用AFLP技术分析紫茎泽兰的遗传多样性

利用AFLP技术,选择EcoRI/MesI这一酶切组合,应用6对E 3/M 3引物组合进行选择性扩增,检测了24个地区的紫茎泽兰种群的基因组DNA多态性。共扩增出509个遗传位点,其中392个多态位点,多态率为77.01%。有效等位基因数(Ne)为1.50,Nei’s基因多样性指数(H)为0.29,Shannon信息指数(I)为0.42。并通过Jaccard的方法将电泳带矩阵转化为遗传相似性系数矩阵,进行了UPGMA聚类分析。结果表明:被检测的24个紫茎泽兰种群遗传多样性丰富;丰富程度与入侵定植时间有关,最早受紫茎泽兰入侵的云南省遗传多样性最丰富,变异最大,大致分为9个类群,紫茎泽兰的遗传基因可能对该省多样的气候条件或生境产生了明显的适应性变化;新入侵地区的则遗传多样性相对较低,并与云南省临近地区具有明显的种源地的地源性亲缘关系。子实的风媒传播是紫茎泽兰入侵的主要途径,样点间均具有明显的地源性亲缘关系;水媒也可能是该草传播的途径之一。紫茎泽兰可能首先从缅甸、越南、老挝经风媒入侵我国云南南部,逐渐扩散入侵中北部,又从云南北部和东部传入四川、贵州、广西、湖北等省。     

外来入侵植物紫茎泽兰不同组织RNA提取方法

从植物组织中提取高质量的RNA是进行cDNA文库构建等分子生物学研究的前提。在苯酚法的基础上,改进并得到了一种适合紫茎泽兰根、茎、叶总RNA快速提取的方法,消除了蛋白质、DNA、多糖等的污染。该方法提取的紫茎泽兰不同组织总RNA纯度高、完整性好,可用于RT-PCR、cDNA文库构建、Northern杂交等分子生物学实验,而且简单、经济、重复性好,适合于多种植物组织总RNA的提取。Northern杂交表明F3’H基因在紫茎泽兰的根、茎、叶等组织中广泛存在,但在叶中的表达量最高,在根中的表达量最低。     

光皮桦AFLP分子标记体系的建立

为了建立光皮桦AFLP分子标记体系,利用SDS法、常规CTAB法和CTAB-硅珠法提取光皮桦(Betula luminifera H.Wink.)嫩叶DNA,并进行检测比较.结果显示,CTAB-硅珠法更适合光皮桦基因组DNA的提取,所得在基因组DNA纯度高OD值在1.8左右,适用于AFLP分析.利用AVLP分子标记技术,采用Mse I-EcoRI酶切组合,从64个引物组合中筛选出51个带型分布均匀、多态性高且分辨能力强的引物组合.同时,通过对比实验确定了光皮桦AFLP反应的最佳模板DNA用量为300ng、酶切时间4h和预扩增产物稀释倍数30倍等,优化了相关AFLP反应体系,为今后利用AFLP分子标记技术研究光皮桦野生居群的遗传多样性分析和分子遗传图谱的构建打下坚实的基础.     

紫茎泽兰中泽兰酮类成分的抗炎活性研究

探讨紫茎泽兰提取物与单体的体外抗炎活性,为寻找紫茎泽兰的抗炎有效成分提供依据。以超临界CO_2提取物(SFE)、超临界CO_2萃取-分子蒸馏提取物(MDE)、甲醇提取物(MSE)及泽兰酮单体9-羰基-10,11-去氢泽兰酮(euptoxA)、9-羰基-10-Hβ-泽兰酮(9-oxoageraphorone)为实验材料,采用体外脂多糖(LPS)诱导小鼠单核巨噬细胞RAW264.7炎症模型,通过对NO以及TNF-α炎症因子的测定,进行紫茎泽兰各提取部位及泽兰酮单体的抗炎活性研究。超临界CO_2提取物、超临界CO_2萃取-分子蒸馏提取物、甲醇提取物及单体euptox A、9-oxoageraphorone均具有抗炎活性;泽兰酮单体euptox A、9-oxoageraphorone均具有显著的抗炎活性,抗炎活性高于各个提取物,其中9-oxoageraphorone的NO抑制率(IC_(50 )=10.09μg/mL)大于euptox A(IC_(50 )=16.23μg/mL)。本实验发现富含泽兰酮类成分的提取物及泽兰酮单体有较好的抗炎活性,泽兰酮类成分是紫茎泽兰的发挥抗炎作用的有效成分之一,为紫茎泽兰的资源开发与利用提供了依据。     

吉富罗非鱼AFLP反应体系的建立与优化

本文以我国水产养殖主导品种吉富罗非鱼(farmed tilapia)为研究材料,进行扩增片段长度多态性(AFLP)反应体系的研究,初步建立了一套适合罗非鱼的AFLP反应体系。对体系中关键环节的优化结果如下:基因组DNA要求无降解和RNA污染,OD260/OD280比值在1.8~2.0之间;基因组DNA EcoRⅠ/MseⅠ37℃双酶切6h、20℃连接20h,连接产物10倍稀释,PCR预扩增产物稀释45倍作为选择性扩增的模板,选择性扩增产物经6%变性丙烯酰胺凝胶电泳检测可获得条带清晰、背景干扰小的图像。该体系的构建为今后进行罗非鱼群体遗传多样性分析、种质鉴定、重要经济性状分子标记的开发及遗传连锁图谱构建等方面研究提供了参考。     

中华稻蝗AFLP反应体系的建立与优化

以中华稻蝗为研究材料,提取到高质量的总DNA.通过优化酶切连接、预扩增、选择性扩增等实验条件,确定了AFLP银染技术方法,从而建立了中华稻蝗AFLP分子标记研究体系,得到了清晰的AFLP指纹图谱,为探讨稻蝗种间遗传学关系提供了新的技术手段.     

紫茎泽兰乙醇提取物对棉铃虫生长发育和繁殖力的影响

采用室内生物测定法研究了紫茎泽兰Eupatorium adenophorum Spreng乙醇提取物对棉铃虫Helicoverpa armigera (Hübner)生长发育和繁殖力的影响。结果表明,当给棉铃虫幼虫饲喂含紫茎泽兰提取物浓度分别为0.088、0.44、2.2和11 g/kg的饲料时,棉铃虫的死亡率显著高于对照,而羽化率和蛹重均显著低于对照（P≤0.05）。在棉铃虫成虫产卵实验中,0.08、0.4和2 g/L紫茎泽兰提取物对产卵有一定引诱作用,而10 g/L则对产卵有驱避作用。取食浓度分别为0.08、0.4、2和10 g/L紫茎泽兰提取物的棉铃虫卵孵化率降低。表明取食紫茎泽兰提取物对棉铃虫的生长发育和繁殖有明显不利影响。     

紫茎泽兰土壤种子库特征及其对幼苗的影响

研究紫茎泽兰(Eupatorium adenophorum)土壤种子库特征及其对紫茎泽兰种子的萌发、幼苗命运的影响, 在综合治理紫茎泽兰入侵危害及防止紫茎泽兰的继续扩散等方面有着重要的意义。该文在攀枝花紫茎泽兰入侵严重的地区, 通过采集果园、放牧灌丛以及禁牧灌丛3种不同生境紫茎泽兰土壤种子库的样本, 以说明不同干扰程度下紫茎泽兰种子在土壤中的分布状况。通过野外调查并结合盆栽实验, 初步研究了紫茎泽兰土壤种子库基本特征以及光照和种子在土壤中的埋藏深度等对紫茎泽兰幼苗的影响。结果表明: 1)果园、放牧灌丛和禁牧灌丛等3种干扰程度不同生境的深层种子量占总种子量的比例分别为56.44%、46.96%和24.86% (p=0.006), 这说明土壤深层种子量大小与干扰成正比, 干扰越大, 深层次紫茎泽兰种子量占总种子量的比重越大。2)播种在0、1和5 cm深度的种子萌发率分别为64.67%、22.67%和13.33%, 即种子埋藏越深, 萌发率越低, 不同层次种子萌发率差异极显著(p=0.00); 幼苗死亡率分别为27.95%、0和0, 表层种子萌发的幼苗有较高的死亡率, 而由埋藏在深层的种子萌发的幼苗没有死亡, 土壤表层发芽的幼苗与不同埋藏深度种子萌发的幼苗之间死亡率差异极显著(p=0.00)。3)在无遮蔽、半遮蔽和全遮蔽3种不同情况下, 紫茎泽兰幼苗的死亡率分别为72.15%、30.38%和4.87%, 定居率分别为6.66%、33.99%和46.67%, 即遮蔽程度越高, 死亡率越低, 定居率越高, 不同处理之间死亡率和定居率差异均极显著(p=0.00)。研究结果暗示, 强光可能是导致紫茎泽兰幼苗死亡的重要原因, 人类活动的干扰可能导致更多的紫茎泽兰种子进入土壤深层, 从而改变了紫茎泽兰土壤种子库的结构。由于土壤深层种子比表层种子具有更强的抵抗强光照射等不良环境因子影响的能力, 所萌发的幼苗成活率高, 表明其具有更高的繁殖效率, 因此人类活动干扰是紫茎泽兰入侵后难以根除的原因之一。     

泽兰实蝇雌成虫的寄主选择行为

【背景】泽兰实蝇是紫茎泽兰的专性寄生天敌,已成为控制紫茎泽兰的重要因子。泽兰实蝇的寄主选择力对其控制紫茎泽兰至关重要,但相关报道还很少。【方法】采用"Y"形嗅觉仪在实验室内测定泽兰实蝇雌成虫对寄主植物——紫茎泽兰及其他植物的选择行为,以及紫茎泽兰受重金属胁迫后泽兰实蝇对其选择的变化。【结果】不同日龄的泽兰实蝇雌成虫对紫茎泽兰的选择性不具有显著差异;交配与否对雌成虫的寄主选择没有显著影响;雌成虫对紫茎泽兰的选择显著高于黄蒿、水葫芦和蓝花鼠尾草等非寄主植物;雌成虫对花期及机械损伤的紫茎泽兰选择更显著;紫茎泽兰经重金属镉(Cd)、铅(Pb)和锌(Zn)单一及复合处理后,泽兰实蝇雌成虫对其选择性显著降低。【结论与意义】泽兰实蝇雌成虫的寄主选择性随植物种类、寄主植物(紫茎泽兰)的生育期、健康状况及重金属胁迫而变化,这有助于更好地利用泽兰实蝇控制紫茎泽兰。     

紫茎泽兰花蕾发育过程中类Beclin1基因的克隆表达分析

以紫茎泽兰不同发育时期花蕾为实验材料,通过光学显微观察、DNA ladder检测表明紫茎泽兰花蕾发育过程中绒毡层有PCD现象发生,自紫茎泽兰花蕾中克隆长为679 bp的Beclin1 cDNA基因部分片段,同烟草叶片的Beclin1基因序列（AY701316）同源性为98%,通过Northern blotting分析,在紫茎泽兰花蕾发育过程中细胞程序性死亡（PCD）相关Beclin1基因表达在花蕾发育第Ⅱ期和Ⅲ期较为旺盛,且在花蕾发育第Ⅱ期最强烈。本研究首次克隆并证实紫茎泽兰花蕾Beclin1基因与PCD有一定的相关性,为分析紫茎泽兰入侵机理提供新的思路和手段。     

Curcuminoids as inhibitors of thioredoxin reductase: A receptor based pharmacophore study with distance mapping of the active site

Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.     

A Unique Protease Cleavage Site Predicted in the Spike Protein of the Novel Pneumonia Coronavirus (2019-nCoV) Potentially Related to Viral Transmissibility

正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases     

Comparative morphology of the carpel in the Liliaceae: Hewardieae, Petrosavieae, and Tricyrteae

The young pistils in the melanthioid tribes, Hewardieae, Petrosavieae and Tricyrteae, are uniformly tricarpellate and syncarpous. They lack raphide idioblasts. All are multiovulate, with bitegmic ovules. The Petrosavieae are marked by the presence of septal glands and incomplete syncarpy. Tepals and stamens adhere to the ovary in the Hewardieae and the Petrosavieae but not in the Tricyrteae. Two vascular bundles occur in the stamens of the Hewartlieae and Tricyrtis latifolia. Ventral bundles in the upper part of the ovary of the Hewardieae are continuous with compound septal bundles and placental bundles in the lower part. Putative ventral bundles occur in the alternate position in the Tricyrteae and putative placental bundles in the opposite. position in the Petrosavieae. The dichtomously branched stigma in each carpel of the Tricyrteae is supplied by a bifurcated dorsal bundle.     

Entwicklungsgeschichte und Ultrastruktur von Pollenkitt und Exine bei nahe verwandten entomophilen und anemophilen Angiospermensippen derAlismataceae,Liliaceae, Juncaceae,Cyperaceae, Poaceae undAraceae

Some closely related members of the monocotyledonous familiesAlismataceae, Liliaceae, Juncaceae, Cyperaceae, Poaceae andAraceae with variable modes of pollination (insect- and wind-pollination) were studied in relation to the ultrastructure of pollenkitt and exine (amount, consistency and distribution of pollenkitt on the surface of pollen grains). The character syndromes of pollen cementing in entomophilous, anemophilous and intermediate (ambophilous or amphiphilous) monocotyledons are the same in principal as in dicotyledons. Comparing present with former results one can summarize: 1) The pollenkitt is always produced in the same manner by the anther tapetum in all angiosperm sub-classes. 2) The variable stickiness of entomophilous and anemophilous pollen always depends on the particular distribution and consistency of the pollenkitt, but not its amount on the pollen surface. 3) The mostly dry and powdery pollen of anemophilous plants always contains a variable amount of inactive pollenkitt in its exine cavities. 4) A step-by step change of the pollen cementing syndrome can be observed from entomophily towards anemophily. 5) From the omnipresence of pollenkitt in all wind-pollinated angiosperms studied one can conclude that the ancestors of anemophilous angiosperms probably have been zoophilous (i.e. entomophilous) throughout.

     

Identification and Characterization of the First Equine Parainfluenza Virus 5

正Dear Editor,Parainfluenza virus 5 (PIV5), known as canine parainfluenza virus in the veterinary field, is a negative-sense,nonsegmented, single-stranded RNA virus belonging to the Paramyxoviridae family (Chen 2018). The virus was first reported in primary monkey kidney cells in 1954 (Hsiung1972), then it has been frequently discovered in various     

Pathogenic Characterization and Full Length Genome Sequence of a Reassortant Infectious Bursal Disease Virus Newly Isolated in Pakistan

<正>Dear Editor,Infectious bursal disease (IBD) is one of the most important diseases of the poultry. The IBD virus (IBDV), a nonenveloped virus belonging to the Birnaviridae family with a genome consisting of two segments of double-stranded RNA (segments A and B), targets B lymphocytes of bursa of Fabricious leading to immunosuppression. In Pakistan,poultry farming is the second biggest industry and IBD is the second biggest disease threating the poultry sector.However, there is limited genome information of IBDV