葡萄碘泡虫、似葡萄碘泡虫和茄形碘泡虫的鉴别及系统发育

葡萄碘泡虫Myxobolus acinosus Nie & Li, 1973、似葡萄碘泡虫Myxobolus pseudoacinosus Guo, et al., 2018和茄形碘泡虫Myxobolus toyamai Kudo, 1917形态非常相似, 有着共同的宿主和相同的寄生部位, 是病原鉴定中容易混淆的种。文章基于形态学和18S rRNA基因信息对三者进行了鉴别和分子系统学研究。成熟孢子形态特征的比较分析显示, 三者形态存在显著差异。葡萄碘泡虫与似葡萄碘泡虫18S rDNA序列相似度为98.4—98.8%, 遗传距离为0.013—0.020; 葡萄碘泡虫与茄形碘泡虫18S rDNA序列相似度为96.1—97.2%, 遗传距离为0.038—0.042; 似葡萄碘泡虫和茄形碘泡虫18S rDNA序列相似度为96.4—97.6%, 遗传距离为0.033—0.040。18S rDNA序列比对显示, 葡萄碘泡虫含有15个关键变异位点, 可将该虫与似葡萄碘泡虫和茄形碘泡虫区分; 似葡萄碘泡虫含有5个关键变异位点, 可将该虫与葡萄碘泡虫和茄形碘泡虫区分; 茄形碘泡虫含有33个关键变异位点可将该虫与葡萄碘泡虫和似葡萄碘泡虫区分。18S rRNA二级结构V4区的E23-2构型可将葡萄碘泡虫与似葡萄碘泡虫和茄形碘泡虫区分, 而V7区的H43构型可将茄形碘泡虫与葡萄碘泡虫和似葡萄碘泡虫区分。以上表明, 三者无论在形态上还是在遗传上均具有独立物种的特征。系统发育分析显示, 葡萄碘泡虫、似葡萄碘泡虫和茄形碘泡虫为系统树中分化较晚的一支。     

淡水鱼类粘孢子虫的18S rDNA分子系统学研究

利用两个通用引物myxoF(5‘-CGCGGTAATTCCAGCTCCAGTAG-3‘)和myxoR(5-ACCAGGTAAGTTTTCCGTGTTGA-3’)成功扩增出圆形碘泡虫、全圆碘泡虫、武汉单极虫、微山尾孢虫和库班碘泡虫5种粘孢子虫的部分18S rDNA序列,其GenBank登录号为：AY165179-AY165183。并结合GenBank其他13个相关序列构建了18个物种的分子系统树。结果表明,碘泡虫,尾孢虫和单极虫较Tetracapsula bryozoides和“PKX”分化晚,它们形成了2个聚类：T．wuhanensis-T.hovorkai-M.rotundatus-M.rotundus-M.bidullatus-M.pellicides-M.pendula-H.salminicola聚类和H.exilis-H.ictaluri-M.spinacurvatura-M.osburni-H.lesteri-H.weishanensis-M.kubanicum的聚类;同一属内,采自国内的种类并没有因为地域关系而形成独立的分支,而是与国外的种类交叉在一起,这意味着粘孢子虫种类之间的地域差别并不大;碘泡虫和尾孢虫在进化上都不是单系的,而且分子数据难以将这两个属的种类分开,因此尾孢虫的尾突可能并不是有效的分类依据,而是和碘泡虫的壳片突起同源的一种附属结构。     

碘泡虫属粘孢子虫分子进化及其与宿主协同关系的研究

以18S rDNA为分子标记,对碘泡虫属Myxobolus粘孢子虫的分子进化规律进行了系统研究。结果显示:碘泡虫属各物种18S rDNA的AT含量(53.2%)高于GC含量(46.8%),表现出一定的AT组成偏向性;各物种18S rDNA遗传距离在0.00~0.35;脑碘泡虫M.cerebralis保守区序列变异介于淡水物种和广盐性物种之间;碘泡虫属18S rDNA分子系统发育树呈现生境相同先聚枝,形态相似后聚枝的规律,并表现出一定的宿主特异性和组织向性。此外,以线粒体COⅠ基因为分子标记,对碘泡虫属相应宿主进行了系统进化分析,结果表明:淡水物种与广盐性物种分别聚在不同的分类枝系中,与寄生于宿主体内的碘泡虫所呈现的支序树有相同的进化趋势,并表现出一定的协同进化关系;海水起源的虹鳟Oncorhynchus mykiss在适应淡水过程中被脑碘泡虫寄生,两者经过长期的演化,最终适应淡水环境。     

田中碘泡虫的重描述及系统发育分析

基于形态学和分子数据(18S rDNA)重新描述了寄生于重庆江津地区金鱼 (Carassius auratus auratus)鳃丝的田中碘泡虫 (Myxobolus tanakai Kato, et al., 2017), 并结合系统发育分析对其近缘种进行了比较研究。形态学特征: 该虫种孢子为后端钝圆的长梨形, 大小为(14.58±0.54) μm×(6.09±0.37) μm; 两个极囊等大; 极丝与极囊长轴垂直缠绕8—10圈; 囊间突起位于极囊前端呈三角形; 具有嗜碘泡。分子特征: 共获得3个分离样本的18S rDNA序列, 其一级结构序列与日本黑川地区鲤 (Cyprinus carpio) 鳃丝寄生的田中碘泡虫的一致性达99.3%—99.6%, 且在系统发生树中聚为一支。结果表明: 本研究种与日本的田中碘泡虫为同种, 此为中国新记录; 金鱼为田中碘泡虫的宿主新记录。18S rDNA的V2、V4和V7区二级结构的比较结果表明: 田中碘泡虫已发生了明显的种内变异, 推测可能因宿主差异和地理隔离同时所致。此外, GenBank中报道的数个野鲤碘泡虫的序列是否为同种有待进一步厘清。     

尼氏单极虫(粘体门,双壳目)重描述及基于18S rDNA系统地位分析

采用常规细胞学方法与18S rDNA序列比较的方法,对采自于重庆市嘉陵江段的粘孢子虫,尼氏单极虫Thelohanellus nikolskii进行了形态学和分子生物学的研究,此为首次在我国长江流域检获尼氏单极虫。本文在补充了尼氏单极虫中国种群18S rDNA序列信息的分析、比较基础上,探讨了该虫与相邻种属的系统关系,佐证了其分类学位置。     

洪湖碘泡虫的再描述及其近缘种的鉴别性研究

研究基于形态特征和18S rDNA序列相似度、遗传距离、变异位点、GC含量和系统发育比较分析,对采自河南龙湖的寄生于异育银鲫鳃部的一种黏孢子虫以及相似性极高且易混淆的黏孢子虫种类(洪湖碘泡 Myxobolus honghuensis Liu,et al. 2012、瓶囊碘泡虫Myxobolus ampullicapsulatus Zhao,et al. 2008、咽碘泡虫Myxobolus pharynae Lu,et al. 2012和吴李碘泡虫Myxobolus wulii (Wu Li,1986) 进行了系统的鉴别研究。研究结果显示: 河南龙湖异育银鲫鳃部所检获的黏孢子虫为洪湖碘泡虫,该种群对所寄生的异育银鲫未造成疾病症状; 咽碘泡虫与洪湖碘泡虫各种群在形态上极相似,两者间18S rDNA序列相似度为99%-100%,遗传距离为0-0.0013,GC含量均为44.31%,变异位点为2个,表明咽碘泡虫与洪湖碘泡虫应为同一物种。     

鲤肠道寄生岳阳碘泡虫新种的形态特征及系统发育分析

在洞庭湖岳阳地区开展鱼类寄生虫调查中,发现一种寄生于鲤Cyprinus carpio L.肠道的黏孢子虫。该黏孢子虫的孢囊呈白色,椭圆形,大小为(1.0±0.2) mm (0.8—1.2 mm)。成熟孢子具有壳瓣,壳面观近似圆形,后端有4—6个“V”形褶皱;缝面观呈纺锤形,缝脊直而粗;孢质均匀,含有一个嗜碘泡;孢子长(9.8±0.6) μm (9.6—10.0 μm),孢子宽(8.2±0.3) μm (8.0—8.5μm),孢子厚(7.3±0.1) μm (7.0—7.5 μm);2个极囊梨形,位于孢子顶端,大小相等,呈“八”字形;极囊长(4.4±0.4) μm (3.8—5.1 μm),宽(2.7±0.2) μm (2.2—3.2 μm),极丝4—5圈。该黏孢子虫与肠膜碘泡虫、丑陋圆形碘泡形态特征非常相似,但其极囊/孢子小于1/2;与文献已报道的鲤肠道寄生北京碘泡虫和鲤肠碘泡虫相比较,其在孢子形态、孢子和极囊大小方面分别存在明显差异。基于该黏孢子虫18S rDNA基因序列(GenBank登录号KY203795)比对分析,该黏孢子虫与山东碘泡虫相似率最高,仅为96%。系统发育分析发现,该黏孢子虫与山东碘泡虫、倪李碘泡虫、住心碘泡虫、Myxobolus encephalicus、Sphaerospora molnari、多涅茨尾孢虫和Henneguya zikaweiensis聚为独立分支,和其他已报道的黏孢子虫亲缘关系较远。综合形态学和18S rDNA基因序列数据,文章报道的鲤肠道寄生黏孢子虫为碘泡虫属一新物种,将其命名为岳阳碘泡虫。     

尖形碘泡虫(黏体门:碘泡虫科)的重描述及其分子系统学研究

研究基于形态和分子信息重描述了寄生于嘉陵江重庆段鲫(Carassius auratus Linnaeus)鳃部和胆囊的尖形碘泡虫(Myxobolus acutus Wu and Chen, 1987),并获得了该虫体的18S rDNA和ITS1 rDNA序列。尖形碘泡虫成熟孢子壳面观呈梨形,前端稍尖,后端钝圆,缝面观呈宽纺锤形。孢子长(13.6±0.9)μm [(11.4—15.3)μm],宽(10.2±0.9)μm [(7.5—12.8)μm],厚(7.6±0.6)μm [(6.9—8.3)μm]。两梨形极囊开口处紧靠并位于孢子前端,极囊大小不等,大极囊长(6.2±0.4)μm [(5.1—7.5)μm],宽(3.8±0.4)μm [(2.8—4.7)μm],极丝盘绕5—8圈,小极囊长(2.7±0.4)μm [(1.7—3.7)μm],宽(1.4±0.2)μm [(0.9—1.9)μm],极丝盘绕2—3圈。基于18S rDNA为分子标记的系统发育分析显示:尖形碘泡虫与中华单极虫(Thelohanellus sinensis)有最近的亲缘关系,两物种形成的进化支与贝壳碘泡虫(M. mu...     

黄颡单尾虫(粘体门,双壳目)的重描述及基于28S rDNA和ITS-5.8S序列的系统地位分析

采用形态分类学方法与以28S rDNA和ITS-5.8S序列为基础的分子系统学研究方法,对采自嘉陵江重庆市磁器口江段的黄颡单尾虫Unicauda pelteobagrusMa,1998进行了形态学和分子生物学的研究。基于28S rDNA数据探讨了黄颡单尾虫以及单尾虫属与相邻种属粘孢子虫间的系统地位;基于5.8S rDNA数据比较分析了粘孢子虫的系统地位。补充了黄颡单尾虫重庆种群形态学信息和28S rDNA、ITS-5.8S rDNA序列的分子信息。     

前沟藻18S rDNA序列克隆和分子鉴定分析

旨在扩增一株未知微藻18S rDNA,并进行序列分析和物种鉴定分析.提取该微藻总DNA,用PCR技术扩增其18S rDNA,对序列进行测序,在GenBank进行序列比对分析,用ClustalX软件构建进化树;用光学显微镜进行形态学分析.经测序其序列长度为1 736 bp,G+C为47％;同源性分析表明,与GenBank中多个强壮前沟藻种的18S rDNA基因序列同源性迭99％;与前沟藻属的其它藻种同源性均迭95％以上.而与裸甲藻属和环藻属的藻株同源性为85％左右.经光学显微镜观察,该藻具有典型的强壮前沟藻形态.结合该藻18S rDNA序列分析和粗略的形态学分析,推断该藻可能为前沟藻属的强壮前沟藻.微藻形体微小,结构简单,需借助电子显微镜才可准确地从形态学上签定其种属,鉴定工作繁琐费时;而利用分子技术,则可能使鉴定工作变得简单快捷.     

Collagen-like triple helix formation of synthetic (Pro-Pro-Gly)10 analogues: (4(S)-hydroxyprolyl-4(R)-hydroxyprolyl-Gly)10, (4(R)-hydroxyprolyl-4(R)-hydroxyprolyl-Gly)10 and (4(S)-fluoroprolyl-4(R)-fluoroprolyl-Gly)10.

For the rational design of a stable collagen triple helix according to the conventional rule that the pyrrolidine puckerings of Pro, 4-hydroxyproline (Hyp) and 4-fluoroproline (fPro) should be down at the X-position and up at the Y-position in the X-Y-Gly repeated sequence for enhancing the triple helix propensities of collagen model peptides, a series of peptides were prepared in which X- and Y-positions were altogether occupied by Hyp(R), Hyp(S), fPro(R) or fPro(S). Contrary to our presumption that inducing the X-Y residues to adopt a down-up conformation would result in an increase in the thermal stability of peptides, the triple helices of (Hyp(S)-Hyp(R)-Gly)(10) and (fPro(S)-fPro(R)-Gly)(10) were less stable than those of (Pro-Hyp(R)-Gly)(10) and (Pro-fPro(R)-Gly)(10), respectively. As reported by B?chinger's and Zagari's groups, (Hyp(R)-Hyp(R)-Gly)(10) which could have an up-up conformation unfavorable for the triple helix, formed a triple helix that has a high thermal stability close to that of (Pro-Hyp(R)-Gly)(10). These results clearly show that the empirical rule based on the conformational preference of pyrrolidine ring at each of X and Y residues should not be regarded as still valid, at least for predicting the stability of collagen models in which both X and Y residues have electronegative groups at the 4-position.     

Synthesis and luminescence properties of a blue-emitting Sr(3.5) Y(6.5) O(2) (PO(4) )(1.5) (SiO(4) )(4.5) :Eu(2+) phosphor

A novel blue‐emitting Sr_3.5Y_6.5O₂(PO₄)_1.5(SiO₄)_4.5:Eu²⁺ phosphor was synthesized via a solid‐state reaction. Powder X‐ray diffraction (XRD) analysis demonstrated that the Sr_3.5Y_6.5O₂(PO₄)_1.5(SiO₄)_4.5 host had a hexagonal crystal structure in the space group P6₃/m and unit cell parameters a = 9.418 Å, c = 6.900 Å. The as‐prepared phosphor showed a blue emission and all the main emission peaks were located at around 466 nm for different excitation wavelengths of 297, 333 and 391 nm. The temperature dependence of the photoluminescence property was investigated in the range 20–250 °C, and the emission intensity decreased to 71% of the initial value at room temperature on increasing the temperature to 150 °C. According to the classical theory of fluorescent thermal quenching, the activation energy (ΔE) for the thermal quenching luminescence of the as‐prepared Sr_3.45Y_6.5O₂(PO₄)_1.5(SiO₄)_4.5:0.05Eu²⁺ phosphor was determined to be 0.20 eV. Copyright © 2011 John Wiley & Sons, Ltd.     

Role of tfdC(I)D(I)E(I)F(I) and tfdD(II)C(II)E(II)F(II) gene modules in catabolism of 3-chlorobenzoate by Ralstonia eutropha JMP134(pJP4)

The enzymes chlorocatechol-1,2-dioxygenase, chloromuconate cycloisomerase, dienelactone hydrolase, and maleylacetate reductase allow Ralstonia eutropha JMP134(pJP4) to degrade chlorocatechols formed during growth in 2,4-dichlorophenoxyacetate or 3-chlorobenzoate (3-CB). There are two gene modules located in plasmid pJP4, tfdC(I)D(I)E(I)F(I) (module I) and tfdD(II)C(II)E(II)F(II) (module II), putatively encoding these enzymes. To assess the role of both tfd modules in the degradation of chloroaromatics, each module was cloned into the medium-copy-number plasmid vector pBBR1MCS-2 under the control of the tfdR regulatory gene. These constructs were introduced into R. eutropha JMP222 (a JMP134 derivative lacking pJP4) and Pseudomonas putida KT2442, two strains able to transform 3-CB into chlorocatechols. Specific activities in cell extracts of chlorocatechol-1,2-dioxygenase (tfdC), chloromuconate cycloisomerase (tfdD), and dienelactone hydrolase (tfdE) were 2 to 50 times higher for microorganisms containing module I compared to those containing module II. In contrast, a significantly (50-fold) higher activity of maleylacetate reductase (tfdF) was observed in cell extracts of microorganisms containing module II compared to module I. The R. eutropha JMP222 derivative containing tfdR-tfdC(I)D(I)E(I)F(I) grew four times faster in liquid cultures with 3-CB as a sole carbon and energy source than in cultures containing tfdR-tfdD(II)C(II)E(II)F(II). In the case of P. putida KT2442, only the derivative containing module I was able to grow in liquid cultures of 3-CB. These results indicate that efficient degradation of 3-CB by R. eutropha JMP134(pJP4) requires the two tfd modules such that TfdCDE is likely supplied primarily by module I, while TfdF is likely supplied by module II.     

Induction of in vitro flowering in Dendrobium Madame Thong-In (Orchidaceae) seedlings is associated with increase in endogenous N(6)-(Delta (2)-isopentenyl)-adenine (iP) and N (6)-(Delta (2)-isopentenyl)-adenosine (iPA) levels

We analysed the endogenous cytokinin levels of Dendrobium Madame Thong-In seedlings grown in vitro during vegetative and flowering-inductive periods. HPLC was used to fractionate the extracts and radioimmunoassay (RIA) was used for assay of zeatin (Z), dihydrozeatin (DZ), N(6)-(Delta(2)-isopentenyl)-adenine (iP) and their derivatives. Coconut water used in experiments was found to contain high level (>136 pmol ml(-1)) of zeatin riboside (ZR). Protocorms and seedlings cultured in medium with coconut water were found to contain 0.5-3.9 pmol g(-1) FW of the cytokinins analysed. Seedlings (1.0-1.5 cm) cultured in flowering-inductive liquid medium containing 6-benzyladenine (BA, 4.4 muM) and coconut water (CW, 15%) contained up to 200 and 133 pmol g(-1) FW of iP and iPA, respectively. These levels were significantly higher than all other cytokinins analysed in seedlings of the same stage and were about 80- to 150-folds higher than seedlings cultured in non-inductive medium. During the transitional (vegetative to reproductive) stage, the endogenous levels of iP (178 pmol g(-1) FW) and iPA (63 pmol g(-1) FW) were also significantly higher than cytokinins in the zeatine (Z) and dihydrozeatin (DZ) families in the same seedlings. Seedlings that grew on inductive medium but remained vegetative contained lower levels of iPA. The importance of the profiles of iP and its derivatives in induction of in vitro flowering of D. Madame Thong-In is discussed.     

The helix-coil transitions of poly (dA)-poly (dT) and poly (dA-dT)-poly (dA-dT)

Helix–coil transition curves are calculated for poly (dA) poly(dT) and poly (dA-dT) poly (dA-dT) using the integral equation approach of Goel and Montroll.⁵ The transitions are described by the loop entropy model with the exponent of the loop entropy factor, k, remaining an arbitrary constant. The theoretical calculations are compared with experimental transition curves of the two polymers. Results indicate that the stacking energies for these two polymers differ by about 1 kcal/mole of base pairs. Also, a fit between theory and experiment was not possible for k > 1.70.     

Biology of Bactrocera (Zeugodacus) tau (Walker) (Diptera: Tephritidae)

The biology of the fruit fly Bactrocera tau, an important horticultural pest, was studied under laboratory conditions at 25°C and 60–70% relative humidity on Cucurbita maxima. The duration of mating averaged 408.03 ± 235.93 min. After mating, the female fly had a preoviposition period of 11.7 ± 4.49 days. The oviposition rate was 9.9 ± 8.50 eggs and fecundity was 464.6 ± 67.98 eggs/female. Eggs were elliptical, smooth and shiny white, turning darker as hatching approached, and measured 1.30 ± 0.07 mm × 0.24 ± 0.04 mm. The chorion has polygonal microsculpturing and is species-specific with polygonal walls. The egg period lasts for 1.3 ± 0.41 days. The duration of the larval period is 1.2 ± 0.42, 1.7 ± 0.48 and 4.0 ± 0.94 days for first, second and third instars, respectively. Pupation occurs in the sand or soil and pupal periods are 7.0 ± 0.47 days. The life cycle from egg to adult was completed in 14.2 ± 1.69 days; the longevity of mated females and males was 130.33 ± 14.18 and 104.66 ± 31.21 days, respectively. At least two to three generations were observed from June 2008 to June 2009.     

Spectroscopic investigations of Er(3+) :CdO-Bi(2) O(3) -B(2) O(3) glasses

This article reports on the optical properties of Er³⁺ ions doped CdO–Bi₂O₃–B₂O₃ (CdBiB) glasses. The materials were characterized by optical absorption and emission spectra. By using Judd–Ofelt theory, the intensity parameters Ω_λ (λ = 2, 4, 6) and also oscillatory strengths were calculated from the absorption spectra. The results were used to compute the radiative properties of Er³⁺:CdBiB glasses. The concentration quenching and energy transfer from Yb³⁺–Er³⁺ were explained. The stimulated emission cross‐section, full width at half maximum (FWHM) and FWHM × values are also calculated for all the Er³⁺:CdBiB glasses. Copyright © 2011 John Wiley & Sons, Ltd.