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DNA条形码技术在田间常见蓟马种类识别中的应用 总被引:2,自引:0,他引:2
蓟马类害虫种类多、 体型小, 传统的形态学鉴定方法难以快速准确识别。本研究利用DNA条形码通用型引物, 以我国田间常见的25种蓟马为靶标扩增其线粒体DNA细胞色素C氧化酶亚基Ⅰ (mitochondrial cytochrome c oxidase subunit Ⅰ gene, mtDNA COⅠ) 基因 (约650 bp), 通过对靶标片段碱基序列的测序及比对分析, 以邻接法 (NJ法) 构建系统发育树, 并以Kimura双参数模型计算种内、 种间遗传距离。结果表明: 聚类分析与形态学鉴定结果一致, 表现为较长的种间分支和较短的种内分支, 每个单系分支对应一个物种, 同一物种不同单倍型的最初分支自展值均为100%。25种蓟马的种内平均遗传距离为0.0027, 种间平均遗传距离为0.2757, 种间遗传距离为种内遗传距离的102.1倍; 而且种内、 种间遗传距离没有重叠区域。结果说明基于COⅠ基因的DNA条形码技术可以用于不同种类蓟马的快速准确鉴别。 相似文献
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DNA条形码是一种快捷高效的分子鉴定新技术,近年来在动物分类学领域中得到迅速的发展和应用。在条形码的研究中有基于进化树、距离和特征3种常用的分析方法:第1种方法需要构建系统发育树,分析样本在树上的聚类情况;第2种方法依赖于物种种内和种间的序列差异;第3种则是通过一系列的诊断特征位点来鉴定物种。本研究扩增了北京百花山地区14种草螟科昆虫88个样本的线粒体细胞色素C氧化酶亚基Ⅰ(mitochondrial cytochrome c oxidase subunitⅠ,COⅠ)基因片段,分别基于进化树、距离和特征方法进行了分析,以探讨不同DNA条形码方法在草螟科物种鉴定中的可行性。结果表明:在使用邻接法(neighbor-joining,NJ)构建的系统发育树上,14个草螟物种各自聚成一个单系,均被成功区分。基于Kimura双参数模型计算遗传距离得出,种内和种内有一个明显的"barcoding gap",且ABGD软件对样本的划分完全符合形态鉴定结果。在所有的草螟物种中都找到了诊断核苷酸位点,基于特征来鉴定草螟物种的成功率为100%。结果显示,这3种方法对于本研究中所涉及的草螟都具有较好的区分,基于COⅠ基因的DNA条形码可以作为一种有效的工具在草螟科昆虫的物种鉴定中进行应用。 相似文献
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基于线粒体COⅠ基因的齿小蠹属昆虫DNA条形码研究 总被引:2,自引:0,他引:2
齿小蠹属(鞘翅目: 小蠹科)昆虫是植物检疫中经常截获的类群, 为探讨线粒体细胞色素C氧化酶亚基Ⅰ(COⅠ)基因的特定区段作为DNA条形码快速准确鉴定齿小蠹种类的可行性, 以齿小蠹属昆虫为研究对象, 测定分析了线粒体COⅠ基因462 bp碱基序列。序列分析结果显示: 变异位点为259个, 保守位点203个, 简约信息位点181个, 自裔位点78个。所有位点中, A, G, C和T碱基平均含量分别为30.7%, 16.5%, 17.0%和35.8%。A+T含量较高, 为66.5%, 明显高于G+C含量, 表现明显的A+T碱基偏嗜, 且A与T含量相当, 符合昆虫线粒体基因碱基组成的基本特征。转换与颠换结果显示: 该段序列未达到饱和, 可以得到准确的进化分析。利用Kimura 2-parameter模型分析遗传距离得到, 同物种间的遗传距离介于0.002~0.007之间, 不同种间的遗传距离介于0.056~0.431间, 平均遗传距离为0.199, 说明该段序列能够区分不同物种。基于COⅠ基因序列构建的邻接法系统发育树(NJ树)显示, 同一物种聚为同一小支, 且分支自展值均为100%; 近缘种能聚集在一起, 且置信度很高(≥97%)。结果表明应用基于COⅠ基因片段的DNA条形码进行齿小蠹属昆虫分类鉴定具有可行性。 相似文献
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P. A. Clancey 《Ibis》1958,100(3):451-454
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R. Wayne Litaker Mark W. Vandersea Steven R. Kibler Kimberly S. Reece Nancy A. Stokes Franois M. Lutzoni Bryan A. Yonish Melissa A. West Megan N. D. Black Patricia A. Tester 《Journal of phycology》2007,43(2):344-355
Dinoflagellate taxonomy is based primarily on morphology and morphometric data that can be difficult to obtain. In contrast, molecular data can be rapidly and cost‐effectively acquired, which has led to a rapid accumulation of sequence data in GenBank. Currently there are no systematic criteria for utilizing taxonomically unassigned sequence data to identify putative species that could in turn serve as a basis for testable hypotheses concerning the taxonomy, diversity, distribution, and toxicity of these organisms. The goal of this research was to evaluate whether simple, uncorrected genetic distances (p) calculated using ITS1/5.8S/ITS2 (ITS region) rDNA sequences could be used to develop criteria for recognizing putative species before formal morphological evaluation and classification. The current analysis used sequences from 81 dinoflagellate species belonging to 14 genera. For this diverse assemblage of dinoflagellate species, the within‐species genetic distances between ITS region copies (p=0.000–0.021 substitutions per site) were consistently less than those observed between species (p=0.042–0.580). Our results indicate that a between‐species uncorrected genetic distance of p≥0.04 could be used to delineate most free‐living dinoflagellate species. Recently evolved species, however, may have ITS p values <0.04 and would require more extensive morphological and genetic analyses to resolve. For most species, the sequence of the dominant ITS region allele has the potential to serve as a unique species‐specific “DNA barcode” that could be used for the rapid identification of dinoflagellates in field and laboratory studies. 相似文献
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以中国蜜环菌(Armillaria mellea (vahl: Fr.) Kummer)生物种B.生物种E分别与欧洲种A. gallica 和A. ostoyae交配,用AP—PCR技术分析了中国五个生物种及欧洲2个种的代表菌株的系统发育关系。根据系统聚类分析的结果,将其分为4个群:生物种B与A.Gallia, A与C,D与E,A. ostoyae 单独为一群。这与交配试验及RAPD图谱直观分析的结果一致,最小支撑树也支持将中国生物种B鉴定为A.Gallica。证明RAPD是研究蜜环菌生物种的进化关系的有用手段。 相似文献
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WILT OF LUCERNE CAUSED BY SPECIES OF VERTICILLIUM 总被引:1,自引:0,他引:1
The incidence and symptoms of a disease of lucerne induced by Verticillium spp. are much less severe immediately after the crop has been first cut than later when the crop has matured. Infection becomes more marked as the season advances and the age of the ley increases, and cuts taken in the third harvest year are proving unprofitable.
In cross-inoculation studies the causal organisms of lucerne wilt, V. albo-atrum and V. dahliae prove to be only mild pathogens to hop, potato, tomato and antirrhinum, and are not pathogenic to sainfoin and clover. The last two may therefore be cultivated as fodder crops in those areas most severely affected by lucerne wilt, but the cultivation of potatoes after lucerne should be discouraged, particularly if lucerne is to be sown in future years, since the potatoes might build up a reservoir of inoculum of the lucerne pathogen. In studies of host-parasite relations it is shown that V. albo-atrum and V. dahliae from lucerne, V. albo-atrum from hop, V. tricorpus from tomato and V. nigrescens and V. nubilum from potato vary in their effects upon lucerne seedlings growing on agar media, V. albo-atrum from lucerne being, as in the field, the most virulent and the only one to reach the xylem vessels under the conditions of the experiments.
The dressings Agrosan-GN (containing mercury) and Fernasan (containing thiram) are toxic both to spores of Verticillium on lucerne seed coats and to infected plant material carried with seed, and although they are also toxic in vitro to Rhizobium mellelotus this did not prevent dressed seeds which had been inoculated with the bacterium from developing into healthy seedlings bearing normal root-nodules. 相似文献
In cross-inoculation studies the causal organisms of lucerne wilt, V. albo-atrum and V. dahliae prove to be only mild pathogens to hop, potato, tomato and antirrhinum, and are not pathogenic to sainfoin and clover. The last two may therefore be cultivated as fodder crops in those areas most severely affected by lucerne wilt, but the cultivation of potatoes after lucerne should be discouraged, particularly if lucerne is to be sown in future years, since the potatoes might build up a reservoir of inoculum of the lucerne pathogen. In studies of host-parasite relations it is shown that V. albo-atrum and V. dahliae from lucerne, V. albo-atrum from hop, V. tricorpus from tomato and V. nigrescens and V. nubilum from potato vary in their effects upon lucerne seedlings growing on agar media, V. albo-atrum from lucerne being, as in the field, the most virulent and the only one to reach the xylem vessels under the conditions of the experiments.
The dressings Agrosan-GN (containing mercury) and Fernasan (containing thiram) are toxic both to spores of Verticillium on lucerne seed coats and to infected plant material carried with seed, and although they are also toxic in vitro to Rhizobium mellelotus this did not prevent dressed seeds which had been inoculated with the bacterium from developing into healthy seedlings bearing normal root-nodules. 相似文献
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WILT OF LUCERNE CAUSED BY SPECIES OF VERTICILLIUM 总被引:2,自引:0,他引:2
IVOR ISAAC 《The Annals of applied biology》1957,45(3):550-558
A wilt disease of lucerne caused by species of Verticillium is described: at twenty-eight disease areas in England and Wales the pathogen was V. albo-atrum , while at one site it was V. dahliae . Both pathogens form superficial conidia on the basal regions of infected stems. It is shown that V. albo-atrum is introduced into a new area in contaminated plant material harvested with the seed from an infected crop. Rapid secondary spread of the disease follows the dissemination of spores from infected stems, and by contact of these and transported fragments of diseased tissues with the wounded surfaces of recently cut lucerne plants. It is recommended that seed should be collected only from healthy crops and that machinery, footwear, etc. should be disinfected before leaving a site of infection.
Manurial trials showed that the incidence of wilt induced by V. albo-atrum was very severe under all soil conditions tested, whereas V. dahliae is a virulent pathogen only to plants in soil rich in superphosphate. Generally the more vigorous the growth of the lucerne–in soils rich in potash and hoof and horn–the more rapid is both the onset of wilt and the resulting secondary spread of the disease throughout the crop. 相似文献
Manurial trials showed that the incidence of wilt induced by V. albo-atrum was very severe under all soil conditions tested, whereas V. dahliae is a virulent pathogen only to plants in soil rich in superphosphate. Generally the more vigorous the growth of the lucerne–in soils rich in potash and hoof and horn–the more rapid is both the onset of wilt and the resulting secondary spread of the disease throughout the crop. 相似文献
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IVOR ISAAC 《The Annals of applied biology》1953,40(4):630-638
An investigation has been made of the spread of Verticillium disease of tomato and antirrhinum by root contact between diseased and healthy plants, and by the growth of the fungi through the soil. V. alboatrum and V. Dahliae spread rapidly by root contact, while V. tricorpus, V. nigrescens and V. nubilum showed no appreciable spread. Where the tomato and antirrhinum hosts wilted and died the causal organism was invariably isolated from the soil near the roots of the dead plants, but where the inoculated host harboured species without showing disease symptoms, as in the case of V. tricorpus and V. nubilum in antirrhinum, the parasite was never obtained from the soil. Moreover, these species in antirrhinum, and V. nigrescens and V. nubilum in tomato would appear to be mild pathogens since when they were already well established V. alboatrum was later actually found to enter and parasitize the respective hosts and bring about its own typical disease symptoms.
Spread through the soil in the absence of hosts was tested by the insertion into it of infected wheat grains followed by later attempts to isolate the fungi at varying distances from the point of infection. V. alboatrum, V. Dahliae and V. tricorpus showed little or no growth through the soil, whereas V. nigrescens and V. nubilum spread saprophytically to some extent, though never attaining sufficient concentration to cause appreciable infection of later planted tomatoes.
The conclusion is reached that spread of disease caused by species of Verticillium takes place mainly by root contact, with a rapidity relatively proportional to that of the death of the host. This finding, together with the lack of spread from antirrhinum plants infected with the non-lethal V. tricorpus and V. nubilum , suggests that the parasites remain in the vascular tracts until the death of the hosts. 相似文献
Spread through the soil in the absence of hosts was tested by the insertion into it of infected wheat grains followed by later attempts to isolate the fungi at varying distances from the point of infection. V. alboatrum, V. Dahliae and V. tricorpus showed little or no growth through the soil, whereas V. nigrescens and V. nubilum spread saprophytically to some extent, though never attaining sufficient concentration to cause appreciable infection of later planted tomatoes.
The conclusion is reached that spread of disease caused by species of Verticillium takes place mainly by root contact, with a rapidity relatively proportional to that of the death of the host. This finding, together with the lack of spread from antirrhinum plants infected with the non-lethal V. tricorpus and V. nubilum , suggests that the parasites remain in the vascular tracts until the death of the hosts. 相似文献
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五种水螅基因组DNA的RAPD多态性研究 总被引:2,自引:1,他引:1
利用RAPD技术对水螅科2属5种水螅进行了基因组DNA多态性研究.80个随机引物筛选出18个引物,共扩增出303个清晰稳定的DNA片段,分子量在0.3~3.0kb之间.以UPGMA聚类法构建树状图,阐明水螅属种的分类关系:大庆水螅Hydra daqingensis、北京水螅H.beijingensis和强壮水螅H.robusta 3种分成两组并聚在一起;不同属的漠河水螅H.mohensis和绿水螅Chlorohydra sp.聚为一支;由此2属5种水螅分为两个支系. 相似文献
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原位复染鉴别细胞的凋亡与坏死 总被引:3,自引:0,他引:3
用10μg/ml去甲斑蝥素作用于人红白血病K562细胞24h,诱导其发生凋亡。采用台盼蓝染液染色,再瑞特-吉姆萨染液原位复染的方法,验证了凋亡细胞与坏死细胞质膜通透性不同的说法。利用此方法可以在普通光学显微镜下把一张涂片上的凋亡细胞、坏死细胞及活细胞区别开来。该实验也表明,常规用台盼蓝染液拒染细胞数来统计活细胞数的方法是值得商榷的。 相似文献