植物特异性DNA探针的制备与应用研究进展

本文简述了植物特异性DNA探针的种类,介绍了特异性探针的制备方法,主要是特异性DNA序列的分离与筛选,总结了植物特异性DNA探针在种质鉴定、外源染色体识别、核型分析和基因组研究等方面的应用,提出了存在的问题与应用前景。 Abstract:In this paper,the types of specific DNA probes from plant were briefly described,and some methods of constructing specific probes were introduced,mainly were the isolation and screening of specific DNA sequences.The applications of the specific probes in the germplasm and foreign chromosome identifications,the karyotype and genome analyses were also summarized and discussed.     

长穗偃麦草中E和E1基因组高分子量麦谷蛋白 基因启动子部分序列的进化分析

通过PCR克隆的方法,获得了分别来自二倍体长穗偃麦草的E基因组和四倍体长穗偃麦草的E1基因组的4个高分子量麦谷蛋白亚基（HMW-GS）基因启动子的部分序列。序列分析表明,它们之间的同源性较高,两个x型亚基启动子序列之间只有1个碱基的差异,而两个y型亚基启动子序列完全相同, x和y型亚基启动子序列之间的长度和部分碱基位点都有差异。推测四倍体长穗偃麦草中的E1基因组可能起源于二倍体的E基因组。与来自小麦族的A、B、D和G基因组部分亚基基因的启动子序列比较表明,小麦族的这一区域在进化上是相当保守的,不同基因组来源的序列同源性都在90%以上。经过对这些序列的聚类分析,表明长穗偃麦草的y型HMW-GS基因与其他亚基基因的进化关系较远,而x型亚基基因与一个来自小麦1B染色体的亚基基因关系最近。Abstract: The partial promoter regions of HMW glutenin subunit genes were cloned form the genomes E (in diploid Agropyron elongatum) and E1 (in tetraploid Agropyron elongatum) by PCR approach. There was only one nucleotide acid difference in the promoter sequences of x-type subunits between the two genomes; moreover, the promoter sequences of the two y-type subunits were completely identical. Although these promoter regions were very similar to each other, differences still existed in sequence size and the kind of nucleotide acid between the x-type and y-type subunits. It was speculated that the E1 genome in tetraploid Agropyron elongatum was probably originated from E genome in diploid species. The comparisons of these subunits with some of those from A, B, D and G genome of Triticeae demonstrated that the sequences of their partial promoter regions were conserved and shared a high homology more than 90%. The phylogenetic analysis based on the sequences in this region indicated that the y-type HMW glutenin subunits of Agropyron elongatum species were different from other subunits, whereas the x-type subunits of them were most closely related to that from the B genome.     

Genetic Relationships Among Five Basic Genomes St, E, A, B and D in Triticeae Revealed by Genomic Southern and in situ Hybridization

The St and E are two important basic genomes in the perennial tribe Triticeae （Poaceae）. They exist in many perennial species and are very closely related to the A, B and D genomes of bread wheat （Triticum aestivum L.）. Genomic Southern hybridization and genomic in situ hybridization （GISH） were used to analyze the genomic relationships between the two genomes （St and E） and the three basic genomes （A, B and D） of T. aestivum. The semi-quantitative analysis of the Southern hybridization suggested that both St and E genomes are most closely related to the D genome, then the A genome, and relatively distant to the B genome. GISH analysis using St and E genomic DNA as probes further confirmed the conclusion. St and E are the two basic genomes of Thinopyrum ponticum （StStE^eE^bE^x） and Th. intermedium （StE^eE^b）, two perennial species successfully used in wheat improvement. Therefore, this paper provides a possible answer as to why most of the spontaneous wheat-Thinopyrum translocations and substitutions usually happen in the D genome, some in the A genome and rarely in the B genome. This would develop further use of alien species for wheat improvement, especially those containing St or E in their genome components.     

大麦基因组中的微卫星标记及其应用

微卫星是以少数几个核苷酸为单位多次串联重复的DNA序列,是一种简单序列重复(simple sequence repeats,SSR),两侧一般是保守序列。由于它具有多态性高、共显性、容易用PCR检测和结果稳定可靠等特点,因此是一种十分理想的分子标记。大麦的微卫星DNA随机分布于基因组中,平均每一个微卫星基因座有3～18个等位基因,最高可达37个。SSR标记已广泛用于分子遗传图谱的构建、遗传多样性研究、种质鉴定、主要性状基因的定位及分子标记辅助选择育种等。大多数SSR标记集中在着丝粒附近区域,1HL、5HL和6HS明显缺乏SSR标记。大麦的SSR标记还有待进一步的开发。 Microsatellite Markers and Applications in the Barley Genome FENG Zong-yun1,2,3,ZHANG Yi-zheng1,LING Hong-qing3 1.College of Life Sciences,Sichuan University,Chengdu 610065,China; 2.College of Agronomy,Sichuan Agricultural University,Ya'an 625014,China; 3.The State Key Laboratory of Plant Cell & Chromosome Engineering,Institute of Genetics & Developmental Biology,Chinese Academy of Sciences,Beijing 100101,China Abstract:Microsatellites,also called simple sequence repeats (SSR),are simple,tandemly repeated DNA sequences with a repeat length of a few base pairs,and are very ideally used as molecular markers because of their abundance,high level of polymorphism,co-dominance and ease of assay with the polymerase chain reaction (PCR) by selecting primers as the conserved DNA sequences flanking the SSRs,as well as better stability.The experiments showed that SSRs are randomly distributed throughout the barley genome,and there are 3~18 alleles at a single SSR locus,up to 37 alleles/locus.SSR markers have being widely applied in the construction of molecular genetic map,the study of genetic diversity,the identification of germplasm,gene mapping for important traits and molecular marker-assisted selection.Meanwhile,most of markers are strongly clustered around the centromeric regions of all seven linkage groups.As a result of the clustering,genome coverage with SSRs remains incomplete with an obvious lack of markers on the long arms of chromosomes 1H and 5H and short arm of chromosome 6H.Therefore,it is very potential and necessary to further develop SSR markers in barley. Key words：barley;microsatellite marker;simple sequence repeats;genetic diversity;molecular mapping     

Journal of Genetics and Genomics

正Founded in 1974Monthly www.j genetgenomics.org Published on behalf of Genetics Society of China and Institute of Genetics and Developmental Biology of Chinese Academy of Sciences Impact Factor:3.585Source from Thomson Reuters 2014 Journal Citation Reports     

Genetic Relationships Among Five Basic Genomes St,E,A,B and D in Triticeae Revealed by Genomic Southern and in situ Hybridization

The St and E are two important basic genomes in the perennial tribe Triticeae (Poaceae). They exist in many perennialspecies and are very closely related to the A, B and D genomes of bread wheat (Triticum aestivum L.). Genomic Southernhybridization and genomic in situ hybridization (GISH) were used to analyze the genomic relationships between the twogenomes (St and E) and the three basic genomes (A, B and D) of T. aestivum. The semi-quantitative analysis of the Southernhybridization suggested that both St and E genomes are most closely related to the D genome, then the A genome, andrelatively distant to the B genome. GISH analysis using St and E genomic DNA as probes further confirmed the conclusion.St and E are the two basic genomes of Thinopyrum ponticum (StStE~eE~bE~x) and Th. intermedium (StE~eE~b), two perennialspecies successfully used in wheat improvement. Therefore, this paper provides a possible answer as to why most of thespontaneous wheat-Thinopyrum translocations and substitutions usually happen in the D genome, some in the A genomeand rarely in the B genome. This would develop further use of alien species for wheat improvement, especially thosecontaining St or E in their genome components.     

Repetitive Sequences in Plant Nuclear DNA:Types,Distribution, Evolution and Function

Repetitive DNA sequences are a major component of eukaryotic genomes and may account for up to 90% of the genome size. They can be divided into minisatellite, microsatellite and satellite sequences. Satellite DNA sequences are considered to be a fast-evolving component of eukaryotic genomes, comprising tandemly-arrayed, highly-repetitive and highly-conserved monomer sequences. The monomer unit of satellite DNA is 150–400 base pairs(bp) in length.Repetitive sequences may be species- or genus-specific, and may be centromeric or subtelomeric in nature. They exhibit cohesive and concerted evolution caused by molecular drive, leading to high sequence homogeneity. Repetitive sequences accumulate variations in sequence and copy number during evolution, hence they are important tools for taxonomic and phylogenetic studies, and are known as ‘‘tuning knobs' ' in the evolution. Therefore, knowledge of repetitive sequences assists our understanding of the organization, evolution and behavior of eukaryotic genomes. Repetitive sequences have cytoplasmic, cellular and developmental effects and play a role in chromosomal recombination. In the post-genomics era, with the introduction of next-generation sequencing technology, it is possible to evaluate complex genomes for analyzing repetitive sequences and deciphering the yet unknown functional potential of repetitive sequences.     

小麦分子与细胞遗传学研讨会论文摘要2000年4月22～28日

培育磷高效小麦品种的遗传学与生理学基础 李振声 （中国科学院遗传研究所植物细胞与染色体工程国家重点实验室,北京 100101） 　　关键词：磷高效;小麦;生理基础 　　中图分类号：Q943,Q945　　　文献标识码：A　　　文章编号：0253-9772(2001)-01-0041-01 Genetic and Physiological basis for breeding Phosphorus Nutrient Efficient Wheat Varieties LI Zhen-sheng (The State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics, The Chinese Academy of Sciences, Beijing 100101, China) 用于小麦染色体工程的蓝粒小麦单体系列材料的创制 李振声 （中国科学院遗传研究所植物细胞与染色体工程国家重点实验室） 关键词：染色体工程;蓝粒小麦 中图分类号：Q943　　　文献标识码：A　　　文章编号：0253-9772(2001)-01- Establishment of a Set of blue Grained Wheat Monosomic Lines for Wheat Chromosome engineering studies LI Zhen-sheng (The State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics, The Chinese Academy of Sciences, Beijing 100101, China) 禾本科植物大小基因组间在基因密度上的共线性与保守性 Beat Keller (Institute of Plant Biology, University of Zürich Zollikerstrasse 107, CH-8008 Zürich, Switzerland) 关键词：禾本科植物;基因组;基因密度;共线性 中图分类号：Q78　　　文献标识码：A　　　文章编号：0253-9772(2001)-01-0042-02 Micro-colinearity and Conservation of High Gene Density in Small and Large Grass Genomes Beat Keller (Institute of Plant Biology, University of Zürich Zollikerstrasse 107, CH-8008 Zürich, Switzerland) 利用栽培一粒小麦的BAC文库精细定位小麦基因 Beat Keller (Institute of Plant Biology, University of Zürich Zollikerstrasse 107, CH-8008 Zürich, Switzerland) 关键词：一粒小麦;BAC文库;小麦基因 中图分类号：Q75　　　文献标识码：A　　　文章编号：0253-9772(2001)-01-0043-02 High Resolution Mapping of Wheat Genes Using a Triticum monococcum BAC Library Beat Keller (Institute of Plant Biology, University of Zürich Zollikerstrasse 107, CH-8008 Zürich, Switzerland) 图位克隆小麦抗叶锈基因Lr1 凌宏清,Beat Keller (Institute of Plant Biology, University of Zurich, Zollikerstrasse 107. CH-8008 Zurich, Switzerland) 关键词：图位克隆;抗叶锈;小麦 中图分类号： Q343.1　　　文献标识码：A　　　文章编号：0253-9772(2001)-01-0044-01 Towards map-based cloning of the leaf rust disease resistance gene Lr1 in wheat LING Hong-qing,Beat Keller (Institute of Plant Biology, University of Zurich, Zollikerstrasse 107. CH-8008 Zurich, Switzerland) 认识和改良中国小麦蛋白质量的遗传基础：策略与现有的研究 王道文,曲乐庆,贾　旭,张相岐,万永芳,李振声 （中国科学院遗传研究所植物细胞与染色体工程国家重点实验室,北京 100101） 关键词：小麦;蛋白质;遗传基础 中图分类号：Q512.1.032　　　文献标识码：A　　　文章编号：0253-9772(2001)-01-0045-01 Understanding and Manipulating the Genetic Basis of Protein Quality in Chinese Wheat:Strategies and Current Experiments WANG Dao-wen, QU Le-qing , JIA Xu , ZHANG Xiang-qi, WAN Yong-fang, LI Zhen-sheng (The State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics, the Chinese Academy of Sciences, Beijing 100101, China) 比较遗传学研究在认识禾本科植物基因组与基因功能中的应用价值 Mike Gale,Katrien Devos,and Graham Moore (John Innes Centre, Norwich, Norwich Research Park,Colney,Norwich.UK) 关键词：比较遗传学;基因组;基因功能 中图分类号：Q943　　　文献标识码：A　　　文章编号：0253-9772(2001)-01-0046-01 Cereal Comparative Genetics-research Opportunities Mike Gale,Katrien Devos,and Graham Moore (John Innes Centre, Norwich, Norwich Research Park,Colney,Norwich.UK) 小麦与环境互作的遗传学基础及其在提高小麦产量中的作用 J W Snape (John Innes Centre, Norwich Research Park, Colney, Norwich,UK) 关键词：小麦;环境互作;遗传学 中图分类号： S521.1　　　文献标识码：A　　　文章编号：0253-9772(2001)-01-0046-02 The Genetics of Adaptation in Wheat and Its Role in Maximising Yield Potential J W Snape (John Innes Centre, Norwich Research Park, Colney, Norwich,UK) John Innes中心禾本科植物系所从事的研究： 为认识小麦的生物学而努力 J W Snape (John Innes Centre, Norwich Research Park, Colney, Norwich, UK) 关键词：小麦;禾本科植物 中图分类号：S512.F　　　文献标识码：A　　　文章编号：0253-9772(2001)-01-0048-02 Towards Understanding the Biology of wheat:Work in the Cereals Research Department at the John Innes Centre, Norwich, UK J W Snape (John Innes Centre, Norwich Research Park, Colney, Norwich, UK) 杀配子染色体的作用机制及其应用 Takashi Endo (Laboratory of Genetics,Faculty of Agriculture, kyoto University Kyoto, 606,Japan) 关键词：杀配子;染色体;作用机制 中图分类号：Q343　　　文献标识码：A　　　文章编号：0253-9772(2001)-01-0049-01 Action and application of gametocidal chromosomes Takashi Endo (Laboratory of Genetics Faculty of Agriculture, kyoto University Kyoto, 606, Japan) 利用杀配子染色体2C诱导大麦染色体产生结构变异 施芳 (Laboratory of Genetics, Faculty of Agriculture, kyoto University Kyoto, 606, Japan) 关键词：杀配子;染色体;大麦 中图分类号：Q343　　　文献标识码：A　　　文章编号：0253-9772(2001)-01-0049-02 Generation of structural changes in barley chromosomes by the gametocidal chromosome 2C SHI Fang (Laboratory of Genetics, Faculty of Agriculture, kyoto UniversityKyoto,606,Japan) 小偃麦附加系Z1和Z2中外源染色体2Ai-2的结构组成 张增燕,辛志勇,陈　孝 (中国农业科学院作物育种栽培研究所,北京 100081) 关键词：小偃麦;附加系;染色体 中图分类号：Q343.2　　　文献标识码：A　　　文章编号：0253-9772(2001)-01-0050-02 Structural Organization of an Alien Group 2 Chromosome (2Ai-2）in Wheat-Thinopyum intermedium Addition Lines Z1 and Z2 ZHANG Zeng-yan, XIN Zhi-yong, CHEN Xiao (Key Lab of Crop Genetics ＆ Breeding of Ministry of Agriculture, Institute of Crop Breeding and Cultivation,CAAS,Beijing 100081, China) 　　The barley yellow dwarf virus（BYDV）resistance lines of Z1 and Z2 were derived from Zhong 5, a partial amphiploid resulted from the cross between Triticum aestivum（wheat） and Thinopyrum intermedium. Genomic in situ hybridization （GISH） was used to analyze the chromosome constitution of Zhong 5 by using genomic DNA of Pseudoregneria strigosa(StSt,2n＝14）as the probe. The GISH results showed that zhong 5 contains 42 wheat chromosomes and l4 Th.intermedium chromosomes composed of 4 St, 4 Js,4 St-J translocation and 2 St-Js Robertsonian translocation chromosomes. The chromosome constitution of Z1 and Z2 was analyzed by GISH using genomic DNA probes from Th.intermedium and Ps.Strigosa. The GISH results indicated that both Z1 and Z2 possess 42 wheat chromosomes and 2 Th.intermedium chromosomes that were identical to a pair of St-J translocation chromosomes in Zhong 5. The Th.intermedium chromosomes,designated as 2Ai-2 chromosome derived from Zhong 5,mostly belong to the St genome except the middle region (about one third of the long arm) belonging to the E（J）genome. A detailed RFLP analysis was conducted for Z1,Z2 and their parents,St and E (J) genomes. The results of RFLP analyses demonstrated that the Th.intermedium chromosomes（2Ai-2,St-J）in Z1 and Z2 are extensively homologous to the Wheat group 2 chromosomes. The results of RFLP analyses on the genome composition of the 2Ai-2 chromosome were in agreement with the GISH results. Presence of psr 928 on 2AS and 2DS but absence on 2Ai-2S suggests some internal structural differences between 2Ai-2 and the wheat group 2 chromosomes. Some RFLP markers specific to the 2Ai-2 chromosome were identified and may be effectively used to select translocation lines with small segment of the 2Ai-2 chromosome and to localize the BYDV resistance gene in wheat background. Key words：wheat;Thinopyrum intermedium;barley yellow dwarf virus （BYDV）;genomic in situ hybridization （GISH）;RFLP;homoeologous group 2 抗大麦黄矮病的小偃麦易位系的创制与鉴定 辛志勇,张增燕,陈　孝,林志珊 (中国农业科学院作物育种载培研究所,北京 100081) 关键词：大麦;小偃麦;易位系 中图分类号：QS512　　　文献标识码：A　　　文章编号：0253-9772(2001)-01-0051-02 Development and Characterization of Common wheat-Thinopyrum intermedium Translocation Lines with Resistance to Barley Yellow Dwarf Virus XIN Zhi-yong,ZHANG Zeng-yan,CHEN Xiao,LIN Zhi-shan, MA You-zhi,XU Hui-jun,XU Qiong-fang,DU Li-pu (Key Lab of Crop Genetics＆Breeding of Agriculture Ministry, Institute of Crop Breeding and Cultivation,CAAS,Beijing,100081,China) 　　Barley yellow dwarf virus（BYDV）,vectored by several aphid species,is the most significant viral pathogen of wheat and other grain cereals．Significant economic losses resulting from BYDV in wheat,barley and oats have been reported in many countries．The most economic means of controlling BYDV is to develop wheat varieties with resistance to BYDV. So far no BYDV resistance has been described in wheat collections except one gene in some cultivars tolerant to BYDV. However,Thinopyrum intermedium,two octoploids Zhong 4 awnless and TAF46,and the disomic addition lines,L1,Z1,Z2 and Z6 all showed resistance to BYDV. We developed several wheat-Th.Intermedium translocation lines, Yw642, Yw443 and Yw243 etc., showing good BYDV resistance from L1 by inducing homologous pairing using CS Ph1 mutant. It was found that their BYDV resistance was controlled by a single dominant gene. Characterization of these wheat lines was carried out by GISH and RFLP analysis. The results of GISH showed that the lines, Yw642, Yw443 and Yw243 etc., were homozygous wheat-Th.intermedium translocation lines containing 20 pairs of wheat chromosomes and 1 pair of wheat-Th.intermedium translocation chromosomes,in which the chromosome segments of Th intermedium were transferred to the distal end of a pair of wheat chromosomes. RFLP analysis indicated that the translocation chromosome of the wheat lines was T7DS*7DL-7XL translocation. The breakpoint of translocation is located on the distal end of 7DL,between Xpsr965 and Xpsr680,about 90-99 cM from the centromere. The BYDV gene is located on the distal end of 7XL around Xpsr680,Xpsr687 and Xwg380．The RFLP markers of psr680,psr687 and wg380 co-segregated with the BYDV resistance and could be used for marker-assisted selection（MAS）in wheat breeding program for BYDV resistance. Key words:Thinopyrum intermedium,BYDV,disease resistance, translocation, GISH, RFLP, homoeologous group 7 多枝赖草DNA导入小麦引起重要农艺性状变化及相应的分子证据 李维琪,等 (中国科学院新疆化学研究所,乌鲁木齐 830000) 关键词：小麦;多枝赖草DNA;农艺性状 中图分类号：Q　　　文献标识码：A　　　文章编号：0253-9772(2001)-01-0052-02 Important Agronomic trait Changes in Wheat Caused by Introduction of Leymus racemoses DNA and Some Molecular Proofs LI Wei-qi,ZHAO Min-an,LIU Yun-ying,MIAO Jun,LIU Min (Xinjiang Institute of Chemistry, The Chinese Academy of Sciences, Urumqi, 830011) QI Jia-hua,ZHANG Mao-yin,WANG Zi-xia,Haizigoli Yang Ke-rui (Institute of Nuclear and Biotechnology, Xinjiang Academy of Agricultural Sciences, Urumqi, 830000) 利用异源双代换系杂交产生染色体易位系 贾　旭 (中国科学院遗传研究所植物细胞与染色体工程国家重点实验室,北京 100101) 关键词：双代换系;染色体;易位系 中图分类号：Q343　　　文献标识码：A　　　文章编号：0253-9772(2001)-01-0053-01 Production of Chromosome Translocation via Crossing Two Different Alien Substitution Lines in Wheat JIA Xu (The State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics, The Chinese Academy of Sciences, Beijing 100101, China) 过去50年中中国小麦品种在Glu-A1, Glu-B1 和Glu-D1位点上等位基因的变化 张学勇,董玉琛 (中国农业科学院品种资源研究所,北京 100081,China) 关键词：小麦;基因 中图分类号： Q943　　　文献标识码：A　　　文章编号：0253-9772(2001)-01-0053-02 Allelic variation of Glu-A1, Glu-B1 and Glu-D1 in Chinese Commercial wheat Varieties in the Last 50 Years ZHANG Xue-yong,DONG Yu-cheng (Institute of Crop Germ plasm Resource,CAAS,Beijing 100081,China) 化学杀雄剂导致小麦雄性不育分子机制的初步研究 张爱民,等 (中国农业大学植物遗传育种系,北京 100094,China) 关键词：小麦;杀雄剂;雄性不育;分子机制 中图分类号：Q　　　文献标识码：A　　　文章编号：0253-9772(2001)-01-0054-02 Preliminary Study of Molecular Basis of Male Sterility Induced by Chemical Hybridizing Agent in Wheat (Tritium aestivum) ZHANG Ai-min,LIU Dong-cheng (Dept.of Plant Genetics and Breeding, China Agri. Uni.,Beijing 100094,China) 利用小偃麦附加系对Agropyron elongatum生化标记进行染色体定位 刘树兵,等 (山东农业大学农学系,泰安　271018) 关键词：小偃麦;附加系;生化标记 中图分类号：Q343　　　文献标识码：A　　　文章编号：0253-9772(2001)-01-0055-02 Chromosomal Location of Agropyron elongatum Markers in Wheat-Agropyron elongatum (2n=14) Addition Lines LIU Shu-bing1, WANG Hong-gang1, ZHOU Rong-hua2,JIA Ji-zeng2 (1.Agronmy Dept., Shandong Agricultural University, Taian 271018,China; 2.Institute of Crop germplasm Resource, CAAS, Beijing, 100081,China) 　　Isoelectric focusing (IFF) technique was used to locate biochemical loci in Agropyron elongatum by using wheat-Agropy-ron elongatum addition lines. There were six loci being located initially in all. The structural genes of Est-E5 and Est-E8 were located in 3EL, β-Amy-1 in 4EL, Per-E1 in 7E, and Per-E4 in 5E. The α-Amy-E1 was relocated in 6EL. Chromosome location of these genes provide evidence of homoeology between wheat groups 3, 4, 6 and Agropyron elongatum chromosome 3E, 4E, 6E, respectively. It also indicated that chromosome rearrangement probably took place between 1E and 7E chromosomes during the evolution of the E genome. Key words:chromosome location, biochemical mark, isoelectric focusing, addition lines 小麦春化相关基因的分子克隆与功能分析 种　康,许智宏,谭克辉 (中国科学院植物研究所,北京 100093) 关键词：小麦;基因;分子克隆 中图分类号：S512.1　　　文献标识码：A　　　文章编号：0253-9772(2001)-01-0056-01 Molecular Cloning and Functional Analysis of Vernalization Related Genes in Wheat CHONG Kang, XU Zhi-hong, TAN Ke-hui (Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China) 中字系列小偃麦遗传材料的培育、遗传分析和利用 孙善澄,白建荣 (山西农业科学院作物遗传研究所,太原 030031) 关键词：小偃麦;遗传分析 中图分类号：Q　　　文献标识码：A　　　文章编号：0253-9772(2001)-01-0056-02 Breeding, Genetic Analysis and Utilization of “Zhong Series” Varieties of Trielytrigia SUN Shan-cheng,BAI Jian-rong (Institute of Crop Genetics. Shanxi Academy of Agricultural Sciences, Taiyuan 030031,China) 紫外线诱导小麦和长穗偃麦草体细胞融合产生可育株及其后代的细胞遗传学分析 夏光敏,周爱芬,等 (山东大学生命科学院,济南 250100) 关键词：小麦;长穗偃麦草;细胞融合 中图分类号：Q343　　　文献标识码：A　　　文章编号：0253-9772(2001)-01-0057-02 Production of Fertile Hybrid Plants Via UV Fusion between Triticum aestivum and Agropyron elongatum and Cytogenetic Analysis of the Resulted Progenies XIA Guang-min,XIANG Feng-ning, ZHOU Ai-fen, LI Zhong-yi,CHEN Hui-min (School of Life Sciences, Shandong University, Jinan 250100,China; *　CSIRO Division of Plant Industry, G.P.O.1600, Australia) 　　Protoplasts of common wheat (Triticum aestivum L.2n=42) cv. Jinan 177 were fused with ultraviolet (UV) irradiated protoplasts of Agropyron elongatum(2n=70) via a PEG method. Sterile hybrid plants were regenerated from the fused product and their ovaries were induced to form calli.From the resulted callus tissue, green plants were differentiated. These somaclonal plants (SF0) were investigated using chromosome and isozyme analysis. The results showed that they contained chromosomes from both donors (I.e.,Triticum aestivum and Agropyron elongatum). Two of the SF0 plants grew to maturity and set seeds. The analysis of phenotype, chromosome constitution, isozyme pattern and RAPD polymorphism of the F1 plants confirmed their hybrid nature. Collectively, these results demonstrated that fertile hybrid plants could be produced from the procedure described above. Three different phenotypes were observed in F2 progenies. The type I and II plants had higher stalks (average 75～85 cm) and big ears and grains, but plants of the former phenotype possessed fewer tillers. Type III plants had short stalks (average 55 cm) but possessed high ability of tillering. Cytogenetical analysis of F1 plants and their successive generations showed that in F1 to F3 generations the chromosome numbers of root tip cells varied in the range of 36～44, and many cells contained 1-4 micro-chromosomes (mc). In PMC MI stage of the F2 plants, the chromosome configuration was mainly 17II-22II with 1-4 additional micro-chromosomes. Comparing to F2, more chromosome configuration of 20II -21II occurred in F3, and over 70% of cells had the chromosome configurations of 21II+1-2 mcs. A large population of the different hybrid lines have been obtained through propagation and selection in successive generations. Their agronomic traits have been studied and will be reported in a separate paper. Key words:Triticum aestivum;Agropyron elongatum;Asymmetric somatic hybridization; Fertile hybrid plants; Cytogenetic analysis 普通小麦F1杂种Glu-1基因表达 过程中的共显性,基因组互作和剂量效应 潘幸来,等 (山西农业科学院棉花研究所,运城 044000) 关键词：小麦;基因表达;基因组 中图分类号：S512.1.035　　　文献标识码：A　　　文章编号：0253-9772(2001)-01-0058-01 Evidence for Co-dominance,Genome Interaction and Dosage Effects in Glu-1 Gene Expression in F1 Seeds of Common Wheat (Triticum aestivum L.) PAN Xing-lai, PAN Qian-ying,ZHANG Gui-yuan,WANG Yong-jie,XIE San-gang (Cotton Research Insritute,Shanxi Agri.Sci.Academy,Yuncheng,Shanxi,China,044000) WANG Dao-wen (State Key Laboratory of Plant Cell and Chromosome Engineering,Institute of Genetics, Chinese Academy of Sciences, Beijing 100101) PAN Deng-kui (Shanxi Agriculture University, Taigu,Shanxi 030801,China) ph1b基因对簇毛麦遗传物质导入普通小麦的影响 陈　静,等 (中国科学院成都生物研究所,成都 610041) 关键词：ph1b基因;小麦;簇毛麦 中图分类号：Q　　　文献标识码：A　　　文章编号：0253-9772(2001)-01-0058-02 Effect of ph1b gene on direct genetic transfer from Haynaldia villosa to Triticum aestivum CHEN Jing,DENG Guang-bing, ZHANG Xiao-pin, MA Xin-rong, YU Mao-qun (Chengdu Institute of Biology, Chinese Academy of Sciences, 610041,China) 　　Hybrid plants between Triticum aestivum var.“Chinese Spring”(CS), its ph1b mutant (CSph1b) and Haynaldia villosa were obtained by immature embryo culture. After selfing, the two types of hybrids showed seed setting rates of 6.67% and 6.25%, respectively. The analysis of chromosome pairing behaviors at meiotic metaphase I showed that, on the average, only 1.61 chromosomes could form bivalent and trivalent in each PMC of the hybrid F1 CS×H. Villosa, with the configuration 2n=28=26.39I+0.79II+0.007III. However, in the hybrid F1 CS ph1b×H. Villosa, 14.43 chromosomes per PMC were involved in bivalent and multivalent formation, with the chromosome configuration of 2n=28=13.55I+5.95II+0.55III=0.22IV, and, in over 56% of the PMCs, 1-4 multivalents (trivalents and quadrivalents) were produced. The observation of meiotic chromosome pairing by using genomic fluorescent in situ hybridization (GISH) revealed three types of chromosomal associations: wheat-wheat, wheat-H. Villosa and H. Villosa-H. Villosa in PMCs for ‘CS ph1b×H.villosa’F1 hybrid. The seed set of the backcross of the‘CS ph1b×H.villosa’F1 with CS was 6.67% and that of‘CS×H.villosa’F1 with CS or CS ph1b was only 0.45%. The chromosome number of BC1 plants varied from 48 to 72. Robertsonian translocation chromosomes consisting of chromosome arms from wheat and H.villosa were detected by mitosis GISH in the BC1 plants from the backcross of‘CSph1b×H.villosa’to CS ph1b. These results led to the conclusion that the ph1b gene induced a higher level of homoeologous chromosome pairing between common wheat and H.villosa.The transfer of desirable genes from H.villosa to common wheat may be facilitated by using the ph1b gene. Key words:genetic transfer, GISH,Haynaldia villosa;ph1b gene;homoeologous pairing;Triticum aestivum 利用分子细胞遗传学方法向小麦中转移和富积优异外源基因 陈佩度 (南京农业大学细胞遗传研究所,南京 210095) 关键词：小麦;细胞遗传学;外源基因 中图分类号：Q343.1　　　文献标识码：A　　　文章编号：0253-9772(2001)-01-0059-02 Utilization of Molecular Cytogenetics Technology in Transferring and Pyramiding Useful Alien Genes into Common Wheat CHEN Pei-du (Cytogenetics Institute, Nanjing Agricultural University, Nanjing 210095,China) 小麦品种复壮30中与抗白粉病基因连锁的一个RAPD标记 王立新,等 (北京市农业科学院植物细胞工程实验室,北京 100089) 关键词：小麦;抗白粉病;基因;RAPD标记 中图分类号：Q343.1　　　文献标识码：A　　　文章编号：0253-9772(2001)-01-0060-01 A RAPD Marker Linked to the Resistance Gene to Powdery Mildew in Wheat Variety-Fu Zhuang 30 WANG Li-xin, SU Ai-lian, XU Min-xin,WANG Xiu-qin,Peter.P.Ueng*,JIA Ji-zheng** (Beijing Plant Cell Bioengineering Laboratory, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100089,China; *　Molecular Plant Pathology Laboratory, USDA-ARS, MD 20705,US **　Key Laboratory of Crop Germplasm and Biotechnology, Institute of Crop Germplasm Resource,China Academy of Agriculture Sciences,Beijing 100081,China) 小麦和簇毛麦体细胞可育杂种植株的产生 周爱芬,夏光敏,等 (山东大学生命科学学院,济南 250100) 关键词：小麦;簇毛麦;体细胞 中图分类号：S512.035.1　　　文献标识码：A　　　文章编号：0253-9772(2001)-01-0060-02 Production of Fertile Somatic Hybrid Plants Between Triticum Aestivum and Haynaldia villosa ZHOU Ai-fen1,XIA Guang-min1,ZHANG Xiang-qi2,CHEN Hui-min,HU Han2 (1.Life Science School, Shandong University, Jinan 250100,China; 2.Institute of Genetics, Chinese Academy of Science, Beijing 100101,China) 小麦基因组中外源染色体片段的检测和小麦基因分子标记的建立 石锐 (哈尔滨师范大学生物系,150080) 关键词：小麦;外源染色体;分子标记 中图分类号：Q75　　　文献标识码：A　　　文章编号：0253-9772(2001)-01-0061-01 Studies on the Detection of Alien Chromosome Fragments and Obtainment of Molecular Markers Linked to Specific Genes in Wheat SHI Rui (Department of Biology,Harbin Normal University,Harbin,Heilongjiang Province 150080,China) 小麦春化发育机制的初步研究 尹　军,等 (山西农业大学,太谷　030801) 关键词：小麦;春化;发育机制 中图分类号：Q344　　　文献标识码：A　　　文章编号：0253-9772(2001)-01-0062-01 Preliminary Study on the Mechanism of the Vernalization Development in Wheat YIN Jun,REN Jiang-ping,DONG Ai-xiang （Shanxi Agricultural University,Taigu 030801,China） 非Robertsonian类型小黑麦易位系的研究 胡　含 (中国科学院遗传研究所植物细胞与染色体工程国家重点实验室,北京 100101) 关键词：小黑麦;易位系 中图分类号：Q343　　　文献标识码：A　　　文章编号：0253-9772(2001)-01-0062-02 About non-Robertsonian wheat-rye chromosome translocation lines HU Han (State Key Laboratory of Plant Cell and Chromosome Engineering,Institute of Genetics CAS,Beijing 100101,China) 小麦6B染色体的微切割与其区域特异性DNA文库的构建 周奕华,王　槐,陈正华 (中国科学院遗传研究所,北京 100101) 关键词：小麦;微切割;染色体;DNA 中图分类号：Q343　　　文献标识码：A　　　文章编号：0253-9772(2000)-01-0063-01 Micro-dissection of Wheat Chromosome 6B and Construction of Its Region Specific DNA Libraries ZHOU Yi-hua,WANG Huai,CHEN Zheng-hua (Institute of Genetics,Chinese Academy of Sciences, Beijing 100101,China) 组织培养诱导外源染色体发生结构 变异及其在小麦易位系创制中的利用 李洪杰1,贾　旭1,楚成才2 (1.中国科学院遗传研究所植物细胞与染色体工程国家重点实验室,北京 100101;2.中国科学院植物研究所,北京 100093) 关键词：小麦;染色体;结构变异;易位系 中图分类号：Q343　　　文献标识码：A　　　文章编号：0253-9772(2001)-01-0064-01 Structural Changes of Alien Chromosomes Arising from Tissue Culture and Their Exploitation in Producing Novel Translocation Wheat Lines LI Hong-Jlie1,2,JIA Xu1,CHU Cheng-chai2 (1.The State Key Laboratory of Plant cell and Chromosome Engineering, Institute of Genetics, The Chinese Academy of Sciences, Beijing 100101, China; 2.Institute of Botany, The Chinese Academy of Sciences, Beijing 100093, China)     

USE OF REPEATED DNA SEQUENCES AS CYTOLOGICAL MARKERS

The majority of DNA that is found in most of the flowering plants appears to be non-coding DNA. Much of this excess DNA consists of nucleotide sequences which exist as multiple copies throughout the genome and are designated as repetitive sequences. Those sequences which are found in moderately high to high numbers of copies are observed to be of the greatest value as cytological markers. Moderately high copies may exist as sequences which are dispersed throughout the chromosomes of some species and not dispersed in other more distantly related species. By taking advantage of this characteristic and the technique of in situ hybridization with biotinylated probes, breakpoints of chromosomal translocations may be observed between species such as wheat and rye. Many of the high copy number repetitive sequences are organized in a tandem fashion in specific loci in the chromosome. Chromosomal identification may be accomplished by using the in situ hybridization technique. Upon in situ hybridization with a repetitive sequence isolated from Aegilops squarrosa, the patterns of the sites of hybridization allowed the D-genome chromosomes to be identified. The sequence was also observed only on the D-genome chromosomes of several polyploid species indicating its usefulness as a genome specific marker. Using this genome specificity, assessment of the orientation of the D-genome chromosomal segments of hexaploid wheat carrying the sequence during interphase and prophase of mitotic root tip cells was possible. Repetitive DNA sequences, therefore, provide cytological markers necessary for studies of chromosomal identification, genome allocation, and genome orientation. The use of biotin-labeled DNA probes allows the technique of in situ hybridization to be performed much more rapidly and with a greater degree of safety and reliability.     

Genome-specific repetitive sequences in the genus Oryza

Summary Repetitive DNA sequences are useful molecular markers for studying plant genome evolution and species divergence. In this paper, we report the isolation and characterization of four genome-type specific repetitive DNA sequences in the genus Oryza. Sequences specific to the AA, CC, EE or FF genome types are described. These genome-type specific repetitive sequences will be useful in classifying unknown species of wild or domestic rice, and in studying genome evolution at the molecular level. Using an AA genome-specific repetitive DNA sequence (pOs48) as a hybridization probe, considerable differences in its copy number were found among different varieties of Asian-cultivated rice (O. sativa) and other related species within the AA genome type. Thus, the relationship among some of the members of AA genome type can be deduced based on the degree of DNA sequence similarity of this repetitive sequence.     

Genus-specific localization of the TaiI family of tandem-repetitive sequences in either the centromeric or subtelomeric regions in Triticeae species (Poaceae) and its evolution in wheat.

The TaiI family sequences are classified as tandem repetitive DNA sequences present in the genome of tribe Triticeae, and are localized in the centromeric regions of common wheat, but in the subtelomeric heterochromatic regions of Leymus racemosus and related species. In this study, we investigated the chromosomal distribution of TaiI family sequences in other Triticeae species. The results demonstrated a centromeric localization in genera Triticum and Aegilops and subtelomeric localization in other genera, thus showing a genus-dependent localization of TaiI family sequences in one or the other region. The copy numbers of TaiI family sequences in species in the same genus varied greatly, whether in the centromeric or subtelomeric regions (depending on genus). We also examined the evolution of TaiI family sequences during polyploidization of hexaploid common wheat. A comparison of chromosomal locations of the major TaiI family signals in common wheat and in its ancestral species suggested that the centromeric TaiI family sequences in common wheat were inherited from its ancestors with little modification, whereas a mixed origin for the B genome of common wheat was indicated.     

Repeated DNA sequences in the microbat species Miniopterus schreibersi (Vespertilionidae; Chiroptera)

Repetitive DNA sequences represent a substantial component of eukaryotic genomes. These sequences have been described and characterized in many mammalian species. However, little information about repetitive DNA sequences is available in bat species. Here we describe an EcoRI family of repetitive DNA sequences present in the species Miniopterus schreibersi. These repetitive sequences are 57.85%, A-T rich, organized in tandem, and with a monomer unit length of 904 bp. Methylation analysis using the isoesquizomer pair MspI and HpaII indicates that the cytosines present in the sequences CCGG are partially methylated. Furthermore, Southern blot analysis demonstrated that these DNA sequences are absent in the genomes of four related microbat species and suggest that it could be specific to the M. schreibersi genome.     

Repetitive sequences: cause for variation in genome size and chromosome morphology in the genus Oryza

Large variation in genome size as determined by the nuclear DNA content and the mitotic chromosome size among diploid rice species is revealed using flow cytometry and image analyses. Both the total chromosomal length (r_0.939) and the total chromosomal area (r_0.927) correlated well with the nuclear DNA content. Among all the species examined, Oryza australiensis (E genome) and O. brachyantha (F genome), respectively, were the largest and smallest in genome size. O. sativa (A genome) involving all the cultivated species showed the intermediate genome size between them. The distribution patterns of genome-specific repetitive DNA sequences were physically determined using fluorescence in situ hybridization (FISH). O. brachyantha had limited sites of the repetitive DNA sequences specific to the F genome. O. australiensis showed overall amplification of genome-specific DNA sequences throughout the chromosomes. The amplification of the repetitive DNA sequences causes the variation in the chromosome morphology and thus the genome size among diploid species in the genus Oryza.     

Isolation of a D-genome specific repeated DNA sequence fromAegilops squarrosa

Three repeated sequence clones, pAS1(1.0 Kb), pAS2(1.8 Kb) and pAS12(2.5 Kb), were isolated fromAegilops squarrosa (Triticum tauschii). The inserts of the three clones did not hybridize to each other. Two of the clones, pAS2 and pAS12, contain repeated sequences which were distributed throughout the genome. The clone pAS1 sequence was more restricted and was located in specific areas on telomeres and certain interstitial sites along the chromosome length. This cloned sequence was also found to be restricted to the D genome at the level ofin situ hybridization. The pAS1 sequence will be useful in chromosomal identification and phylogenetic analysis. All three clones will allow assessment of genome plasticity inAegilops squarrosa. Nuclear DNA content varies over a range of 10,000 fold among all organisms (Nagl et al., 1983). Among angiosperms, at least a 65-fold range in genome size occurs in diploid species (Sparrow, Price and Underbrink, 1972; Bennett, Smith and Heslop-Harrison, 1982). This DNA variation has been reported within families, genera, and species (Rothfels et al., 1966; Rees and Jones, 1967; Miksche, 1968; Price, Chambers and Bachmann, 1981). Much of the interspecific variation in genome size among angiosperms appears to be due to amplification and/or deletion of DNA within chromosomes. The variation in genome size does not appear to result in changes in the number of coding genes (Nagl et al., 1983). While the number of coding genes, with the exception of rDNA in specific examples, appears to remain constant, the remaining non-coding regions are quite flexible. This non-coding DNA encompasses over 99% of the plant genome and consists of sequences that exist as multiple copies throughout the genome and are identified as repeated DNA sequences (Flavell et al., 1974). Flavell et al. (1974) have reported that increasing genome size in higher plants is associated with increasing repetitive DNA amounts. Subsequent reports have substantiated this correlation (Bachmann and Price, 1977; Narayan, 1982). In various cereals, heterochromatin, which has been demonstrated to be correlated with the location of specific repeated DNA sequences, has been positively correlated with genome size (Bennett, Gustafson and Smith, 1977; Rayburn et al., 1985). Furuta, Nishikawa and Makino (1975) found significant DNA content variation among different accessions ofAegilops squarrosa L. This species contains the D genome, a pivotal genome in several polyploid species and also found in hexaploid wheat (AABBDD). The importance of this genome to the study of bread wheat genomes makes the mechanism(s) of this genomic plasticity of particular interest. In order to determine which sequences are varying, one must first have a way to identify specific types of chromatin and/or DNA. Specific types of chromosome banding such as C- and N-banding have been used to identity types of chromatin in previous studies. C-banding of the D genome results in very lightly staining bands whose pattern is somewhat indistinct. N-banding alternatively has been shown to be useful in identifying certain chromosomes of hexaploid wheat but is limited by the lack of major bands in the D genome (Endo and Gill, 1984). Specific DNA sequences have been isolated fromTriticum aestivum cultivar “Chinese Spring” (hexaploid wheat). However, these sequences are representatives of the A and/or B genomes of hexaploid wheat and are not found in significant quantities in the D genome (Hutchinson and Lonsdale, 1982). Various other repeated DNA sequences have been successfully isolated from rye (Bedbrook et al., 1980) and identified on rye chromosomes (Appels et al., 1981; Jones and Flavell, 1982). Certain of these sequences are found in wheat genomes, but the sequences are representative of only a minor fraction of the D genome (Bedbrook et al., 1980; Rayburn and Gill, 1985). The purpose of this report is to describe three distinct repeated DNA sequences isolated fromA. squarrosa (D genome). Two clones appear to be distributed throughout the total genome, and the third clone is restricted to specific sites along the chromosomes. This latter clone will prove useful in cytologically defining the D genome chromosomes. These sequences appear representative of two types of repeated DNA genome organization: 1) sequences distributed throughout the genome and 2) specific arrays of repeated sequences. The availability of such repeated DNA sequence clones along with the known intraspecific DNA content variation inA. squarrosa will allow the study of genomic plasticity of this species.     

The nuclear genome of Brachypodium distachyon: analysis of BAC end sequences

Due in part to its small genome (~350 Mb), Brachypodium distachyon is emerging as a model system for temperate grasses, including important crops like wheat and barley. We present the analysis of 10.9% of the Brachypodium genome based on 64,696 bacterial artificial chromosome (BAC) end sequences (BES). Analysis of repeat DNA content in BES revealed that approximately 11.0% of the genome consists of known repetitive DNA. The vast majority of the Brachypodium repetitive elements are LTR retrotransposons. While Bare-1 retrotransposons are common to wheat and barley, Brachypodium repetitive element sequence-1 (BRES-1), closely related to Bare-1, is also abundant in Brachypodium. Moreover, unique Brachypodium repetitive element sequences identified constitute approximately 7.4% of its genome. Simple sequence repeats from BES were analyzed, and flanking primer sequences for SSR detection potentially useful for genetic mapping are available at . Sequence analyses of BES indicated that approximately 21.2% of the Brachypodium genome represents coding sequence. Furthermore, Brachypodium BES have more significant matches to ESTs from wheat than rice or maize, although these species have similar sizes of EST collections. A phylogenetic analysis based on 335 sequences shared among seven grass species further revealed a closer relationship between Brachypodium and Triticeae than Brachypodium and rice or maize. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. N. Huo and G.R. Lazo contributed equally to this work.     

A novel repetitive DNA sequence in the genus Oryza

Summary Repetitive DNA sequences in the genus Oryza (rice) represent a large fraction of the nuclear DNA. The isolation and characterization of major repetitive DNA sequences will lead to a better understanding of rice genome organization and evolution. Here we report the characterization of a novel repetitive sequence, CC-1, from the CC genome. This repetitive sequence is present as long tandem arrays with a repeat unit 194 bp in length in the CC-diploid genome but 172 bp in length in the BBCC and CCDD tetraploid genomes. This repetitive sequence is also present, though at lower copy numbers, in the AA and BB genomes, but is absent in the EE and FF genomes. Hybridization experiments revealed considerable differences both in copy numbers and in restriction fragment patterns of CC-1 both between and within rice species. The results support the hypothesis that the CC genome is more closely related to the AA genome than to the BB genome, and most distantly related to the EE and FF genomes.     

The efficacy of Cot-based gene enrichment in wheat (Triticum aestivum L.).

We report the results of a study on the effectiveness of Cot filtration (CF) in the characterization of the gene space of bread wheat (Triticum aestivum L.), a large genome species (1C = 16,700 Mb) of tremendous agronomic importance. Using published Cot data as a guide, 2 genomic libraries for hexaploid wheat were constructed from the single-stranded DNA collected at Cot values > 1188 and 1639 M x s. Compared with sequences from a whole genome shotgun library from Aegilops tauschii (the D genome donor of bread wheat), the CF libraries exhibited 13.7-fold enrichment in genes, 5.8-fold enrichment in unknown low-copy sequences, and a 3-fold reduction in repetitive DNA. CF is twice as efficient as methylation filtration at enriching wheat genes. This research suggests that, with improvements, CF will be a highly useful tool in sequencing the gene space of wheat.