Water-Pretreatment Squash Technic: A New and Simple Practical Method for the Chromosome Study of Animals

To simplify the staining of animal chromosomes (especially in insect testes) the authors have borrowed (with necessary modifications) the squash technic of plant cytology. The method has four steps: (1) Water pretreatment. This step requires only about 5-10 minutes either in water at room temperature or in water kept at about 38°C. in a water bath. (2) Fixation. Ordinarily only 5 minutes in 10-15% aqueous solution of glacial acetic acid is necessary. (3) Staining. The fixed tissue is rinsed in two or three changes of distilled water and then placed in a solution of basic fuchsin: either 1% in 30% ethyl alcohol, or 0.2-0.4% in 5-10% lactic acid. In the former solution the staining period should be about 2 minutes: in the latter, 5-20 minutes. The time is not critical. (4) Squashing. The material is rinsed in several changes of distilled water, placed on a clean slide and squashed under a cover glass. Such preparations last 4-5 weeks, and a technic is described for removing the cover glass in order to mount in Euparal and to make them permanent. The authors list various species of vertebrates as well as invertebrates in which the technic has given good chromosome staining, as shown by illustrations.     

The chorion genes of the medfly. II. DNA sequence evolution of the autosomal chorion genes s18, s15, s19 and s16 in Diptera

We present a total of approximately 15 kb of DNA sequences, encompassing four chorion genes Ccs18, Ccs15, Ccs19, Cc16 and their flanking DNA in the medfly C. capitata. Comparison of coding regions, introns and intergenic sequences in five Dipteran species, D. melanogaster, D. subobscura, D. virilis, D. grimshawi and C. capitata documented an extensive divergence in introns and coding regions, but few well conserved elements in the proximal 5′ flanking regions in all species. These elements are related to conserved regulatory features of three of the genes, including tissue- and temporal regulation. In the fourth, gene s15, significant alterations in the 5′ flanking region may be responsible for its changed temporal regulation in C. capitata. One long intergenic sequence, located in the distal 5′ flanking region of gene s18, is homologous to ACE3, a major amplification control element and contains an 80-bp A/T-rich sequence, known to stimulate strong binding of the origin recognition complex (ORC) in D. melanogaster. Analysis of the nucleotide composition of all chorion genes in C. capitata and D. melanogaster showed that C. capitata exhibit less biased representation of synonymous codons than does D. melanogaster.     

Genetic analysis of mortality in murine angiostrongyliasis costaricensis using SMXA recombinant inbred mouse strains

The SMXA recombinant inbred mouse strain set was produced by systematic inbreeding from the F2 generation of a cross between two progenitor inbred strains, A/J and SM/J, which differed markedly with respect to the patterns of infection with Angiostrongylus costaricensis. We have applied this set to genetic analysis of mouse susceptibility to this nematode infection. The mortality was variable among substrains of the SMXA RI strains, indicating the involvement of multiple genes. Linkage analysis showed several chromosomal regions closely linked to mortality; chromosome 6 (D6Rik86, 87; P0.001), 10 (D10Rik66–D10Mit12; P=0.0058), 13 (D13Rik79, 80; P=0.0096) and 17 (D17Mit28–D17Rik76; P=0.0088).     

金钟藤入侵群落的种间联结及生态位特征

金钟藤(Decalobanthus boisianus)是林业有害植物, 其暴发生长和扩散对森林生态系统造成了严重破坏。本文以海南岛48个金钟藤典型分布群落为研究对象, 用方差比率法和贡献定律法探究群落的稳定性; 用χ ²统计量、联结系数(AC)、共同出现百分率(PC)、Ochiai指数和Dice指数分析金钟藤与伴生物种的种间联结关系; 用生态位宽度、生态位相似性系数和生态位重叠指数研究群落中各物种的生态位特征, 以期为金钟藤生物防治的植物物种筛选提供借鉴。结果表明: (1)金钟藤所在48个群落共有156种伴生植物, 其中大戟科、茜草科、桑科、无患子科和樟科植物占优势; (2)群落中优势物种呈正联结关系, 植物种类累积倒数百分比与累积相对频度交点坐标为(44.53, 55.47), 远离稳定交点坐标(20, 80), 说明群落处于不稳定状态; (3)金钟藤与芳槁润楠(Machilus suaveolens)、黄椿木姜子(Litsea variabilis)、岭南山竹子(Garcinia oblongifolia)、显脉杜英(Elaeocarpus dubius)、鸭脚木(Schefflera octophylla)和银柴(Aporusa dioica)都紧密关联, 说明金钟藤与这些物种的资源利用方式较相似; (4)金钟藤的生态位宽度最大, 与伴生物种间的生态位重叠度较高, 但伴生物种间的生态位重叠度较低。金钟藤的入侵导致群落处于不稳定状态, 并与伴生物种间存在激烈的竞争关系。因此, 建议在金钟藤已入侵的群落中大量栽种芳槁润楠、黄椿木姜子、显脉杜英、鸭脚木和银柴, 以遏制其蔓延; 大量栽种翻白叶树(Pterospermum heterophyllum)、海南菜豆树(Radermachera hainanensis)、九节(Psychotria rubra)和肉实树(Sarcosperma laurinum)用于金钟藤入侵群落的植被恢复。     

三峡库区不同消落带下中华蚊母树群落特征及其与土壤环境因子的关系

中华蚊母树(Distylium chinense)是华中地区特有种, 在三峡库区局部地段占优势形成群落。对三峡库区不同生境下中华蚊母树群落特征及其与土壤环境因子的关系进行研究可为其保护及消落带植被恢复提供科学依据。运用数据分类和排序等方法, 对中华蚊母树群落物种组成、植物区系、物种多样性及其与土壤环境因子的关系进行研究。结果表明, 共发现维管植物56种, 隶属于36科54属, 植物区系以世界分布、泛热带分布和北温带分布为主, 生活型以草本和灌木为主。自然消落带灌木层重要值最大的是中华蚊母树, 草本层重要值最大的是藤本植物地果(Ficus tikoua), 主要伴生种为具有一定水淹耐受性的灌木和多年生草本, 如小梾木(Swida paucinervis)、白茅(Imperata cylindrica)等; 反季节消落带灌木层仅中华蚊母树一种, 草本层重要值最大的物种是狗牙根(Cynodon dactylon), 主要伴生种为苍耳(Xanthium sibiricum)、苘麻(Abutilon theophrastic)等一年生草本。双向聚类分析将调查的6个样地分为四大类群: 中华蚊母树+小梾木群落、中华蚊母树+地果群落、中华蚊母树+细叶水团花(Adina rubella)群落、中华蚊母树+狗牙根群落。典范对应分析表明, 海拔高度和土壤pH是影响中华蚊母树群落物种分布的主要环境因子, 土壤有机质、全氮、全磷和速效钾含量是次要因素, 但它们是影响群落物种多样性指数的主要因子, 其中土壤氮是群落的限制因子。因此, 在反季节消落带植被重建中, 对中华蚊母树群落进行构建时, 除考虑细叶水团花、小梾木等作为高海拔消落带伴生物种外, 还应增加土壤氮和钾的供给, 使中华蚊母树群落保持较高的物种多样性, 维持消落带生态系统稳定。     

Daphnia development and reproduction: Responses to temperature

1. 1.|The development times and reproduction were measured for Daphnia pulex and Daphnia magna from 5 to 30°C at 5°C increments.

2. 2.|The general trends for D. pulex and D. magna were for the duration of all juvenile instars to be less than that of adults and for the last juvenile (or adolescent) instar to be longer than all previous juvenile instars.

3. 3.|The number of juvenile instars both species pass through before adulthood is influenced by temperature with increasing numbers occurring at temperature extremes.

4. 4.|Duration of development time decreased over the entire range of increasing temperatures measured for D. pulex but increased for D. magna at 30°C in relation to 25°C.

5. 5.|Quadratic models were less desirable than simple linear logarithmic transformations of the form ln Y = ln a + b ln X for describing the temperature/development relationship.

6. 6.|The greatest young production occurs at 15 and 20°C with significant decreases occurring at temperatures above and below these.

7. 7.|The observed temperature-dependent phenomena an the ecological relationships for the two species are discussed.

东太平洋公海茎柔鱼饵料中鱼类和头足类组成初步分析

茎柔鱼(Dosidicus gigas)广泛分布于东太平洋海域,在海洋生态系统中扮演着重要角色。本研究根据2019年6—12月我国鱿钓生产船在东太平洋公海采集的62尾茎柔鱼样本,对其胃含物残留的4131个耳石和75个角质颚样本进行鉴定分析。结果表明: 茎柔鱼主要摄食鱼类10种、头足类4种。从摄食饵料的出现频率和数量百分比来看,主要摄食的鱼类饵料为荧串光鱼、朗明灯鱼和墨西哥尾灯鱼,荧串光鱼在不同胴长组的茎柔鱼胃内均有出现,是茎柔鱼最重要的饵料;摄食的头足类主要有茎柔鱼、鸢乌贼和爪乌贼。随着胴长的增大,茎柔鱼摄食的饵料种类不断增加,头足类和灯笼鱼饵料比例增加,且摄食的饵料尺寸增大,摄食营养结构发生改变,摄食营养水平随之提高。研究结果可为评估不同饵料生物在茎柔鱼食性转变过程中的食物贡献提供基础资料。     

温度对新菠萝灰粉蚧生长发育和繁殖的影响

新菠萝灰粉蚧是近年在我国新发现的一种重要外来入侵害虫,温度是决定新菠萝灰粉蚧能否建立稳定种群的最基本因素.本研究以南瓜作为寄主,探索恒温条件对新菠萝灰粉蚧生长发育和繁殖的影响.在17、20、23、26、29和32 ℃,光周期14L∶10D,相对湿度(75±5)%的实验室条件下,测定了新菠萝灰粉蚧各虫态的发育历期、发育速率、存活率和繁殖力,组建了新菠萝灰粉蚧的实验种群生命表.结果表明: 在20～29 ℃,各虫态的发育历期均随温度的升高而缩短,在20 ℃下雌、雄若虫期的发育历期最长,分别为46.95和50.26 d;29 ℃时雌虫若虫期的发育历期最短,为20.28 d;而雄虫若虫期在32 ℃时最短,为20.70 d.各虫态的温度与发育速率的关系均符合二次回归关系.此外,温度显著影响新菠萝灰粉蚧的存活率,在29 ℃时雌、雄若虫期的存活率均最高,分别为70.3%和69.3%;雌虫世代的发育起点温度为13.80 ℃,有效积温为491.50日·度;雄虫世代的发育起点温度为11.61 ℃,有效积温为388.85日·度.新菠萝灰粉蚧的产卵前期和成虫寿命随着温度的升高而缩短;成虫产卵量在29 ℃时最高,达每雌442.2粒;最小为20 ℃,仅为每雌111.8粒;29和20 ℃下种群趋势指数分别为168.2和19.1,且在17和32 ℃时,新菠萝灰粉蚧1龄若虫和3龄若虫均表现为生长停滞.说明温度过高或过低均不利于其生长发育.温度对新菠萝灰粉蚧的生长发育、存活、繁殖及种群增长有显著的影响,23～29 ℃是最适宜新菠萝灰粉蚧生长发育和繁殖的温度范围.     

Permanent Mounting of Unstained and Aceto-Orcein Stained Cells in the Water-Soluble Medium, Abopon

For cellular morphology, mammalian cells were grown on cover slips in Leigh ton tubes, fixed in 1% osmic acid vapor for 2 min, decolorized with 30% H₂O₂ in 5% ammonium oxalate solution (1:7) for 2 min, then washed thoroughly, and finally mounted in a water-soluble medium consisting of a saturated solution of Abopon in 0.2 M phosphate buffer, pH 7.0. For chromosomal analysis of similarly cultured cells, aceto-orcein preparations were made by conventional methods, with the following minor modifications: following pretreatment with colchicine and hypotonic expansion, the cells on the cover slips were fixed in acetic-alcohol (1:3), air dried, incubated at 37° C for 15 min in 2% orcein in 45% acetic acid, rinsed in 45% acetic acid, washed several times in distilled water, and finally mounted in Abopon mounting medium. Both kinds of preparations were allowed to harden for 24 hr before being handled. Such slides will keep for years at room temperature. Studies requiring frequent comparisons of cellular and chromosomal morphology of cultured cells can thus be extended over long periods of time.     

Thermal avoidance and preference in Daphnia magna

1. Water fleas (Daphnia magna) bred at 23°C were non-responsive to temperatures between 13 and 25°C.

2. At the lower (11°C) and upper limits (30°C) their klinokinetic avoidance behaviour showed a larger intraindividual than interindividual variation.

3. Thermal sensitivity for avoidance responses in D. magna was about 1.5°C.

4. For D. magna bred for one parthenogenetic generation at 14°C heat avoidance temperature was about 8°C lower, and cold avoidance temperature was about 1°C higher than in D. magna from 23°C.

5. In group experiments the animals showed some preference for the acclimation temperature.

6. Cold induced stenothermy and warm induced eurythermy in D. magna were related to the mode of reproduction.

铁棍山药微型块茎遗传转化体系的建立

怀山药(Dioscorea opposita)遗传转化是对其进行基因功能分析和遗传改良的基础, 但目前国内外尚未见相关报道。以怀山药优良品种铁棍山药(D. opposita cv. ‘Tiegun’)的微型块茎为受体材料, 对影响遗传转化的因素进行优化, 建立了由根癌农杆菌介导的山药遗传转化体系。过表达质粒载体pCAMBIA1301-DoSERK2含GUS标记基因和潮霉素(Hyg)抗性筛选基因, 沉默质粒载体pART27-DoSERK2含卡那霉素(Kan)抗性筛选基因。根癌农杆菌抑制剂特美汀(Tim)的最佳浓度为500 mg·L ^-1; 再生芽和生根时, Hyg的最佳浓度分别为15和20 mg·L ^-1, Kan的最佳浓度分别为120和160 mg·L ^-1。对转化植株进行PCR和GUS组织化学检测, 结果显示外源基因已整合到铁棍山药转基因株系的基因组中并在细胞中表达。该研究建立了一套取材便利的铁棍山药遗传转化方法, 对其它品种山药的转化也具有参考价值。     

Aggression among sea urchins on Caribbean coral reefs

The existence and nature of intra- and interspecific aggression were examined for five species of sea urchins inhabiting Caribbean coral reefs. The studies took place in the San Blas Islands of Panama and involved the following species: Echinometra lucunter (Linnaeus), E. viridis Agassiz, Diadema antillarum Phillipi, Lytechinus williamsi Chesher, and Eucidaris tribuloides (Lamarck). An intruder was placed next to an undisturbed resident and the behaviors and responses of both individuals were followed. All pairwise combinations of resident species and intruder species were tested except for combinations involving D. antillarum as intruder.

For E. viridis and E. lucunter, agonistic interactions occurred commonly (46–79% of trials) between conspecifics and congeners. Almost all of the agonistic encounters involved pushing. Additionally, biting occurred in 8–25 % of the trials. Residents were most often the aggressors and usually succeeded in retaining their location. Intruders only succeeded in forcing residents out of their positions if the intruder was equal to or larger in size than the resident. Surveys of an undisturbed population of E. viridis during the daytime indicated that 16% of the individuals were engaged in intraspecific agonistic interactions at any one time.

D. antillarum exhibited biting behavior against both species of Echinometra in 23–24% of the trials. Biting attacks against L. williamsi and E. tribuloides occurred rarely. L. williamsi only once demonstrated pushing behavior and never was observed biting another sea urchin. E. tribuloides occasionally exhibited pushing and biting behaviors, both as residents and as intruders, and was twice observed biting Echinometra.

These studies suggest that two kinds of agonistic interactions may commonly occur among Caribbean reef-dwelling sea urchins: (1) intraspecific and interspecific aggression among Echinometra, and (2) predatory/aggressive attacks against Echinometra by D. antillarum. The former may result in greater dispersion of Echinometra relative to food and shelter resources and control the spatial distribution and concentration of Echinometra grazing pressure within an area. The attacks by D. antillarum may result in the restriction to, or higher densities of, Echinometra in crevices and rugose microhabitats that provide shelter from the larger-bodied D. antillarum.     

黄土丘陵区两种丛藓科植物展叶时长及关键影响因子

藓类植物是变水植物,展叶时长是影响其生长发育的关键。目前,干旱半干旱区藓类植物野外生长发育动态及其影响因子的研究鲜见报道。本研究以黄土丘陵区常见的土生对齿藓和扭口藓为对象,研究了9月5日—11月25日野外自然条件下两种藓类的展叶情况。结果表明: 9—10月,两种藓类叶片表现出有规律的“展开-闭合-展开”的日动态变化,并且土生对齿藓叶片在上午闭合时间较扭口藓平均提前0.68 h,下午叶片展开时间则推后了1.79 h。两种藓类在雨季时展叶时间较长,土生对齿藓展叶时长为251 min,较扭口藓(361 min)低30.4%。近地表大气相对湿度是影响藓株展叶时长的关键因子,藓种的形态结构也会影响展叶时长;相比于土生对齿藓,扭口藓植株较为低矮,中肋占叶片比例大,茎皮部细胞镶嵌结构更为突出,其展叶时的湿度阈值(54.3%)低于土生对齿藓(60.1%)。展叶时长主要受湿度的影响,扭口藓对环境的适应性更强。     

Indirect Immunofluorescence and FISH for Enumerating Contaminated Site Sulfate-Reducers

A new species of sulfate-reducer, Desulfosporosinus meridiei, was recently isolated from gasoline-contaminated anaerobic groundwater in which degradation of toluene and other hydrocarbons occurred. Ground-water from inside (three sites) and outside (one site) the contaminant plume was probed with specific polyclonal antibodies raised against two strains of D. meridiei (strains T2 and S6). Molecular 16S rRNA probes designed to hybridize with cells of D. meridiei were also used. Cell counts using antistrain T2 antibodies (specific for all strains of D. meridiei and two strains of D. orientis) were similar (10³ cells/mL) both inside and outside the plume as were total DAPI counts (10⁶ cells/mL). The numbers of cells stained with antibodies specific for Group B strains of D. meridiei varied between locations. The molecular probes DSP477A and DSP477B were designed for, and were effective on, pure cultures of D. meridiei and were able to distinguish this species from Desulfovibrio desulfuricans in hybridization experiments. No cells were seen to hybridize with probe DSP477B in groundwater samples. Cell counts in groundwater using the universal eubacterial probe, EUB338, were only 8% to 29% of DAPI counts. Fluorescence intensity was poor and auto fluorescence of particles made counting difficult. This study showed that molecular probing using techniques commonly employed in many laboratories was of little use for evaluating microbial populations in this groundwater. Polyclonal antibodies were considerably more useful for identifying populations of specific cells. The lack of difference in cell numbers between contaminated and nearby uncontami-nated groundwater suggests that cell counts will not always be useful as indicators of intrinsic remediation.     

基于模式产地的分子证据澄清南平鳞毛蕨的分类学地位

地球上有多少物种一直以来都是人们极度关注的问题, 而存疑物种是造成该问题难以解决的重要阻碍之一。目前, 模式标本是存疑物种得以确定的唯一凭证标准, 然而, 因其特殊的分类学地位而难以进行有效的观察和研究。因此, 前往模式产地寻找存疑物种的原始信息成为最有效的替代途径。随着分子生物学, 特别是DNA条形码技术在物种分类与鉴定中的广泛应用, 准确而快速鉴定存疑物种成为可能。以存疑物种——南平鳞毛蕨(Dryopteris yenpingensis)为例, 利用来自模式产地的分子数据进行DNA条形码分析, 并结合形态学特征澄清了南平鳞毛蕨的分类学地位, 确定南平鳞毛蕨与荔波鳞毛蕨(D. liboensis)为同一物种, 同意将荔波鳞毛蕨作为其异名。研究结果和方法可为今后澄清更多存疑物种的分类学地位提供参考。     

Comparative analysis of complete plastid genome reveals powerful barcode regions for identifying wood of Dalbergia odorifera and D. tonkinensis (Leguminosae)

Dalbergia odorifera T. C. Chen (Leguminosae), a rare and endangered tree species endemic to Hainan Island of China, produces the most expensive and rarest wood in China. The wood characteristics of D. odorifera are remarkably similar to those of D. tonkinensis (a much less sought-after species from Vietnam), and the DNA from wood is often highly degraded, making it very difficult to identify the two species using anatomical features or DNA barcoding based on regular DNA markers. To solve the confusion of identifying wood reliably from the two species, we built and analyzed the plastome library of 26 samples from 18 Dalbergia species, of which 12 samples from eight closely related species of D. odorifera are newly sequenced in this study. Phylogenomic analysis suggested that the relationships among the 26 samples are mostly well resolved, and conspecific individuals from different populations of D. odorifera and D. tonkinensis clustered together. Between the plastid genomes of the two species, we identified 129 indels and 114 single nucleotide polymorphisms. By assessing a subset of 20 nucleotide polymorphisms and 10 indels using 37 population-level samples (20 samples of D. odorifera and 17 samples of D. tonkinensis), we recovered eight species-specific barcode regions that could be suitable for identifying the wood D. odorifera and D. tonkinensis. To examine their utility in wood identification, we amplified the eight DNA barcodes using six wood samples and recovered an amplification success rate of 83.3%, demonstrating a reliable method for precise wood identification of the two species.     

新疆翠雀花属二新种

描述了自新疆维吾尔族自治区发现的毛茛科翠雀花属二新种,腺梗翠雀花和巩留翠雀花,并分别给出了此二种与其近缘种的区别特征。     

Chloroplast DNA and the determination of species status in the Disa tripetaloides complex (Orchidaceae), and its relationships to three species Racemosae, section Disa

Disa cardinalis and three populations within the D. tripetaloides species complex contain variation in their chloroplast DNA (cpDNA) variability. All four taxa possessed unique cpDNAs and sequence divergence values ranged from 0.34 to 1.03%. A phylogeny of these genomes was reconstructed, along with the genomes of three other species, D. racemosa, D. uniflora and D. venosa, all of which are also section Disa and series RAcemosae, to determine the relationship of these closely related species to the D. tripetaloides complex. A phylogeny of the taxa using morphological data was also reconstructed. Outgroup comparison was made with D. sagittalis, a member of section Coryphaea. Although the molecular and morphological data were not completely congruent, both data types revealed D. cardinalis, rather than D. tripetaloides ssp. aurata, to be more closely allied with D. tripetaloides ssp. tripetaloides, suggesting that D. tripetaloides ssp. aurata should be elevated to species rank. Additionally, the high sequence divergence observed between the Natal and Cape populations, coupled with their geographical isolation and alternate flowering seasons, suggests that these two D. tripetaloides ssp. tripetaloides populations may, in fact, be more appropriately ranked as subspecies.