福建黄兔线粒体基因组全序列测定与分析

福建黄兔是中国优良的小型兼用品种，从线粒体水平分析福建黄兔在兔形目动物中的系统发育地位，对探究其起源、进化和种群分类具有重要意义。利用PCR高保真扩增技术和生物信息学软件对福建黄兔线粒体基因组进行获取、拼接与注释。结果发现福建黄兔线粒体基因组全长17 000 bp(Genbank No.MN518689)，主要包括tRNA基因（22个）、rRNA基因（2个）、蛋白编码基因（13个）和控制区（1个）4 部分，基因排列紧密。除控制区，共存在37个编码基因，有9个基因由轻链（L链）编码，其余28个编码基因均由重链（H链）编码。22个tRNA基因中，除 tRNA-Ser(AGY)基因由于缺失DHU臂不能形成三叶草结构外，其余21种tRNA基因均可折叠形成经典的三叶草结构。控制区含有3种结构域：延长终止序列区（ETAS1和ETAS2）、中央保守区、保守序列区（CSB1、CSB2和CSB3），其中，CSB1和CSB2间存在200 bp的短重复序列，CSB3和tRNA-Phe间存在306 bp的长重复序列。基于线粒体基因组控制区序列构建10 种兔形目动物的系统进化树，结果表明福建黄兔起源于穴兔，支持将福建黄兔划归为穴兔属。     

青贮玉米蠕形菌的分离鉴定及其致病性分析

本研究为初步明确引起青贮玉米(Zea Mays L.)叶斑病的蠕形菌种类,采集疑似病斑叶片样本,对病原菌进行分离纯化,形态特征观察,ITS-rDNA序列和GADPH基因序列系统发育分析及致病性测定。形态学和分子生物学鉴定结果表明分离得到的142株病原菌为3属6种真菌,其中麦根腐平脐蠕孢(Bipolaris sorokiniana)和玉米生平脐蠕孢(B.zeicola)为优势病原菌,分别为77株和48株;其次为大斑病菌(Exserohilum turcicum)9株,疣状弯孢(Curvularia verruculosa)2株,新月弯孢(C.lunata)3株,穗状弯孢(C.spicifera)3株。致病性测定发现麦根腐平脐蠕孢和玉米生平脐蠕孢对青海省8个主栽青贮玉米品种均有致病性,其余4种病原菌的致病性各有差异。本研究初步明确引起青贮玉米叶斑病的蠕形菌种类及其致病性,为后续开展病害诊断及其综合防控奠定了基础。     

基于叶绿体基因组探讨地黄属及其近缘类群的系统发育关系

利用叶绿体基因组序列对地黄属及其近缘类群的系统发育关系进行研究。基于GenBank公共数据库,下载地黄属及相关类群的叶绿体全基因组序列52条(50个物种),包括广义玄参科物种23种、近缘类群17种和外类群8种(木犀科和苦苣苔科),分别采用最大似然法及贝叶斯推断进行系统发育分析。结果表明,叶绿体全基因组数据能够显著提高系统发育分析的支持率;广义玄参科不是一个单系;除外类群外,我们共得到5个分支,分别为列当科、泡桐属、肉果草属、玄参属、车前科。地黄属形成一个单系,为列当科其它属的基部类群。列当科内物种间叶绿体基因组大小及所包含的基因数目均差异显著。本研究结果支持APG系统对相关科属的处理,包括将婆婆纳属、腹水草属、毛地黄属从广义玄参科移出至车前科;火焰草属、钟萼草属、马先蒿属、地黄属移出至列当科;泡桐属独立为泡桐科。     

Phylogenetic relationships in the North African genus <Emphasis Type="Italic">Hedysarum</Emphasis> as inferred from ITS sequences of nuclear ribosomal DNA

Sequences of the internal transcribed spacers (ITS) of nuclear ribosomal DNA were analysed to precise their length (637–643 bp) and resolve phylogenetic relationships among eight Mediterranean species of the genus Hedysarum (Fabaceae). The infra-specific variability levels of the ITS sequences of spontaneous population of H. coronarium proved a lack of polymorphism both in the length and in the sequences examined in this species. Hence, a consensus ITS sequence characterising each Hedysarum species has been investigated for analysis of inter-specific polymorphisms. The level of variation of ITS sequence was high enough to make the ITS1 and ITS2 a useful tool for phylogenetic reconstruction. However, ITS2 seems to be relatively more polymorphic and more informative than ITS1 regarding length or GC percent. The phylogenic relationships in the genus Hedysarum based on ITS1 and ITS2 sequences taken independently or together, are discussed in the context of current work in molecular biosystematics. Results exhibited the distinctiveness of the two H. spinosissimum subspecies (i.e. H. spinosissimum ssp. capitatum and H. spinosissimum ssp. spinosissimum). In addition, the great similarity of the ITS sequences between H. coronarium (the only cultivated species of the genus) and H. carnosum suggests the usefulness of the latter in selection programmes to improve pastoral production in semi-arid areas.     

胨冻样芽孢杆菌(Bacillus mucilaginosus YNUCC0001)絮凝剂生产、絮凝特性及其系统发育学分析

对BacillusmucilaginosusYNUCC000116SrDNA的1481bp片段与GenBank中最相似的16个分类单位进行了比较。UPGMA,NJ,ME和MP方法构建的系统发育树显示B.mucilaginosusYNUCC0001与B.mucilaginosusHSCC1605T、B.mucilaginosus1480D及Paenibacillussp.NBT形成一个单系群分支。在50L全自动发酵罐中30℃发酵52h后,菌株YNUCC0001产生的胞外生物多聚絮凝剂(EBF)达到最大产率(粘度:3420C.P.)。在pH4.0、用量为0.25mlL-1的条件下,这种EBF对高岭土悬浊液的絮凝活性最大(99.8%);121℃高压灭菌60min后絮凝活性维持在98.6%。Hg2+,Ca2+,Mg2+,K+,Zn2+对其絮凝活性有促进作用,而Fe3+、Al3+、EDTA和Cu2+则有强烈抑制。     

利用ITS序列和trnL-F序列探讨赖草属6个物种的系统发育关系

本研究克隆了赖草属6个物种Leymus karelinii,L.angustus,L.racemosus,L.arenarius,L.triticoides和L.ambiguus的ITS序列和trnL-F序列,并选用3个新麦属物种的ITS序列和27个小麦族二倍体物种的trnL-F序列,均以Bromus catharticus为外类群,用最大简约法构建系统发育树。结果发现:不同组的赖草属物种分别聚在一起;赖草属植物与新麦草属植物亲缘关系较近;新麦草属植物是这6个赖草属物种的母本来源。     

Molecular phylogeny in Indian Citrus L. (Rutaceae) inferred through PCR-RFLP and trnL-trnF sequence data of chloroplast DNA

The taxonomy and phylogeny of Indian Citrus is revisited using PCR-RFLP of the trnD-trnT and rbcL-ORF 106 regions as well as sequence data analysis of the trnL-trnF intergenic spacer region of cpDNA. The study was based on 50 accessions of Citrus genotypes, collected from wild, semi-wild and domesticated stocks. Of the 13 restriction enzymes (RE) used for restriction digestion of the polymerase chain reaction (PCR) amplicons, four (Hinf I, Msp I, Alu I, Hae III) generated 47 restriction fragments, of which 24 (51%) were polymorphic. PCR-RFLP data showed a genetic distance ranging from 0 to 0.79 among 50 accessions of Citrus, and a cluster analysis, based on Neighbor-Joining (NJ) method, placed all the accessions in eight major clusters. Analysis of trnL-trnF sequences from 23 representative accessions of Citrus showed a pair-wise sequence divergence rate in the range of 0–0.064. NJ, minimum evolution (ME) and maximum parsimony (MP) analyses of trnL-trnF sequences produced phylogenetic trees, which placed all the 23 accessions in five clusters. PCR-RFLP analysis resulted in a well resolved phylogenetic tree with branches supported by moderate to high bootstrap values, while the trnL-trnF sequence-based trees showed only moderate to low bootstrap support for the internal tree branches, indicating uncertain origin of some Citrus genotypes. This study shows that the trnL-trnF spacer sequence data can detect genetic variation in Indian Citrus genotypes, but the utility of the data in inferring phylogeny at intra and inter-specific levels is limited probably by factors such as hybridization, bud mutations, apomixis and polyploidy. However, PCR-RFLP and trnL-trnF data supported the recognition of C. maxima, C. medica, and C. reticulata as the basal species of edible Citrus.     

Discovery of a new avian bornavirus genotype in estrildid finches (Estrildidae) in Germany

Avian bornaviruses (ABV) are known to be the causative agent of proventricular dilatation disease (PDD) in parrots and their relatives (Psittaciformes). A broad range of ABV genotypes has been detected not only in psittacine birds, but also in other avian species including canary birds (Serinus canaria forma domestica) and Bengalese finches (Lonchura striata f. dom.), which are both members of the order songbirds (Passeriformes).During this study 286 samples collected from captive and wild birds of various passerine species in different parts of Germany were screened for the presence of ABV. Interestingly, only three ABV-positive samples were identified by RT-PCR. They originated from one yellow-winged pytilia (Pytilia hypogrammica) and two black-rumped waxbills (Estrilda troglodytes) from a flock of captive estrildid finches in Saxony. The ABV isolates detected here were only distantly related to ABV isolates found in passerine species in Germany and Japan and form a new genotype tentatively called ABV-EF (for “estrildid finches”).     

伏牛白山羊mtDNAD—loop序列多态性和系统进化分析

为研究河南省伏牛白山羊的遗传多样性和系统进化,试验测定了该品种8个个体的线粒体控制区全序列,结果表明,山羊控制区线粒体控制全序列长度为1212bp或1213bp,A T含量占60.1%,其中40个核苷酸位点存在变异(约占3.30%),核苷酸多样度为1.562%,这些差异共定义了7种单倍型,单倍型多样性为0.964,表明中国山羊品种遗传多样性丰富。根据伏牛白山羊序列和GENBANK两条野山羊序列构建了NJ分子系统树,聚类表明,伏牛白山羊和角骨羊单独聚在一枝上,二者亲缘关系较近,伏牛白山羊可能起源于角骨羊。