Expression analysis of <Emphasis Type="Italic">RUS1</Emphasis> and construction of <Emphasis Type="Italic">RUS1</Emphasis> plant expressing vector

RUS1 was one of the disease resistance gene analogs obtained from Setaria italica Beauv. Semi-quantitative RT-PCR analysis result showed that RUS1 gene could be induced by Uromyces setariae-italicae and had relation to the resistance response of Setaria italica Beauv. against Uromyces setariae-italicae infection. Promoter sequence of RUS1 was obtained by the method of Genome Walking, and its length was 675 bp. RUS1 promoter and pCAMBIA1300 vector were fused to construct RUS1::GUS vector. GUS histochemical staining result showed that promoter could activate gene expression. RUS1 gene (including the promoter sequence) was obtained through PCR amplification and then fused with pCAMBIA1300 vector to construct pCAMBIA1300:RUS1 plant expressing vector. The research laid a foundation for gene functional identification of RUS1.     

安徽3株柔嫩艾美耳球虫cox1基因部分序列的比对分析

为研究柔嫩艾美耳球虫不同地理株的线粒体细胞色素c氧化酶第1亚基(cox1)基因与球虫种群之间的遗传关系,以安徽3个地区的3株柔嫩艾美耳球虫为研究对象,通过卵囊的分离与纯化获得纯种虫株,然后应用PCR技术对柔嫩艾美球虫3个地理株的cox1序列进行扩增,并与GenBank上登录的鸡柔嫩艾美耳球虫、巨型艾美耳球虫、布氏艾美耳球虫和堆形艾美耳球虫虫株的相应序列进行比对分析。结果显示,每个虫株都成功扩增出800 bp左右的cox1部分有效序列(pcox1),与柔嫩艾美耳球虫pcox1序列种内无差异,pcox1相应序列种间的差异程度为9.9%～14.9%。表明柔嫩艾美耳球虫的cox1序列可作为艾美耳球虫种间遗传变异研究的标记,为进一步研究艾美耳球虫的群体遗传学特性和耐药性奠定了基础。     

Elymus wawawaiensis编码乙酰辅酶A羧化酶Acc-1基因序列的分析及系统学意义

质体乙酰辅酶A羧化酶(ACCase)催化长链脂肪酸的合成。编码小麦质体ACCase的基因(Acc-1)已用于一年生小麦的系统与进化研究,但还未有多年生物种相应的研究报道。本研究发现,Elymus wawawaiensis的Acc-1基因第8外显子和第7内含子与其他已知该序列相比除核苷酸替代外,多了176个碱基,从而与现有已知的一年生小麦相应序列有显著差异。在NCBI中对该序列和已报道的Acc-1基因相应序列blast比对后,结果表明与Hordeum vulgare Acc-1具有87.15%的最高相似性。以Panicum virgatum、Zea mays和Lolim rigidum为外类群,最大简约法(MP)和邻接法(NJ)对该基因和已报道的Acc-1基因分别构建系统进化树,其中E.wawawaiensis和H.vulgare形成一个分支,自展支持率100%。研究同时表明该基因在小麦族多年生物种系统与进化研究中有重要的应用前景。     

Bioinformatics analysis of tyrosinase-related protein 1 gene (<Emphasis Type="Italic">TYRP1</Emphasis>) from different species

As one member of the tyrosinase-related family directly involved in the production of melanin, TYRP1 is involved in not only melanogenesis but also prevention of melanocyte death, stabilizing tyrosinase and helping determine the shape of melanosomes, etc. Multi-species sequence comparisons showed that there were two evolutionally conserved non-coding regions (from −1306 to −733 and from −642 to −515 according to AL138753) upstream of translational initiation sites, representing putative regulatory regions subject to subsequent experimental tests. Coding sequence length variation and genetic diversity analysis showed that Felis catus, Homo sapiens and Canis familiaris had more genetic diversities than the other species for TYRP1, especially Felis catus that could be a better choice for studying the TYRP1-associated genetic basis underlying the color diversity. As a 75 kDa type-1 transmembrane glycoprotein, mature TYRP1 possesses about 17 kDa modifying components, whose function predominantly depends on the existing glycosyl-groups and the Cu components. In addition, the mutated amino acids within species and the highly conserved amino acids among species were listed in our paper.     

普通小麦济麦21的LOX1基因cDNA片段克隆及分析

以普通小麦品种济麦21为材料,根据GenBank中已发表的LOX1基因序列设计引物,利用RTPCR技术获得一个小麦LOX1的基因片段,并对该片段进行测序和生物信息学分析。结果表明:克隆到的基因片段为384bp(GenBank登录号JX126806),GC含量为64.84%,推导氨基酸残基为127个,分子质量为11.15ku,等电点(pI)为4.93,与杜伦小麦、大麦、高粱、水稻的同源性分别为99.7%、94.8%、73.7%、56.2%。进化树分析表明,六倍体普通小麦LOX1基因与杜伦小麦LOX1基因(GenBank登录号HM126468)亲缘关系较近,先聚为一支,然后再与大麦的LOX1基因(GenBank登录号L35931.1)、高粱的LOX1基因(GenBank登录号GQ369443)聚为一类,最终与亲缘关系较远的水稻LOX1基因(GenBank登录号EU267789)聚类,这与物种亲缘关系的远近基本一致,表明所得LOX1基因片段可作为研究物种亲缘关系或遗传距离的参考标记之一。     

Monosomic Addition Lines of Flowering Chinese Cabbage （B. campestris L. ssp. chinensis var. parachinensis L. H. Bailey）-Chinese Kale （B. oleracea L. var. alboglabra L. H. Bailey）

Interspecific alien addition lines have played significant roles in gene mapping, intergenomic gene transfer and chromosomal homoeological identification between closely related species. Selection of alien addition lines was conducted by karyotype analysis and morphological observation with the reference of parents. Triploid interspecies hybrid （AAC, 2n = 3x = 29） was obtained from Brassica campestris ssp. chinensis var. parachinensis Qinglu 9601 （tetraploid, AAAA, 2n = 4x = 40） x B. oleracea var. alboglabra Baihua 9705 （diploid, CC, 2n = 2x = 18） by immature hybrid embryo culture in vitro. Five different alien monosomic addition lines （AA ＋ C2, AA ＋ C3, AA ＋ C4, AA ＋ C6, AA ＋ C7） were obtained from the backcross progenies of AAC x AA. Each alien monosomic addition line has some specific morphological characters. It is feasible to obtain alien addition lines from the progenies of AAC × AA by karyotype analysis and morphological observation based on the reference of parents     

两型豆属叶绿体基因组特征及密码子偏好性分析

为明确锈毛两型豆的叶绿体基因组结构和两型豆属叶绿体基因组密码子使用偏性及影响因素,以亚热带中、南部地区具有广阔开发利用前景的豆科草种—锈毛两型豆(Amphicarpaea ferruginea)为试验材料,利用高通量测序技术对锈毛两型豆进行叶绿体基因组测序、组装和注释,对其叶绿体基因组结构、基因组成进行分析。同时利用CodonW 1.4.2软件和CUSP在线程序等软件分析锈毛两型豆和两型豆的基因密码子使用偏性参数和核苷酸组成。结果显示：锈毛两型豆叶绿体基因组全长为152 531 bp,包含83 364 bp的大单拷贝(LSC)区、17 935 bp的小单拷贝(SSC)区和25 616 bp的1对反向重复序列,为典型四分体结构,GC含量为35.44%;叶绿体基因组共编码130个基因,包括85个蛋白质编码基因、37个tRNA基因和8个rRNA基因;叶绿体基因组共检测出73个简单重复序列(SSRs),单、二、三、四、五和六核苷酸SSRs的数目分别为41、28、3、1、0和0。从锈毛两型豆和两型豆叶绿体基因组中筛选到适用于密码子使用偏好性分析的CDS基因共48条,两种植物叶绿体基因组具有相似的...     

赖草属林下组植物GBSSI基因序列的系统发育与分子进化

【目的】探讨赖草属林下组植物的种间系统关系与GBSSI基因的分子进化式样。【方法】对赖草属林下组3个类群的GBSSI基因序列进行多克隆测序,并将其与来自小麦族8个代表Ns,St,P,F,Ee和Eb基因组的二倍体植物的GBSSI序列进行序列与系统比较分析。【结果】基因序列多态性与系统发育分析表明:①L.duthiei和L.duthiei var.longearistatus亲缘关系密切;②不同Ns基因组供体参与多倍化物种形成可能导致L.komarovii与L.duthiei和L.duthiei var.longearistatus发生遗传分化;③赖草属林下组植物的GBSSI基因序列在Ns基因组拷贝上的多态性水平低于新麦草属植物Ns基因组序列多态性。【结论】支持L.duthiei和L.duthiei var.longearistatus互为种与变种的分类等级。复制基因GBSSI在多倍体中多态性水平不平等。     

瘤背石磺低频听力相关基因OrNXN克隆及表达分析

从瘤背石磺不同频率声音基因转录组中将有显著差异表达的基因与IMPC人类听觉感知和耳聋相关基因对比发掘出与听力相关的基因NXN。利用cDNA末端快速扩增(RACE)技术克隆获得OrNXN基因cDNA全长序列,并对其序列进行生物信息学分析;利用实时定量PCR(qPCR)技术分析OrNXN在瘤背石磺不同组织、不同声音频率及不同潮汐节点的表达水平。结果显示,OrNXN基因cDNA全长为1 593 bp,开放阅读框长度为432 bp,共编码143个氨基酸,相对分子质量为15.57 ku;结构域预测分析表明,OrNXN含有Thioredoxin家族典型的硫氧还蛋白结构;多序列比对以及进化树构建结果表明,OrNXN与其他物种的相似度较高,保守性较强;荧光定量结果表明,在瘤背石磺7个组织中OrNXN在肠、神经节、性腺和腹足中均有表达,在肠中表达量最高,其次为神经节。不同声音频率刺激下,OrNXN在200和50 Hz处有差异表达,在不同潮汐节点,OrNXN表达量均不同,表达量基本与潮汐水位变化一致。研究表明,OrNXN可能在瘤背石磺参与感知潮汐节率时对潮汐发出的低频声音产生了响应。     

鹅观草属、披碱草属、猬草属和仲彬草属植物的RAPD分析及其系统学意义

利用随机扩增多态性DNA(RAPD)技术分析了鹅观草属 ( 7种 )、披碱草属 ( 3种 )、猬草属 ( 2种 )和仲彬草属 ( 3种 ) 4个属的 15种植物。对 34个OPRON公司十聚体随机引物进行多态性筛选 ,2 7个 ( 79 4% )能产生多态性。 16个引物产生的 2 13个DNA片断 ,用于计算种间Nei氏遗传相似性系数分析 ,在NTSYS程序中利用UPGMA构建系统发育树状图。分析结果表明 :①这 4个属的物种各自聚类在一起 ,在相似系数 0 6水平上 ,明显分为 4组 ,分别代表了Hystrix、Elymus、Roegneria和Kengyilia 4个属 ;②Hystrix的 2个物种与Roegneria和Kengyilia物种的亲缘关系较远 ,与同具StH基因组的Elymus也有较大的RAPD变异 ;③Roegneria与Elymus亲缘关系较近 ;④Roegneria中StY和StYH基因组组合的物种亲缘关系很近 ,StY基因组的物种间存在着RAPD遗传变异 ;⑤Kengyilia与Roegneria亲缘关系较近 ;⑥RAPD结果与形态学和细胞学等分析结果一致 ,支持将它们独立作为属分类等级来处理。本文还对RAPD分析方法在小麦族系统学研究中的应用进行了讨论     

小麦远缘杂交现状、抗病基因转移及利用研究进展

小麦近缘植物中含有丰富的抗病、抗逆和抗虫等基因,是小麦育种的优异基因源。通过远缘杂交可以将近缘植物优异基因转移给小麦,创制包括双二倍体或部分双二倍体、附加系、代换系和易位系等在内的小麦-近缘植物异染色体系。这些含小麦近缘植物血缘的异染色体系是研究物种染色体行为与进化、基因定位与作图的重要素材,也是拓宽小麦的遗传基础、抵御小麦重要病虫害、增加小麦产量和提升小麦品质的重要物质基础。为了更加清晰地了解小麦远缘杂交概况及小麦近缘植物抗病基因向小麦的转移,也为今后小麦远缘杂交研究和种质资源的开发利用提供参考,文中对小麦族物种分类、小麦远缘杂交的定义与意义、小麦族山羊草属、黑麦属、偃麦草属、簇毛麦属、冰草属、大麦属、披碱草属、赖草属、新麦草属以及旱麦草属物种与小麦远缘杂交现状和异染色体系创制情况进行了概括,并对来源于小麦近缘植物被正式命名的17个抗条锈病基因、35个抗叶锈病基因、30个抗秆锈病基因、41个抗白粉病基因、3个抗赤霉病基因、1个抗麦瘟病基因、1个抗叶枯病基因、1个抗颖枯病基因、4个抗褐斑病基因、2个抗眼斑病基因、1个抗梭条花叶病基因、2个抗线条花叶病基因和2个抗禾谷类黄矮病基因向小麦的转移情况及其所在染色体的位置信息进行了归纳。小麦-黑麦1RS·1BL易位系、1RS·1AL易位系和小麦-偏凸山羊草2NS/2AS易位系等抗病优良种质的育成与利用在世界小麦育种史上做出了突出贡献,然而,这仅仅得益于对少数抗病基因的利用。与目前已经被命名的基因数量相比,被利用到小麦育种中的抗病基因相对较少。文中分析了当前已命名抗病基因利用情况比例偏低的原因,并对今后如何利用这些抗病基因提出了建议。同时,还列举了已克隆的源自小麦近缘植物的抗病基因,并对克隆这些基因的方法以及今后可能的研究热点进行了分析,认为加强无遗传累赘的小麦-近缘植物易位系的创制与应用仍可能是今后小麦育种材料创新与新品种培育的一个重要发力点。     

Study on the origin and taxonomic status of yak (Poephagus) using cytochrome b gene of mitochondrial DNA

There still exists a remarkable difference on the yak’s taxonomic status in Bovinae. Primers designed according to the mitochondrial gene sequences of Bos taurus reported were used to amplify and sequence the yak’s cytochrome b gene, and the whole sequence of cytochrome b gene was finally obtained. Using Ovis aries as outgroup taxa, the phylogeny about the representative species of Bovinae was analyzed. Results showed that among the different species, the ratio of transition/transversion (Ts/Tv) of Cytochrome b gene was 4.9, suggesting that the mutation was not saturation. The percentage nucleotide sequence divergence between yak and Bovinae was 8.0%–8.6%, which was higher than that of yak and Bison bison. Phylogeny analysis found that Poephagus grunniens and Poephagus mutus clustered first before gathering with Bison bison, indicating higher genetic comparability than that of Bos. The results sustained the idea that Poephagus grunniens and Poephagus mutus shared one ancestor—the primitive yak. The approximate divergence time between these two species was 0.55 million years. The data also supported the viewpoint that the yak is classified into Poephagus of Bovinae, including two species of Poephagus grunniens and Poephagus mutus. Translated from Acta Veterinaria et Zootechnica Sinica, 2006, 37(11): 1128–1123 [译自: 畜牧兽医学报]     

Molecular markers in identification of sunflower pollen fertility restorer genes

The presence of the Rf1 gene (pollen fertility restorer of CMS PET1) in genotypes of most restorer lines is confirmed by molecular and hybridological analysis methods. Molecular markers of the Rf1 locus are identified for the first time in genotypes of autofertile lines of the collection of the Vavilov All-Russian Research Institute of Plant Industry (VIR) as well as in genomes of perennial and annual wild species of the genus Helianthus. The possibility of introgression of the fertility restorer gene into the genome of the cultivated sunflower during distant hybridization is shown.     

Cloning of TaCYP707A 1 Gene that Encodes ABA 8′-Hydroxylase in Common Wheat（Triticum aestivum L.）

The plant hormone abscisic acid （ABA） regulates many important physiological and developmental processes in plants. The objective of this study was to clone the ABA 8′-hydroxylase gene in common wheat. In the present study, we used the eDNA sequence of barley HvCYP707A1 gene （GenBank accession no. AB239299） as a probe for BLAST search against the common wheat （Triticum aestivum L.） EST database in GenBank. All wheat ESTs sharing high similarity with the reference gene were subjected to contig assembly. Primers were designed based on the constructed contigs to clone the wheat CYP707A1 gene, designated as TaCYP707A1. The genomic DNA sequence of TaCYPTO7A1 gene comprised five exons and four introns, with a size of 2225 bp. The corresponding cDNA sequence of TaCYP707A1 was 1737 bp, containing an open reading frame （ORF） of 1431 bp, a 42-bp 5′-untranslated region （UTR） and a 264-bp 3′UTR, with 94.9% of identical sequences to HvCYP707A1 gene （AB239299）. The neighbor joining tree indicated that the deduced amino acid sequences of TaCYP707A1 gene was highly similar to those of barley and rice. The TaCYP707A1 gene was located on chromosome 6BL using a set of Chinese Spring nullisomic-tetrasomic lines and ditelosomic line 6BS. These results will be of high importance in understanding of molecular mechanism of ABA catabolism.     

Molecular Phylogenetic Analysis of the Main Lineages of Nymphalinae （Nymphalidae： Lepidoptera） Based on the Partial Mitochondrial COI Gene

The phylogenetic relationships of the subfamily Nymphalinae （sensu Chou 1994） were analyzed based on 1 488 bp of mtDNA cytochrome oxidase subunit I （COI） gene sequence data obtained from 24 individuals, along with those of eight species obtained from GenBank. The base compositions of this COI fragment varied among the individuals as follows： T 39.9%, C 14.6%, A 32.2%, and G 13.4%, with a strong AT bias （72.1%）, as usually found in insect mitochondrial genomes. The A ＋T contents of the third, second, and first codon positions of the COI fragments in this study was 92.4, 62.2, and 61.4%, respectively. The phylogenetic trees were reconstructed by neighbor-joining （NJ）, maximum likelihood （ML）, and Bayesian methods by using Byblia anvatara as outgroup. Phylogenetic analyses based on the COI gene sequence data created very similar topologies, which were producing trees with two main clades A and B, and five subclades. The data indicated that the tribes Nymphalini and Hypolimni （sensu Chou 1994） are not monophyletic groups, and the genus Junonia should be removed from Nymphalini to Hypolimni （=Junoniini）. On the basis of the data, the Symbrenthia and Araschnia had a relative distant relationship with the rest of Nymphalini. The relationships of species in the Nymphalini were confirmed via the NJ, ML, and Bayesian methods, namely （（（（Nymphalis ＋ Kaniska） ＋ Polygonia） ＋Aglais） ＋ Vanessa） ＋ （Symbrenthia ＋Araschnia）. This investigation provides a little novel information for Chinese researches of butterflies.     

DNA markers of the <Emphasis Type="Italic">R1</Emphasis> and <Emphasis Type="Italic">R3</Emphasis> genes as predictors of potato late blight resistance

To investigate whether the presence of R-genes is associated with higher late blight resistance of potato cultivars, we modified an earlier described SCAR marker for the R1 gene and developed a new SCAR marker for the R3 gene. We demonstrated the significant input of R1 transferred from Solanum demissum and probably S. stoloniferum into foliage and tuber resistance; the effect of R3 introgression was less conclusive.     

伊洛河流域外来草本植物分布格局

外来生物入侵及其防治已经成为生态学关注的重点和热点问题。目前的研究主要集中在外来入侵种上,然而入侵种仅占外来种中很少一部分,因此,研究外来种现有分布格局对研究生物入侵及其防治有重要意义。以伊洛河流域草本植物群落中的外来种为对象,沿河从河源地到入黄河口选取典型样地,在调查流域内草本植物群落中物种组成的基础上选取外来种,并对外来种种类组成及其分布格局进行研究。结果表明:流域内有外来草本植物27种,分属于15科,种类较多的科为菊科、苋科和豆科;引入方式以有意引种为主。流域横向不同生境间,河滩地在水流的养分富集、季节性洪水物理干扰及人为活动扰动作用下,呈现出受外来种分布较多,而受人类活动扰动最强且营养丰富的农田分布较小的分布格局;纵向环境梯度下,上游河源山地属于自然植被区,人为干扰较轻,且受外来种影响较小;中游丘陵区从自然生态系统向农业生态系统的过渡区域,人类活动的扰动有所加强;下游平原农业区,人类活动强烈,区域内以人工生态系统为主,群落物种组成简单但受外来种影响最大,受自然环境和人类活动的双重影响。不同物种在不同生境间差异明显,其中,小蓬草、钻叶紫菀和反枝苋广泛分布于3种生境中。总体上,伊洛河外来草本植物分布格局在自然因素的基础上强烈受人为因素的影响,呈现出从上游到下游逐渐增多的趋势。     

马转录因子TBX3基因的密码子使用偏好性分析

为了解马TBX3基因的密码子使用特性,为选择该基因最佳的受体以及合适的异源表达系统提供依据,利用CHIPS、CUSP、Codon W等软件对NCBI公布的马TBX3基因(GenBank登录号为XM_014742147.1)的密码子使用情况进行分析,并将其与马的10个肢体生长和肢体形态形成相关基因、模式生物基因组以及其他物种TBX3基因进行比较。结果表明:马TBX3基因偏好使用G/C结尾的密码子,存在19种偏好使用的密码子(RSCU1.20),其中:GCC、CTG、AGC和GTG偏好性较强(RSCU2.00);10个马相关基因在20种偏好密码子中偏好性较强的只有CTG,17种密码子偏好G/C结尾;通过比较30种动物的TBX3基因密码子偏好性,发现其TBX3基因表达水平一般,并且密码子偏好G/C结尾;基于RSCU值和CDS序列的聚类分析发现,奇蹄目的家马与食肉目的家猫、北极熊聚在一起,可见系统进化树更接近这30个物种的真实系统分类;在密码子的使用频率上,小鼠更适合作为马TBX3基因的外源表达宿主。本研究可为TBX3基因在动物遗传育种改良中选择适合的宿主动物、筛选最佳的外源表达系统以及提高其表达水平提供依据。     

小麦族猬草属和近缘属物种醇溶蛋白分析

利用酸性聚丙烯酰胺凝胶电泳(A-PAGE)对小麦族猬草属、赖草属、新麦草属、拟鹅观草属、披碱草属和Lophopyrum共6属19份材料进行醇溶蛋白遗传分析。结果表明:①19份材料共出现19种不同的醇溶蛋白图谱,共分离出56条带纹,多态性高达100%,说明属间和种间具有丰富的醇溶蛋白遗传多态性;②Hystrix duthieissp.duthiei与H.duthieissp.longearistata的亲缘关系较近,它们与H.patula的亲缘关系较远;Psathyrostachys juncea和Psa.huashanica亲缘关系较远;Lophopyrum elongatum和Lo.bessarabicum的亲缘关系较远;③含相似染色体组的物种能各自聚类在一起,它们具有较近的亲缘关系;④醇溶蛋白揭示的亲缘关系与形态学、细胞学研究结果基本一致,表明醇溶蛋白能够用于小麦族多年生物种系统亲缘关系分析。     

通过多基因组比较的方法在5种绿球蓝细菌中识别基因组岛(英文)

为探索水平基因转移在绿球蓝细菌基因组多样性中的作用,5个高光适应性绿球蓝细菌基因组被比较.先用BLASTP两两最佳匹配搜索和基因位置保守性的方法预测直系同源基因,并进一步计算每个基因组中所有基因在其他4种绿球蓝细菌基因组中直系同源基因的数量,从而达到识别外源基因的目的;而基因组岛是指由1个或几个连续外源基因组成的区域.结果表明:在5个绿球蓝细菌基因组中共识别出了16至52个不等的基因组岛,它们中的许多基因在病毒基因组或富含病毒的环境样本中都有同源基因.以上这些结果显示,在绿球蓝细菌中,病毒可能介导了基因的水平转移,从而导致了基因组的多样性.