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RAPD技术在芦荟属植物分类研究中的应用 总被引:6,自引:0,他引:6
采用CTAB法提取11个芦荟材料的基因组DNA为模板,以50个随机引物进行RAPD分析。结果表明:大部分引物可以在不同模板上扩增出条带,但仅有6个引物可以同时在11个芦荟材料的DNA上扩增出条带,对11个芦荟种变种的RAPD结果进行聚类分析,结果表明基本符合传统分类观点。对RAPD技术在芦荟属植物分类研究中的问题进行了讨论。 相似文献
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香蕉RAPD分析初步研究 总被引:12,自引:1,他引:11
比较了不同提取方法对香蕉植株不同部位组织提取 DNA的质量及其 PCR扩增结果 ,对香蕉 RAPD分析中引物种类和浓度 ,复性温度 ,d NTPs,Taq DNA聚合酶浓度 ,热循环数等因素进行了比较影响分析。结果表明 ,虽然改良的 SDS法、CTAB法和 PVP法提取的植株嫩叶和吸芽 DNA提取量和纯度各不相同 ,但其 PCR扩增结果基本相同 ;相同克隆不同植株的 DNA其 PCR扩增结果也基本相同 ;建立了适合香蕉大规模 DNA多态性分析 RAPD反应体系 :2 5 μL反应液中 ,含 1倍缓冲液 ,0 .2 m Md NTPs,0 .3 2 p M随机引物 ,IUTaq酶 ,2 0 ng模板 DNA;反应循环数为 45 ,热循环条件为 94°C,1 min;3 7°C,1 min;72°C,1 .5 min;之前为 94°C,5 min;之后为72°C,1 0 min。在筛选的 2 49个随机引物中 ,有 1 8个在 7个品种上都能扩增出 3~ 1 0条比较清晰条带。 相似文献
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高粱抗丝黑穗病基因的RAPD初步分析 总被引:1,自引:0,他引:1
目的:对高粱抗丝黑穗病的基因进行分析。方法:以高粱2381恢复系(抗病)、矮四恢复系(感病)、7050B保持系(抗病)、TX622B保持系(感病)为材料,采用CTAB提取DNA的方法提取高粱基因组DNA,然后应用RAPD分子标记技术对DNA进行多态性扩增。结果:初步建立了高粱丝黑穗病的RAPD反应体系;从RAPD反应中所用的48个随机引物中筛选出28个适宜引物,其余20个引物没有扩增出谱带,被淘汰;共扩增出114条谱带,其中引物OPM-05300和OPM-13450扩增出了差异谱带,分别命名为OPM-05300和OPM-13450。结论:在该反应体系下找到了高粱抗丝黑穗病抗感品种间基因组差异的两条差异谱带。 相似文献
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共生在水生蕨类植物满江红(又称“红萍”:Azolla)叶腔中的固氮兰细菌曾被认为是一个种满江红鱼腥藻(Anabaena azollae Strasb.).但由于现存的满江红在分类学上有2个亚属,7个种1,对不同满江红叶腔中鱼腥藻的鉴别,多年来引起人们的重视.80年代,鱼腥藻的单克隆抗体和RFLP的研究,发现了它与宿主分类上一定程度的对应关系2,3.90年代以后,Van Coppenolle和Eskew分别以RAPD-PCR和DAF-PCR对从满江红共生体中提取的DNA进行了分析4,5,但后者发现了共生体中藻的DNA对整个PCR产物的干扰作用会影响满江红属本身系统分类.
相似文献
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从16个代表不同种属或地域来源的满江红样本中分离出共生藻并通过处理获得无藻的满江红宿主,对二者同步进行了RAPD扩增,分别得到了大量DNA多态片段。通过建立满江红鱼腥藻及其宿主的UPGMA聚类关系图,看出二者在遗传分支上存在着一定程度的协同对应关系。但在种内的不同品系间,这种协同性有所减少,发现有的品系的共生藻发生了明显的变异。
Abstract:Symbiotic Anabeana azollae and its host plant Anabeana-free Azolla were isolated from 16 Azolla accessions representing different Azolla species or geographic origins.DNA polymorphic fragments were obtained by simultaneous RAPD amplification of both symbiont and host.The UPGMA clusters of Anabeana azollae and its host Azolla were established separately based on Dice coefficient caculation and a coordinated relationship was shown between Anabeana azollae and its Azolla host along both individual genetic divergence,but this genetic homology was reduced among different strains within Azolla species while the obvious mutants of Anabeana azollae were detected in some Azolla tested strains collected from different geographic area in the same host species. 相似文献
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冬虫夏草(Cordyceps sinensis)的随机扩增多态DNA及其遗传分化 总被引:16,自引:0,他引:16
本文对来自青藏高原3个区域5个具有代表性地方的13个冬虫夏草(Cordyceps sinensis[Berk.] Sacc.)样本进行随机扩增多态DNA(RAPD)分析。19个随机引物获得的RAPD谱带清晰并呈现多态,单个引物获得的RAPD片段数在3~10个之间。该19个引物在每个样本中扩增的RAPD片段总数平均约为65个。基于遗传距离分析,受试的13个冬虫夏草样本中,来自同一地方的样本间遗传差异甚微,同一区域不同地方的样本间遗传差异较大,不同区域的样本间遗传差异最大。这说明冬虫夏草地理群体间存在着遗传分化。应用UPGMA和NJ方法构建的分子系统树显示,来自5个地方的冬虫夏草实际上可以归并为显著不同的3个组,对应于样本来源的3个区域,提示RAPD标记在冬虫夏草群体中有显著的地区特异性。我们的结果还表明,冬虫夏草地理群体间的遗传差异度与地理距离呈正相关。因此,RAPD作为有效的遗传标记,可用于研究冬虫夏草的遗传多样性、起源以及系统演化等。 相似文献
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直接从人工老化的菜心干种子中提取基因组DNA用于RAPD分析 总被引:7,自引:0,他引:7
利用 4 0℃、1 0 0 %RH对菜心种子进行人工加速老化处理获得了不同活力的种子批 ,利用平衡酚_氯仿法直接从人工老化的菜心干种子中提取基因组DNA ,并对提取的基因组DNA进行了RAPD扩增。结果表明 ,所提取的基因组DNA量多 ,而且比较整齐一致。引物S2 0 8扩增所获得的基因组DNA指纹图谱上的DNA带清晰、明亮 ,从而表明利用本方法从人工老化菜心干种子中直接提取的基因组DNA完全可以用于RAPD分析。 相似文献
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Genetic diversity among and within cultured cyanobionts of diverse species of <Emphasis Type="Italic">Azolla</Emphasis> 总被引:1,自引:1,他引:0
The cyanobionts isolated from 10 Azolla accessions belonging to 6 species (Azolla mexicana, A. microphylla, A. rubra, A. caroliniana, A. filiculoides, A. pinnata) were cultured under laboratory conditions and analyzed on the basis of whole cell protein profiles and molecular marker dataset generated using repeat sequence primers (STRR(mod) and HipTG). The biochemical and molecular marker profiles of the cyanobionts were compared with those of the free-living cyanobacteria and symbiotic Nostoc strains from Anthoceros sp., Cycas sp. and Gunnera monoika. Cluster analysis revealed the genetic diversity among the selected strains, and identified 3 distinct clusters. Group 1 included cyanobionts from all the 10 accessions of Azolla, group 2 comprised all the symbiotic Nostoc strains, while group 3 included the free-living cyanobacteria belonging to the genera Nostoc and Anabaena. The interrelationships among the Azolla cyanobionts were further revealed by principal component analysis. Cyanobionts from A. caroliniana-A. microphylla grouped together while cyanobionts associated with A. mexicana-A. filiculoides along with A. pinnata formed another group. A. rubra cyanobionts had intermediate relationship with both the subgroups. This is the first study analyzing the diversity existing among the cultured cyanobionts of diverse Azolla species through the use of biochemical and molecular profiles and also the genetic distinctness of these free-living cyanobionts as compared to cyanobacterial strains of the genera Anabaena and Nostoc. 相似文献
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The structures of photosynthetic apparatuses such as leaves, chloroplasts and
symbiotic cyanobacterum (blue-green algae) in Azolla-Anabaena azollae associations (Azolla
imbricata (Roxb) Nakai) which occur in paddy fields of China were examined using light,
scanning and transmission electrn microscopy. Some comparisons were made with A.
filiculoides, A. japonica, A. caroliniana, A. pinnata and A. mexicana.
Cross sections of A. imbricata were observed by light microscopy and the symbiotic association between the eukaryotic water fern and its prokaryotic blue-green algal symbiont, an
Anabaena, was studied. The symbiotic cyanobacterum cells occur not only in a mature leaf
cavity, but also in early stages of leaf development, around leaf primordia, and even in macrospores.
Under scanning electron microscopy (SEM) it is possible to see stomata and nipples on
the surface of dorsal lobes of the fern. The species in the subgenus Euazolla (i.e.A. filiculoides, A. japonica, A. caroliniana and A. mexicana) have rounded nipples, but those in the
subgenus Rizosperma (i.e.A. imbricata and A. pinnta) prolate ones. This morphological
character is first reported to be related to the taxonomic system.
The result of the observation with transmission electron microscopy (TEM) shows
that A. filiculoides contains more thylakoides in chloroplasts than A. imbricata does, and the
grana lamellae have more stacks in the former than in the latter. The differences are in agreement with the differentiation of the two species in photosynthetic capacity. This may be
one of the differences between the two subgenera.
The ultrastructures of the symbiotic cyanobacterum are similar to those of free-living
Anabaena. The vegetative cells show a typical bilayered cell wall and the heterocysts have a
thikened wall. The thylakoid membranes in both heterocysts and vegetative cells are oftenseen forming whirls. During the division of vegetative cells, their contents aggregate and then redistribute. 相似文献
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满江红属系统学研究的新进展 总被引:2,自引:0,他引:2
本文从植物系统学角度对满江红属的研究进展作了全面综述,近年来从细胞水平提出了将满江红属重新划分为Azolla和Tetrasporocarpia2个亚属,经典的三膘(Azolla)和九膘(Rhizosperma)亚属作为新的Shbgen,Azolla中的2个组(Section),而原有九膘亚属中的A.nilotica则为Subgen,Tetrasporocarpia的一个种,遗传育种和同工酶的研究证据对A.mexicana,A.microphylla和A.caroliniana能否在Sect.A-zolla中占据3个种的分类地位提出疑问,但要完全解决满江红属的系统学问题,分子生物学技术是重要手段,由于满江红为蕨-藻共生体,其样本的异质性影响了现有分子标记研究结果的可靠性。 相似文献
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By investigating the variance of Azolla leaf colour of F1 generation obtained from negative and positive crossing of Azolla between two species (Azolla filiculoides × A. microphylla, A. filiculoides × A. mescicana) and two subgenus (A. filiculoides × A. imbricata), it was revealed that the albinism of the hybrid F1 generation was variation resulting from maternal cytoplasmic inheritance, when A. filiculoide was used as female parent. Electron microscopic observation demonstrated abnormal development of plastid in the albino sporeling. The cell of light green seedling contained both normal and abnormal plastid. Both were probably related to variation in the plastid genotype. Significant difference occurred in the degree and freguency of albinism from various crossing forms, and such change had its vegularity in accord with the variation of the nuclear genotype. The results speculated that albinism were also closely related to the nuclear genotype. 相似文献
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The occurrence and development of the hair ceils on the shoot tips and in the leaf cavities of A. filiculoides, A. microphylla, A. pinnata and their algae-free cultures were examined by means of scanning electron microscopy with microdissect technique. The patterns of Anabacna moving into the leave cavities from the shoot tips were investigated on three species of Azolla during their vegetative growth. The results showed that the patterns of symbiotic Anabaena infecting the leaf cavities are similarity among three species of Azolla and may be divided to the four phases which are summarized as follows: 1. occurrence of primary branched hair and adhesion of Anabaena; 2. development of primary branched hair and spreding of Anabaena; 3. building of hair bridge and entrance of Anabaena into the cavities; 4. formation of secondary simple hair and transference of Anabaena within the cavity. These observations resulted in a hypothesis that hair induces and leads its partner. It is suggested that the hair cell is likely to be a structure of Azolla for attracting and recognizing its symbiont in addition to transport substance between fern and algae. 相似文献
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Symbiotically associated cyanobacteria from Azolla mexicana and Azolla pinnata were isolated and cultured in a free-living state. Morphological analyses revealed differences between the free-living isolates and their symbiotic counterparts, as did restriction fragment length polymorphism (RFLP) analyses with both single-copy glnA and rbcS gene probes and a multicopy psbA gene probe. RFLP analyses with Anabaena sp. strain PCC 7120 nifD excision element probes, including an xisA gene probe, detected homologous sequences in DNA extracted from the free-living isolates. Sequences homologous to these probes were not detected in DNA from the symbiotically associated cyanobacteria. These analyses indicated that the isolates were not identical to the major cyanobacterial symbiont species residing in leaf cavities of Azolla spp. Nevertheless, striking similarities between several free-living isolates were observed. In every instance, the isolate from A. pinnata displayed banding patterns virtually identical to those of free-living cultures previously isolated from Azolla caroliniana and Azolla filiculoides. These results suggest the ubiquitous presence of a culturable minor cyanobacterial symbiont in at least three species of Azolla. 相似文献
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Identification of a common cyanobacterial symbiont associated with Azolla spp. through molecular and morphological characterization of free-living and symbiotic cyanobacteria. 总被引:1,自引:0,他引:1 下载免费PDF全文
Symbiotically associated cyanobacteria from Azolla mexicana and Azolla pinnata were isolated and cultured in a free-living state. Morphological analyses revealed differences between the free-living isolates and their symbiotic counterparts, as did restriction fragment length polymorphism (RFLP) analyses with both single-copy glnA and rbcS gene probes and a multicopy psbA gene probe. RFLP analyses with Anabaena sp. strain PCC 7120 nifD excision element probes, including an xisA gene probe, detected homologous sequences in DNA extracted from the free-living isolates. Sequences homologous to these probes were not detected in DNA from the symbiotically associated cyanobacteria. These analyses indicated that the isolates were not identical to the major cyanobacterial symbiont species residing in leaf cavities of Azolla spp. Nevertheless, striking similarities between several free-living isolates were observed. In every instance, the isolate from A. pinnata displayed banding patterns virtually identical to those of free-living cultures previously isolated from Azolla caroliniana and Azolla filiculoides. These results suggest the ubiquitous presence of a culturable minor cyanobacterial symbiont in at least three species of Azolla. 相似文献
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P. J. ASHTON R. D. WALMSLEY 《Botanical journal of the Linnean Society. Linnean Society of London》1984,89(3):239-247
The three Azolla species which occur in southern Africa, A. pinnata var. pinnata, A. nilotica and A. filiculoides are identified and their distributions illustrated. The endosymbiotic cyanobacterium Anabaena azollae occurred in dorsal leaf lobe cavities of all three species. The first two species are indigenous to southern Africa, whilst the third, A. filiculoides , is an introduced species from North America. Possible means whereby this species could have been introduced are discussed. 相似文献
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本文从植物系统学角度对满江红属的研究进展作了全面综述。近年来从细胞研
究水平提出了将满江红属重新划分为〖WTBX〗Azolla和Tetrasporocarpia 2个亚
属,经典的三膘(Azolla)和九膘(Rhizosperma)〖WTBZ〗亚属作为新的Subgen Azolla中的2个组(Section),而原有九膘亚属中的A.nilotica
则成为Subgen.Tetrasporocarpia〖WTBZ〗的一个种。遗传育种和同工酶
的研究证据对A.mexicana,A.microphylla和A.caroliniana能否在Sect.
Azolla中占据3个种的分类地位提出疑问。但要完全解决满江红属的系统
学问题,分子生物学技术将是重要手段。由于满江红为蕨 藻共生体,其样本的异质性影响了现有分子标记研究结果的可靠性。 相似文献
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本文报道满江红(Azolla imbricata)、卡洲满江红(A. caroliniana)和蕨状满江红(A. filiculoides)的光合与固氮(乙炔还原)活性对温度的反应差异。根区培养液温度在7—40℃范围内满江红与卡洲满江红的光合活性在30℃达到峰值,高于或低于30℃均急剧下降。而蕨状满江红在7—25℃的根区温度范围内光合活性差异不大,高于25℃则下降(图2)。0℃24小时或40℃2小时暗预处理对三种满江红光合活性与乙炔还原活性表现出不同影响(图3,4)。本文报道的方法可用于满江红生理生态参量的比较研究。 相似文献