基于GISH的甘蔗与斑茅F1染色体遗传与核型分析

斑茅在甘蔗育种的利用是现代甘蔗育种种质创新的热点,育种者期望把斑茅中优异的特性通过杂交渗透到甘蔗中。甘蔗与斑茅的F1是斑茅利用研究的难点也是基础。本研究利用基因组原位杂交技术(GISH)分析甘蔗与斑茅的F1染色体构成和核型,探讨甘蔗与斑茅F1染色体的遗传行为。GISH结果表明,甘蔗与斑茅杂交F1的染色体众数68~69条,其中40条来自甘蔗热带种Badila,28~29条来自海南斑茅,未发现有染色体的交换或易位现象。参试材料大部分染色体都属于中部着丝点(m)的染色体,少数为近中部着丝点(sm),YCE95-41核型属2B型,其余的核型都为1B型。甘蔗与斑茅的染色体按n+n的方式传递给F1,本研究结果为斑茅种质在甘蔗育种中的利用及其杂交后代染色体细胞遗传研究提供参考依据。     

New Karyological and Morphometric Data on Poorly Known Bufo surdus and Bufo luristanicus in Comparison with Data of Diploid Green Toads of the Bufo viridis Complex from South of Iran

Previous studies on the Bufo viridis complex, which is distributed broadly across Iran, are incomplete and restricted to a few regions or a few samples. In this paper a new detailed study on the B. viridis complex in southern of Iran （from West to East） is presented. The analysis of 18 morphometric characters with univariate and multivariate methods reveals significant differences between three members of the B. viridis complex namely B. variabilis, B. luristanicus, and B. surdus distributed in southern part of Iran. Our result help to resolve an old taxonomic problem about B. surdus subgroup （taxa closely related to B. surdus） confirming that B. luristanicus and B. surdus are distinct species. Moreover, for the first time we report and describe karyotype details of B. luristanicus and B. surdus which confirmed that they are diploid. Karyological studies demonstrate that all toads from three mentioned species have 2n = 22 chromosomes. These chromosomes are arranged into two groups. First group has six large chromosomes and the second group is composed of five small chromosomes. These chromosomes are metacentric or submetacentric. The number of submetacentric chromosomes is different in three mentioned species of B. viridis complex. Neither sexual heteromorphism, nor secondary constriction was observed in any pairs of chromosomes.     

核型多角体病毒侵染及其与宿主免疫系统互作的研究进展

核型多角体病毒(Nucleopolyhedrovirus,NPV)应用广泛,已被开发成微生物杀虫剂和用于重组蛋白表达等.NPV具有两种病毒颗粒:包埋型病毒粒子(occlusion-derived virus,ODV)和芽生型病毒粒子(budded virus,BV),两者的构成和组装存在差异.病毒包涵体在肠道中溶解后释放出ODV进行初级感染,子代核衣壳通过质膜出芽产生BV引发全身性的次级感染.病毒感染昆虫后能引发宿主的免疫反应,同时NPV病毒已进化出多种策略抑制或逃避宿主的免疫反应,如免疫信号通路、黑化和凋亡等过程.本文将总结鳞翅目特异性NPV与宿主的互作研究,着重介绍NPV的侵染机制、NPV与宿主免疫系统互作的研究进展.     

杆状病毒凋亡抑制基因IAP1促进家蚕CyclinB的核内积累

[目的]家蚕核型多角体病毒(Bombyx mori nucleopolyhedrovirus,BmNPV)是生产上危害最严重的病原之一。BmNPV感染BmN-SWU1细胞将细胞周期阻滞于G2/M期。CyclinB是调控细胞周期G2期向M期转换的重要细胞周期蛋白。因此,研究BmNPV感染后CyclinB变化对解析病毒调控细胞周期的机制具有重要意义,同时探究这个过程中与CyclinB互作的病毒蛋白,可为构建家蚕转基因品系提供分子靶标。[方法]qRT-PCR检测BmNPV感染后BmCyclinB的表达变化;免疫荧光观察病毒感染前后BmCyclinB的定位变化,通过细胞质细胞核蛋白分离实验验证。免疫共沉淀钓取与BmCyclinB互作的病毒蛋白。BmNPV感染期间敲除BmNPV IAP1观察BmCyclinB的入核比例。[结果]BmNPV感染后BmCyclinB转录水平下调。BmNPV感染前BmCyclinB主要定位于细胞质,而感染后主要定位于细胞核。BmNPV感染BmN-SWU1细胞后促进BmCyclinB在核内积累。共钓取了7个与BmCyclinB互作的病毒蛋白,免疫共沉淀和细胞共定位证明BmNPV IAP1与BmCyclinB之间存在相互作用。敲除BmNPV IAP1后BmCyclinB进入细胞核的数量显著减少。[结论]BmNPV IAP1可通过与BmCyclinB互作,促进BmCyclinB在核内积累。     

6个香雪兰品种的染色体核型及聚类分析

以香雪兰（Freesia hybrida）6个品种的根尖为研究材料进行核型分析以揭示其核型特征,进一步通过聚类分析品种间的相似性并探讨可能的亲缘关系。结果表明：香雪兰6个品种均为四倍体,染色体数目都为44。6个品种的核型公式不同：‘Gold River’为2n=4x=44=33m+11sm,‘White River’为2n=4x=44=1M+34m+9sm,‘Castor’为2n=4x=44=35m+9sm,‘上农红台阁’为2n=4x=44=32m+12sm,‘Pink Passion’为2n=4x=44=38m+6sm,‘Red River’为2n=4x=44=37m+7sm。核型参数因品种而异,其中,‘Castor’在相对长度范围和臂比范围上均为最广,‘上农红台阁’核不对称系数最大。6个品种核型不对称系数为57.00%~60.07%,核型类型均为2B,进化程度中等,处于由对称至不对称的过渡阶段,其核型进化的趋势大致为：‘Pink Passion’→‘Red River’→‘Gold River’→‘White River’→‘Castor’→‘上农红台阁’。聚类结果显示,遗传距离为15时可将所有品种分为两类,第一类包含‘Gold River’‘上农红台阁’和‘Castor’; 第二类包含‘Pink Passion’‘Red River’和‘White River’,其核型参数的差异可作为香雪兰品种分类和鉴定的辅助依据,为其细胞学研究和育种实践提供理论参考。     

Construction of the Bac-to-Bac System of Bombyx mori Nucleopolyhedroviru

To construct the Bac-to-Bac expression system of Bombyx mori nucleopolyhedrovirus(BmNPV),a transfer vector was constructed which contained an Escherichia coli(E.coli)mini-F replicon and a lacZ:attTN7:lacZ cassette within the upstream and downstream regions of the BmNPV polyhedrin gene.B.mori larvae were cotransfected with wild-type BmNPV genomic DNA and the transfer vector through subcutaneous injection to generate recombinant viruses by homologous recombination in vivo.The genomic DNA of budded viruses extracted from the hemolymph of the transfected larvae was used to transform E.coli DH10B.Recombinant bacmids were screened by kanamycin resistance,PCR and restriction enzyme(REN)digestion.One of the bacmid colonies,BmBacJS13,which had similar REN profiles to that of wild-type BmNPV,was selected for further research.To investigate the infectivity of BmBacJS13,the polyhedrin gene was introduced into the bacmid and the resultant recombinant(BmBacJS13-ph)was transfected to BmN cells.The budded viruses were collected from the supernatant of the transfected cells and used for infecting BmN cells.Growth curve analysis indicated that BmBacJS13-ph had a similar growth curve to that of wild-type BmNPV.Bio-assays indicated that BmBacJS13-ph was also infectious to B.mori larvae.     

福建大头蛙五个地理居群核型多样性

采用骨髓细胞蒸汽固定法制备染色体标本,研究了产于安徽、浙江、江西、福建和湖南5个地理居群福建大头蛙的核型。结果表明：5居群样本的细胞染色体数均为2n=22,NF=44,核型模式为7＋4。福建居群11对染色体全为中部着丝粒染色体;其它4个居群中的NO．3为亚中部着丝粒染色体,其余为中部着丝粒染色体;各居群中NO．3和NO．7的相对长度和臂比值存在显著或极显著差异。显示不同地理居群福建大头蛙的核型具有丰富的多样性。     

淫羊藿属十二个种的核型

用细胞压片法对12 个淫羊藿属( Epimedium) 植物的根尖有丝分裂进行了观察, 并对其核型进行了比较研究。结果表明: 这12 种淫羊藿在核型上基本相似, 都有1 对中间随体染色体, 而且都为对称核型。粗毛淫羊藿( E. acuminatum) 、印江淫羊藿( E . yinjiangense )、单叶淫羊藿( E. simplicifloum) 、巫山淫羊藿( E. wushanense) 、光叶淫羊藿( E. myrianthum) 、宝兴淫羊藿( E. davidi) 、罗甸淫羊藿( E. luodianense) 、木鱼坪淫羊藿( E. franchetii ) 、箭叶淫羊藿( E. sagittatum) 、E. pubigerum、高山淫羊藿( E. alpinum) 等11 个种的核型公式均为2n = 2x= 6m (2SAT ) + 6sm, 而黔岭淫羊藿( E. leptorrhizum) 的核型公式为2n = 2x = 8m ( 2SAT ) + 4sm。结果显示12 种淫羊藿的体细胞染色体数目均为2n = 12 , 基数X = 6; 都属于对称核型、都有1 对中间随体; 染色体均为m、sm 两种类型。     

合欢核型的分析

应用根尖压片法对含羞草科合欢属合欢的染色体数目和核型进行了研究,结果表明:合欢体细胞染色体数目为2n=26,核型为:2n=26=18m+6sm+2st,属于“2B”类型。染色体相对长度组成为2n=26=4L+8M2+12M1+2S。从染色体水平证明合欢在系统演化上属于较进化的种类。