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61.
同工酶技术已经被广泛应用于虾类群体遗传学及标记辅助选择育种等研究方面.同工酶电泳显示对虾有相应的亚群差异,暖水性虾类的遗传多样性水平高于冷温性种类;连续分布的且在不同生活史阶段所处的栖息地环境变化幅度大的种类往往表现出较低的遗传多态性,虾类群体的多态位点比例和杂合度都较低,养殖群体比野生群体更低.但同工酶电泳技术往往检测不出一些"隐性"或"中性"变异而低估了遗传多样性水平.随着分子生物学技术的发展,从20世纪90年代开始,虾类遗传多样性的研究逐渐向线粒体DNA和核DNA的多态性方向发展.限制性片断长度多态性(RFLP)在虾类线粒体多倍型、物种标记以及比较野生和养殖群体线粒体COI 基因的多态性比较等方面应用普遍.随机扩增多态性(RAPD)技术主要应用于虾类不同地理群体遗传多态性调查,养殖群体与野生群体之间、养殖群体世代之间的遗传多态性比较,该技术得到的虾类遗传多样性水平高于同工酶电泳技术.扩增片断长度多态性(AFLP)技术在虾类中主要应用于遗传连锁图谱的构建,有关虾类的AFLP遗传多样性分析的报道不多.利用AFLP技术对连续选育群体的遗传多样性研究显示,随着选育世代的增加,选育群体的遗传多样性呈现下降趋势,但随着选育时间的延长,群体之间的分化逐渐降低,群体的遗传结构开始趋于稳定,成为一个品系.单一重复序列(SSR)技术主要用于标记种群遗传特异性、不同地理群体间以及野生和养殖群体间的遗传多样性差异.DNA序列分析技术特别是线粒体序列分析技术近年来在虾类遗传多样性研究中的应用逐渐增多.mtDNACOI、mtDNA12S rRNA和mtDNA16S rRNA的基因序列差异被应用于形态相近物种的鉴别、亚属分类、确定个体间亲缘关系和野生与养殖群体间的遗传多样性差异等.为了实现对虾类种群遗传差异的更深入了解,人们把种群遗传结构的研究和系统地理学相结合,即用分子系统地理学的方法来研究遗传谱系空间分布的历史特征,通过种群遗传结构的分析来探讨种内系统地理格局的形成机制、系统发育关系以及现有分布特征,并结合种群的地理分布状况来发现和验证与其相关的地质事件,追溯和揭示种群的进化历程.从育种、养殖以及资源保护的角度出发,人们开始研究野生群体的种质基因库,希望根据群体遗传结构确定合理的增殖策略,制定科学的保护措施,确保虾类养殖业的持续发展.  相似文献   
62.
采用Chelex-100树脂法制备了26个物种(分别属于腔肠动物门、海绵动物门和软体动物门)的共生藻以及一株纯培养共生藻的DNA样品。通过PCR扩增叶绿体23S rRNA基因片段获得了26个有效序列,测序比对结果显示获得的序列与扩增目的基因相符。该方法不仅样品消耗量小、操作时间短、设备需求低廉,而且可以避免使用有害化学试剂。这是一种高效、环保的DNA提取方法,可为共生藻分子研究提供技术参考。  相似文献   
63.
In order to thoroughly investigate the diversity of glacier microorganisms, four DNA extraction methods with different lysis patterns were tested and two screened methods (the Bosshard-Bano method and the Zhou method) were optimized for the most effective form of the filter membrane (cut vs. uncut), the DNA extraction method, and the precipitation method. The two optimized methods were then compared with the commercial Mo-Bio DNA extraction kit, and the results showed that the kit was generally suitable for extraction of microorganism DNA from glacier surface snow. Procedurally, it was found that a modified Bosshard- Bano method (i.e., cutting the filter membrane into pieces, using a specific lysis pattern [lysozyme (5 mg/mL)-protease K (1 mg/mL)-CTAB (1%)-SDS (1%)], performing the extraction only once by chloroform-isoamyl alcohol (24:1), and conducting DNA precipitation by pure ethanol) was also an effective and less expensive method for extraction of microorganism DNA from glacier surface snow.  相似文献   
64.
A study has been made using a variable temperature flowing afterglow Langmuir probe technique (VT-FALP) to determine the equilibrium temperature dependencies of the dissociative electron-ion recombination of the protonated cyanide ions (RCNH+, where R=H, CH3 and C2H5) and their symmetrical proton-bound dimers (RCNH+NCR). The power law temperature dependencies of the recombination coefficients, αe, over the temperature range 180 to 600 K for the protonated ions are αe(T)(cm3 s−1)=3.5±0.5×10−7 (300/T)1.38 for HCNH+, αe(T)=3.4±0.5×10−7 (300/T)1.03 for CH3CNH+, and αe(T)=4.6±0.7×10−7 (300/T)0.81 for CH3CH2CNH+. The equivalent values for the proton-bound dimers are αe(T)(cm3 s−1)=2.4±0.4×10−6(300/T)0.5 for (HCN)2H+ to αe(T)=2.8±0.4×10−6(300/T)0.5 for (CH3CN)2H+, and αe(T)=2.3±0.3×10−6(300/T)0.5 for (CH3CH2CN)2H+. The relevance of these data to molecular synthesis in the interstellar medium and the Titan ionosphere are discussed.  相似文献   
65.
洱海沉积物中有机质和DNA的分布特征   总被引:7,自引:0,他引:7  
测定了洱海沉积物中有机碳、氮,提取了沉积物中的DNA。有机碳氮呈现相同的变化趋势降解主要发生在表层10cm;7cm前后氧化-还原条件变化,微生物电子受体改变,所以有机质的降解方式不同,出现不同的特征。利用分子生物学和地球化学相结合的方法,能更全面的认识和理解早期成岩过程。  相似文献   
66.
本研究建立了一种高效、经济的非损伤性扇贝DNA提取方法,在不影响扇贝个体生存状态的前提下,采用擦拭法和室温裂解相结合的方式在20 min内即可获得个体基因组DNA。以虾夷扇贝(Patinopecten yessoensis)、栉孔扇贝(Chlamys farreri)和海湾扇贝(Argopecten irradians)为实验对象,研究了不同擦拭材料(棉签、滤纸)和不同擦拭部位(外套膜、鳃丝、内脏团)的核酸提取效果,评估了本方法在贝类基因分型领域应用的可行性。研究表明,用棉签、滤纸2种材料擦拭扇贝的鳃丝、内脏团或外套膜组织均能获得主带清晰完整的基因组DNA,且不影响扇贝的存活状态。在3种扇贝中采用棉签擦拭法的DNA得率均高于滤纸法,以这2种方式获得的DNA为模板进行SSR和SNP标记分析,能获得均一稳定的扩增条带,SSR标记分型结果准确可靠。本研究建立了简便、快速进行扇贝基因型分析的非损伤性DNA提取方法,为贝类全基因组选育和珍稀贝类资源的种质保护开发提供了新的技术手段。  相似文献   
67.
Marine ecosystems provide a wide variety of diverse habitats that frequently promote migration and ecological adaptation. The extent to which the geographic distribution of marine organism has reshaped by human activities remains underappreciated. The limitations intrinsic to morphology-based identification systems have engendered an urgent need for reliable genetic methods that enable the unequivocal recognition of fish species,particularly those that are prone to overexploitation and/or market substitution. In the present study, however, an attempt has been taken to identify two locally adapted fish species, Siganus sutor(Valenciennes, 1835) and Seriolina nigrofasciata(Rüppell, 1829) of order Perciformes, which happens to be the first record in Odisha coast,Bay of Bengal. The diagnostic characteristics of Siganus sutor are: dorsal fin XIII-10, anal fin VII-9, pectoral fin 15,pelvic fin II-3, while that of Seriolina nigrofasciata dorsal fin VI-I-35, anal fin I-17, pectoral fin 16, pelvic fin 5. All COI barcodes generated in this study were matched with reference sequences of expected species, according to morphological identification. Bayesian and likelihood phylogenetic trees were drawn based on DNA barcodes and all the specimens clustered in agreement with their taxonomic classification at the species level. The phylogeographic studies based on haplotype network and migration rates suggest that both the species were not panmitic and the high-frequency population distribution indicates successful migration. The result of this study provides an important validation of the use of DNA barcode sequences for monitoring species diversity and changes within a complex marine ecosystem.  相似文献   
68.
DNA甲基化参与调节动物配子发生和胚胎发育过程,TET基因负责DNA主动去甲基化,在基因印迹去除和细胞全能性获得中起重要作用。为了解海洋贝类配子发生和胚胎发育过程中DNA去甲基化如何发生,本研究以虾夷扇贝为研究对象,鉴定了其TET基因,并分析了该基因在性腺和胚胎幼虫发育中的表达变化。结果表明,虾夷扇贝基因组中含有1个TET基因(PyTET),该基因长39127 bp,包含10个外显子,编码1592个氨基酸,其蛋白具有完整的2OGFeDO超家族的加氧酶结构域。在性腺发育过程中,PyTET基因表达峰值出现在休止期精巢和增殖期卵巢,原位杂交结果显示其在精原细胞、精母细胞中均有表达,在卵母细胞中的表达明显高于卵原细胞;在早期发育过程中,其峰值出现在囊胚期。以上结果提示虾夷扇贝配子发生和胚胎发育过程中均发生了DNA主动去甲基化,这将有助于系统了解表观调控在贝类发育中的作用。  相似文献   
69.
Bohai Bay, a representative semi-enclosed bay of China, has been severely impacted in recent decades by rapid human development. Although the mantis shrimp Oratosquilla oratoria plays an important role in the Bohai Bay ecosystem and fishery, its feeding ecology and the impact of habitat changes on its feeding habits are poorly known. In this study, we sought to identify its feeding strategy through the separation of stomach contents, and to describe its trophic ecology during maturation in the Tianjin coastal zone of Bohai Bay by the DNA barcoding method using a fragment (about 690 bp) of the mitochondrial cytochrome oxidase I (CO I) gene. A total of 594 specimens were collected from March to July 2018, and 347 (58.42%) stomachs were found to have food remains. More than half of the O. oratoria individuals had poor feeding activity, and the degree of feeding of females was higher than that of males, but there was no significant difference in the visual fullness index and the fullness weight index (FWI) between sexes for each month. In addition, the feeding activity of O. oratoria was consistent across the months, suggesting in artificial breeding programs it is not worried too much about the accessional diet supplied to the female bloodstocks except maintaining their normal food. A total of 167 retrieved prey items yielded 187 readable sequences; 86.63% of these sequences could be assigned to the species level. Consequently, 25 different prey taxa covering all readable sequences were identified, showing O. oratoria is a generalist predator. In terms of percent composition by frequency of occurrences (%N), crustaceans and fishes accounted for the same and highest percentage (35.83%) in the mantis shrimp diet, followed by mollusks (24.59%) and the remaining marine organisms (3.73%) including Polychaeta (2.67%), Sagittoidea (0.53%), and Chromadorea (0.53%). The result shows the benthic fish Oxudercidae (family) appears to be an especially important food supply for O. oratoria during maturation. The degree of cannibalism decreased with an increase in diet species and the frequencies of cannibalism in this study was noticeably higher than that in previous studies, suggesting that cannibalism may be a significant feeding strategy in the mantis shrimp O. oratoria in the Tianjin coastal zone of Bohai Bay, and because of the frequently cannibalistic behavior, the high stocking density of broodstocks should be avoided.  相似文献   
70.
高通量测序技术广泛应用于环境真核微生物多样性的研究,可检获较之传统形态学方法更高的多样性。然而,检获的高分子多样性与基于形态的物种多样性在构成上的差异仍然不明。本研究首次对比分析了基于形态学的南黄海沉积物中的纤毛虫物种多样性与基于核糖体18S DNA和c DNA高通量测序的多样性异同。结果表明:形态鉴定获得8纲、20目、30科、36属共97种纤毛虫;DNA测序检获10纲、28目、55科、76属共174个OTUs;而通过c DNA测序获取的纤毛虫多样性最高,获10纲、31目、68科、99属共284个OTUs。研究发现,形态学方法检获的纤毛虫均为底栖生纤毛虫;两种分子手段检获的多样性更高,群落结构更为近似,覆盖了形态鉴定所获的绝大部分类群。但DNA测序还检获了序列比例高达90%的浮游类群,可能源于包囊或死亡沉降的物种;而c DNA测序检获了约7%的浮游纤毛虫序列。较之DNA测序,c DNA法检获的底栖纤毛虫在群落结构上与形态学结果更为接近。本研究表明,分子手段有助于更全面地揭示沉积物中的纤毛虫多样性,DNA测序可同时揭示休眠包囊、胞外DNA及过去群落的信息,而c DNA测序在研究活动纤毛虫多样性上更有优势。  相似文献   
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