中国啤酒大麦品种RAPD标记的遗传多样性分析

采用RAPD技术对中国38个啤酒大麦品种的遗传资源进行了聚类分析。结果表明：从筛选出的28个有多态性的随机引物中,共扩增出153条谱带,其中91条谱带具有多态性,占59．4％。每个引物可扩增出1～8条多态性谱带,平均3．3条。聚类分析表明,在遗传距离GD值0．27水平上38个啤酒大麦品种可聚成两大类,下分5个亚类。品种间遗传距离GD变幅为0．00952～0．37846。RAPD标记揭示出这38个啤酒大麦品种遗传变异较小,遗传基础比较狭窄。     

Genetic Relationships within Tripsacum as Detected by RAPD Variation

Genetic diversity within species of Tripsacum was surveyed basedon randomly amplified polymorphic DNA (RAPD) variation, as detectedwith the polymerase chain reaction (PCR). Thirteen of the 16Tripsacum species, including both temperate and tropical species,were included in this study using 56 decamer oligonucleotideprimers. All of the 56 primers generated repeatable RAPD profilesand 53 of them detected polymorphic bands among the Tripsacumspecies. These 53 primers generated 350 repeatable bands rangingin size from 150–1600 bp, each primer generating an averageof seven scoreable bands. Cluster analysis of polymorphic RAPDsindicated four major clusters. Cluster 1 consists of North AmericanTripsacum species, cluster 2 consists of South American Tripsacumspecies, cluster 3 includes T. zopilotense and T. latifoliumfrom Mexico, and cluster 4 consists of Mesoamerican Tripsacumspecies. Cluster analysis does not reveal the division of twotaxonomic sections (Fasciculata and Tripsacum). Copyright 1999Annals of Botany Company Tripsacum, RAPD, genetic relationships, genetic diversity.     

6个中外猪品种的RAPD分析

应用RAPD技术分析了梅山猪、淮南猪、通城猪、八眉猪、合作猪、大白猪的遗传变异。用276个随机引物对6个猪种混合DNA扩增筛选,24个引物产生多态性。此24个引物经重新优化反应条件体系后,对276个个体DNA进行扩增。结果6个猪种的遗传多样性指数分别为0.178672、0.17781、0.15995、0.14549、0.16949和0.14157,群体内平均遗传多样性指数为0.16216,总群体遗传多样性指数为0.2534。基于Rogers公式计算的遗传距离采用NJ和UPGMA法进行聚类分析,结果6个猪种的亲缘关系与它们的地理分布基本一致。 Abstract:The genetic variation of Meishan,Huainan,Tongcheng,Bamei,Hezuo and Largewhite pigs were analyzed by RAPD markers.Twenty-four single polymorphic primers were selected out of 276 primers by amplifying six pool DNA.The index of Shannon were 0.178672,0.17781,0.15995,0.14549,0.16949,0.14159 respectively;Hpop was 0.16216,Hsp was 0.2534.The phylogenetic tree was constructed using NJ and UPGMA.The results indicated that the phenylogenetic relationship of the six pig breeds was consistent with their distribution.     

Climatic factors influencing spore production in Alternaria brassicae and Alternaria brassicicola

Sporulation in A. brassicae and A. brassicicola on naturally-infected leaf discs of oilseed rape and cabbage required humidities equal to or higher than 91.5% and 87% r.h. respectively. The optimum temperatures for sporulation were 18–24°C for A. brassicae and 20–30°C for A. brassicicola at which temperatures both fungi produced spores in 12–14 h. Above 24°C sporulation in A. brassicae was inhibited. At sub-optimal temperatures sporulation times for A. brassicicola were significantly longer than for A. brassicae with the differences increasing with decrease in temperature. Interrupting a 16-h wet period at 20°C with a period of 2 h at 70% or 80% r.h. did not affect sporulation in either fungus but a dry interruption of 3–4 h inhibited sporulation in both. Exposure of both fungi to alternating wet (18 h at 100% r.h., 20°C) and dry periods (6 or 30 h at 5565% r.h., 20°C) did not affect the concentration of spores produced in each wet period. Sporulation times were not affected by either the host type of the age of the host tissue. White light (136 W/m²) inhibited sporulation in A. brassicae with the degree of inhibition increasing with increasing light intensity. The effect of light on sporulation in A. brassicicola was not tested.     

Survival and perpetuation of Alternaria brassicae causing Alternaria blight of oilseed crucifers

Modes of survival and perpetuation of Alternaria brassicae attacking rapeseed and mustard were examined. The pathogen remained viable in diseased plant debris and seeds of infected plants which served as primary sources of inoculum. The pathogen was internally seed-borne and infected seeds resulted in damping-off of seedlings. Air-borne conidia produced on aerial parts of the infected plants were responsible for secondary spread of the disease.     

Ultrastructural Changes in Brassica Leaves Caused by Alternaria brassicae and Destruxin B

Alternaria blight in Brassica spp (caused by Alternaria brassicae) is characterized by dark brown to blackish necrotic lesions surrounded by chlorotic areas on the leaves. Similar chlorotic lesions were mimicked by a chlorotic toxin (destruxin B) purified from the culture filtrate. Ultrastructural studies were performed to study and compare the changes caused by A. brassicae inoculation in vivo and its host selective toxin under in vitro conditions. Electron microscopy of healthy, chlorotic and necrotic portions of B. campestris leaves naturally infected with A. brassicae revealed considerable differences at ultrastructural level. The necrotic lesions showed plasmolysis with total disruption of cell organelles. The chlorotic lesions had normal plasma membrane but swollen mitochondria with reduced number of cristae and vesiculation of the envelope. Chloroplasts showed degeneration of granal fretwork with an increase in the number of plastoglobuli. Chlorotic lesions due to foliar application of destruxin B induced indentical canges in leaves.     

用RAPD技术研究我国苹果果实轮纹病菌的遗传变异(英文)

为了从分子水平上明确近年来我国苹果生产上发生的A、B、D和F 4种不同症状型苹果果实轮纹病的遗传分化现象及其不同致病菌的遗传变异程度。本研究对来自我国三大苹果产区的26个相关病原菌株的DNA指纹图谱进行了分析(Table 1)。结果表明 (1) 上述4种不同症状型轮纹病原菌株在基因组DNA水平上具有明显的遗传差异(Fig.1&Table 2),并在所分析的11个多态性引物中有4个引物的扩增图谱(Table 4)均可使上述4种不同致病型菌株相互区别(Table 5)。统计分析结果显示,这4种不同致病菌相互之间的遗传距离为0.1314~0.2541(Table 6);(2) 相同症状型的不同来源致病菌之间具有较高的遗传相似性 (88.4~100%)(Table 3),由此说明苹果果实轮纹病菌的遗传分化与地理环境条件差异无明显的相关性。 以上研究结果对于进一步阐明苹果果实轮纹病害的遗传演化规律及该病害监测与防治技术的改进具有重要的理论与实际意义。     

香果树组织培养过程中遗传变异的RAPD分析

用RAPD分子标记方法,从DNA水平上分析野生型香果树以及通过器官发生途径和体细胞胚胎发生途径得到的香果树再生植株以及体细胞胚胎发生过程中不同继代次数的培养物之间的遗传变异。筛选了100个随机引物,其中有75条随机引物能够扩增出条带,从中选取11个引物进行PCR扩增的结果显示：香果树体细胞胚胎无性系中有RAPD多态性位点,在胚性愈伤组织中也检测到少数RAPD变异位点。表明RAPD分子标记方法可以鉴定香果树组织培养过程中的遗传变异。     

Genetic Variation in Hippophae rhamnoides ssp. sinensis (Elaeagnaceae) Revealed by RAPD Markers

Hippophae rhamnoides ssp. sinensis is endemic to China, and it is a dioecious, outcrossing plant. Although many studies have been undertaken mainly on its agricultural, nutritional, medical, and ornamental value, little is known about its population genetics. This study uses random amplified polymorphic DNA to investigate the genetic diversity and population genetic structure of 13 natural populations of the subspecies sinensis. Fifteen primers amplified 107 reproducible bands, with 95 (88.79%) being polymorphic. The gene diversity within population was 0.168, considerably lower than that of tree species and most perennial, outcrossing species, but higher than that of annual or short-lived, selfing species. The Gst value showed that 18.3% of the total genetic variation resided among populations, a little lower than that of outcrossing species. The present results are quite similar to those previously reported in another subspecies, H ssp. . rhamnoides rhamnoides. The low genetic differentiation among populations in ssp. sinensis may be attributed to the long-distance dispersal of seeds facilitated by birds, in addition to its characteristics of outcrossing, wind pollination, and widespread distribution. No association between genetic distance and geographical distribution was found. The population relationships revealed by the UPGMA dendrogram parallel this result, in that genetic distance did not increase with geographic separation. This pattern of population differentiation may imply the adaptation of ssp. s populations to the local environment, given that its habitats vary greatly across its distribution.     

Detection of Somaclonal Variations in Tissue Culture-Derived Date Palm Plants Using Isoenzyme Analysis and RAPD Fingerprints

Isoenzyme analysis and activities of peroxidase (PER), polyphenol oxidase (POD) and glutamate oxaloacetate (GOT) and randomly amplified polymorphic DNA (RAPD) fingerprints were used to analyze somaclonal variations in tissue culture-derived date palm plants. The frequency of somaclonal variations was found to be age dependent. Similar isoenzyme patterns for PER and GOT were detected in all analyzed plants. However, variations in activities of the three enzymes studied and in POD isoenzymes were detected. RAPD analysis showed genetic variations in approximately 4 % of the analyzed plants (70 regenerants). The genetic variations were only detected in 6- and 12-months-old cultures. It was observed also that all morphologically abnormal shoots showed genetic variations at the molecular level.     

Survival of Alternaria brassicae and Alternaria brassicicola on crop debris of oilseed rape and cabbage

Alternaria brassicae and A. brassicicola lesions present on infected leaves of oilseed rape and cabbage placed outdoors on soil produced viable spores for as long as leaf tissues remained intact. For oilseed rape this was up to 8 wk and for cabbage up to 12 wk. On leaves exposed in November and January spore concentrations decreased with time but on leaves exposed between April and June spore concentrations increased up to 9-fold in the first 4–6 wk and then declined. On stem sections of seed plants of oilseed rape and cabbage similarly placed on the soil, the fungi produced viable spores for up to 23 wk with spore concentrations increasing up to 11-fold in the first 6–8 wk after harvest. These results indicate that infected debris of brassica crops remaining on the ground after harvest may provide a source of dark leaf spot infection which may be implicated in the spread of the disease within and between crops.     

云南灯盏花遗传变异的RAPD分析

运用RAPD (randomamplifiedpolymorphicDNA)技术对云南灯盏花 (Erigeronbrevis capus) 6个居群进行分析 ,研究其遗传变异及遗传结构 ,用外类群紫菀对比分析其亲缘关系。随机挑选 16个引物在 6个居群中共产生 2 33条DNA片段 ,其中 192条带具有遗传多态性 ,约占 82 4 0 %。多态位点百分比PPB、等位基因数A、有效等位基因数Ae、Nei’s基因多样性指数H和Shannon多样性指数I在物种水平分别为 82 4 0 %、 1 82 4 0、 1 30 0 5、0 1896、 0 30 2 1;居群的平均值分别为 5 4 2 3%、 1 5 40 1、 1 2 6 91、 0 16 0 7、 0 2 4 6 0。灯盏花的遗传多样性较丰富 ,基因分化系数Gst值为 0 346 0 ,有 6 5 4 0 %的变异来自居群内 ,6个居群的遗传一致度较高 ,在 0 90 37～ 0 972 3之间 ,平均为 0 94 10。利用UPGMA对 6居群聚类分析 ,结果表明 :丘北居群 (YB)和文山居群 (YX)亲缘关系最近 ,小哨居群 (Y )和新平居群 (YA)次之 ,丘北居群 (YB)和腾冲居群 (ZA)亲缘关系最远。遗传关系与各居群地理分布区间的距离大致成正相关     

基于RAPD标记的丽江云杉遗传多样性及谱系地理

由于青藏高于独特的地理环境,第四纪冰期反复的退缩和扩展对目前该地区生物物种的地理分布及居群结构产生重要的影响。本文对这一地区特有分布物种丽江云杉(Picea likiangnsis)3个变种丽江云杉变种(var. likiangensis)、川西云杉变种(var. rubescens)和林芝云杉变种(var. linzhiensis)共11个种群228个个体进行RAPD分析,研究其遗传多样性及谱系地理。结果表明,该种具有较高的总体遗传多样性,多态位点达85.42%;然而在群体水平上却保持了相对低的多态性,种群间差异不显著,种群平均多态条带百分率为62.31%,Nei’s平均遗传多样性指数(H_E)为0.250,Shannon’s多样性指数(H_pop)从0.267到0.421 1。Nei’s基因多样性(G_ST=0.256)和AMOVE分析(Phi_st=0.236)表明,群体间有较高的遗传分化,群体间基因流有限(N_m=1.453 2),远远低于已报道的其它松科植物。UPGMA聚类分析表明3个形态上分离的变种没有单个聚成一枝,形成单系群。本研究认为,在第四纪冰期丽江云杉这一种在南部可能存在至少3个不同的避难所,北边和西边的居群应该是南方避难所里的居群经过不同的回迁路线而产生的,有可能是由于种内亚种间的反复杂交造成了目前的种群分布模式。     

Detection of Low Genetic Variation in a Critically Endangered Chinese Pine, Pinus squamata, Using RAPD and ISSR Markers

With only 32 individuals in the northeastern corner of Yunnan Province, China, Pinus squamata is one of the most endangered conifers in the world. Using two classes of molecular markers, RAPD and ISSR, its very low genetic variation was revealed. Shannon's index of phenotypic diversity (I) was 0.030, the mean effective number of alleles per locus (A_e) was 1.032, the percentage of polymorphic loci (P) was 6.45, and the expected heterozygosity (H_e) was 0.019 at the species level based on RAPD markers. The results of ISSR were consistent with those detected by RAPD but somewhat higher (I = 0.048, A_e = 1.042, P = 12.3, H_e = 0.029). The genetic variation of the subpopulation on the southwest-facing slope was much higher than that of the subpopulation on the northeast-facing slope, which may be attributed to the more diverse environment on the southwest-facing slope. The genetic differentiation between the two subpopulations was very low. The between-subpopulation variabilities, Φ_ST, calculated from RAPD and ISSR data were 0.011 and 0.024. Because of the lack of fossil records and geological historical data, it was difficult to explain the extremely low genetic diversity of the species. We postulate that this ancient pine might have experienced strong bottlenecks during its long evolutionary history, which caused the loss of genetic variation. Genetic drift and inbreeding in post-bottlenecked small populations may be the major forces that contribute to low genetic diversity. Human activities such as logging may have accelerated the loss of genetic diversity in P. squamata.     

应用RAPD指纹探讨黄麻属种间遗传多样性及其亲缘关系

采用RAPD标记构建了黄麻属(Corchorus)植物10个种27份材料的指纹图谱,从119个随机引物中筛选出清晰且多态性高的25个引物,共扩增出329条：DNA片段,分子量在0．3～3．0kb之间,其中294条谱带具有遗传多态性,多态比率(PPB)为89．36％,平均每个引物扩增出13．16条带。用Biol D^ 数据分析软件,计算Nei氏相似性系数,建立了UPCMA聚类图。结果表明：(1)供试黄麻属15份野生种和12份栽培种具有丰富的遗传多样性,遗传相似系数在0．49～0．98;(2)在聚类分析中,当L1聚值水平D=0．785时,可将两个栽培种及其近缘野生种(C．capsulris和C．olititorius)与原始黄麻野生种划分为3个不同类群。反映出栽培种及其近缘野生种与原始野生种间有明显的遗传差异;(3)当L2取值水平D=0．850时,可将供试27份材料划分为10个以物种为单元的亚类群或个类,①即假黄麻C．aestuans(3份),②三齿种C．tridens,③梭状种C．fascicularis,④假长果种C．psendo-olitorius,⑤假圆果种C．pseudo-capsularis,⑥三室种C．tilacularis,⑦甜麻(新种未定名),⑧圆果种C．capsularis(9份),⑨长果种C．olitorius(7份),⑩荨麻叶种C．uriticifolius,结果与10个种的经典分类相吻合,揭示了种间的遗传差异性。其中圆果种C．cap—sularis与长果种C．olitorius两个种亲缘关系较近,与荨麻叶种C．uriticifolius种间亲缘关系较远;(4)非洲荨麻叶种C．uriticifolius和中国的甜麻(新种)、假黄麻C．aestuans3个种与其他种间的遗传差异较大,处在分子聚类树较基础的地位,为较原始的黄麻野生种;(5)非洲的三室种21C为三室种一个生态型亚种;采集于中国的黄麻野生种河南南阳的粘粘菜、福建漳浦的麻菜,云南开远的猪菜为3个不同生态类型的假黄麻;海南野生圆果为圆果黄麻栽培种的近缘野生种;漳浦野生长果、河南野生长果、马里野生长果为长果栽培种的近缘野生种,种内不同材料间遗传相似性较高,亲缘关系较密切;(6)在两个栽培种中,品种基因型相似性系数圆果栽培种高于长果栽培种。     

Induced resistance against Alternaria brassicae blight of mustard through plant extracts

An experiment was conducted to study the effect of plant extracts on soluble sugar, soluble phenol and defence-related enzymes response against Alternaria blight in mustard crop. The efficacy of six selected plant extracts (5 and 10%) used as foliar sprays at 60 and 70 days after sowing and mustard leaves was used for investigation. The results indicate that soluble phenol and sugar content in mustard leaves significantly increases in response to spraying of Azadirachta indica seed kernel, Calotropis procera and A. indica leaf extracts. The soluble protein, viz. peroxidase, polyphenol oxidase and phenylalanine ammonia lyase content, was higher in mustard leaves sprayed with C. procera leaves extract, A. indica seed kernel and Allium sativum bulb extract.     

Chitinase-mediated inhibitory activity of Brassica transgenic on growth of Alternaria brassicae

Chitinase, capable of degrading the cell walls of invading phytopathogenic fungi, plays an important role in plant defense response, particularly when this enzyme is overexpressed through genetic engineering. In the present study, Brassica plant (Brassica juncea L.) was transformed with chitinase gene tagged with an overexpressing promoter 35 S CaMV. The putative transgenics were assayed for their inhibitory activity against Alternaria brassicae, the inducer of Alternaria leaf spot of Brassica both in vitro and under polyhouse conditions. In in vitro fungal growth inhibition assays, chitinase inhibited the fungal colony size by 12-56% over the non-trangenic control. The bioassay under artificial epiphytotic conditions revealed the delay in the onset of disease as well as reduced lesion number and size in 35S-chitinase Brassica as compared to the untransformed control plants.     

Detection of Intra-Clonal Genetic Variability in Vegetatively Propagated Tea Using RAPD Markers

The role of random amplified polymorphic DNA (RAPD) markers in detecting intra-clonal genetic variability in vegetatively propagated UPASI-9 clone of tea (Camellia sinensis) was studied. Twenty five decamer primers were used, of which three did not amplify, three gave single bands and the rest of nineteen primers generated upto twelve bands (an average of 6.3 bands per primer). Twenty one primers exhibiting amplified products gave monomorphic banding patterns. Only one primer (OPE-17) gave a unique extra band of similar size in four plants.     

Detection of Drug-Drug Interactions by Modeling Interaction Profile Fingerprints

Drug-drug interactions (DDIs) constitute an important problem in postmarketing pharmacovigilance and in the development of new drugs. The effectiveness or toxicity of a medication could be affected by the co-administration of other drugs that share pharmacokinetic or pharmacodynamic pathways. For this reason, a great effort is being made to develop new methodologies to detect and assess DDIs. In this article, we present a novel method based on drug interaction profile fingerprints (IPFs) with successful application to DDI detection. IPFs were generated based on the DrugBank database, which provided 9,454 well-established DDIs as a primary source of interaction data. The model uses IPFs to measure the similarity of pairs of drugs and generates new putative DDIs from the non-intersecting interactions of a pair. We described as part of our analysis the pharmacological and biological effects associated with the putative interactions; for example, the interaction between haloperidol and dicyclomine can cause increased risk of psychosis and tardive dyskinesia. First, we evaluated the method through hold-out validation and then by using four independent test sets that did not overlap with DrugBank. Precision for the test sets ranged from 0.4–0.5 with more than two fold enrichment factor enhancement. In conclusion, we demonstrated the usefulness of the method in pharmacovigilance as a DDI predictor, and created a dataset of potential DDIs, highlighting the etiology or pharmacological effect of the DDI, and providing an exploratory tool to facilitate decision support in DDI detection and patient safety.     

玉米自交系遗传变异的RFLP分析

利用ＲＦＬＰ标记研究了１３个玉米（Ｚｃａ ｎａｙｓ Ｌ．）自交系的遗传变异。从３０对探针／酶组合中筛选出杂交带型清晰、稳定、重复性好的２４对组合,在１３个自交系中获得８５个等位基因杂交片段,平均每个位点为３．３条,平均多态性指数为０．４９９。１３个自交系之间的遗传相似系数为０．５２３－０．８０２,平均为０．６４９。ＵＰＧＭＡ聚类分析表明,供试自交系共分为５个类群,分群结果与其系谱关系基本吻合;表明