利用多基因序列探讨稻属药稻复合体二倍体物种的系统发育关系

稻属药稻复合体Oryza officinalis complex中有5个二倍体物种,涉及B、C和E3个染色体组,分布在亚洲、非洲和大洋洲。其中,O.australiensis是稻属中惟一含E染色体组的物种;O.punctata则是惟一含B染色体组的物种。虽然O.officinalis、O.rhizomatis和O.eichingeri都含C染色体组,但由于它们间断分布在亚洲和非洲,可能在稻属异源多倍体成种中发挥了不同作用。染色体组B、C和E之间,以及上述二倍体物种之间的系统发育关系及其在稻属多倍体物种形成     

一些水稻的重复DNA顺序在稻属和其它禾本科植物中的多态性

重复DNA顺序是真核生物基因组的特征,很多重复DNA顺序已从小麦、拟南芥菜、燕麦、水稻、玉米等植物的基因组中克隆出来,还发现有一些重复DNA顺序具有基因组特异性,用它们作探针可以分析同属或同科物种的起源和亲缘关系,并建立系统进化树。小卫星DNA或微小卫星DNA所产生的指纹图谱可作为一种遗传学标志来研究系统进化、染色体的精细结构和物种的鉴定。一些中度重复DNA序列还可以作为组织培养株系和细胞杂交筛选的分子标志。稻属已发现并定名的有22个种,根据杂交亲和性、细胞遗传学和生理生化等将它分为6个二倍体组型(AA、BB、CC、DD、EE和FF)和2个四倍体组型(BBCC和CCDD)。现多把禾本科分作5个亚科:竹亚科、稻亚科、早熟禾亚科、画眉草亚科和     

燕麦属细胞遗传学研究进展

整理燕麦属(Avena L.)细胞遗传学研究文献,总结相关研究进展。燕麦属有7组29种植物,分属5个基因组类型(A、C、AB、AC、ACD)。基于荧光原位杂交技术和种间杂交实验表明,A、C基因组染色体结构差异较大,A基因组二倍体物种具有等臂染色体,C基因组二倍体物种具有不等臂染色体。燕麦属植物D基因组和A基因组间分化程度较小,B基因组有可能是A基因组的变型——A′基因组。普遍观点认为A基因组二倍体物种可能是燕麦属六倍体物种母系亲本,砂燕麦(A.strigosa)为该属多倍体物种A基因组祖先的假说备受争议,有学者认为加那利燕麦(A.canariensis)可能是多倍体物种A或D基因组的供体。燕麦属多倍体物种基因组互换及染色体重排事件,增加燕麦属种间亲缘关系、多倍体物种基因组起源研究的困难。结合基因组学、分子细胞遗传学技术,有望为上述问题提供新证据。     

合成六倍体小麦的遗传育种

普通小麦是由四倍体小麦栽培类型与野生二倍体节节麦远缘杂交形成的异源六倍体.普通小麦保持了四倍体小麦的高产潜力,D基因组的加入丰富了食品加工产品类型、增强了环境适应能力.与二倍体作物不同,普通小麦有3个亚基因组,存在大量重复基因,基因组缓冲性、可塑性强,单个基因拷贝可能对育种改良的效果有限.小麦3个亚基因组的遗传多样性是...     

稻属三种植物叶片下表皮的形态比较

稻属有20多种,其中3种和其他物种明显不同.Oryza eichingeri被认为在亚洲和非洲间断分布;O. officinalis和O punctata被证明同时含有二倍体和四倍体.为了识别种内微形态差异,利用光镜和扫描电镜,对这3个物种的成熟叶片下表皮进行了观察.结果表明,O. eichingeri的叶脉长细胞乳突形状在种内是不一致的,除了斯里兰卡地理宗具有的瘤状乳突外,乌千达地理宗还具有长条状的乳突;同样,种内多样性也存在于O. officinalis和O.punctata内,特别存在于它们不同倍性的细胞型间.在D officinalis中,四倍体有粒状气孔器外围乳突和脉带表皮毛较少,二倍体细胞型具有三角形拱状排列的气孔器外围乳突和较密集的脉带表皮毛;而在O. punctata中,和四倍体相比,二倍体具有小的长细胞表面乳突和低密度分布的气孔.这些稳定的差异为3种植物的种内鉴定和分类修订提供了新的证据.     

芸薹属多倍体植物基因组进化的RAPD分析

多倍化是促进高等植物发生进化的重要力量。为了更清楚地了解多倍体在形成之后其基因组是如何进化的,利用38个随机引物对芸薹属Brassica L.禹氏三角（U’Triangle）中的多倍体物种及其祖先二倍体物种进行了研究。根据扩增出的273条带计算了遗传距离,并用UPGMA法进行了聚类分析。结果发现,二倍体物种B.campestris（AA）与B.oleracea（CC）的亲缘关系比与B.nigra（BB）的要近;异源多倍体B.napus（AACC）比起其二倍体祖先之一B.campestris（AA）与另一个     

芸薹属蔬菜Chs基因序列多态性

为探讨我国芸薹属蔬菜的起源及遗传多样性,克隆、测序芸薹属不同种的Chs基因序列。A基因组二倍体、A基因组多倍体、B基因组多倍体和C基因组二倍体的Chs基因突变位点数分别为120、172、194和25个,Chs基因多态性表现为:B基因组多倍体A基因组二倍体A基因组多倍体C基因组多倍体。Tajima'D、Fu and Li'D和Fu and Li'F检验表明A基因组二倍体、C基因组二倍体Chs基因是中性进化基因。HKA平衡检验及误配分析表明A基因组多倍体和B基因组多倍体Chs基因进化中存在选择作用。A基因组和B基因组间存在较低的共有差异和较高的共有多态性,C基因组与A、B基因组存在较高的共有差异和较低的共有多态性。系统发育树将芸薹属Chs基因序列分成4个亚支、10个支系。网状分析表明,白菜可能是四倍体A基因组的供体,黑芥可能是四倍体B基因组的供体,甘蓝可能是四倍体C基因组的供体。中国芸薹属蔬菜在Chs基因位点有较高的遗传多态性,不同基因组间分化程度不一样,B基因组分化较大,A和C基因组分化较小。A和B基因组的亲缘关系较A和C基因组以及B和C基因组更为接近。建议根据基因组的不同将中国芸薹蔬菜分成白菜组、芥菜组和甘蓝组,研究结果支持芸薹属进化的禹式三角模型。     

山羊草属异源多倍体植物基因组进化的ＲＡＰＤ分析

和２４个随机引物对山羊草属（Ａegilops L.)异源多倍体物种对其祖先二倍体物进行ＲＡＰＤ分析,对扩增出的３１３条带进行聚类分析发现,含Ｄ基因组的多倍体与二倍体祖先Ａe.squarrosa(DD)在聚类图上聚为一支;除Ａe.juvenalis(DDMMUU)聚到上一支外,含Ｕ基因组的多倍 与二倍体祖先Ae.umbellulata(ＵＵ）在聚类图上聚为另一支;多倍体与其他二倍体均不聚在一起,表明多倍体分别与Ａe.squarrosa(DD)、Ae.umbellulata(UU)具有较近的亲缘关系,这说明多倍体开之后,Ｄ和Ｕ基因组变化较小,而其他基因组则发生了较大的变化。     

水稻CMS相关基因在稻属AA基因组中的分布及其单核苷酸多态性

水稻线粒体基因组嵌合基因orf79 和 orfH79分别被认为与BT-型和HL-型水稻CMS有关, 两者具有98%的同源性, 并且其DNA序列只存在4核苷酸的差异。对于这两个嵌合基因, 前者来源于栽培稻(Oryza. sativa L.), 而后者则来源于普通野生稻(O. rufipogon Griff.)。这意味着orf79/ orfH79可能在广泛分布于稻属AA基因组中。为了调查orf79/ orfH79在稻属物种中的分布和变异, 190份栽培稻品系[包括156份亚洲栽培稻(O. sativa var. landrace)和34份非洲栽培稻(O. glaberrima)]以及104份稻属AA基因组野生稻品系(包括O. rufipogon、O.nivara、O. glumaepatula、O. barthii、O. longistaminata和O. meridionalis 6个种), 被用于PCR扩增检测。31份具有控制粤泰A和笹锦A的特异片段的稻属AA基因组水稻品系被检测出。所有特异片段均被回收并测序, 基于DNA 序列的聚类结果显示31份水稻材料被分成了两组, 分别代表为BT-型和HL-型水稻不育细胞质组群。结果也进一步表明: HL-型水稻CMS胞质主要分布于一年生的O. nivara中; BT-型水稻CMS胞质可能来源于栽培稻变种或多年生野生稻O. rufipogon。     

异源四倍体鲫鲤、三倍体湘云鲫和它们父母本线粒体DNA 12S rRNA基因遗传变异的分析

采用质粒克隆测序方法,获得了异源四倍体鲫鲤5个个体、异源四倍体鲫鲤雌核发育二倍体后代2个个体、三倍体湘云鲫2个个体及红鲫、湘江野鲤和日本白鲫各1个个体的线粒体DNA 12S rRNA基因的全序列。经对比发现,异源四倍体5个个体共享2种单元型,异源四倍体鲫鲤雌核发育二倍体后代2个个体、三倍体湘云鲫2个个体以及红鲫、湘江野鲤和日本白鲫各1个个体分别共享1种单元型。用MEGA 1.0 软件分析了它们的碱基组成和核苷酸序列差异,用邻接法构建系统进化树。它们间的序列同源性在95%~99%之间,异源四倍体鲫鲤、三倍体湘云鲫和它们母本（分别为红鲫和日本白鲫）之间的序列同源性大于异源四倍体鲫鲤、三倍体湘云鲫和它们父本（分别为湘江野鲤和异源四倍体鲫鲤）之间的序列同源性,结果表明：异源四倍体鲫鲤和三倍体湘云鲫在线粒体DNA 12S rRNA基因上具有母性遗传特征。本研究另一值得注意地方的是异源四倍体鲫鲤经过9代(F3-F11)繁殖后,在5个个体中发现了2种单元型,说明在四倍体基因库中存在遗传多样性,为四倍体基因库的繁殖、保护和种群复壮提供了一些有价值的信息。     

Cytoplasmic and genomic effects on meiotic pairing in brassica hybrids and allotetraploids from pair crosses of three cultivated diploids

Interspecific hybridization and allopolyploidization contribute to the origin of many important crops. Synthetic Brassica is a widely used model for the study of genetic recombination and "fixed heterosis" in allopolyploids. To investigate the effects of the cytoplasm and genome combinations on meiotic recombination, we produced digenomic diploid and triploid hybrids and trigenomic triploid hybrids from the reciprocal crosses of three Brassica diploids (B. rapa, AA; B. nigra, BB; B. oleracea, CC). The chromosomes in the resultant hybrids were doubled to obtain three allotetraploids (B. juncea, AA.BB; B. napus, AA.CC; B. carinata, BB.CC). Intra- and intergenomic chromosome pairings in these hybrids were quantified using genomic in situ hybridization and BAC-FISH. The level of intra- and intergenomic pairings varied significantly, depending on the genome combinations and the cytoplasmic background and/or their interaction. The extent of intragenomic pairing was less than that of intergenomic pairing within each genome. The extent of pairing variations within the B genome was less than that within the A and C genomes, each of which had a similar extent of pairing. Synthetic allotetraploids exhibited nondiploidized meiotic behavior, and their chromosomal instabilities were correlated with the relationship of the genomes and cytoplasmic background. Our results highlight the specific roles of the cytoplasm and genome to the chromosomal behaviors of hybrids and allopolyploids.     

Identification of Chromosomes from Multiple Rice Genomes Using a Universal Molecular Cytogenetic Marker System

To develop reliable techniques for chromosome identification is critical for cytogenetic research, especially for genomes with a large number and smaller-sized chromosomes. An efficient approach using bacterial artificial chromosome （BAC） clones as molecular cytological markers has been developed for many organisms. Herein, we present a set of chromosomal arm-specific molecular cytological markers derived from the gene-enriched regions of the sequenced rice genome. All these markers are able to generate very strong signals on the pachytene chromosomes of Oryza sativa L. （AA genome） when used as fluorescence in situ hybridization （FISH） probes. We further probed those markers to the pachytene chromosomes of O. punctata （BB genome） and O. officinalis （CC genome） and also got very strong signals on the relevant pachytene chromosomes. The signal position of each marker on the related chromosomes from the three different rice genomes was pretty much stable, which enabled us to identify different chromosomes among various rice genomes. We also constructed the karyotype for both O. punctata and O. officinalis with the BB and CC genomes, respectively, by analysis of 10 pachytene cells anchored by these chromosomal arm-specific markers.     

Genetic structure and genealogy in the Sphagnum subsecundum complex (Sphagnaceae: Bryophyta)

Allopolyploidy is probably the most extensively studied mode of plant speciation and allopolyploid species appear to be common in the mosses (Bryophyta). The Sphagnum subsecundum complex includes species known to be gametophytically haploid or diploid, and it has been proposed that the diploids (i.e., with tetraploid sporophytes) are allopolyploids. Nucleotide sequence and microsatellite variation among haploids and diploids from Newfoundland and Scandinavia indicate that (1) the diploids exhibit fixed or nearly fixed heterozygosity at the majority of loci sampled, and are clearly allopolyploids, (2) diploids originated independently in North America and Europe, (3) the European diploids appear to have the haploid species, S. subsecundum, as the maternal parent based on shared chloroplast DNA haplotypes, (4) the North American diploids do not have the chloroplast DNA of any sampled haploid, (5) both North American and European diploids share nucleotide and microsatellite similarities with S. subsecundum, (6) the diploids harbor more nucleotide and microsatellite diversity than the haploids, and (7) diploids exhibit higher levels of linkage disequilibrium among microsatellite loci. An experiment demonstrates significant artifactual recombination between interspecific DNAs coamplified by PCR, which may be a complicating factor in the interpretation of sequence-based analyses of allopolyploids.     

Identification of genome constitution of Oryza malampuzhaensis, O. minuta,and O. punctata by multicolor genomic in situ hybridization

Multicolor genomic in situ hybridization (McGISH) was applied to identify the genomic constitution of three tetraploid species (2n = 4x = 48) in the Oryza officinalis complex of the genus Oryza, i.e. Oryza malam-puzhaensis, Oryza minuta, and Oryza punctata. The genomic probes used were from three diploids, i.e. Oryza officinalis (CC), Oryza eichingeri (CC) and Oryza punctata (BB), respectively. The results indicated that all three tetraploids are allotetraploid with the genomic constitution of BBCC, and among them the genome constitution of O. malampuzhaensis was verified for the first time. Restoration of the independent taxonomic status of O. malampuzhaensis is suggested. One pair of satellite chromosomes belonging to the B genome was identified in O. malampuzhaensis, but no such satellite chromosomes were found in either O. minuta or the tetraploid O. punctata. The average chromosome length of the C genome was found to be slightly larger than that of the B-genome chromosomes of O. minuta, but not in the tetraploids O. punctata and O. malampuzhaensis. McGISH also revealed that the B genome of O. minuta and the B genome of diploid O. punctata showed clear differentiation from each other. Therefore, the suggestion was proposed that the B genome in diploid O. punctata was not the source of the B genome of O. minuta. The present results proved that multicolor GISH had high resolution in identifying the genomic constitution of polyploid Oryza species. Received: 14 February 2000 / Accepted: 13 November 2000     

稻属不同染色体组着丝粒BAC克隆的分离和鉴定

着丝粒在真核生物有丝分裂和减数分裂染色体正常的分离和传递中起着重要的作用。通过构建5个稻属二倍体野生种的基因组BAC文库, 采用菌落杂交和FISH技术, 筛选和鉴定了各染色体组着丝粒克隆, 并且分析了这些克隆在不同基因组间的共杂交情况, 结果表明: (1) C染色体组的野生种O. officinalis 和F染色体组的野生种O. brachyantha具有各自着丝粒特异的卫星DNA序列, 并且O. brachyantha着丝粒还具有特异的逆转座子序列; (2) A、B和E染色体组的野生稻O. glaberrima、O. punctata和O. australiensis着丝粒区域都含有与栽培稻着丝粒重复序列CentO和CRR同源的序列; (3) C染色体组野生稻O. officinalis的2条体细胞染色体着丝粒具有CentO的同源序列, 同时也发现其所有着丝粒区域都包含栽培稻CRR的同源序列。这些结果对克隆稻属不同染色体组的着丝粒序列、研究不同染色体组间着丝粒的进化关系和稻属不同着丝粒DNA序列与功能之间的关系均具有重要意义。     

Photosynthetic characteristics in <Emphasis Type="Italic">Oryza</Emphasis> species

Photosynthetic rate (P _N), SPAD value, specific leaf area (SLA), flag leaf area (FLA), and nitrogen content (LN) of genus Oryza were investigated and their correlation was analyzed to assess some of the main photosynthetic traits among different species in the genus Oryza. The results revealed wide variation in these traits. The species O. rufipogon and O. australiensis exhibited maximum photosynthetic rate. Comparison of different types of genomes (diploid: 2n=2x=24; tetraploid: 2n=4x=48) and growth habit (shade- or sun-grown) showed the species of diploid (with genome symbol EE; 2n=2x=24) genomes, with perennial and sun-grown species, had high apparent photosynthesis compared to others. The species with BB/BBCC, shade-grown and the tetraploids showed high SPAD value, and the flag leaf in sun-grown species and diploids were thicker (low SLA) compared with others. However, no significant difference could be noticed among the different types of genomes. Higher leaf area was noticed among the species of CC/CCDD genome, perennial shade-grown species and tetraploids than in others. The variety IR 36 exhibited highest leaf nitrogen concentration. Correlation analysis showed a strong relationship between P _N and leaf nitrogen concentration while no marked relationships were observed among other characteristics. It implies that the species with thick and small leaves with high nitrogen concentration and high photosynthesis evolved better than others. O. rufipogon, with the same genome as O. sativa, could be one of the wild rice resources for elite crop improvement.     

Phylogenetic analysis of Oryza species, based on simple sequence repeats and their flanking nucleotide sequences from the mitochondrial and chloroplast genomes

Simple sequence repeats (SSR) and their flanking regions in the mitochondrial and chloroplast genomes were sequenced in order to reveal DNA sequence variation. This information was used to gain new insights into phylogenetic relationships among species in the genus Oryza. Seven mitochondrial and five chloroplast SSR loci equal to or longer than ten mononucleotide repeats were chosen from known rice mitochondrial and chloroplast genome sequences. A total of 50 accessions of Oryza that represented six different diploid genomes and three different allopolyploid genomes of Oryza species were analyzed. Many base substitutions and deletions/insertions were identified in the SSR loci as well as their flanking regions. Of mononucleotide SSR, G (or C) repeats were more variable than A (or T) repeats. Results obtained by chloroplast and mitochondrial SSR analyses showed similar phylogenetic relationships among species, although chloroplast SSR were more informative because of their higher sequence diversity. The CC genome is suggested to be the maternal parent for the two BBCC genome species (O. punctata and O. minuta) and the CCDD species O. latifolia, based on the high level of sequence conservation between the diploid CC genome species and these allotetraploid species. This is the first report of phylogenetic analysis among plant species, based on mitochondrial and chloroplast SSR and their flanking sequences.     

Phylogeny of the StH genome species in Triticeae (Poaceae): Evidence from chloroplast trnL-F and mitochondria COXII intron sequences

The StH genome species in Triticeae exhibit different morphological variations and extensive geographic distribution. To estimate the phylogenetic relationship of the StH genome species in Triticeae, mitochondria COXII intron and chloroplast trnL-F sequences of 16 StH genome species were analyzed with those of four Pseudoroegneria species (St) and four Hordeum species (H). Sequence diversity and genealogical analysis suggested that (1) the trnL-F and COXII sequence may evolve faster in the polyploid species than in the diploids; (2) the COXII intron has a high evolutionary rate compared to trnL-F sequence and would provide potentially useful phylogenetic analysis in the StH genome species; (3) different Pseudoroegneria species might serve as the maternal donor during the polyploid speciation of the StH genome species; (4) phylogenetic relationships of the StH genome species may be not linked with the inter-continental disjunction between Eurasian and North American.