瓯江彩鲤线粒体DNA的限制性内切酶分析

用 13种限制性内切酶对瓯江彩鲤的线粒体DNA(mtDNA)进行RFLP分析 ,结果表明 :(1)共产生 18种限制性态型 ,其中 5种酶产生限制性片段长度多态性 (RFLPs) ,归结为 5种基因单倍型。 (2 )瓯江彩鲤mtDNA大小为 16 .6 0± 0 .15kb ,单倍型间的基因多样性指数和群体核苷酸多样性指数分别为 0 .75 17、0 .0 2 86 ,遗传多样性较丰富     

上海地区实验用兔线粒体DNA-RFLP分析

提取3个品种实验用兔肝组织中的线粒体DNA,并用ApaI等21种限制性内切酶进行消化。分析上海地区实验用兔mtDNA的多态性。酶切产物经琼脂糖凝胶电泳检测,共检出8种不同的限制性单倍型,少数个体出现的差异,可能来源于几个位点发生偶然突变,但不同兔群的限制性无规律性。结果表明上海地区实验用兔群体mtDNA的遗传多样性贫乏,遗传结构单一,提示所研究的兔种可能有共同祖先,或人工饲养繁殖出现血缘杂交现象。     

籼稻品种遗传变异性初探

 选用水稻RFLP图谱上的50个DNA探针，分析了25个水稻早、中籼品种的RFLP，其中26个能在研究材料间检测到多态性，共检测到73条多态性带。根据各品种之间的等长片段比值，分析了各品种的相似性程度。结果表明，现代早、中籼矮秆品种及其主要原始亲本的遗传背景比较单一，但广东省育成的品种的遗传背景尚较丰富，各地的稻瘟病抗源材料和抗性品种的遗传变异性则相当丰富。本文还讨论了品种抗性与遗传背景的可能联系以及籼稻品种遗传单一性的原因。     

Effect of fungicide thiram on microbial communities of rice seed surface estimated by culture-dependent and culture-independent (PCR-RFLP) methods

Coating of rice seeds with fungicide Thiram improved the seed germination capability over a long period of time (11 weeks) under low temperature conditions (4 and 8°C), which simulated the sowing of rice seeds in the winter season (the farmer's slack season). To analyze the effect of Thiram on the community structure of microorganisms on the rice seed surface, culture-dependent and culture-independent (PCR-RFLP) methods were applied. PCR-RFLP patterns of 16S rDNA showed that the bacterial communities on the rice seed surface were different between coated and uncoated treatments under 8°C conditions, but that they were very similar under 4°C conditions. PCR-RFLP patterns of 18S rDNA revealed the remarkable effect of Thiram on the fungal community structure under both 4 and 8°C conditions. Although the fungal communities were quite different between coated and uncoated seeds at the beginning of incubation, the fungal communities on the coated seed surface became similar to those of uncoated seeds along with the duration of the incubation period. As the dominance percentage of Fusarium spp. among the isolates increased with the duration of the incubation period for both coated and uncoated seeds, Fusarium was considered to be a responsible for the poor germination of rice seeds that were sown in the winter season.     

氮肥缺失对旱地土壤细菌群落多样性的影响

本文旨在研究氮肥缺失对旱地土壤细菌群落多样性的影响.采用直接提取土壤微生物总DNA的方法,对不施肥(CK)、适量施肥(F1)、和缺氮施肥(F2)3种不同施肥水平土样DNA进行提取,扩增细菌16S rDNA基因片段,建立克隆文库.用限制性内切酶Hha I和Rsa I进行PCR-RFLP分析,分别得到146、187、11个酶切类型.采用α多样性的测度对试验结果进行分析统计结果表明,不同处理间土壤细菌的多样性(H′、Ds和Dg)和物种丰富度(dMa、R2和E)均为F1>CK>F2;λ、dMa、E和H′指数在不同施肥处理间的变异系数达到56.96%～163.1%,尤其Simpson指数λ是非常敏感的指标,处理间的差异最大,表明氮肥缺失严重影响土壤细菌群落多样性,合理施肥有利于土壤细菌的多样性.     

绒山羊瘤胃甲烷菌mcrA基因的RFLP分析(英文)

[Objective] The aim of this study is to understand the population composition of methanogens in rumen fluid of grazing Inner Mongolian cashmere goat. [Method] Total DNAs of various bacteria in rumen fluid were isolated for PCR amplification using the specifically designed primers based on conservative mcrA sequence of methanogens; then mcrA specific clone library was accordingly established. The restriction fragment length polymorphism(RFLP) of the library was further analyzed by digestion of restriction enzyme Taq I. [Result] One hundred and five randomly selected specific colonies were classified into six RFLP types, among which the dominant type accounts for 38%, and other types account for 27%, 18%, 5.5% and 4.5%, respectively. [Conclusion] There are at least six different methanogens in rumen fluid of grazing Inner Mongolian cashmere goat.     

中国不同地区亚洲韧皮杆菌遗传多样性分析

 柑桔黄龙病是最严重的柑桔病害之一,现已威胁世界柑桔产业。已被证实在中国南部该病害通过嫁接和木虱传播。因此黄龙病的流行病学研究就显得尤为重要。本研究中,基于外膜蛋白(omp)基因的PCR-RFLP的方法被用来研究中国南部各柑桔主产区的亚洲韧皮部杆菌的遗传多样性。研究采用不同的限制性内切酶消化各地区的亚洲韧皮部杆菌的omp基因产生了不同的RFLP指纹图谱,并对各地亚洲韧皮部杆菌的omp基因进行克隆测序然后构建系统发育树。结果显示:中国不同地理区域的亚洲韧皮部杆菌具有一定的遗传多样性,即便在某一特定的地区也有一定的差异。序列分析显示中国地区的亚洲韧皮部杆菌有很高的同源性。这些结果对揭示中国柑桔黄龙病的流行病学及制定科学有效的防治策略具有一定的指导意义。     

Molecular mapping of a new gene for resistance to rice blast (Pyricularia grisea Sacc.)

A single dominant blast resistance gene conferring resistance to a Korean rice blast isolate was identified in rice variety `Suweon 365'. We report the chromosomal localization and molecular mapping of this blast resistance gene designated as Pi-18, which confers resistance to Korean isolate `KI-313' of the blast pathogen. To know whether there is a relationship among genes conditioning resistance to location-specific isolates of the blast pathogen and thereby to identify linked markers to resistance gene for isolate KI-313 collected in Korea, RFLP markers previously reported to be linked to major blast resistance genes in different rice germplasm and other markers mapped to nearby regions were surveyed for polymorphism between a resistant (`Suweon 365') and a susceptible (`Chucheongbyeo') parent. Linkage associations of the RFLP markers with the resistance gene were verified using an F2 and F3 segregating population of known blast reaction. RFLP analysis showed that Pi-18 was located near the end of chromosome 11, linked to a single copy clone RZ536 at a distance of 5.4 centiMorgans (cM) and that this gene was different from Pi-1(t). An allelism test revealed that this gene was also different from Pi-k. Currently, a combination of RAPD and microsatellite primers is being employed to find additional markers in this region. Tightly linked DNA markers will facilitate selection for resistant genotypes in breeding programs and provide the basis for map based cloning of this new blast resistance gene. This revised version was published online in July 2006 with corrections to the Cover Date.     

Comparisons of RFLP and PCR-based markers to detect polymorphism between wheat cultivars

Previously chromosome 3A of wheat (Triticum aestivum L.) was reported to carry genes influencing yield, yield components, plant height, and anthesis date. The objective of current study was to survey various molecular marker systems for their ability to detect polymorphism between wheat cultivars Cheyenne(CNN) and Wichita (WI), particularly for chromosome3A. Seventy-seven `sequence tagged site' (STS), 10simple sequence repeat (SSR), 40 randomly amplified polymorphic DNA (RAPD) markers, and 52 restriction fragment length polymorphism (RFLP) probes for wheat homoeologous group 3 chromosomes, were investigated. Three (3.9%) STS-PCR primer sets amplified polymorphic fragments for the two cultivars, of which one was polymorphic for chromosome 3A. Sixty percent of SSR markers detected polymorphism between CNN and WI of which 50% were polymorphic for chromosome 3A. Twenty percent of RAPD markers detected polymorphism between CNN and WI in general, but none of these detected polymorphism for chromosome 3A. Of the fifty-two RFLP probes, 78.8% detected polymorphism between CNN and WI for group 3 chromosomes with one or more of seven restriction enzymes and 42% of the polymorphic fragements were for chromosome 3A. These high levels of RFLP and SSR polymorphisms between two related wheat cultivars could be used to map and tag genes influencing important agronomic traits. It may also be important to reconsider RFLP as the most suitable marker system at least for anchor maps of closely related wheat cultivars. This revised version was published online in July 2006 with corrections to the Cover Date.     

RFLP markers are an effective tool for the identification of creeping bentgrass (Agrostis stolonifera L.) cultivars

The potential use of RFLP molecular markers for the identification of four creeping bentgrass (Agrostis stolonifera L.) cultivars and one variety of A. tenuis Sibth. used as control, was investigated. Seven probes out of the total 44 screened were able to differentiate all five cultivars at once. On the basis of their genetic similarity the varieties bred at the Pennsylvania State University were grouped closely together, whereas the variety Prominent and the A. tenuis control were more distantly related.