紫花苜蓿复合体(Medicago sativa complex)叶片形态特征及数量分类研究

运用比较形态学和比较解剖学方法,使用扫描电镜和光学显微镜对紫花苜蓿复合体(Medicago sativa complex)6个分类群的叶片形态特征和叶片解剖结构进行了观察和比较,并以15个叶片表征形态性状为基础,采用聚类分析法(UPGMA)和主成分分析方法(PCA)对6个分类群进行了数量分类研究.观察结果表明:各分类群叶片的上、下表皮多为不规则形细胞;垂周壁呈深浅不一的波状;气孔器为不规则型,具有蜡质气孔盖,气孔密度有一定差异.6个分类群的叶片均为薄纸质型,厚度130～170 μm,表皮细胞切面近圆形或近长方形;栅栏组织细胞1～2层,厚度41～68 μm,细胞排列紧密;海绵组织厚度32～75 μm,细胞排列疏松;不同分类群叶片的组织疏松度和组织紧缩度有一定的差异,大花苜蓿(M. trautvetterii Sumnev. )叶片的组织疏松度最高,紫花苜蓿叶片的组织紧缩度最高.UPGMA结果显示:在结合线1.53处可将6个分类群划分为2支,其中,黄花苜蓿(M. falcata L. )独立为一支,其余5个分类群聚成另一支;在结合线1.18处,第2支又被分成2个亚支,其中一个亚支包含紫花苜蓿和天山苜蓿(M. tianschanica Vassilcz. ),另一个亚支则包含西锡金苜蓿(M. schischkinii Sumnev. )、座垫苜蓿(M. rivularis Vassilcz. )和大花苜蓿.PCA结果表明:对紫花苜蓿复合体而言,叶片表皮细胞形状、垂周壁式样、轴性分化特征、组织疏松度和气孔密度等特征具有较好的分类价值;基于主成分分析的Q分布图与聚类分析结果也具有较高的一致性.根据本研究结果及前人的研究结果,认为国产的紫花苜蓿复合体应包含3个分类群,即紫花苜蓿、黄花苜蓿及多变苜蓿(M. varia Martyn).此外,西锡金苜蓿、座垫苜蓿、天山苜蓿和大花苜蓿等杂交后代分类群的性状分化不稳定,应属于多变苜蓿的同种异名植物.     

苜蓿属和胡卢巴属植物的形态特征及数量分类研究

为研究苜蓿属(Medicago L.)与胡卢巴属(Trigonella L.)间的亲缘关系,寻找两属间的分属依据,解决中间过渡类群的分类归属问题,对苜蓿属与胡卢巴属种类的种皮结构和花粉形态特征进行了观察和测量,并以苜蓿属和胡卢巴属植物的40个形态性状为依据、应用数量分类方法对供试的16个种类进行了聚类分析(UPGMA)和主成分分析(PCA).此外,还根据具有重要分类学意义的形态性状,编写了供试的苜蓿属和胡卢巴属16个种类的分类检索表.观察结果表明,供试的两属植物(11种)的种皮结构基本相同,均由栅栏组织细胞、厚壁细胞和薄壁细胞组成,但胡卢巴属种皮的帽状结构和栅栏组织的厚度均小于苜蓿属.供试的两属植物(9种)花粉形态特征相似,均为椭球形或近球形,赤道面观为椭圆形,极面观为三裂圆形;极轴和赤道轴长分别为22.67～34.67 μm和17.34～31.60 μm;花粉表面纹饰较简单,呈小穴状.种皮结构和花粉特征显示两属的亲缘关系较近,仅种皮结构中的帽状结构差异具有一定的属间分类学意义.聚类分析结果表明,在欧氏距离10.85处,供试的苜蓿属(11种)和胡卢巴属(5种)的16个种类被划分为2组,A组均由胡卢巴属种类组成,B组均由苜蓿属种类组成;在欧氏距离9.32处,B组可被进一步划分为3个亚组,所包含的种类分别为苜蓿属的阔荚亚属(Subgen. Platycarpos D. F. Cui)、镰荚亚属(Subgen. Medicago Tutin)和天蓝苜蓿亚属(Subgen. Lupulina Gressh.)的种类.主成分分析结果表明,前3个主成分的累积贡献率较低,仅57.50%;在前3个主成分中,果实和小叶的长宽比、花萼长和被毛状况、茎和小叶的被毛状况、小叶尖形状、花序长等相关性状的绝对权重值均在0.6以上,具有重要的分类学意义;基于前2个主成分所做的二维散点图中供试的16个种类组成4个表征类群,与聚类分析结果一致.     

草木樨属根瘤菌的16S rDNA-RFLP分析

采用16S rDNA-RFLP分析方法对草木樨属根瘤菌进行了遗传多样性及系统分类研究。结果表明,3种限制性内切酶(HaeⅢ、HinfⅠ、MspⅠ)对所有供试菌株的酶切图谱类型组合只有2种。类型I的代表菌株CC-NWSX0003-1与豌豆根瘤菌(R.leguminosarum)的模式菌株USDA2370的序列相似性达到99.8%,在分类地位上属于根瘤菌属(Rhizobium)分支;类型Ⅱ的代表菌株CCNWGS0006与草木樨中华根瘤菌(Sinorhizobium meliloti)的16S rDNA相似性为100%,属于中华根瘤菌属(Sinorhizobium)分支。     

基于28S rDNA序列构建侧耳属系统发育树

通过对侧耳属18个分类单元的28S rDNA序列进行分析,构建了侧耳属较为完整的系统发育树。分子系统学资料显示:Coremiopleurotus组和侧耳属内单、二系菌丝系统分别为多系起源的;Pleurotus组单系菌丝种类、被划分在Tuberregium组的具核侧耳和Lentodiellum组的P. levis能够分别与侧耳属内其他成员进行区分;红侧耳、P. calyptratus、P. opuntiae三者关系密切,而金顶侧耳应作为白黄侧耳的种下分类单元。     

基于28SrDNA序列构建侧耳属系统发育树

通过对侧耳属18个分类单元的28S rDNA序列进行分析,构建了侧耳属较为完整的系统发育树。分子系统学资料显示:Coremiopleurotus组和侧耳属内单、二系菌丝系统分别为多系起源的;Pleurotus组单系菌丝种类、被划分在Tuberregium组的具核侧耳和Lentodiellum组的P. levis能够分别与侧耳属内其他成员进行区分;红侧耳、P. calyptratus、P. opuntiae三者关系密切,而金顶侧耳应作为白黄侧耳的种下分类单元。     

西北地区天蓝苜蓿根瘤菌16S rDNA RFLP分析

利用RFLP和序列测定方法,对分离自西北地区的67株天蓝苜蓿根瘤菌16S rDNA进行了分析研究。结果表明:所有供试菌株分别归属于中华根瘤菌属(Sinorhizobium)、根瘤菌属(Rhizobium)和土壤杆菌属(Agrobac-terium)。以CCNWNX0042-2为代表的大部分天蓝苜蓿根瘤菌属于草木樨中华根瘤菌(Sinorhizobium meliloti),其余菌株在分群上表现出了较为明显的地域特征。     

合欢、金合欢和银合欢根瘤菌系统发育研究方法比较

利用位于染色体不同位点的多个基因序列进行分析是原核生物分类与系统发育研究的一个热点.本文采用atpD和glnⅡ两个持家基因的部分序列对9株分离自我国合欢、金合欢和银合欢的根瘤菌进行系统发育研究,并与以16S rDNA基因序列构建的系统发育树进行比较.结果表明三者在属水平上基本一致,CCBAU43060、CCBAU61139位于Rhizobium-Agrobacterium系统发育分支内;CCBAU51471、CCBAU35220、CCBAU51276和CCBAU61158属于Mesorhizobium,CCBAU35234、CCBAU61178和CCBAU35085位于Bradyrhizobium系统发育分支;在属内种间个别菌株(CCBAU61158、CCBAU43060、CCBAU61178)的系统发育地位存在差异,表明属内种间存在较广泛的基因交流.因此利用16S rDNA确定属水平的分类地位比较可靠,但利用系统发育的方法研究属内种间亲缘关系应采用多个功能保守的持家基因同时进行分析才能得出比较可靠的结论.     

中国的被孢霉种类

被孢霉属(Mortierella Coemans)是接合菌纲(Zygomycetes)、毛霉目(Mucorales)、被孢霉科(Mortierellaceae)中的一个大属,目前已知约有90种;主要存在于土壤、植物残体、动物粪便等基物中。我国过去对被孢霉的研究不多,在《中国真菌总汇》(1979)中记录了8个种。本研究从全国22个省、市、自治区采集的2000多号样品中,分离到约220个被孢霉菌株。本研究主要采用Gams (1970, 1977)的分类系统进行分类鉴定,并对该系统进行了修改。在属下分3个亚属(Micromucor, Mortierella和Gamsiella), 8个组(Actinomortierella, Alpina,Hygrophila, Mortierella, Schmuckii, Simplex, Spinosa和Stylospora),单囊霉(Haplosporangium)被承认为独立的一个属。本研究鉴定出22个种和3个变种,包括一个新种(武夷山被孢霉Mortierella wuyishanensis sp. nov.)和一个新变种(极细无色被孢霉Mortierella hyalina(Harz) W. Gams var. subtilissima var. nov.), 14个中国新纪录。这14个新纪录为:产芽胞被孢霉(Mortierella. gemmifera M. Ellis)、园圃被孢霉(M. horticola Linnem.)、矮小被孢霉(M. humilis Linnem.)、无色被孢霉(M. hyalina(Harz) W. Gams)、印度被孢霉(M. indica B.S. Mehrotra)、英杜被孢霉(M. indohii C.Y. Chien),詹金氏被孢霉(M. jenkinii (A.L. Sm.) Naumov)、可疑极小被孢霉(M. minutissima Tiegh. var. dubia Linnem.)、易变被孢霉(M. mutabilis Linnem.)、微孢被孢霉(M. parvispora Linnem.)、角胞拉曼被孢霉(M. ramanniana(Moller) Linnem. var. angulispora (Naumov) Linnem.)、网孢被孢霉(M. reticulata Tiegh.& G. Le Monn.)、多疣被孢霉(M. verrucosa Linnem.)、轮枝被孢霉(M. verticillata Linnem.)。文中讨论和评价了一些分类性状,还列出分亚属、分组、分种和变种的检索表.每个分类单元都有描述和讨论以及线条图、并列出分布地区。     

后木虱属二新种(同翅目:木虱科)

后木虱属(Metapsylla)是木虱亚科(Psyllinae)里的一个小属,桑山茂(Sh.Kuwayama)1907年成立此属时是以日本的M.nigra为模式种的;同时还发表了另一个新种M.ma-rginata,则为我国台湾高雄的,只有一雌。宫武赖夫(Y.Miyatake)1963年研究了保存在北海道大学昆虫研究所的M.marginata模式标本,认为应将marginata移到Euphalerus属去,同时重新描述了M.nigra Kuwayama;并记述了日本的第二个新种M.uei Miyatake。我国此属的记载除上述误置者外,余凤麟(Yu Feng-ling)1956 年记述了痘点木虱 M.granulasa Yu,标本14个雌雄是马骏超1940—Ⅳ-20采自福建崇安县武夷山的。他同时指出日本的 M.robinae Shinji(1938)应隶他属;后来宫武(1963)也认为该种需从Metapsylla属中去掉,并判断应移至Arytaina属中。七十余年来此属共发表过五个种,而实际只剩下日本2种和我国一个种。     

淡水球状绿藻一新种——汾河麦可藻

麦可属(Mychonastes)是一种分布广泛的超微型球状绿藻,是微藻能源生产、水质净化方面的潜力藻种。该研究对采自山西省太原市汾河公园的水样分离得到的一株球状绿藻(FHGY-19)进行藻株形态显微观察,并进行18S rDNA系统发育与ITS2二级结构分析鉴定,以明确FHGY-19的分类位置以及生态利用潜力。结果表明:(1)FHGY-19藻株为单细胞,球形,直径1.5~2.5μm,无黏质被膜,杯状叶绿体1个,周生,不具蛋白核,以2~4个似亲孢子进行繁殖,常见2个似亲孢子包被于母细胞壁内。(2)18S rDNA系统发育分析表明,藻株FHGY-19位于麦可属分支内部,与麦可属内的其他成员共同形成环藻目内一独立的单系分支,且FHGY-19与麦可属分支中的Mychonastes sp.5C3和Mychonastes sp.2C1亲缘关系较近,表明藻株FHGY-19为麦可属一成员。(3)ITS2 rDNA系统发育分析表明,FHGY-19与同是单细胞类型的Mychonastes frigidus、同球麦可藻(M.homosphaera)、M.pusillus、M.rotundus和M.ovahimbae聚为一支,且FHGY-19与5个藻种在ITS2二级结构上的总CBCs(碱基补偿替换)和保守区域HelixⅢ上的CBCs数分别为16个/7个、13个/6个、12个/4个、11个/4个和14个/5个,但藻株FHGY-19与系统发育树另一支的6个物种在ITS2序列长度、二级结构中总CBCs及HelixⅢ上的CBCs数均不同,表明FHGY-19的ITS2二级结构不同于已描述的11个种,为麦可属一新种。研究鉴定确认FHGY-19藻株为麦可属一新种,命名为汾河麦可藻(Mychonastes fenhensis sp.nov.)。FHGY-19藻株保存于太原师范学院淡水藻种库。     

Phylogeny and character evolution in Medicago (Leguminosae): Evidence from analyses of plastid trnK/matK and nuclear GA3ox1 sequences

? Premise of the study: The genus Medicago, with about 87 species, includes the model legume species M. truncatula, and a number of important forage species such as M. sativa (alfalfa), M. scutellata (snail medic), and M. lupulina (black medic). Relationships within the genus are not yet sufficiently resolved, contributing to difficulty in understanding the evolution of a number of distinguishing characteristics such as aneuploidy and polyploidy, life history, structure of cotyledons, and number of seeds per fruit. ? Methods: Phylogenetic relationships of 70-73 species of Medicago and its sister genus Trigonella (including Melilotus) were reconstructed from nucleotide sequences of the plastid trnK/matK region and the nuclear-encoded GA3ox1 gene (gibberellin 3-β-hydroxylase) using maximum parsimony and Bayesian inference methods. ? Key results: Our results support certain currently recognized taxonomic groups, e.g., sect. Medicago (with M. sativa) and sect. Buceras. However, other strongly supported clades-the "reduced subsection Leptospireae clade" that includes M. lupulina, the "polymorpha clade" that includes M. murex and M. polymorpha and the "subsection Pachyspireae clade" that includes M. truncatula-each of which includes species presently in different subsections of sect. Spirocarpos, contradict the current classification. ? Conclusions: These results support the hypothesis that some characters considered important in existing taxonomies, for example, single-seeded fruits that have arisen more than once in both Medicago and Trigonella, are indeed homoplastic. Others, such as the 2n = 14 chromosome number, have also arisen independently within the genus. In addition, we demonstrate support for the utility of GA3ox1 sequences for phylogenetic analysis among and within closely related genera of legumes.     

Simple sequence repeat DNA markers in alfalfa and perennial and annual Medicago species.

Simple sequence repeat (SSR) or microsatellite DNA markers have been shown to function well in plant and mammalian species for genetic map construction and genotype identification. The objectives of the work reported here were to search GenBank for the presence of SSR-containing sequences from the genus Medicago, to assess the presence and frequency of SSR DNA in the alfalfa (Medicago sativa (L.) L. &L.) genome, and to examine the function of selected markers in a spectrum of perennial and annual Medicago species. The screening of an alfalfa genomic DNA library and sequencing of clones putatively containing SSRs indicated approximately 19 000 (AT)n + (CT)n + (CA)n + (ATT)n SSRs in the tetraploid genome. Inheritance was consistent with Mendelian expectations at four selected SSR loci with different core motifs. Additionally, genotypes of a range of Medicago species, including 10 perennial subspecies of the M. sativa complex and other perennial and annual Medicago species, were analyzed at each of the loci to ascertain the presence, number, and size of SSR alleles at each locus in each genotype. These studies indicate that SSR markers can function in alfalfa for the construction of genetic maps and will also be useful in a range of Medicago species for purposes of assessing genetic relatedness and taxonomic relationships, and for genotype identification.     

Genome size and base composition in Medicago sativa and M. truncatula species.

The genome size (1C value) and base composition of 14 ecotypes of two species of tetraploid and diploid Medicago have been assessed by flow cytometry. These parameters vary both between and within species. The diploid annual Medicago truncatula Gaertn. had the smallest genome of the group studied (which also covered M. sativa L. subsp. sativa, M. sativa L. subsp. caerulea (Less. ex Ledeb.) Schmalh., M. sativa L. subsp. quasifalcata Sinsk., M. sativa L. subsp. x varia (Martyn) Arcangeli; however, its ecotypes revealed substantial intraspecific variation. The smallest M. truncatula genome observed was ecotype 108-1 with 1C = 0.49 pg and 38.1% GC and the largest was Jemalong with 1C = 0.57 pg and 38.6% GC. The degree of polysomaty in these Medicago was low, although in some tissues the frequency of cells with 4C nuclei reached 50%.     

Effect of field inoculation with Sinorhizobium meliloti L33 on the composition of bacterial communities in rhizospheres of a target plant (Medicago sativa) and a non-target plant (Chenopodium album)-linking of 16S rRNA gene-based single-strand conformation polymorphism community profiles to the diversity of cultivated bacteria

Fourteen weeks after field release of luciferase gene-tagged Sinorhizobium meliloti L33 in field plots seeded with Medicago sativa, we found that the inoculant also occurred in bulk soil from noninoculated control plots. In rhizospheres of M. sativa plants, S. meliloti L33 could be detected in noninoculated plots 12 weeks after inoculation, indicating that growth in the rhizosphere preceded spread into bulk soil. To determine whether inoculation affected bacterial diversity, 1,119 bacteria were isolated from the rhizospheres of M. sativa and Chenopodium album, which was the dominant weed in the field plots. Amplified ribosomal DNA restriction analysis (ARDRA) revealed plant-specific fragment size frequencies. Dominant ARDRA groups were identified by 16S rRNA gene nucleotide sequencing. Database comparisons indicated that the rhizospheres contained members of the Proteobacteria (alpha, beta, and gamma subgroups), members of the Cytophaga-Flavobacterium group, and gram-positive bacteria with high G+C DNA contents. The levels of many groups were affected by the plant species and, in the case of M. sativa, by inoculation. The most abundant isolates were related to Variovorax sp., Arthrobacter ramosus, and Acinetobacter calcoaceticus. In the rhizosphere of M. sativa, inoculation reduced the numbers of cells of A. calcoaceticus and members of the genus Pseudomonas and increased the number of rhizobia. Cultivation-independent PCR-single-strand conformation polymorphism (SSCP) profiles of a 16S rRNA gene region confirmed the existence of plant-specific rhizosphere communities and the effect of the inoculant. All dominant ARDRA groups except Variovorax species could be detected. On the other hand, the SSCP profiles revealed products which could not be assigned to the dominant cultured isolates, indicating that the bacterial diversity was greater than the diversity suggested by cultivation.     

Development of S-SAP markers based on an LTR-like sequence from Medicago sativa L.

The Sequence-Specific Amplification Polymorphism (S-SAP) method, recently derived from the Amplified Fragment Length Polymorphism (AFLP) technique, produces amplified fragments containing a retrotransposon LTR sequence at one end and a host restriction site at the other. We report the application of this procedure to the LTR of the Tms1 element from Medicago sativa L. Genomic dot-blot analysis indicated that Tms1 LTRs represent about 0.056% of the M. sativa genome, corresponding to 16 x 10(3) copies per haploid genome. An average of 66 markers were amplified for each primer combination. Overall 49 polymorphic fragments were reliably scored and mapped in a F(1) population obtained by crossing diploid M. falcata with M. coerulea. The utility of the LTR S-SAP markers was higher than that of AFLP or SAMPL (Selective Amplification of Microsatellite Polymorphic Loci) markers. The efficiency index of the LTR S-SAP assay was 28.3, whereas the corresponding values for AFLP and SAMPL markers were 21.1 and 16.7, respectively. The marker index for S-SAP was 13.1, compared to 8.8 for AFLP and 9.5 for SAMPL. Application of the Tms1 LTR-based S-SAP to double-stranded cDNA resulted in a complex banding pattern, demonstrating the presence of Tms1 LTRs within exons. As the technique was successfully applied to other species of the genus Medicago, it should prove suitable for studying genetic diversity within, and relatedness between, alfalfa species.     

Physical mapping of rRNA genes in Medicago sativa and M. glomerata by fluorescent in situ hybridization

Fluorescent in situ hybridization (FISH) was applied to diploid and tetraploid subspecies of alfalfa (Medicago sativa L.) to investigate the distribution of rRNA genes and to utilize the sites of 18S-5.8S-25S rDNA and 5S rDNA sequences as markers for studying the genome evolution within the species. Medicago glomerata Balb., the species considered to be the ancestor of alfalfa, was included in this study in order to obtain more information on the phylogenetics of alfalfa. Simultaneous in situ hybridization was performed with the probes pTa71 and pXVI labeled with digoxigenin and biotin, respectively. In the diploid taxa, M. glomerata, M. sativa ssp. coerulea Schmalh and ssp. falcata Arcangeli, the 18S-5.8S-25S rDNA sequences were mapped to two sites corresponding to the secondary constrictions of the nucleolar chromosome pair, while 5S rDNA appeared to be distributed in two pairs of sites. Chromosomes carrying 5S loci could be distinguished on the basis of their morphological characteristics. The number of rDNA sites detected in the tetraploid M. sativa ssp. falcata and ssp. sativa (L.) L. & L. were twice the number found in the respective diploid ssp. falcata and ssp. coerulea. The results of this study show that the distribution of ribosomal genes was maintained during the evolutionary steps from the primitive diploid to the cultivated alfalfa. Modifications of the number of rRNA loci were not observed. The importance of in situ hybridization for improving karyotype analysis in M. sativa L. is discussed.     

Characterization of Rhizobia from Ineffective Alfalfa Nodules: Ability to Nodulate Bean Plants [Phaseolus vulgaris (L.) Savi.

This study was initiated to characterize Rhizobium isolates obtained from root nodules of ineffectively nodulated, field-grown alfalfa (Medicago sativa L.) plants. The purpose was to determine if these isolates possessed characteristics which would explain either their ineffectiveness in N(2) fixation or their apparent ability to tolerate the moderately acid soil conditions from which they originated. Isolates were characterized by analysis of growth rate, 39 degrees C tolerance, acid production on conventional media, and symbiotic performance. All isolates were ineffective in N(2) fixation on alfalfa, and they contained one or more anomalous characteristics. These included either slow growth rate, lack of 39 degrees C tolerance, or lack of acid production on conventional media. Infectiveness tests on a broad range of legumes revealed that the isolates formed root nodules on M. sativa, Medicago lupulina L., and Phaseolus vulgaris (L.) Savi. (common bean). These results provide evidence that, in some situations, ineffective nodulation of M. sativa in the field may be due to the presence of promiscuous, native Rhizobium species.     

AFLP-based assessment of genetic diversity among nine alfalfa germplasms using bulk DNA templates.

Improving commercial utilization of perennial Medicago collections requires developing approaches that can rapidly and accurately characterize genetic diversity among large numbers of populations. This study evaluated the potential of using amplified fragment length polymorphism (AFLP) DNA markers, in combination with DNA bulking over multiple genotypes, as a strategy for high-throughput characterization of genetic distances (D) among alfalfa (Medicago sativa L.) accessions. Bulked DNA templates from 30 genotypes within each of nine well-recognized germplasms (African, Chilean, Flemish, Indian, Ladak, Medicago sativa subsp. falcata, Medicago sativa subsp. varia, Peruvian, and Turkistan) were evaluated using 34 primer combinations. A total of 3754 fragments were identified, of which 1541 were polymorphic. The number of polymorphic fragments detected per primer combination ranged from 20 to 85. Pairwise D estimates among the nine germplasms ranged from 0.52 to 1.46 with M. sativa subsp. falcata being the most genetically dissimilar. Unweighted pair-group method arithmetic average (UPGMA) analysis of the marker data produced two main clusters, (i) M. sativa subsp. sativa and M. sativa subsp. varia, and (ii) M. sativa subsp. falcata. Cluster-analysis results and D estimates among the Chilean, Peruvian, Flemish, and M. sativa subsp. varia germplasms supported the hypothesis that Peruvian was more similar to original Spanish introductions into Central and South America than Chilean. Hierarchical arrangement of the nine germplasms was supported by their respective geographic, subspecific, and intersubspecific hybrid origins. Subsets of as few as seven highly informative primer pairs were identified that produced comparable D estimates and similar heirarchical arrangements compared with the complete dataset. The results indicate that use of primer-pair subsets for AFLP analysis of bulk DNA templates could serve as a high-throughput system for accurately characterizing genetic diversity among large numbers of alfalfa populations.     

6个紫花苜蓿材料的核型及其亲缘关系分析

运用染色体压片法对陇东野生紫花苜蓿(Medicago sativa L.)等6个紫花苜蓿材料进行核型分析,并通过核型似近系数对供试材料进行聚类分析.结果显示:(1)全部供试材料皆为2n=32的四倍体,核型进化由高到低依次为:[陇东野生紫花苜蓿、新疆大叶紫花苜蓿(M.sativa,Xinjiangdaye)]2B＞[皇后2000紫花苜蓿(M.sativa,Regina 2000)、陇东紫花苜蓿(M.sativa,Longdong)、Pick 8925紫花苜蓿(M.sativa,Pick 8925)]1B＞[肇东紫花苜蓿(M.sativa,Zhaodong)]1A.(2)供试材料间的核型似近系数(λ)均在0.900 0以上,并在进化距离(De)0.073 8处分为两类,陇东野生紫花苜蓿和新疆大叶紫花苜蓿归为一类,其它供试材料归为另一类.研究表明,陇东野生紫花苜蓿的染色体较不对称,与其它供试材料相比,亲缘关系远,系统演化快.     

Comparison of nucleotide diversity and symbiotic properties of Rhizobium meliloti populations from annual Medicago species

Abstract: Forty-three isolates of Rhizobium meliloti were trapped from soil with five annual species of Medicago (M. polymorpha, M. truncatula, M. rigidula, M. orbicularis and M. minima ) and one perennial species of Medicago (M. sativa) . The annual species were growing naturally near the soil sampling site, and the commonly studied perennial species was used for comparison. Each R. meliloti was characterized by PCR-RFLP methods applied to two DNA regions nested between 16S rRNA and 23S rRNA genes and between nif D and nif K genes. They fell into two highly divergent groups (groups I and II), separated at a genetic distance of 0.024 by rDNA-amplified pattern analysis (profiles R1 and R2) and at 0.029 by nif -amplified pattern analysis (profiles N1-N2 and N3). These two groups were consistent with some cross-nodulation and -fixation results: rhizobia with the R1 genetic background elicited rudimentary nodules and could not fix nitrogen on M. polymorpha , while they were able to nodulate the five other species of Medicago . In contrast, rhizobia with an R2 profile were highly effective on M. polymorpha and poorly nodulated M. rigidula species, but were able to nodulate efficiently the other species. The striking phenotypic traits on M. polymorpha were also shared by reference strains: strains genetically closed to R2 type triggered typical and efficient nodules on M. polymorpha while those close to R1 type elicited rudimentary and non-efficient ones. Our results suggest that the presence of R. meliloti with R2 genetic backgrounds could be favoured by the distribution of M. polymorpha species.