禾本科植物内生真菌研究 5.中国产部分epichloё内生真菌的遗传多样性

Epichloё内生真菌(epichloё endophytes)是禾本科植物的共生真菌,开展其多样性研究对菌株的生物学特性和进化规律的了解以及开发利用都有着十分重要的意义。本研究筛选了12条随机引物对24株epichloё内生真菌进行RAPD扩增,探讨真菌种类、宿主种类、采集地和菌株间的遗传多样性。结果表明,宿主为拂子茅Calamagrostis sp.、雀麦Bromus sp.、披碱草Elymus sp.、小颖羊茅Festuca parvigluma Steud.的菌株分别独立聚为一个分枝;而宿主为鹅观草Roegneria spp.的Neotyphodium属菌株聚类情况较为复杂;宿主为早熟禾Poas pp.的菌株分别在三个不同的分枝,表现出丰富的遗传多样性。以上结果反应了我国具有丰富的epichloё内生真菌资源,它们和其宿主之间有着比较明显的相互关系。本研究还显示,采集于南京的拂子茅与其内生真菌共生可能发生在较久远的年代。     

禾本科植物内生真菌研究5. 中国产部分epichlo?内生真菌的遗传多样性

Epichlo?内生真菌（epichlo? endophytes）是禾本科植物的共生真菌,开展其多样性研究对菌株的生物学特性和进化规律的了解以及开发利用都有着十分重要的意义。本研究筛选了12条随机引物对24株epichlo?内生真菌进行RAPD扩增,探讨真菌种类、宿主种类、采集地和菌株间的遗传多样性。结果表明,宿主为拂子茅Calamagrostis sp.、雀麦Bromus sp.、披碱草Elymus sp.、小颖羊茅Festuca parvigluma Steud.的菌株分别独立聚为一个分枝;而宿主为鹅观草Roegneria spp.的Neotyphodium属菌株聚类情况较为复杂;宿主为早熟禾Poa spp.的菌株分别在三个不同的分枝,表现出丰富的遗传多样性。以上结果反应了我国具有丰富的epichlo?内生真菌资源,它们和其宿主之间有着比较明显的相互关系。本研究还显示,采集于南京的拂子茅与其内生真菌共生可能发生在较久远的年代。     

中国产禾本科植物内生真菌的遗传多样性及rDNA-ITS系统发育分析

采用RAPD技术对分离自我国的13株Epichlo?spp.和9株Neotyphodiumspp.的禾本科植物内生真菌进行了遗传多样性分析,同时对其中的4株进行了rDNA-ITS序列分析及系统发育研究。RAPD分析结果显示:原产自我国的菌株与原产自欧洲的菌株N.uncinatum之间亲缘关系较远;我国的21个菌株之间也存在一定的遗传多样性。rDNA-ITS序列分析表明:原产自我国的菌株聚为一枝,表明我国的Neotyphodium属真菌很有可能直接由我国的Epichlo?属真菌演化而来的新的类群;我国的Epichlo?属真菌有可能和国外推测的一个未确定的Neotyphodium属真菌的杂交进化起源(LAC)有关。     

禾本科植物内生真菌资源及其物种多样性

植物内生真菌是当代微生物资源研究的一个热门话题,对禾本科植物内生真菌资源研究进行了总结。与冷季型禾本科植物共生的麦角菌科epichlo内生真菌是目前的研究重点,包括偶尔在宿主植物体表形成子座的Epichlo属真菌和几乎完全不形成症状的Neotyphodium属真菌。具体介绍该真菌类群的物种多样性及其地理分布多样性,概括了它们在全球各大区域的分布特征。对近年来中国迅速发展的epichlo内生真菌的研究作了总结,最后展望了国际国内epichlo内生真菌资源探索的发展方向。特别指出:冷季型禾本科植物内生真菌在南美洲和亚洲,暖季型禾本科植物内生真菌在热带亚热带地区的研究还相对薄弱,值得今后继续加强探索和挖掘。     

中国禾本科植物内生真菌研究4.Epichlo(e) yangzii的人工杂交

作者首先研究Epichloe yangzii植株上的人工杂交,明确了供试菌株的交配型.然后将分离自无子座鹅观草属植物的23株"Neotvphodium属"真菌孢子分别与E.yangzii的子座杂交,其中发现有21株与E yangzii(mat-1,mat-2)杂交不亲和,有2株与E yangzii(mat-1)杂交亲和.利用tubB基因片段对8株"Neotyphodium属"真菌菌株进行系统发育分析,结果表明与E yangzii杂交亲和的2个菌株和E.yangzii聚为一枝,而其它6个菌株形成独立的分枝,进一步证实了这2个菌株足有时在宿主植物上不形成子座的Eyangzii.这说明了在宿主植物上的人工杂交是区别有时不产子座的E yangzii和Neotyphodium属真菌的有效手段.     

内蒙古中东部草原羽茅内生真菌的遗传多样性

该文从形态学和基因水平上对内蒙古中东部草原4个羽茅(Achnatherum sibiricum)种群所含的27个内生真菌(Neotyphodium)菌株的遗传多样性进行了研究,结果表明羽茅内生真菌在形态学和基因水平上均表现出较高的遗传多样性。在PDA培养基上,根据菌落颜色、质地、生长速率和分生孢子外观等特征,可分为4个形态型;西乌旗、霍林郭勒和定位站3个种群所含的内生真菌菌落具有相似的生长特性和外观,而羊草(Leymus chinensis)样地种群无论从菌落外观和质地的多样性,还是遗传多样性指数均高于其它3个样地。用20个随机引物对这27株内生真菌基因组DNA进行RAPD分析,共检测到463个位点,其中461个为多态位点,多态位点百分率达99.6%,特有位点93个,占20.1%。Nei基因多样性指数h和Shannon多样性指数I分别为 0.238 0和0.387 0;多数菌株间的遗传一致度较低而遗传距离较大,在DNA水平上存在较显著的遗传变异。基于Nei无偏遗传距离和UPGMA法的聚类分析结果将27株内生真菌分为7组,并且与形态型的划分基本一致,RAPD分析方法提高了内生真菌类群划分的可靠性。     

内生真菌与禾本科植物之间的关系及其生态学意义

1　前　言内生真菌是一类生长在植物体内的真菌 ,至今已在许多高等植物中发现了内生真菌的存在 ,但以禾本科植物中尤为常见。这些真菌包括子囊菌纲、麦角菌科 (Ｃｌａｖｉｃｉｐｉｔａｃｅａｅ)、瘤座菌族 (Ｂａｌａｎ ｓｉｅａｅ)的许多种类 ,也包括与瘤座菌族关系密切的一些半知菌类 ,如Ａｃｒｅｍｏｎｉｕｍ[1 5] 。与菌根真菌的一个显著不同是 ,它们仅存在于植物地上部分的组织内而不是根的组织中。至少有 80个属、几百种禾本科植物被确认含内生真菌[1 6] ,但到目前为止 ,大量的研究都集中在两个有重要经济意义的植物种 ,即高羊茅 (Ｆｅ…     

中国禾本科植物内生真菌研究4. Epichloё yangzii的人工杂交

作者首先研究Epichloё yangzii在植株上的人工杂交,明确了供试菌株的交配型。然后将分离自无子座鹅观草属植物的23株"Neotyphodium属"真菌孢子分别与E.yangzii的子座杂交,其中发现有21株与E.yangzii(mat-1,mat-2)杂交不亲和,有2株与E.yangzii(mat-1)杂交亲和。利用tubB基因片段对8株"Neotyphodium属"真菌菌株进行系统发育分析,结果表明与E.yangzii杂交亲和的2个菌株和E.yangzii聚为一枝,而其它6个菌株形成独立的分枝,进一步证实了这2个菌株是有时在宿主植物上不形成子座的E.yangzii。这说明了在宿主植物上的人工杂交是区别有时不产子座的E.yangzii和Neotyphodium属真菌的有效手段。     

中国禾本科植物内生真菌研究4. Epichlo? yangzii的人工杂交

作者首先研究Epichlo? yangzii在植株上的人工杂交,明确了供试菌株的交配型。然后将分离自无子座鹅观草属植物的23株“Neotyphodium属”真菌孢子分别与E. yangzii的子座杂交,其中发现有21株与E. yangzii（mat-1,mat-2）杂交不亲和,有2株与E. yangzii（mat-1）杂交亲和。利用tubB基因片段对8株“Neotyphodium属”真菌菌株进行系统发育分析,结果表明与E. yangzii杂交亲和的2个菌株和E. yangzii聚为一枝,而其它6个菌株形成独立的分枝,进一步证实了这2个菌株是有时在宿主植物上不形成子座的E. yangzii。这说明了在宿主植物上的人工杂交是区别有时不产子座的E. yangzii和Neotyphodium属真菌的有效手段。     

卷柏内生真菌多样性研究

为探究蕨类植物内生真菌的多样性,本研究采用可培养方法分离江西龙虎山和广东丹霞山的岩壁常见蕨类植物——卷柏的内生真菌,结合形态特征和ITS序列进行菌株鉴定、研究多样性。结果表明：从1 080个组织块中,共分离出234株菌株,卷柏内生真菌的总定殖率为18.98%,总分离率为21.67%。根据分离频率,毛壳菌属Chaetomium（26.50%）、青霉属Penicillium（18.38%）、棒束孢属Isaria（11.97%）、曲霉属Aspergillus（9.42%）为优势属。不同居群地春季卷柏的内生真菌多样性存在差异,龙虎山的分离率为34.72%,而丹霞山仅为6.39%,其优势属也不尽相同。对龙虎山不同季节的卷柏内生真菌多样性研究表明,春季的分离率为34.72%,高于冬季的23.89%,春、冬两季之间的真菌类群相似性系数仅为0.39,差异显著。研究结果将有助于进一步揭示丹霞地貌地区蕨类植物内生真菌物种组成及其影响因素。     

禾本科植物内生真菌研究20：基于NRPS5基因的我国东部早熟禾亚科植物内生真菌系统发育学分析

【背景】冷季型禾草-Epichloae真菌内生共生体在我国广泛存在,Epichloae内生真菌具有遗传多样性。【目的】选取来自江苏、山东和安徽等6省共8地的早熟禾亚科植物中的21个Epichlo?sp.菌株,检测菌株的NRPS5基因;分析不同宿主内Epichlo?属内生真菌NRPS5基因多样性。【方法】提取Epichlo?spp.菌株DNA,利用特异性引物对菌株中NRPS5基因进行PCR检测和序列分析,并基于NRPS5基因构建最大简约法系统发育树。【结果】NRPS5基因可能广泛分布于我国东部冷季型禾本科植物Epichloae内生真菌中。在同种内生真菌和同一宿主(除鹅观草)中的不同种内生真菌中NRPS5基因相对保守,但不同宿主的内生真菌其碱基序列差异大,与宿主植物采集地点、采集时间和子座的有无均无相关性,NRPS5基因的差异主要与宿主植物有关。这说明NRPS5基因序列比较保守,推测属于NRPS蛋白的功能核心区。【结论】NRPS5基因序列相对保守,可作为真菌遗传多样性分析的一种辅助手段。     

Genetic (RAPD) diversity in Peromyscus maniculatus populations in a naturally fragmented landscape

We assessed the effects of long-term habitat fragmentation on genetic (random amplified polymorphic DNA) diversity in 11 Peromyscus maniculatus populations in the Lake Superior watershed. We analysed genetic structure at two spatial scales and the effect of island size and isolation on genetic diversity. At the regional scale, island populations differed from mainland populations (FST = 0.36), but mainland populations did not differ from each other (FST = 0.01). At the local scale, populations of the main island of Isle Royale differed from adjacent islet populations (P < 0.001; Monte Carlo approximation of Fisher's exact test), but not from each other (combined P = 0.63). Although geographical distance and genetic distance were positively correlated (P < 0.01; Mantel test), cluster analysis revealed some inconsistencies. Finally, genetic diversity was inversely related to isolation (P = 0.01), but had an unexpectedly negative relationship with island area (P = 0.03). The genetic structure of P. maniculatus populations in portions of the Lake Superior watershed appears to have been affected by long-term habitat fragmentation.     

内蒙古中部小驼嗡蜣螂七个种群的遗传多样性

采用RAPD分子标记技术分析了内蒙古中部地区小驼嗡蜣螂（Onthophagus gibbulus）7个种群的遗传多样性及遗传分化。16条引物共检测到363条带,总多态位点比率为100％,Nei基因多样度为0．280,总的Shannon信息指数为0．433,结果表明,小驼嗡蜣螂种群遗传多样性丰富。在总的遗传变异中,12．57％的变异存在于种群间,87．43％的变异存在于种群内,种群间的遗传分化程度较低,种群间遗传分化系数（Gst）在0．074～0．203,基因流（Nm）在4．569～9．791,表明种群间存在广泛的基因交流。在本研究涉及的地理范围内,小驼嗡蜣螂种群的遗传距离与地理距离不相关,说明小驼嗡蜣螂种群间的分化与其生境的异质性有关。     

大白菜细胞质雄性不育的分子生物学研究

细胞质雄性不育(cytoplasmic male sterility,CMS)是作物杂种优势利用的一 条重要途径。对其机制的研究不仅具有重要的理论意义,而且具有重要的应用价值。对大白菜而言,利用CMS最关键的环节是要育成不育系和其相应的保持系。利用不育系和其它材料杂交产生F1代种子;用不育系和保持系杂交来繁殖不育系;保持系自交繁殖种子。但采用常规的方法来选 育不育系和其相应的保持系周期长、见效慢,远不能满足生产发展的需要。现代生物技术为创造新的不育系和保持系提供了便利。本论文对大白菜细胞质雄性不育的分子机理和 RAPD分析的有关问题进行了研究。     

利用RAPD标记评价小豆种质遗传多样性

本研究利用10个RAPD引物对180份小豆种质的基因组DNA进行扩增,共扩增出44条带,其中35条具有多态性,比例为79.5%,平均每个引物扩增出3.5条多态性带;平均遗传距离为0.274,变异幅度为0.05～0.60,平均遗传多样性指数为0.692;基于RAPD标记,把180份小豆种质聚类划分为4个组群,该组群的划分与小豆的生态地域性似乎不存在明显的相关性.     

Spatio-temporal dynamics of endophyte diversity in the canopy of European ash (Fraxinus excelsior)

Leaf-inhabiting endophytic fungi of Fraxinus excelsior growing in a floodplain forest were isolated during 2008 to investigate vertical community structure, species richness and seasonal variation. The analysis of 848 fungal endophytes from 213 leaves resulted in 50 different species. In the understorey, infection density and species richness were higher than in the crowns of mature trees throughout the whole vegetation period. Within tree crowns, sun-exposed leaves of the top canopy exhibited the lowest infection rates. Most species were rare or absent in spring and in the light crowns and frequent in autumn and the understorey. However, some species, especially the two most frequent, Alternaria infectoria and A. alternata, deviated from these patterns. Young leaves were nearly free of endophytes. Apparently, the subsequent infection and establishment of fungi strongly depend on microclimatic parameters and leaf characters, which create highly variable spatial and temporal colonisation patterns within an individual tree.     

Grass species and endophyte effects on survival and development of fall armyworm (Lepidoptera: Noctuidae)

Grass selections including 10 zoysiagrasses, 18 paspalums, 34 Bermuda grasses, tall fescue, creeping red fescue, and perennial ryegrasses with and without endophyte were evaluated for potential resistance to fall armyworm, Spodoptera frugiperda (J. E. Smith), larvae. Laboratory evaluations assessed the degree of antibiosis among >70 grass lines to first-instar fall armvworms. When all parameters measured were considered, the trend in resistance to fall armyworm among endophyte-infected (E+) and endophyte-free (E-) cool season grasses from greatest to least was: 'Dawson' E+ > APR 1234 > 'Dawson' E- > 'Rosalin' E+ > Lp 5425, 'Rosalin' E-, ATF 480 > 'Tulsa' or: E+ slender creeping red fescue > E+ turf- type perennial ryegrass > E- slender creeping red fescue > E+ forage-type perennial ryegrass > E- forage-type perennial ryegrasses, and E+ tall fescue > E- turf-type tall fescue. Among warm season grasses larval weight gain was reduced on all zoysiagrasses. Larval weight gain also was lower on the Bermuda grasses 'Tifsport', 'Tifgreen', 97-4, 97-14, 97-22, 97-28, 97-39, 97-40,97-54, 98-15, 98-30, and 98-45 than when larvae were fed 'Tulsa' tall fescue or the diet control. Only APR1234 and 'Dawson' creeping red fescue reduced larval survival to the same extent that was observed for zoysiagrasses. Survival on Bermuda grasses was least on 97-8. Seashore paspalums were only rarely less susceptible to fall armyworm than tall fescue, although pupal weights were consistently lower on 'Temple 1' and 'Sea Isle 1' paspalums than that on 'Tulsa' tall fescue. Genetic resistance to key grass pests can reduce insecticide use and simplify management of these cultivars.     

Genetic diversity and heavy metal tolerance in populations of Silene paradoxa L. (Caryophyllaceae): a random amplified polymorphic DNA analysis

Metal-contaminated sites can occur naturally in serpentine outcrops or as consequence of anthropogenic activities, such as mining deposits, aerial fallout from smelters and industrial processes. Serpentine outcrops are characterized by high levels of nickel, cobalt and chromium and present a typical vegetation which includes endemisms and plants which also live in uncontaminated soils. These latter metal-tolerant populations provide the opportunity to investigate the first steps in the differentiation of plant populations under severe selection pressure and to select plants to be used in the phytoremediation of industrially contaminated soils. In this report eight populations of Silene paradoxa L. (Caryophyllaceae) growing in copper mine deposits, in serpentine outcrops or in noncontaminated soil in central Italy, were analysed using random amplified polymorphic DNA (RAPD) markers to investigate the pattern of genetic variation. The genetic diversity observed in populations at copper mine deposits was found to be at least as high as that of the neighbouring serpentine populations. Analysis of molecular variance (AMOVA) of the RAPD markers gave high statistical significance to the groupings of populations according: (i) with their geographical location; and (ii) with the metals present in the soil of origin (copper vs. nickel), indicating that RAPD markers detected a polymorphism related to the soil contamination by copper. Finally, two RAPD bands exclusive to copper-tolerant populations were identified.