分开。构建了百棉系列棉花自交系品种（系）的SSR数字指纹代码。对棉花指纹图谱构建的供试材料和核心引物进行了分析和讨论。 相似文献
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12.
Variations of the chloroplast DNA (cpDNA) from three of the four cultivated species of cotton (Malvaceae); Gossypium barbadense L., Gossypium hirsutum L., Gossypium arboreum L. and its synonym Gossypium nanking Meyen., were analyzed. Using specific set of primers, the whole circular cpDNAs from the four test species were amplified.
These were subsequently digested with the use of seven restriction enzymes. The amplified fragments of the whole cpDNAs of
the diploid cultivated cotton G. arboreum and its synonym G. nanking did not show any differences. However, the allotetraploid cultivated cottons G. barbadense and G. hirsutum, showed some fragment length differences directly visible after amplification and two types of restriction fragment length
polymorphism (RFLP), the first appeared as slightly lengthened bands and the other as gain or loss of a restriction site.
The results also showed that the chloroplast genomes of the allotetraploid cultivated cottons are highly similar to the diploid
cultivated cottons tested in terms of length and digestion patterns. The detected amplified length differences, RFLPs and
the restriction sites can be considered as species specific markers for the allotetraploid cultivated cottons, which could
be a useful tool for future studies of the cpDNA of the genus Gossypium L. 相似文献
13.
为给葱的染色体的识别提供新标记,建立葱的分子细胞遗传学核型,本研究采用去壁火焰干燥法制备了分散且形态良好的葱中期染色体,并进行了CPD（PI和DAPI组合）染色和45S rDNA荧光原位杂交（FISH）,根据葱染色体的形态特征,结合CPD染色和FISH结果,对葱进行了核型分析。CPD染色结果:葱所有染色体臂末端都显示CPD带。FISH结果：有一对45S rDNA位点（在第5对染色体上）。葱的核型公式：2n=2x=16=2sm+12m+2st(SAT)。研究表明：利用CPD染色和45S rDNA FISH,不仅能为染色体识别提供新标记,还能了解染色体GC丰富区的分布,为葱属植物的物种鉴定、系统分类与进化等研究提供DNA分子方面的证据。 相似文献
14.
The karyotypes and meiotic behaviour of six Chrysantheminae species are investigated. Pyrethrum coccineum (Willd.) Worosch. is a tetraploid, while the other five species (Argyranthemum frutescens (L.) Sch.-Bip., Opisthopappus taihangensis (Ling) shih, Crossostephium chinense (L.) Makino, Tanacetum vulgare L. and P. parthenium (L.) Sm.) are all diploids. Their karyotypes consist mainly of median centromere and submedian centromere chromosomes, although
C. chinense also has two terminal centromere chromosomes, and P. parthenium two subterminal centromere chromosomes. No satellited chromosomes are observed in any of the six species. All the species
have a 2A type karyotype, except for P. coccineum which is 3A. The meiotic behaviour in the pollen mother cells (PMCs) of all six species is essentially normal, with metaphase
I chromosome pairing configurations in the diploid species predominantly showing 9II, while P. coccineum shows 18II. A low frequency of quadrivalents (and trivalents) and chromosome bridges/lagging chromosomes is observed. 相似文献
15.
转基因抗虫棉产量相关性状QTL的分子标记及定位 总被引：1，自引：0，他引：1
采用亚洲棉渐渗的纤维强度突出的陆地棉优质新品系0-153与陆地棉转基因抗虫新品系sGK9708为亲本,构建了F2及F2∶3分离群体。利用3869对SSR引物筛选亲本,得到125对多态性引物。进一步对183个F2群体单株分析得到150个多态性标记位点,其中100个标记位点连锁,构建20个连锁群,共覆盖660 cM,占棉花总基因组的14.67%,每个连锁群平均包含5个标记位点,标记间平均相距6.6 cM,其中13个连锁群确定了对应的染色体。利用F2和F2:3数据,通过复合区间作图,共检测到28个产量及相关因素的QTLs。这些控制产量性状的QTLs只存在于5个连锁群上,成簇分布。与皮棉产量性状有关的2个QTLs,均与其它多个产量相关性状的QTLs在同一个连锁区段内,增效基因遗传效应方向一致,有必要研究其在标记辅助选择中的效果。本研究没有检测到在多世代表现稳定的QTL。因此,需要培育重组自交系,进一步明确产量性状有关QTL的遗传效应。 相似文献
16.
In order to introgress the ‘glandless-seed and glanded-plant’ trait from Gossypium sturtianum Willis (2n= 2x= 26, C1 genome) into the cultivated upland cotton Gossypium hirsutum L. (2n= Ax= 52 (AD), genome), two trispecific hybrids have been created using either Gossypium thurberi Torado (2n= 2x= 26, D1 genome) or Gossypium raimondii Ulbrich (2n= 2x= 26, D5 genome) as bridge species. The cross of both trispecific hybrids by G. hirsutum produced the first backcross progenies (BCl). Cytogenetic analysis showed that the trispecific hybrids had 52 chromosomes, their chromosome configurations at metaphase I (Ml) being 15.071 + 15.3411 + 0.93III + 0.69IV + 0.26VI in G. thurberi×G. sturtianum×G. hirsutum (TSH) and 14.421 + 17.0311 + 0.82III + 0.15IV + 0.07VI in G. hirsutum × G. raimondii ×. G. sturtianum (HRS), respectively. Among six BCl plants analysed, the only plant expressing the ‘glandless-seed and glanded-plant’ trait had 52 chromosomes and a meiotic configuration of 5.261 + 20.61II + 0.69III + 0.77IV at MI. Pollen fertility was 2.90% in TSH, 8.97% in HRS, and ranged from 0% to 40.28% in the BCl progenies. The introgressed BCl plant is perennial in growth habit. It can be used in breeding programmes aiming at the introgression of the ‘glandless-seed and glanded-plant’ trait into a cultivar of upland cotton. 相似文献
17.
First-division restitution in hybrids of Langdon durum disomic substitution lines with rye and Aegilops squarrosa 总被引：3，自引：0，他引：3
Durum wheat ‘Langdon’(LDN) caused a high frequency of first‐division restitution (FDR) and partial fertility in hybrids with rye, Secale cereale L., and Aegilops squarrosa L. In order to determine the genetic control of FDR, a complete set of 14 Langdon durum D‐genome disomic substitution lines (LDNDS) was crossed with ‘Gazelle’ rye and one accession (RL5286) of A. squarrosa. The microsporogenesis and fertility of the hybrids were studied. The results showed that most of the hybrids expressed a high frequency of FDR and partial fertility. However, the hybrids of 2D(2A), 4D(4A), 5D(5B) and 6D(6B) crossed with both rye and A. squarrosa, as well as 1D(1A) with A. squarrosa, had either little or no FDR and were completely sterile. These hybrids had different types of first meiotic divisions compared with LDN control hybrids. The hybrids with 2D(2A), 4D(4A) and 6D(6B) had a high frequency of random segregation of chromosomes at the first division. The hybrids with 5D(5B), as expected, showed high homoeologous pairing. The hybrid of 1D(1A) with A. squarrosa had a high frequency of equational division at first division. These results suggest that the reduced or absent FDR in such hybrids might be related to the substitution of chromosomes with an FDR gene and poor compensation ability of the D‐genome chromosomes for their homoeologous A‐ or B‐ genome chromosomes. Cytological analysis suggested that chromosome 4A in LDN most likely carries a gene for high frequency of FDR in hybrids. In addition, some monads were observed at the end of meiosis in the hybrids of 3D(3A) and 6D(6A) crossed with rye. They were formed from FDR cells that failed to divide at second division, suggesting that the LDN 3A and 6A chromosomes might carry genes for normal second division of FDR cells in the rye crosses. 相似文献
18.
Summary Grain size in wheat is the most stable yield component and has a favorable effect on flour yield. To identify the chromosomes associated with the large grains of line G603-86, (grain weight over 60 mg and grain length of about 9 mm), F3 lines, extracted from F2 populations obtained from F1 monosomics of crosses between G603-86 (P1) and the monosomic set of Favorit (P2) were tested in the field. ANOVA showed significant differences among parents for grain weight and grain length, but not for grain width or the factor expressing the difference in grain form and density. Homoeologous groups had significant effects on grain weight and on all components of grain weight, while genomes were not significantly different for any of these characters. Grain weight was significantly increased by chromosomes 6D and 4A of G603-86. Grain length was significantly increased by chromosomes 4A, 4B, 2B, 3A and 1B, grain width by chromosomes 1A and 1B, and the factor form-density by chromosomes 6D and 6A. The high grain size in G603-86 results from the effects of genes located on many chromosomes which affect grain dimensions, form and density. 相似文献