从顽拗植物荔枝中提取基因组DNA技术的研究

针对荔枝体内含有大量影响ＤＮＡ提取质量的多酚等次生代谢物质的特点,在常规ＳＤＳ和ＣＴＡＢ提取方法的基础上做了技术改进,即在核裂解之间先破碎细胞,将存在于细胞质中次生物质除去后再裂解细胞核,同时加入活性炭以吸咐杂质反复清洗几次后加入核型解液释放基因组ＤＮＡ,纯化之后经外观和电泳检测,以及Ｄ２６０ｎｍ和Ｄ２８０ｎｍ比值的测定,限制性内切酶反应,ＰＣＲ扩增的结果表明,用改进方法提取的ＤＮＡ无论在纯度上还     

彗星电泳法测定双酚A对雄性幼龄小鼠脑细胞的DNA损伤

双酚A是一种日常生活中无处不在的环境雌激素,具有生殖和神经毒性,但低剂量长期暴露对发育期青少年的危害性常常被低估或忽视。本研究以4周龄雄性清洁级小鼠为实验对象,以茶油作为溶媒对照,分别以双酚A浓度为0μg·m L-1、0.1μg·m L-1、10μg·m L-1和1 000μg·m L-1的茶油灌胃小鼠8周,然后利用彗星电泳法检测各组小鼠脑细胞的DNA损伤。结果显示,不同浓度双酚A暴露8周后,彗星电泳图像显示小鼠脑细胞DNA出现不同程度的损伤,随着暴露剂量的增加,带有彗尾的脑细胞比率从对照组小鼠的9.5%分别升高到暴露组小鼠的34.5%、36.0%和50.5%,细胞总体的尾部DNA含量、尾长和尾矩也都逐渐增加,而且各双酚A暴露组小鼠与溶媒对照组小鼠脑细胞都具有显著性差异(P0.01),这说明中长期双酚A暴露(包括低浓度环境暴露)会导致雄性幼龄小鼠脑细胞的DNA损伤。     

Use of genetic,climatic, and microbiological data to inform reintroduction of a regionally extinct butterfly

Species reintroductions are increasingly used as means of mitigating biodiversity loss. Besides habitat quality at the site targeted for reintroduction, the choice of source population can be critical for success. The butterfly Melanargia russiae (Esper´s marbled white) was extirpated from Hungary over 100 years ago, and a reintroduction program has recently been approved. We used museum specimens of this butterfly, mitochondrial DNA data (mtDNA), endosymbiont screening, and climatic‐similarity analyses to determine which extant populations should be used for its reintroduction. The species displayed 2 main mtDNA lineages across its range: 1 restricted to Iberia and southern France (Iberian lineage) and another found throughout the rest of its range (Eurasian lineage). These 2 lineages possessed highly divergent wsp alleles of the bacterial endosymbiont Wolbachia. The century‐old Hungarian specimens represented an endemic haplotype belonging to the Eurasian lineage, differing by one mutation from the Balkan and eastern European populations. The Hungarian populations of M. russiae occurred in areas with a colder and drier climate relative to most sites with extant known populations. Our results suggest the populations used for reintroduction to Hungary should belong to the Eurasian lineage, preferably from eastern Ukraine (genetically close and living in areas with the highest climatic similarity). Materials stored in museum collections can provide unique opportunities to document historical genetic diversity and help direct conservation.     

Characterization of the genes involved in nitrogen cycling in wastewater treatment plants using DNA microarray and most probable number-PCR

To improve nitrogen removal performance of wastewater treatment plants (WWTPs), it is essential to understand the behavior of nitrogen cycling communities, which comprise various microorganisms. This study characterized the quantity and diversity of nitrogen cycling genes in various processes of municipal WWTPs by employing two molecular-based methods:most probable number-polymerase chain reaction (MPN-PCR) and DNA microarray. MPN-PCR analysis revealed that gene quantities were not statistically different among processes, suggesting that conventional activated sludge processes (CAS) are similar to nitrogen removal processes in their ability to retain an adequate population of nitrogen cycling microorganisms. Furthermore, most processes in the WWTPs that were researched shared a pattern:the nirS and the bacterial amoA genes were more abundant than the nirK and archaeal amoA genes, respectively. DNA microarray analysis revealed that several kinds of nitrification and denitrification genes were detected in both CAS and anaerobic-oxic processes (AO), whereas limited genes were detected in nitrogen removal processes. Results of this study suggest that CAS maintains a diverse community of nitrogen cycling microorganisms; moreover, the microbial communities in nitrogen removal processes may be specific.

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Level of environmental threat posed by horticultural trade in Cactaceae

Ornamental horticulture has been identified as an important threat to plant biodiversity and is a major pathway for plant invasions worldwide. In this context, the family Cactaceae is particularly challenging because it is considered the fifth most threatened large taxonomic group in the world; several species are among the most widespread and damaging invasive species; and Cactaceae is one of the most popular horticultural plant groups. Based on the Convention on International Trade in Endangered Species of Wild Flora and Fauna and the 11 largest online auction sites selling cacti, we documented the international cactus trade. To provide an in‐depth look at the dynamics of the industry, we surveyed the businesses involved in the cactus trade in South Africa (a hotspot of cactus trade and invasions). We purchased seeds of every available species and used DNA barcoding to identify species to the genus level. Although <20% of this trade involved threatened species and <3% involved known invasive species, many species were identified by a common name. However, only 0.02% of the globally traded cacti were collected from wild populations. Despite a large commercial network, all South African imports (of which 15% and 1.5% were of species listed as threatened and invasive, respectively) came from the same source. With DNA barcoding, we identified 24% of the species to genus level. Based on our results, we believe that if trade restrictions are placed on the small proportion of cacti that are invasive and there is no major increase in harvesting of native populations, then the commercial trade in cactus poses a negligible environmental threat. However, there are currently no effective methods for easily identifying which cacti are traded, and both the illicit harvesting of cacti from the wild and the informal trade in invasive taxa pose on‐going conservation challenges.     

重金属Cr(VI)、Pb及Cu胁迫对双齿围沙蚕体腔细胞的DNA损伤

为探讨重金属Cr(VI)、Pb以及Cu对沙蚕体腔细胞DNA的毒性效应,以双齿围沙蚕为受试动物,重金属按不同剂量水平,Cr(VI):10、100和200 mg·L~(-1),Pb:5、50和100 mg·L~(-1),Cu:1、10和20 mg·L~(-1),分别胁迫沙蚕24 h,以不加任何重金属离子的海水为对照,采用单细胞凝胶电泳技术,检测其体腔细胞DNA损伤程度。结果表明,与空白对照组相比,3种重金属离子的各浓度组都能引起沙蚕体腔细胞DNA损伤,且3种重金属胁迫浓度与细胞DNA损伤程度之间存在显著的剂量-效应关系。双齿围沙蚕可以作为单细胞凝胶电泳的实验材料用于重金属所致环境污染的生物监测指示生物。     

基于环境DNA宏条形码的太湖流域底栖动物监测与生态健康评价

大型底栖动物是生态环境监测和评估的主要目标生物类群之一。环境DNA宏条形码技术的发展为提高底栖动物多样性监测的通量、精准性、标准化程度提供了新的机遇,但该方法在我国尚未有流域尺度的应用先例,其结果的可靠性和对生态环境健康状况的指示性有待检验。率先将环境DNA宏条形码技术用于太湖流域65个点位的底栖动物监测和流域生态健康评价,并与同步进行的形态学监测结果进行了比较。结果表明:①环境DNA方法能检出更多的底栖动物类群,在科、属、种水平上检出的分类单元数分别是形态学监测结果的106%、132%、155%;②基于环境DNA技术的检测方法能够很好地识别形态学监测结果中的优势物种,检出的科级、属级分类阶元能够覆盖形态学监测结果中90%以上的生物量和个体数,同时包含60%以上的物种数;③两种方法对同一物种的检出频次显著相关(R²>0.7,P<0.0001),总体检出一致率达72.3%;④在底栖动物完整性指数(B-IBI)方面,环境DNA方法与形态学方法的计算结果显著相关(R²=0.235,P<0.0001),94%的点位的B-IBI等级划分误差在1级以内,且两种方法的计算结果在底栖动物完整性的流域空间格局描绘上高度重合。综上所述,环境DNA宏条形码技术在太湖流域底栖动物群落监测和评价中的整体应用结果表明,环境DNA监测方法结果可靠,将其进一步规模化应用有望显著提高我国水生态系统生物监测结果的准确性和生态健康评价的技术水平。     

Frequency and timing of extrapair fertilisation in the polyandrous red phalarope (Phalaropus fulicarius)

In sequentially polyandrous bird species, where females mate with different males in succession during a single breeding season, sperm stored by females can occasionally lead to extrapair fertilisations (EPFs) in clutches cared for by the second and subsequent males. Thus, we predicted that in red phalaropes (Phalaropus fulicarius) – a sex-role-reversed, sequentially polyandrous, arctic breeding shorebird – EPFs would occur more frequently in clutches laid later in the breeding season. We used multilocus DNA profiling to examine the frequency and timing of EPFs in a population of red phalaropes breeding in the Canadian high arctic. Using a technique to determine parentage without maternal DNA, we inferred that 6 of 70 chicks in 18 broods resulted from EPFs – one extrapair chick in each of 6 broods. These results were supported by a further analysis using microsatellite DNA. As predicted, broods containing EPFs hatched from clutches laid significantly later in the season than did broods containing no EPFs. The difference in median hatch dates between broods with and without EPFs was 9.0 days, or 38% of the entire egg-laying period in that season. For the whole breeding season, we estimated that 6.5% of chicks were sired by extrapair males, which is similar to extrapair paternity estimates for other sex-role-reversed birds, but relatively low compared to the majority of socially monogamous species studied so far. Received: 24 August 1998 / Received in revised form: 18 January 1999 / Accepted: 24 January 1999     

Low paternity in the hornet Vespa crabro indicates that multiple mating by queens is derived in vespine wasps

Queen mating frequency was studied in the European hornet, Vespa crabro, by analyzing four DNA microsatellite loci in 20 workers from each of 14 nests. Queens were found to be predominantly singly mated (9/14), although double (4/14) and triple mating (1/14) also occurred. For most multiply mated queens, paternity was significantly biased with the majority male fathering on average 80% of the female offspring. The population-wide effective mating frequency was therefore low (1.11), and sister-sister relatedness high (0.701 ± 0.023 SE). Low effective mating frequency in Vespa, in combination with data from other vespines, suggests that high paternity frequency is derived in the group. Some problems with the non-detection of fathers, where the queen was not sampled or shared alleles with males, are analyzed. Received: 16 November 1998 / Received in revised form: 29 March 1999 / Accepted: 12 April 1999     

Relatedness and avoidance of inbreeding in the lizard, Tiliqua rugosa

Two potential mechanisms for reducing the level of inbreeding, sex-biased dispersal and kin avoidance, were examined in the Australian sleepy lizard, Tiliqua rugosa. The home range centres, and the genotypes at four polymorphic microsatellite DNA loci were determined for adult lizards in a 70-ha study area near Mount Mary, South Australia. From estimates of genetic relatedness, females were as closely related to other females as they were to males, both within the whole study area, and within home ranges. Similarly, males were as closely related to other males as they were to females. This suggests that dispersal in the population is not sex-biased. Sleepy lizards form monogamous pairs during the spring. Partners were less closely related to each other than to other potential partners in the home range area. This suggests active choice of unrelated partners. The mechanism for recognising related from unrelated individuals is unknown, but the behaviour could reduce inbreeding. Received: 7 November 1998 / Accepted: 30 May 1999