富含多糖的转基因石斛基因组DNA提取方法(英文)

在石斛兰转基因研究中,需通过PCR和Southern杂交等手段检测外源基因是否转入并整合到转化植株基因组.由于转化石斛植株生长缓慢,且含有多糖,因此从转化植株中提取高质量的基因组DNA以尽快对转基因苗进行分子检测存在较大困难.本研究旨在通过改良现有的DNA提取法(CTAB法或SDS法1,克服转化石斛植株基因组DNA提取中碰到的产量低,纯度不高而导致难以进行PCR或酶切等问题.在本研究所用的3种改良法中,方法Ⅱ能从少量的转化石斛苗中提取出高产量和高纯度的基因组DNA.研究结果表明方法Ⅱ提取的基因组DNA完全适用于转基因石斛的外源基因PCR扩增,限制性酶切和Southern杂交分析.     

中国88个马铃薯审定品种SSR指纹图谱构建与遗传多样性分析

为对马铃薯品种鉴别、优良杂交组合选配提供分子水平上的依据,利用SSR标记构建了中国2000-2007年审定的88个马铃薯品种的指纹图谱并进行了遗传多样性分析。以138对SSR引物对16份遗传差异较大的马铃薯材料的基因组DNA进行了扩增,筛选出10对多态性高、谱带清晰的引物。利用10对SSR引物对全部供试材料进行扩增及电泳检测,共检测到135个等位位点,其中133个为多态性位点,多态性比率达98.52%。每对SSR引物扩增出的等位位点数7~22个,平均13.5个,多态性信息量变化范围为0.7604~0.9375,平均0.8501。通过对电泳检测结果的统计分析,利用S180、S25、S7、S151、S184及S192等6对引物构建了88份供试材料的SSR指纹图谱。聚类分析表明,在相似系数0.620处,所有供试材料被被聚为一类,在相似系数0.652处,81.8%的材料仍然聚在一起,从分子水平上表明供试材料遗传基础非常狭窄。聚类分析结果与供试材料系谱来源有较好一致性,同一栽培区域育成的品种在不同程度上聚在一类。     

DNA profiling and genetic diversity of Korean soybean (<Emphasis Type="Italic">Glycine max</Emphasis> (L.) Merrill) landraces by SSR markers

Approximately 7,000 accessions of Korean soybean (Glycine max (L.) Merrill) landraces, largely composed of three collections, the Korea Atomic Energy Research Institute’s soybean (KAS), the Korean Crop Experiment Station’s soybean (KLS) and the Korean Agricultural Development and Technology Center’s soybean (KADTC) collections, have been conserved at the Rural Development Administration (RDA) genebank in Korea. The accessions within collections were classified based on their traditional uses such as sauce soybean (SA), sprouted soybean (SP), soybean for cooking with rice (SCR), and OTHERS. A total of 2,758 accessions of Korean soybean landraces were used to profile and to evaluate genetic structure using six SSR loci. A total of 110 alleles were revealed by at the six SSR loci. The number of alleles per SSR locus ranged from 9 to 39 in Satt187 and Satt_074, respectively. The number of alleles ranged from 87 in the KADTC collection to 96 in the KLS collection, and from 63 in the SCR group to 95 in the SP group. Nei’s average genetic diversity ranged from 0.68 to 0.70 across three collections, and 0.64 to 0.69 across the usage groups. The average between-group differentiation (G _st) was 0.9 among collections, and 4.1 among the usage groups. The similar average diversity among three collections implies that the genetic background of the three collections was quite similar or that there were a large number of duplicate accessions in three collections. The selection from the four groups classified based upon usage may be a useful way to select accessions for developing a Korean soybean landrace core collection at the RDA genebank. DNA profile information of accessions will provide indications of redundancies or omissions and aid in managing the soybean collection held at the RDA genebank. The information on diversity analysis could help to enlarge the genetic diversity of materials in breeding programs and could be used to develop a core collection.     

Recent advances in molecular genetics of forest trees

The use of molecular markers has greatly enhanced our understanding of the genome structure of forest trees. Conifers, in particular, have a relatively large genome, containing a very high proportion of repeated DNA, consisting of tandemly repetitive and dispersed repetitive DNA sequences. The nature of highly conserved tandemly repetitive rRNA genes has been investigated in a number of tree species, and their sites mapped on specific chromosomes by fluorescent in situ hybridization (FISH). Different families of retrotransposons (IFG, and TPE1) have been isolated and characterized from the dispersed repetitive DNA of pines. Genome maps have been constructed in a number of forest tree genera: Pinus, Picea, Pseudotsuga, Cryptomeria, Taxus, Populus, and Eucalyptus. EST databases have been established from cDNA clones of pines and poplars. The structure and maternal or paternal modes of inheritance of organelle genomes have been investigated in forest trees. Comparative mapping in conifers has shown that gene families are conserved across genera. Due to lack of polyploidy in conifers, the evolution of this group of trees may have occurred primarily by duplication and dispersal of genes, probably by retrotranspositions, to form complex gene families. The evolution of angiosperm tree species has presumably involved both gene duplication as well as genome duplication (polyploidy). Application of genetic engineering has shown that genes from phylogenetically unrelated organisms can be introduced and expressed in trees, thus offering prospects of genetic improvement of forest trees. This revised version was published online in August 2006 with corrections to the Cover Date.     

Assessment of clonal fidelity of micropropagated gerbera plants by ISSR markers

True-to-type clonal fidelity is one of the most important pre-requisites in micropropagation of crop species. Genetic fidelity of in vitro raised 45 plants of gerbera (Gerbera jamesonii Bolus) derived from three different explants, viz., capitulum, leaf and shoot tips, was assessed by 32 ISSR markers, for their genetic stability. Out of 32 ISSR markers, 15 markers produced clear, distinct and scorable bands with an average of 5.47 bands per marker. The markers designed from AG motif amplified more number of bands. The markers anchored at 3′ ends produced high number of consistent bands than unanchored markers. Fifteen ISSR markers generated a total of 3773 bands, out of which 3770 were monomorphic among all the clones. The Jaccard's similarity coefficient revealed that out of 45 clones derived from different explants, 44 were grouped into a single large cluster alongwith the mother plant with a similarity coefficient value of 1.00, whereas one clone (C38) remained ungrouped. The clones derived from capitulum and shoot tip explants did not show any genetic variation, whereas, one of the leaf-derived clones exhibited some degree of variation.     

DNA序列分析在植物分子系统学研究中的应用现状——-以蔷薇科为例

 近20年来, DNA序列已被广泛应用于植物各分类阶元的系统学研究中, 为解决长期有争议 的和亟待解决的系统进化问题提供了有力的证据。现以蔷薇科为例, 概述了应用DNA片段进行植物分子系统研究的现状, 详细剖析了应用DNA序列进行植物系统发育研究时常见的问题及其原因, 提出了对存在多倍化、杂交起源和快速分化等复杂进化史的植物类群进行系统学分析时选用DNA序列的策略和注意事项。     

流式细胞术在水生生物DNA含量和倍性分析中的应用

利用流式细胞仪对鲤鱼、银鲫、黄颡、牙鲆等淡水和海水鱼类进行了DNA含量测定或倍性分析。通过几百个样品的测定和分析,认为流式细胞仪在DNA相对含量测定和倍性分析上结果比较稳定一致,可以用于大量样品的测定分析。同时对待测样品的保存、处理及上样量等进行了一系列比较研究,找出最好的保存方法、最简便的样品处理方法和最少的上样量。流式细胞仪在水生生物的遗传育种、种群分化等方面有很重要的应用价值。     

Ecology,physiology and genetics of a phycodnavirus infecting the noxious bloom-forming raphidophyte <Emphasis Type="Italic">Heterosigma akashiwo</Emphasis>

Abstract:   Heterosigma akashiwo virus (HaV) is a large icosahedral virus (∼0.2 μm) harboring a double-stranded DNA (dsDNA) genome (∼294 kbp). The virus is the only member of the genus Raphidovirus in the family Phycodnaviridae. Since its first discovery, a number of ecologic, physiologic and genetic studies about HaV have been conducted; especially, the relationship between H. akashiwo and HaV in nature was studied and viral infection is now regarded as a significant factor influencing the dynamics and termination of H. akashiwo blooms. HaV infection has considerable impacts on H. akashiwo populations in both aspects of fluctuation in biomass (quantity) and changes in clonal composition (quality). Partial sequencing of the HaV genome revealed that a number of genes showed considerable similarity to those of other protist-infecting viruses; still, the phylogenetic position of HaV suggested a number of enigmas in host–virus coevolution. Here are summarized the ecology, physiology and genetics of HaV especially from the viewpoint of the host–virus relationship.     

Direct evidence of multiple paternities in natural population of viviparous Japanese surfperch by allelic markers of microsatellite DNA loci

This study was performed to obtain information on the occurrence of multiple paternities in three species of viviparous Japanese surfperch using allelic markers of microsatellite DNA loci. Direct evidence for multiple fertilizations was established by reconstructing paternal genotypes from the progeny of gravid females. Multiple paternities were ascertained in five of 10 broods of Ditrema temmincki and in three of nine broods of Neoditrema ransonneti, but not in Ditrema viride. The number of patrilines detected in the progeny of D. temmincki and N. ransonneti females were two or three, respectively, as determined by the GERUD v2.0 algorithm for reconstructing parental genotypes from half-sib progeny arrays.