Chromosomal Distribution of the 18S-5.8S-26S rDNA Loci and Heterogeneity of Nuclear ITS Regions in Thinopyrum intermedium （Poaceae： Triticeae）

Fluorescent in situ hybridization (FISH) was used to investigate the chromosomal location of 18S-5.8S-26S rDNA loci in Thinopyrum intermedium (Host) Barkworth et Dewey (2n=6x=42). In all accessions and individuals studied, 3 or 4 pairs of major loci were detected. Subsequent genomic in situhybddization (GISH) analyses revealed that one pair was located on the ends of the short arms of one pair of homologous chromosomes of the St genome, while the other 2 or 3 pairs of major loci were located in the E genomes (including the E^o and E^b). It is suggested that 2 to 3 pairs of major loci were probably lost during the evolution of this hexaploid species. The variation in rDNA positions and copy numbers between the diploid donors and Th. interrnedium, as well as the diversity among the accessions of Th. intermedium confirmed that the rDNA gene family conveyed the characters of DNA mobile elements. The internal transcribed spacer (ITS) regions of the rDNA in Th. intermedium were also investigated. Sequence data of seven positive clones from one individual suggested high degree of individual heterogeneity exists among ITS repeats. Phylogenetic analyses showed that there were two distinct types of ITS sequences in Th. intermedium, one with homology to that of Pseudoroegneria species (St genome) and the other to that of the E genome diploid species. This showed that the ITS paralogues in Th. intermedium have not been uniformly homogenized by concerted evolution. The limitation of using the chromosomal location of rDNA loci for phylogenetic analysis is discussed.     

植物远缘杂交中的染色体行为及其遗传与进化意义

远缘杂交与多倍体化在高等植物的进化中起着重要的作用。但矛盾和令人费解的现象是“自然界在合成多倍体方面取得了巨大的成功,而人类在这方面却收效甚微”。其原因一方面可能是自然多倍体是长期自然选择和进化的产物,人类难以在短期内重复和完成这一过程;另一方面可能对不同的染色体组结合后的遗传与互作机制还不太了解。故多倍体化后的遗传和表观遗传成了目前多学科研究的重点。在有些有性和体细胞杂种内亲本染色体在细胞内分开排列,但此染色体行为的遗传和生物学意义还不太清楚。在植物远缘杂交中出现的假配生殖、半配生殖、染色体消除和亲本染色体组分开等异常染色体行为,也反映出不同物种在配子和染色体水平上的不亲和。需对植物远缘杂交中的染色体行为和遗传进行不同层次与系统的研究,才可能深入了解杂交后新种的形成及进化机制。     

染色体分选技术在植物学研究中的应用

介绍了染色体分选技术的基本原理和样品处理的基本流程,对根尖分生区采用同步化处理,制备染色体悬浮液,最后通过流式细胞仪的分析与收集获得纯度高、数量多的目标染色体.综述了染色体分选技术在植物学研究中的主要应用,包括物理图谱的构建、DNA分子标记的开发、以及复杂多倍体植物的基因组测序等.通过染色体分选技术的不断完善与发展,应...     

Effects of Nitrogen Source and Bacterial Elicitor on Isoflavone Accumulation in Root Cultures of Albizzia kalkora （Roxb.） Prain

Changes in cellular isoflavone （daidzein and genistein） contents were monitored in root cultures of Albizzia kalkora （Roxb.） Prain after feeding different ratios of NH4^＋/NO3^- and treatment with a biotic elicitor （three strains of Rhizobium sp.）. The NH4^＋/NO3^- ratio appears to be positively correlated with daidzein content in the roots and shows a negative correlation with genistein. Among the three different strains of Rhizobium used, the strain ATCC 15834 caused a 35% increase in daidzein production by infection. In the case of genistein, maximum production （94%） was obtained when cultures were treated on Day 6 by the strains ATCC 15834 and KCTC 1541. The biosynthetic pathway of the two isoflavones apparently reacts differently to the same culture conditions and the same strains of Rhizobium. Therefore, the present data suggest that the production of daidzein and genistein could be modulated by changing the NH4^＋/NO3^- ratio and the application of Rhizobium.     

白鲫（♀）×红鲫（♂）异源四倍体鱼的倍性操作及其生殖力的研究

采用热休克调控技术诱导出103尾第一代白鲫（♀）×红鲫（）异源四倍体鱼,并对其生殖力进行了研究。1或2龄的四倍体雄鱼能产生精子,而雌鱼不能产生正常的卵。将异源四倍体雄鱼与二倍体白鲫雌鱼交配产生倍间三倍体鱼,染色体检查证明是整三倍体（3N＝150）,但其受精率很低（11．4—51．3％,平均32．4％）．网箱养殖结果表明,倍间三倍体白鲫的生长速度比白鲫快30％以上,雌雄均不育．并用冷休克处理回收异源四倍体4N（）×白鲫2N（♀）受精卵的第二极体产生了新的四倍体鱼．文中还对第一代异源四倍体鱼的批量生产、生殖能力以及异源三倍体鱼的生产应用进行了讨论．     

由鲤(Cyprinus carpio)和鲫(Carassius auratus)染色体组叠加构建的 两个单性人工多倍体克隆

具有天然雌核发育的多倍体杂种鱼可防止杂种优势的分离并保持其后代的杂种优势. 由于假设诱发的多倍体鱼类的生殖模式是天然雌核发育的, 我们进行了鲤鲫杂种的多倍体诱发, 目的是描述经染色体组叠加由有性鲤鲫二倍体转化为异源三倍体及异源四倍体克隆谱系. 鲤鲫杂种产生未减数而具有两亲本染色体组杂种卵子, 未减数的雌核可与入卵的雄核融合叠加形成三倍体合子. 鲤鲫异源三倍体胚胎发育正常, 部分异源三倍体雌性个体可产生未减数的、仍保留母本的三套染色体的成熟卵子. 绝大部分鲤鲫异源人工三倍体个体的成熟卵子的雌核不与入卵的雄核融合, 具有天然雌核发育特性. 异源三倍体卵子在入卵精子的激动下由雌核发育产生全雌后代, 并形成一个单性克隆系, 后代保留异源三倍体母本的形态特征, 并靠雌核发育的生殖方式形成异源三倍体克隆系. 极少数异源三倍体个体的成熟卵子的雌核可与入卵的雄核融合, 再通过染色体组叠加形成鲤鲫异源四倍体. 所有异源四倍体的雌性产生未减数的、含有4个染色体组的成熟卵子. 异源四倍体的成熟卵子保持雌核发育特性, 在近类的精子诱发下产生单性后代, 形成一个异源四倍体单性克隆.     

Construction of a full bacterial artificial chromosome （BAC） library of Oryza sativa genome

We have constructed a full BAC library for the superior early indica variety of Oryza sativa,Guang Lu Ai 4.The MAX Efficiency DH10B with increased stability of inserts was used as BAC host cells.The potent pBelo BACII with double selection markers was used as cloning vector.The cloning efficiency we have reached was as high as 98%,and the transformation efficiency was raised up to 10^6 transformants/μg of large fragment DNA.The BAC recombinant transformants were picked at random and analyzed for the size of inserts,which turned out to be of 120 kb in length on average.We have obtained more than 20,000 such BAC clones.According to conventional probability equation,they covered the entire rice genome of 420,000 kb in length.The entire length of inserts of the library obtained has the 5-to 6-fold coverage of the genome.To our knowledge,this is the first reported full BAC library for a complex genome.     

Karyotype and cytogeography of the genus Heracleum （Apiaceae） in the Hengduan Mountains

In the present study, the karyotypes of 34 populations belonging to 11 species and one variety of Heracleum from the Hengduan Mountains in China were examined. Chromosome numbers and the karyotypes of three species （H. souliei, H. la＇ngdoni, and H. wenchuanense） are reported for the first time, as are the karyotypes of H. moellendorffii and H. henryi （tetraploid）. Populations of H. candicans, H. franchetii, and H. kingdoni in the Hengduan Mountains were found to consist of a mixture of diploid and tetraploid plants. Except for four species of Heracleum, namely H. candicans, H. franchetii, H. henryi, and H.kingdoni, which have both diploid and tetraploid karyotypes, all other species of Heracleum are were found to be diploid. All karyotypes were found to belong to the 2A type of Stebbins, with the exception ofH. candicans var. obtusifolium, which belongs to 2B, and H. hemsleyanum and H. franchetii （Mt. Dujuan, Daocheng, Sichuan, China）, which belong to 1A. There was only a slight difference in the karyotype asymmetry index, which suggests a close kinship for species of Heracleum and that the entire phylogenetic development of Heracleum is relatively primitive. Species that exhibited advanced morphological features were also more advanced in karyotype structure, with the order ofkaryotype evolution being 1A→2A→2B. This phenomenon indicates that the species distributed in the Hengduan Mountains have not diverged completely and that the Hengduan Mountains are a relatively young and active area for the evolution of Heracleum. Polyploidization in Heracleum may be an important evolutionary mechanisms for some species, generating diversity. The biological attributes, distribution range, and the geological history of the genus have all played a part in accelerating the evolution through polyploidization or aneuploidization. It is known that as the distribution latitude of Heracleum decreases from north to south, the chromosome number, ploidy level, and asymmetry structure appear to increase. In the Hengd     

Contribution of Root Respiration to Total Soil Respiration in a Leymus chinensis （Trin.） Tzvel, Grassland of Northeast China

The loss of carbon through root respiration Is an Important component of grassland carbon budgets. However, few data are available concerning the contribution of root respiration to total soil respiration in grasslands in China. We Investigated seasonal variations of soil respiration rate, root blomaaa, microbial blomaaa C and organic C content of the soil In a semi-arid Leymus chinensis （Trin.） Tzvel. grassland of northeast China during the 2002 growing season （from May to September）. The linear regression relationship between soil respiration rate and root blomaaa was used to determine the contribution of root respiration to total soil respiration. Soil respiration rate ranged from 2.5 to 11.9 g C/m^2 per d with the maximum in late June and minimum In September. The microbial blomaaa C and organic C content of the soil ranged from 0.3 to 1.5 g C/m^2 and from 29 to 34 g C/kg respectively. Root blomaaa had two peaks, In early June （1.80 kg/m^2） and mid-August （1.73 kg/m^2）. Root respiration rate peaked In mid-August （6.26 g C/m^2 per d）, whereas microbial respiration rate peaked In late June （7.43 g C/m^2 per d）. We estimated that the contribution of root respiration to total soil respiration during the growing season ranged from 38% to 76%.     

Heterozygosity of Knob-Associated Tandem Repeats and Knob Instability in Mitotic Chromosomes of Zea （Zea mays L. and Z. diploperennis Iltis Doebley）

Knobs are blocks of heterochromatin present on chromosomes of maize （Zea mays L.） and its relatives that have effects on the frequency of genetic recombination, as well as on chromosome behavior. Knob heterozygosity and instability in six maize inbred lines and one Z. diploperennis Iltis Doebley line were investigated using the fluorescence in situ hybridization （FISH） technique with knob-associated tandem repeats （180 bp and 350 bp （TR- 1）） as probes. Signals of seven heterozygous knobs containing 180- bp repeats and of one heterozygous knob containing TR- 1 were captured in chromosomes of all materials tested according to the results of FISH, which demonstrates that the 180-bp repeat is the main contributor to knob heterozygosity compared with the TR- 1 element. In addition, one target cell with two TR- 1 signals on one homolog of chromosome 2L, which was different from the normal cells in the maize inbred line GB57, was observed, suggesting knob duplication and an instability phenomenon in the maize genome.     

Timeline of Genomics （1977-2004）

Frederick Sanger (1980 Noble Prize Laureates for Chemistry) explores the technology of DNA sequencing. Sanger and colleagues use the technique to determine the sequence of all 5,375 nucleotides of the bacteriophage phi-X174, the first completede termination of the genome of an organism.     

Molecular Cytogenetic Characterization of a Wheat - Leymus mollis 3D（3Ns） Substitution Line with Resistance to Leaf Rust

Leymus mollis （Trin.） Pilger （NsNsXmXm, 2n = 28）, a wild relative of common wheat, possesses many potentially valuable traits that could be transferred to common wheat during breeding programs. In this study, the karyotypic constitution of a wheat - L. mollis 3D（3Ns#1） disomic substitution line isolated from the F5 progeny of octoploid Tritileymus M842-16 x Triticum durum cv. D4286, which was designated as 10DM57, was determined using genomic in situ hybridization （GISH）, fluorescent in situ hybridization （FISH）, SSR markers, and EST- STS markers. Screening of mitosis and meiosis showed that 10DM57 had a chromosome karyotype of 2n = 42 =21Ⅱ. GISH indicated that 10DM57 was a line with 40 chromosomes from wheat and two of the Ns chromosomes from L. mollis, which formed a ring bivalent in pollen mother cells at metaphase I. FISH analysis showed that the chromosome 3D may be replaced by 3Ns#1 in 10DM57. DNA markers, including SSR and EST-STS primers, showed that the pair of wheat chromosome 3D in 10DM57 was substituted by the pair of chromosome 3Ns#t from L. mollis. Evaluation of the agronomic traits showed that, compared with its common wheat relative 7182, 10DM57 was resistant to leaf rust while the spike length and number of spikes per plant were improved significantly, which correlated with a higher wheat yield. The new germplasm, 10DM57, could be exploited as an intermediate material in wheat genetic and breeding programs.     

基因组原位杂交技术及其在园艺植物基因组研究中的应用

基因组原位杂交(GISH)技术可以鉴定植物多倍体物种起源、杂种亲本染色体来源和组成,分析栽培种与其近缘野生种的亲缘关系,研究减数分裂染色体行为等。基因组原位杂交包括多色基因组原位杂交、比较基因组原位杂交和自身基因组原位杂交等。基因组原位杂交技术的关键步骤是染色体制片、探针制备及长度优化、探针与封阻的浓度比例和杂交后洗脱强度。该文对近年来国内外有关基因组原位杂交技术的发展及其在园艺植物基因组研究中的应用现状进行了综述,并指出随着多种园艺植物全基因组的测定,未来应从基因组信息中寻找更多的染色体特异性标记,结合荧光显带及荧光原位杂交技术,为深入研究园艺植物的起源以及遗传关系鉴定等提供技术支持。     

Mating system and genetic diversity of a rare desert legume Ammopiptanthus nanus （Leguminosae）

Ammopiptanthus nanus is an endangered evergreen shrub endemic to the deserts of central Asia and plays an important role in delaying further desertification. We examined allozyme variation and AFLP diversity in A. nanus populations and investigated the mating system of this species using progeny arrays assayed for polymorphic allozyme loci. Mating system analysis in the Keyi＇eryongke＇er population showed low levels of outcrossing, and strong inbreeding depression. Low levels of genetic variation were detected at both population （allozyme, Pp=14.0%, A=l.14, He=0.031; AFLP, Pp=14.5%, Shannon＇s information index I=0.063） and species （allozyme, Pp=21.1%, A=1.21, He=0.040; AFLP, Pp=20.9%, I=0.083） levels; while moderate genetic differentiation existed among populations, as indicated by allozymes （Gsa-=0.081） and AFLP （GST=0.151-0.193）. Founder effect, bottlenecks in evolutionary history, the mixed mating system and co-ancestry may have influenced the level of genetic diversity in A. nanus. Markers of both types provide new insights for conservation management, indicating that the Biao＇ertuokuoyi and Keyi＇eryongke＇er populations should be given priority for in situ conservation and regarded as seed sources for ex situ conservation.     

染色体研究的进展与植物分类学（上）

染色体研究的进展与植物分类学（上）徐炳声,张芝玉,陈家宽,洪德元（中国科学院植物研究所系统与进化植物学开放研究实验室北京１０００９３）（第二军医大学药学院上海２００４３３）（武汉大学生命科学学院武汉４３００７２）关键词染色体,植物分类学ＡＤＶＡＮＣＥ...     

黑麦（Secal cereale L.）联会复合体核型与染色体核型的比较研究

国内首次以“胜利”黑麦品系为材料,在电镜下观察联会复合体并分析了联会复合体核型。在同染色体核型比较后,认为“胜利”黑麦存在着两个可能的易位：１Ｒ长臂的一小段易位到６Ｒ短臂;４Ｒ长臂向２Ｒ长臂易位三单位长度,使４Ｒ长臂变成短臂。从而判断“胜利”黑麦为易位杂合体。这个研究方法可以在植物实验分类学研究中得到应用。     

Phylogenetic origin of Beckmannia （Poaceae） inferred from molecular evidence

The phylogenetic origin of Beckmannia remains unknown. The genus has been placed within the Chlorideae, Aveneae （Agrostideae）, Poeae, or treated as an isolate lineage, Beckmanniinae. In the present study, we used nuclear internal transcribed spacer （ITS） and chloroplast trnL-F sequences to examine the phylogenetic relationship between Beckmannia and those genera that have assumed to be related. On the basis of the results of our studies, the following conclusions could be drawn： （i） Beckmannia and Alopecurus are sister groups with high support; and （ii） Beckmannia and Alopecurus are nested in the Poeae clade with high support. The results of our analysis suggest that Beckmannia should be placed in Poeae.     

A revision of Clematis sect. Aspidanthera s.l. （ Ranunculaceae ）

对毛茛科铁线莲属Clematis L.的单性铁线莲组sect. Aspidanthera Spach s.l.进行了全面修订,确定此组共含72种和15变种,这些种被归类于6个亚组中,其中包括首次描述的1系、5种和2变种,以及做出的2新等级.对单性铁线莲组的分类学简史和地理分布做了介绍.写出了组、亚组、系的形态特征和地理分布,分亚组检索表及各亚组的分种检索表;以及各种植物的形态描述、地理分布、生长环境等,并附有多幅插图.在研究了此组全部种类的标本之后,观察到此组几个重要形态特征的演化趋势(1)萼片在数目上由4枚演变到8枚,在开展方向上由平展到直上展,在形状上由长圆形到狭条形,在长度上由比雄蕊稍长到2-4倍长于雄蕊,其卷叠式由镊合状到次生的覆瓦状;(2)花药药隔顶端由不具突起到具短或长的突起;(3)雌花退化雄蕊的数目由多数到定数、少数,以至完全消失;(4)花序自当年生枝的叶腋发生演变到与数叶同自老枝的一腋芽中发出.根据上述演化趋势推断本组各亚组间的亲缘关系如下subsect. Dioicae (34种,广布于北美洲和南美洲)的花构造(萼片4,镊合状排列,平展,通常呈长圆形,稍长于雄蕊或近等长,外面边缘被短绒毛;雄蕊花药呈长圆形,药隔顶端无突起;雌花具多数退化雄蕊;花序自当年生枝的叶腋发出)与具两性花的威灵仙组欧洲铁线莲亚组sect. Clematis subsect. Clematis 的花极为相似,区别主要在于本亚组的花为单性,由此判断subsect. Dioicae可能是单性铁线莲组sect. Aspidanthera的原始群,源出于欧洲铁线莲亚组.本组的第二亚组subsect. Lasianthae(2种,分布于北美西南部)与subsect. Dioicae 在亲缘关系上极为相近,区别只在于其花序或具花当年生枝与数叶一同由老枝的一个腋芽中发出,此亚组当是从subsect. Dioicae衍生出的一个小群.第三亚组subsect. Microphyllae(7种,特产澳大利亚)也与subsect. Dioicae相近缘,但此亚组的萼片变狭长,多呈条形或狭条形,常2-4倍长于多少变短的雄蕊,雌花的退化雄蕊数目变少,16-2枚,根据这些进化特征,推测此亚组也源出于subsect. Dioicae.第四亚组subsect. Aristatae (16种,分布于澳大利亚、新几内亚及邻近岛屿)与subsect. Microphyllae 在亲缘关系上相近,但本亚组的花药药隔顶端具短或长的突起而不同,根据此进化特征,推测本亚组系由后者演化而出.第五亚组subsect. Hexapetalae (11种,特产新西兰)的花构造与分布于澳大利亚的subsect. Microphyllae相似,但其萼片为覆瓦状排列,外面边缘不被短绒毛,在多数种多于4枚,为5-8枚而不同.据Godley 的研究,本亚组中的C. afoliata的花有4枚萼片,排成2轮,每轮的2枚萼片均为近镊合状排列;另外,在C. paniculata花的6枚萼片中,4枚为覆瓦状排列,其他2枚有时内向镊合状排列.从上述情况可见此亚组的萼片覆瓦状卷叠式可能是由镊合状卷叠式演变而来的一种次生现象,并由此推测,此亚组可能与subsect. Microphyllae相同,也源自美洲的subsect.Dioicae.最后一个亚组subsect. Insidiosae(2种,特产马达加斯加)的雌花萼片直上展,退化雄蕊完全消失,具有这些进化特征,当是单性铁线莲组的进化群,可能源出自具定数或少数退化雄蕊的subsect. Microphyllae.     

Molecular Genetic Variation in a Clonal Plant Population of Leymus chinensis （Trin.） Tzvel.

Randomly amplified polymorphic DNA （RAPD） analysis was used to investigate the genetic variation among populations, between populations, and within populations, relationships between genetic distance and geographic distance, and the molecular variation and population size. The effects of geographic and genetic distances, as well as of genetic differentiation and population size, on genetic variations of Leymus chinensis （Trin.） Tzvel. are discussed. The present study showed that there was significant RAPD variation between the Baicheng region population and the Daqing region population, with a molecular variance of 6.35% （P 〈 0.04）, and for differentiation among area populations of the Daqing region, with a molecular variance of 8.78% （P 〈 0.002）. A 21.06% RAPD variation among all 16 populations among two regions was found （P 〈 0.001）, as well as 72.59% variation within populations （P 〈 0.001）. Molecular variation within populations was significantly different among 16 populations.