用荧光素显带技术识别柑橘杂种染色体的多样性

应用色霉素A3（Chromomycin A3，CMA）荧光素显带技术，得到了清晰的文旦柚（Citrus grandis[L．]Osb）染色体构成类型：土佐文旦（Tosa—Buntan Pummelo）为1A＋1B＋5C＋2D＋9E；水晶文旦（Sisho-Buntan Pummelo）为3A＋3C＋3D＋9E．对杂交F1代38株文旦柚的染色体组进行了观察，发现了共13种染色体构成类型．荧光素显带技术对染色体构成多样性的高度识别，可成为植物配子体形成过程中的减数分裂、基因重组等机理分析的有效方法．     

普通栽培稻与非洲栽培稻杂交的可交配性研究

以二倍体普通栽培稻(Oryza sativa，2n=2x=24)为母本，以非洲栽培稻(Oryza globerrima，2n=2x=24)为父本进行远缘杂交配组。研究结果表明，二倍体普通栽培稻与非洲栽培稻杂交表现出一定的可交配性，但结实率很低(0．32％～1．97％)。对其杂交结实的胚胎学观察发现，非洲栽培稻的花粉能在二倍体普通栽培稻的柱头上萌发，萌发率仅为0．18％～0．56％，花粉管在花柱内的伸长速度比较慢，发生的受精频率比较低。以同源四倍体水稻(Oryza sativa，2n=4x=48)为母本，以非洲栽培稻为父本杂交配组。研究结果表明，两间比较容易杂交，其结实率达到5．70％～8．86％。观察其胚胎学表现，非洲栽培稻的花粉在同源四倍体水稻的柱头上萌发频率高达12．81％～14．56％，花粉管在柱头内伸长速度较快，发生的受精频率比较高。利用同源四倍体水稻为桥梁，更容易将非洲栽培稻的遗传物质引入普通栽培稻。     

内蒙古棘豆属Oxytropis细胞分类学研究

本文是对内蒙古１４种棘豆属植物的染色体研究，结果如下：染色体基数ｘ＝８．有二倍体和四倍体两个倍性，核型组成由中部和近中部着丝点染色体组成，最后一对（或二对）具中部随体，核型公式有３种类型：２ｎ＝２ｘ＝１６＝８ｍ（２ＳＡＴ）＋８ｓｍ，２ｎ＝２ｘ＝１６＝１０ｍ（２ＳＡＴ）＋６ｓｍ，２ｎ＝２ｘ＝１６＝１２ｍ（２ＳＡＴ）＋４ｓｍ，其中ＯｘｙｔｒｏｐｉｓａｃｉｐｈｙｌｌａＯ．ｏｃｈｒａｎｔｈａ，Ｏ．ｇｒａｃｉｌｌｉｍａ，Ｏ．ｓｑｕａｍｍｕｌｏｓａ，Ｏ．ｄｉｖｅｒｓｉｆｏｌｉａ，Ｏ．ｌａｔｉｂｒａｃｔｅａｔａ，Ｏ．ｌｅｐｔｏｐｈｙｌ－ｌａ，Ｏ．ｈｉｒｔａ属首次报道。     

马尾松不同种源染色体荧光带型的研究

分析了我国南方主要树种马尾松7个种源染色体的两种荧光带型，色霉素A3（简称CMA带）和4，6－二脒基－2－苯基咪哚（简称DAPI带）。结果表明：（1）马尾松染色体的CMA带型中存在着5种不同类型，它们分别是1对为着丝粒和臂间处均有荧光带的中间着丝粒染色体（A类型）；6对为臂间处有荧光带的中部着丝粒染色体（B类型）；1对为着丝粒处有荧光带的中部着丝粒染色体（C类型）；3对为无荧光带的近中部中部着丝粒染色体（D类型）；1对为着丝粒处有荧光带的近中着丝粒染色体（E类型）。其CMA带型公式2n=24=2A 12B 2C＋6D＋2E。（2）根据CMA的带型状况，将马尾松的7个种源分为2种类群区，第一类为江西吉安，福建漳平，贵州贵阳，四川古蔺和湖南宜樟为中部区；第二类为广东高州和广西宁明为南部区。（3）比较马尾松7种源的DAPI带型，南部区种源的染色体上的荧光带纹较中部区种源染色体上荧光带纹要多些，西部区种源染色体上的荧光带纹较东部区种源染色体上的荧光带纹亦要多些。     

菜心多倍体诱变及其细胞学观察

报道了用秋水仙素水溶液处理菜心种子及幼苗生长点获得多倍体的方法,并对获得的四倍体与二倍体（ＣＫ）进行了形态学、细胞学以及结籽率等的比较观察。试验结果表明：（１）在１０～１５℃的条件下,用０．１％秋水仙素水溶液处理幼苗生长点４８小时的诱变效果最好,诱变率可达２６．６７％,浸泡种子效果不佳,最高诱变率仅为４．０％。（２）对四倍体植株分别进行人工自交均获得了种子,但结籽率较二倍体（ＣＫ）明显降低,平均达４２．３％,而八倍体植株几乎是完全不育的。（３）与二倍体（ＣＫ）比较,四倍体植株生长旺盛,花薹的产量和品质均有所提高。（４）随着染色体组的增加,花粉粒大小和叶片保卫细胞中的叶绿体数均显著增加,可作为鉴定植株倍性的一个指标     

棉花45S rDNA内转录间隔区的克隆与分析

45S rDNA的内转录间隔区(Internal transcribed space, ITS)序列变异性强，是被广泛应用于物种间进化关系研究的序列标签.通过克隆艾克棉(Gossypium ekmanianum,AD₆)、斯蒂芬斯棉(Gossypium stephensii,AD₇)、异常棉(Gossypium anomalum,B₁)、戴维逊氏棉(Gossypium davidsonii,D_3-d)、克劳茨基棉(Gossypium klotzschianum,D_3-k)、旱地棉(Gossypium aridum,D₄)6个棉种的ITS序列，首次获得新鉴定的2个四倍体棉种艾克棉和斯蒂芬斯棉的ITS区序列信息，补充了异常棉、克劳茨基棉和戴维逊氏棉3个棉种ITS区序列的未知碱基.基于ITS序列信息构建了涵盖A,B,C,D,E,F,AD 7个基因组的棉属系统发育树，确定了新鉴定的2个四倍体棉种进化地位，探讨了棉属亲缘关系.研究结果补充和完善了棉属ITS序列信息，...     

不同倍性鲤科鱼CyclinB基因内含子克隆与进化分析

运用PCR扩增、克隆、测序等技术,分别获得三倍体鲫鲂、四倍体鲫鲂、五倍体鲫鲂和团头鲂、草鱼、鲢鱼、鳙鱼等鲤科鱼CyclinB基因部分DNA序列．结合异源四倍体鲫鲤及其原始亲本红鲫和鲤鱼CyclinB基因对应DNA序列,对不同倍性鲤科鱼类CyclinB基因DNA片段序列进行比较分析．结果表明：由相同引物扩增的染色体数为48的鲂、草、鲢、鳙鱼,只扩增出1条DNA片段,片段长度分别为750bp,950bp,720bp和720bp,而染色体数目加倍的红鲫、鲤鱼、异源四倍体鲫鲤、异源四倍体鲫鲂,扩增出2条DNA片段（1200bp和900bp）,三倍体和五倍体鲫鲂扩增出3条DNA片段（1200bp,900bp和750bp）,每个DNA片段均含CyclinB基因第2,3内含子和第2,3,4外显子．序列分析表明,四倍体鲫鲤、三倍体鲫鲂、四倍体鲫鲂和五倍体鲫鲂与其母本1200bp片段同源性分别为99．5％,98．9％,99．5％和88．7％,与母本900bp片段同源性分别为97．5％,94．6％,94．2％和89．99／6,说明四倍体鲫鲤与多倍体鲫鲂1200bp和900bpCyclinB基因片段与母本红鲫同源性高,在进化上具有偏母性遗传特性．而三倍体和五倍体鲫鲂的750bpCyclinB基因片段-9父本同源性分别高达98．6％和98．2％,说明其来自父本团头鲂．在两性可育的异源四倍体鲫鲤和异源四倍体鲫鲂中存在各自父本CyclinB基因片段序列消除现象．此外,用CyclinB基因内含子3序列构建不同倍性鲤科鱼的系统进化树,结果表明,由CyclinB基因内含子序列构建的系统进化树可以正确反映亲缘关系较近的鲤亚科属间鱼类系统进化关系,而不适合亲缘关系较远的亚科间杂交鱼类系统进化分析．     

贵州四种稻蝗核型和C带的比较研究

研究了贵州小稻蝗（Oxya hyia-intricata）,山稻蝗（O．agarisa）,中华稻蝗（O．chinensis）扣上海稻蝗（O．shanghaiensis）等4种稻蝗的核型扣C带。结果显示：4种稻蝗的染色体数目均为2n（♂）=22＋XO=23,全部是端部着丝粒染色体;核型公式：2n（♂）=2X=23t;NF=23;都可按相对长度分成L、M、S3组;都具着丝粒C带,但端带和居间带不是每种每条染色体都有;异染色质总量是山稻蝗的（22．01％）最多,上海稻蝗的（20．13％）最少。     

樱属植物核型参数及种间亲缘关系分析

统计了18份樱属植物的核型数据,借助DPS软件用最大距离法进行聚类分析,探讨了属内系统结构和种间亲缘关系。结果表明:樱属染色体基数x=8,染色体数目有2n=16、2n=24以及2n=32;染色体类型有4种,正中部着丝点染色体(M),中部着丝点染色体(m),近中部着丝点染色体(sm),近端部着丝点染色体(st);随体常见,核型类型有2A、2B、1B 3种;野生类群多为二倍体,栽培品种多倍化现象显著;大叶早樱(Cerasus subhirtella)、‘十月樱’(C.subhirtella‘Autumnalis’)、东京樱花(C.yedoensis)、山樱花(C.serrulata)聚为一类,支持黑果组(C.sect.Sargentella)成立;二倍体酸樱桃(C.vulgaris)、甜樱桃(C.avium)聚为一类,支持芽鳞组(C.sect.Cerasus)成立;虽然红山樱(C.jamasakura)与山樱花表型相似,但聚类结果显示二者遗传距离较远,前者与钟花樱(C.campanulata)遗传距离较近;本属栽培种类多倍化程度高于野生种类且有多个倍性,与其二倍体祖先表现出较远的遗传距离;聚类分析难以解释栽培种的起源。与传统的经典分类结果不同,矮生樱亚属(C.subg.Microcerasus)与典型樱亚属(C.subg.Cerasus)在聚类图上没有明显地分开。     

葡萄气孔、花粉等与倍性的关系及倍性判别分析

测定了二倍体和四倍体葡萄的气孔、花粉、枝叶等11个外部性状,结果表明,染色体倍性与其中8个性状呈极显著相关,其中气孔密度与倍性呈极显著负相关。将这些相关性状作二倍体与四倍体显著性测定表明差异显著,可用二倍体与四倍体的各性状建立判别公式进行未知品种的倍性分析鉴定。     

Evolutionary analysis of allotetraploid hybrids of red crucian carp × common carp, based on ISSR, AFLP molecular markers and cloning of cyclins genes

The allotetraploid hybrids of red crucian carp x common carp are the first reported artificially cultured polyploid fish with bisexual fertility and stable inheritance in vertebrate. Using ISSR and AFLP markers and the cyclins genes, the genomes and cyclin gene sequence changes were analyzed between the allotetraploid hybrids and their parents. The results indicated that the allotetraploids inherited many genetic characteristics from their parents and the genetic characteristics were stable after 15 generations. However, the allotetraploids had a closer genetic relationship with their original female parents and represented a bias toward the maternal progenitor. DNA fingerprinting analysis showed that the allotetraploids had undergone sequences deletion from their original parents and that the deleted sequences were mostly from the male parent＇s genome. Some non-parental bands were found in the allotetraploid hybrids. Sequences analysis of the cyclin A1 and B1 genes showed nonsynonymous substitutions of single nucleotides in codons that were different from their original parents, leading to non-parental amino acid loci. We speculate that the non-additivity in the allotetraploids, compared with their progenitors, could be an adjustment to the genomic shock from heterozygosity and polyploidy, allowing maintenance of genetic stability.     

Comparative analysis of A, B, C and D genomes in the genus Oryza with Cot-1 DNA of C genome

Fluorescence in situ hybridization （FISH） was applied to somatic chromosomes preparations of Oryza officinalis Wall. （CC）, O. sativa L. （AA）xO. officinalis F1 hybrid （AC）, backcross progenies BC1 （AAC and ACC）, O. latifolia Desv. （CCDD）, O. alta Swallen （CCDD） and O. punctata Kotschy （BBCC） with a labelled probe of Cot-1 DNA from O. officinalis. In O. officinalis, the homologous chromosomes showed similar signal bands probed by Cot-1 DNA and karyotype analysis was conducted based on the band patterns. Using no blocking DNA, the probe identified the chromosomes of C genome clearly, but detected few signals on chromosomes of A genome in the F1 hybrid and two backcross progenies of BC1. It is obvious that the highly and moderately repetitive DNA sequences were considerably different between C and A genomes. The chromosomes of C genome were also discriminated from the chromosomes of Dand B-genome in the tetraploid species O. latifolia, O. alta and O. punctata by Cot-1 DNA-FISH. Comparison of the fluorescence intensity on the chromosomes of B, C and D genomes in O. latifolia, O. alta, and O. punctata indicated that the differentiations between C and D genomes are less than that between C and B genomes. The relationship between C and D genomes in O. alta is closer than that of C and D genomes in O. latifolia. This would be one of the causes for the fact that both the genomes are of the same karyotype （CCDD） but belong to different species. The above results showed that the Cot-1 DNA had a high specificity of genome and species. In this paper, the origin of allotetraploid in genus Oryza is also discussed.     

Studies of new EST-SSRs derived from Gossypium barbadense

Existing cotton EST-SSR markers are mostly derived from Gossypium arboreum and Gossypium hir-sutum, but EST-SSR markers from Gossypium barbadense are scarce. One hundred and nineteen EST-SSRs were developed based on 98 unique ESTs from a cDNA library constructed in our laboratory using developing fibers from G. barbadense cv. Pima3-79. Among the SSRs, trinucleotide AAG appeared at a high frequency of 11.76%. 36 accessions (consisting of 13 diploids of the A genome, 11 diploids of the D genome and 12 allotetraploids of the AD genome) were employed to test new EST-SSRs. 76 EST-SSRs were successfully amplified, and 313 polymorphic fragments were yielded, with an average of 4.11 fragments per primer pair. The PIC ranged from 0.17 to 0.95 with an average of 0.53. Based on Jaccard’s genetic similarity coefficient, these 36 accessions were clustered into three groups. 21 EST-SSRs exhibited polymorphisms in BC1 population ((Emian22 × Pima3-79) × Emian22), 24 polymor- phic loci were generated, while 22 of the 24 polymorphic loci were integrated with our interspecific BC1 backbone genetic linkage map, and anchored in 12 chromosomes. This study effectively proved that EST-SSRs from G. barbadense are valuable for genetic diversity analysis and genetic mapping.     

模式生物的外显子、内含子和基因间序列的识别

基于核酸序列在剪切位点上保守性、组分的不同和编码序列阅读框架的3周期性,模式生物全基因组序列分为外显子、内含子和基因间序列三类.三个标准离散源分别由64个三联体在整条序列上的概率和4个碱基序列首尾(剪切位点附近)共30个位点上的概率共同构成.某条序列的类型就由该序列的离散量同相应区间上三个标准离散量的离散增量确定.结果表明:具有184个信号参数的离散量预测比只有64个三联体参数的结果要高出5%,总体预测成功率:线虫为87.37%,拟南芥为91.08%,果蝇为92.28%,原核生物大肠杆菌的二种序列预测率为92.88%,酵母菌为94.88%.     

Patterns of DNA cytosine methylation between haploids and corresponding diploids in rice

In higher plant, about 30% cytosines are methy-lated[1], among which about 90% methylated sites lie in CpG dinucleotide and CpNpG trinucleotide[2]. The me-thylated DNA has inducing and epigenetic effects on cell biological procedures such as gene differen…     

onc sequences (v-fes) of Snyder-Theilen feline sarcoma virus are derived from noncontiguous regions of a cat cellular gene (c-fes)

Type C sarcoma viruses are genetic recombinants containing portions of replication-competent helper viruses linked to sarcoma virus-specific sequences (generically designated onc genes) which are thought to be required for acute fibroblast transformation. The onc elements of different avian and mammalian sarcoma viral isolates are each homologous to subsets of cellular DNA sequences which have no well-defined role in normal cells. Because of the lack of significant homology between helper viral genes and cellular onc sequences, the recombinational mechanisms which facilitate the formation of sarcoma viral genomes remain unclear. In Moloney murine sarcoma virus, viral onc (or v-mos) and cellular onc (or c-mos) sequences exhibit complete and uninterrupted homology as determined by heteroduplex and restriction enzyme analyses of molecularly cloned DNA. By contrast, the cellular counterparts of the onc elements of Rous sarcoma virus (G. Cooper and R. Parker, personal communication), avian erythroblastosis virus (B. Vennstrom, personal communication), Abelson leukaemia virus (D. Baltimore, personal communication), Harvey sarcoma virus (E. Scolnick, personal communication) and simian sarcoma virus (R. Gallo, personal communication) are now known to contain intervening sequences which do not appear in the respective viral genomes. Here we report the use of the Southern blot technique to examine cat cellular DNA sequences (c-fes) homologous to the onc gene (v-fes) of Snyder-Theilen feline sarcoma virus (ST-FeSV). We used cloned DNA 'probes' containing defined portions of the ST-FeSV genome to show that v-fes sequences originate from at least four noncontiguous sequences in cat cellular DNA, separated from each other by intervening sequences.     

Spontaneous forward mutation versus reversion frequencies for maize Adh1 in pollen

Reliable quantitative data on spontaneous, specific gene mutation frequencies in higher plants and animals are few. Detergents include low natural frequencies and difficulty in obtaining in excess of 10(6) scorable organisms or gametophytes. The alcohol dehydrogenase-1 gene (Adh1 gene; ADH enzyme EC 1.1.1.1.), as expressed in pollen grains, is among the exceptionably suitable; much is known about the maize ADHs, Adh1 function is totally dispensible in an aerobic environment and maize pollen is a trinucleate gametophyte which expresses much of its haploid genome, including Adh1. In particular, there have been two recent methodological advances. First, I am able to cytochemically stain pollen, before or after in vitro germination, for the presence of above 5% normal ADH activity. And second, ADH1- pollen grains survive allyl alcohol (C=C-C-OH) vapour concentrations which kill ADH1+ grains; this selection scheme was developed for yeast by Megnet. My genetic resolution is approximately one mutant (Adh1+-->ADH-) per 10(7) chemically selected, viable gametophytes, and one (phenotypic) revertant (Adh1--->ADH+) per 10(8) unselected gatetophytes. In this note, I compare spontaneous forward mutant frequency with previously published revertant frequencies for one naturally occurring and six ethyl methanesulphonate-induced Adhl-deficient (Adh1-) alleles.     

Cytogenetics of intergeneric hybrids between Brassica species and Orychophragmus violaceus

In the sexual intergeneric hybrids between the cultivated Brassica species and Orychophragmus violaceus, both complete separation and partial separation of the parental genomes were found to occur during mitosis and meiosis under genetic control. The cytogenetics of these hybrids was species-specific for Brassica parents. The different chromosome behavior of hybrids with three Brassica diploids ( B. campestris , B. nigra and B. oleracea ) might contribute to the different cytogenetics of hybrids with three tetraploids ( B. napus, B. juncea and B. carinata). Owing to the parental genome separation, Brassica homozygous plants and aneuploids with various chromosome constitutions were identifiable in the progenies of these hybrids, which were valuable for the study of the structure and evolution of Brassica genome and for the breeding of Brassica crops.     

The grapevine genome sequence suggests ancestral hexaploidization in major angiosperm phyla

The analysis of the first plant genomes provided unexpected evidence for genome duplication events in species that had previously been considered as true diploids on the basis of their genetics. These polyploidization events may have had important consequences in plant evolution, in particular for species radiation and adaptation and for the modulation of functional capacities. Here we report a high-quality draft of the genome sequence of grapevine (Vitis vinifera) obtained from a highly homozygous genotype. The draft sequence of the grapevine genome is the fourth one produced so far for flowering plants, the second for a woody species and the first for a fruit crop (cultivated for both fruit and beverage). Grapevine was selected because of its important place in the cultural heritage of humanity beginning during the Neolithic period. Several large expansions of gene families with roles in aromatic features are observed. The grapevine genome has not undergone recent genome duplication, thus enabling the discovery of ancestral traits and features of the genetic organization of flowering plants. This analysis reveals the contribution of three ancestral genomes to the grapevine haploid content. This ancestral arrangement is common to many dicotyledonous plants but is absent from the genome of rice, which is a monocotyledon. Furthermore, we explain the chronology of previously described whole-genome duplication events in the evolution of flowering plants.