黄瓜种质资源遗传多样性的RAPD鉴定与分类研究

利用29个RAPD引物对66份来源和类型不同的黄瓜种质基因组DNA的分析,共扩增出了253条带,其中195条为多态性带,比例为77.08%.不同引物所揭示的种质多样性差异很大.供试种质间平均期望杂合度为0.388.中国种质的平均期望杂合度为0.348,略高于国外引进种质.长江以南黄瓜种质的遗传多样性高于长江以北,华南型种质的遗传多样性高于华北型种质.长江流域以南可能是黄瓜的较早或主要的演化地.供试种质依RAPD标记被分为8个组群.西双版纳黄瓜明显地与其他栽培种质分开了,国外引入的绝大多数种质被聚到一起或分属单独组群.除西双版纳外其他中国栽培种质的遗传关系与形态特征和地域分布虽然存在一定的相关性,但不总是一致,表明地区间的引种交流可能导致了某些基因在不同种质间的渗入.     

羊肚菌栽培菌株遗传多样性分析及种特异性RAPD-SCAR标记开发

近年来中国的羊肚菌Morchella spp.栽培技术取得了长足进步,但基础研究薄弱影响其稳产和高产,国内外尚无羊肚菌栽培菌株种质资源遗传多样性的研究报道。本文对来自全国12省份的36个羊肚菌栽培菌株进行了ITS系统发育分析,并采用RAPD进行了遗传多样性评价。结果表明,结合有效的参考菌株序列,通过ITS序列分析可以将供试菌株进行区分和鉴定,在36个菌株中,26个菌株属于梯棱羊肚菌Morchella importuna,其他10个菌株属于六妹羊肚菌M. sextelata;将自40条RAPD引物中筛选出的14条用于供试菌株遗传多样性分析,共扩增出124条多态性条带;UPGMA聚类可将供试菌株分为两大类群,分别对应于ITS系统发育分析中的梯棱羊肚菌和六妹羊肚菌两个物种,梯棱羊肚菌种内菌株多态性高于六妹羊肚菌。OPA17引物和OPA18引物分别在AA02和AA15菌株中扩增出具有唯一性的特征条带,对两个特征条带进行回收测序后,设计出两个特异性SCAR的引物,它们能有效地从36个供试菌株群体中将菌株AA02和AA15鉴别出来。本文首次全面系统地采用ITS分析鉴别了我国羊肚菌栽培菌株的种性,采用RAPD分子标记系统地评价了羊肚菌栽培菌株的遗传多样性,并验证了RAPD分子标记转化为菌株特征性SCAR标记的可行性。     

木耳栽培菌株酯酶同工酶的酶谱多样性研究

采用聚丙烯酰胺凝胶电泳技术对木耳（Ａｕｒｉｃｕｌａｒｉａａｕｒｉｃｕｌａ（Ｈｏｏｋ）Ｕｎｄｅｒｗ）１０个栽培菌株酯酶同工酶的酶谱多样性进行了研究。１０个供试菌株的幼龄菌丝（７ｄ）中仅检测到２５条酶带,各个菌株分别具有２～３条酶带,１０个菌株仅有２种酶谱类型;老龄菌丝（７２ｄ）中共检测到４４条酶带,各个菌株分别具有３～６条酶带,１０菌株共有８种酶谱类型。研究表明,木耳双核体菌丝中某些酯酶同工酶基因位点在一定的发育时期才开始表达,老龄菌丝酯酶同工酶酶谱在木耳菌株鉴别和遗传育种研究中具有更大的应用价值。     

苎麻疫霉群体的RAPD分析*

利用从126个RAPD(RandomAmplifiedPolymorphicDNAs)随机引物中筛选到的可扩增出清晰条带、主带明显、稳定的8条引物,对采集自江苏、安徽和江西不同寄主的45个Phytophthoraboehmeriae菌株进行全基因组DNARAPD标记遗传多样性分析。选用引物共标出DNA指纹图带68条,其中多态性条带20条,多态性检测率为29.4%,表明该种内不同地区和寄主来源的菌株间变异较小。利用Popgene软件计算供试菌株间的遗传距离并绘制聚类树状图,供试菌株被划分为2个遗传聚类组。菌株间的遗传相似性与菌株的寄主来源有一定的相关性,来自江苏、江西和安徽棉花上的27个菌株被划分在同一遗传聚类组内,而分离自构树、枫杨和苎麻的18个菌株被划分在另一个遗传聚类组。结果还表明菌株间遗传相似性与其地区来源无直接相关性。     

苎麻疫霉群体的RAPD分析

利用从126个RAPD(RandomAmplifiedPolymorphicDNAs)随机引物中筛选到的可扩增出清晰条带、主带明显、稳定的8条引物,对采集自江苏、安徽和江西不同寄主的45个Phytophthoraboehmeriae菌株进行全基因组DNARAPD标记遗传多样性分析。选用引物共标出DNA指纹图带68条,其中多态性条带20条,多态性检测率为29.4%,表明该种内不同地区和寄主来源的菌株间变异较小。利用Popgene软件计算供试菌株间的遗传距离并绘制聚类树状图,供试菌株被划分为2个遗传聚类组。菌株间的遗传相似性与菌株的寄主来源有一定的相关性,来自江苏、江西和安徽棉花上的27个菌株被划分在同一遗传聚类组内,而分离自构树、枫杨和苎麻的18个菌株被划分在另一个遗传聚类组。结果还表明菌株间遗传相似性与其地区来源无直接相关性。     

江汉平原及其周边地区花生根瘤菌的遗传多样性

采用RAPD分析技术和16S－23S rRNA间隔区段（IGS）RFLP分析,分别对分离自江汉平原及其周缘地区的花生根瘤菌进行了遗传多样性和系统发育研究。结果表明,全部供试验菌分别在48％和50％的相似性水平分为Ⅰ、Ⅱ两群,供试花生根瘤菌与参比菌株B.japonicum和B.elkanii聚在群I,参比菌株Rhizobium Sinorhizobium,Mesorhizobium和Agrobacterium聚在群Ⅱ。供试花生根瘤菌的遗传多样性及其在系统发育中的地位主要受地域因素的影响,来自江汉平原中心地带天门和潜江的菌株在76％以上的相似性水平上聚在一起,处于周边地带的武汉和荆州,由于其特定的地理因素的影响。菌株的多样性更为丰富,部分菌株在分类上与其它地域的菌株相互融合,并在较高的相似水平存在一定摆动性,来自外缘随州的菌株,表现了明显的地理分隔作用,其在系统演化中的地位相对独立,总体上从平原腹地到外缘地区。根瘤菌地理分隔作用逐渐明显,在平原外缘的交接地带,根瘤菌的多样性最为丰富。     

苎麻疫霉群体的RAPD分析

利用从126个RAPD(Random Amplifled Polymorphic DNAs)随机引物中筛选到的可扩增出清晰条带、主带明显、稳定的8条引物,对采集自江苏、安徽和江西不同寄主的45个Phytophthora boehmeriae菌株进行全基因组DNA RAPD标记遗传多样性分析。选用引物共标出DNA指纹图带68条,其中多态性条带20条,多态性检测率为29．4％,表明该种内不同地区和寄主来源的菌株间变异较小。利用Popgene软件计算供试菌株间的遗传距离并绘制聚类树状图,供试菌株被划分为2个遗传聚类组。菌株间的遗传相似性与菌株的寄主来源有一定的相关性,来自江苏、江西和安徽棉花上的27个菌株被划分在同一遗传聚类组内,而分离自构树、枫杨和苎麻的18个菌株被划分在另一个遗传聚类组。结果还表明菌株间遗传相似性与其地区来源无直接相关性。     

设施化专用香菇种质资源挖掘

以香菇L808菌株基因组序列为参照,开发了300份插入/缺失(InDel)标记。通过5个菌株的初筛,选出均匀分布于基因组的82份多态标记对42份香菇种质资源进行遗传背景分析。同时在设施化栽培条件下考察了种质资源的生育期、菌棒转色、菌棒硬度、现蕾期和产量等表型。结果表明：供试香菇种质资源的遗传多样性丰富,栽培菌株与野生菌株的遗传距离较远,遗传相似性系数平均值为0.51。群体结构分析将种质资源分为6个亚群,与基于遗传距离的系统聚类结果较为一致。亚群间菌株具有较远的亲缘关系,遗传分化指数平均值为0.290,适合用于杂交育种。表型结果显示,设施化栽培条件下的种质资源农艺性状分化程度高,亚群间菌株的生育期、现蕾期和产量均有显著差异。种质资源多样性分析结果为香菇杂交育种工作奠定了基础。     

毛木耳种质资源的RAPD分析

利用22个随机引物对来源不同的56个木耳菌株进行了RAPD分析。结果表明,所有引物的扩增产物DNA片段均表现出明显的多态性,供试菌株总共扩增出164条多态性片段,占总扩增片段的99%;供试菌株两两间的遗传相似系数变化较大(平均GS值0.2143￣0.8764)。采用系统聚类法中的类平均法,对供试的所有菌株两两间相似系数进行聚类,可将它们分为四大类,各大类的类间和类内菌株的遗传变异程度较大,以IV类内各菌株间的最高(平均GS值0.3891),II和III类间的最低(平均GS值0.5887),表明遗传变异也较丰富(总平均GS值0.4918)。将RAPD技术应用于不同菌株间遗传差异的研究,具有反应迅速、不受外界环境条件影响、能从DNA分子水平上揭示菌株间遗传差异等优点,是一种快速准确评估木耳种质资源的有效方法。     

抗风桐(Pisonia grandis)的生态生物学特征

对收集于广西桂林的17份野生毛葡萄种质和24份栽培葡萄种质,分别使用12条ISSR和12条SCoT引物进行了遗传多样性和亲缘关系检测。结果表明：两种分子标记均能产生较丰富的多态性片段,可有效应用于葡萄的遗传多样性检测,但在聚类分析结果上表现出一定的差异性,SCoT分子标记能更好地区分野生种质和栽培品种,说明SCoT分子标记在葡萄遗传多样性检测和系统进化研究上可能更有优势。从SCoT聚类结果上看,广西植物研究所收集的3个野生毛葡萄种质zws1、zws2和zws3相对其它野生种质而言,更偏向于与栽培种质聚为一类,说明这一类野生毛葡萄可能是这些栽培品种的原始亲本来源之一。不同的野生种质聚为多个类群,并表现出明显的地域特性,但遗传距离相对较远,说明桂林野生毛葡萄资源具有丰富的遗传变异。栽培品种没有明显的聚类特点,可能因为所选用的栽培品种的地域代表性并不是很强,也可能是因为栽培品种在不断的人工杂交选育过程中,遗传背景趋向一致,遗传多样性降低。该研究证明SCoT分子标记在葡萄遗传多样性研究上具有一定的优势。该研究结果为桂林毛葡萄资源的保护、利用和品种选育提供了理论依据,也为葡萄的系统进化研究提供了参考。     

黑龙江黑木耳优良菌株筛选研究

在2012年春季初筛和秋季复筛基础上,对选出的4个野生黑木耳菌株和5个栽培品种,以及在多个木耳基地分离获得的7个菌株进行了进一步栽培比较试验。基于菌丝生长速度、长势、拮抗试验、抗菌试验、栽培农艺性状和产量等因素,最终从16个菌株中筛选出适合黑龙江栽培的优良菌株6个,其中野生菌株2个：A14和A15;栽培菌株4个：M23、M43、M44和M46。     

东北地区木耳"白毛菌病"的病原菌

"白毛菌病"是近年来东北地区木耳栽培中一种常见病害.其病原菌在盛夏伏天侵染木耳子实体,在木耳耳片腹面生长一层白色网状霉层,对木耳生长及商品形态造成严重危害.从黑龙江省尚志地区染病耳片上分离得到两株病原菌,经对它们进行形态学特征观察、rDNA ITS序列分析和致病性测定,证明这两株病菌为尖孢镰孢和厚垣镰孢,二者均对栽培木...     

SCAR标记在黑木耳栽培菌株分类鉴定中的应用

应用序列特异性扩增区域（SCAR）标记技术分析50株黑木耳栽培菌株的遗传多样性。在SRAP标记分析过程中发现1条1 200 bp的特异性条带,经回收克隆测序转为SCAR标记。根据序列设计出1对特异性引物,经PCR可以扩增出1 000 bp大小的片段,说明成功构建出了＂黑931＂的指纹图谱。     

木耳栽培种质资源的数量分类研究

在木耳栽培种质资源农艺性状调查的基础上,应用数量分类学中的Q型聚类分析法对20个木耳菌株进行分类研究,并对14个农艺性状进行R型聚类分析和主成分分析。结果表明：Q型聚类将20个木耳菌株在欧氏距离6.29处依据子实体朵型性状分为簇生型菌株和菊花型菌株两大类群,菊花型类群在欧式距离4.79处依据生育期性状的原基发生类型划分为分散型和集中型两个亚群;R型聚类表明菌丝体性状（1个）、生育期性状（2个）、子实体性状（8个）等11个农艺性状间相关性较强;主成分分析中,发现子实体背面皱褶、耳片数、原基发生时间、子实体朵型、干耳背面颜色等5个性状是14个农艺性状的第1主成分,贡献率高达62.26%,把第1主成分命名为朵型-生育期构成因子,作为种质评价的指标。     

菌糠二次利用栽培黑木耳试验研究

为充分利用营养没有得到充分吸收的栽培过食用菌的废弃菌糠、明确在牡丹江地区再利用食用菌菌糠生产黑木耳的适宜比例,以灵芝和鲍鱼菇的废弃菌糠为试验材料,进行不同配方的对比试验。试验结果表明：黑木耳培养料中灵芝菌糠体积分数为30%时栽培效果最佳,显著优于参入鲍鱼菇菌糠的各种培养料配方（P〈0.01）。因此,可以利用灵芝菌糠废料作为替代料栽培黑木耳使用。     

Strain-typing of Lentinula edodes in China with inter simple sequence repeat markers

To validate strain typing by inter simple sequence repeat (ISSR) analysis in Lentinula edodes cultivars, 17 Chinese L. edodes strains including 15 cultivated strains cultivated on a large scale and two wild strains were analyzed with the ISSR technique. With the use of two ISSR primers, a total of 32 DNA products were detected, of which, 31 DNA products (96.9% of the detected products) were polymorphic between two or more strains. The profiles of those two primers could be employed to differentiate all of the tested strains. A cluster analysis based on ISSR data revealed that the 17 strains could be classified into two distinct groups. One group consisted of eight strains in which the cultivated strains were H (high-temperature)-type or B (broad-temperature)-type, and the other group comprised cultivated strains that were of the L (low-temperature)-type or M (medium-temperature)-type. In contrast to the two wild strains, the genetic diversity of 15 cultivated strains was very rich based on a similarity coefficient analysis.     

Comparative analysis on the diversity of <Emphasis Type="Italic">Auricularia auricula</Emphasis>-<Emphasis Type="Italic">judae</Emphasis> by physiological characteristics,somatic incompatibility and TRAP fingerprinting

Phenotypic traits (physiological characteristics and somatic incompatibility) and genotypic traits (target region amplification polymorphism TRAP) were used to study the diversity of 32 main cultivars of Auricularia auricula-judae in China. Twenty seven important and stable physiological indexes were evaluated; somatic incompatibility test (SIT) reaction was described from three aspects: type, pigment, and intensity; 16 pairs of TRAP primer combinations produced 535 unambiguous and reproducible DNA fragments, of these 524 (97.9%) were polymorphic. Dendrograms were constructed by Unweighted Pair-group Method with Arithmetic Averages (UPGMA) method, and the principal coordinate analysis (PCO) of the three methods (physiological characteristics, SIT intensity and TRAP) exhibited similar clustered patterns, revealing that all the tested strains could be divided into three distinct groups, each of which was correlated with different geographical regions. Most strains originated from the same area were with a narrow genetic basis and could possibly be domesticated from the local wild-type strains, some strains were suspected to be synonymous. The grouping information obtained in the present work provides significant information for further genetic improvement of A. auricula-judae.     

木耳杂交菌株对绿色木霉菌的抗性测定

木耳（Auricularia auricula-judae）在菌丝体阶段极易感染绿色木霉（Trichoderma vider）等多种杂菌。本研究以木耳176个杂交子为供试菌株,采用木耳与绿色木霉菌丝体对峙培养,发酵液相互抑制菌丝体生长,以及调查木耳菌袋田间污染率,比较了杂交子抗绿色木霉的能力,初步建立了木耳抗霉菌污染能...     

Group Q Streptococci I. Ecology, Serology, Physiology, and Relationship to Established Enterococci

The group Q streptococci possess unique serological and physiological characteristics which differentiate them from established enterococci. The group Q antigen was not demonstrable in all strains; however, all possessed the group D antigen. All group Q strains were physiologically similar regardless of whether or not they possessed the group Q antigen. These strains differed from the established enterococcal species, as they neither hydrolyzed arginine nor initiated growth in 1.0% methylene blue-milk. They also differed radically in the fermentation of various carbohydrates, especially the polyhydric sugar alcohols. The results indicate that the group Q streptococci constitute a unique taxonomic entity; the species designation Streptococcus avium sp. n. is suggested, owing to their characteristic occurrence in chicken fecal specimens.     

New available SCAR markers: potentially useful in distinguishing a commercial strain of the superior type from other strains of <Emphasis Type="Italic">Lentinula edodes</Emphasis> in China

At present, more than 100 strains of Lentinula edodes are cultivated on a commercial scale in China. A simple, reliable, and effective method to distinguish some commercial strains of the superior type from other commercial strains is very important for the Lentinula industry. In this study, 23 commercial strains of L. edodes cultivated widely in China at present were collected and analyzed with randomly amplified polymorphic DNA (RAPD) technique. Three informative dominant sequence characterized amplified region (SCAR) markers were developed by designing three pairs of specific SCAR primers from three sequenced differential RAPD bands, respectively. Based on the three SCAR markers, three different multiplex polymerase chain reaction (PCR) phenotypes were detected among the 23 studied commercial strains and in which a multilocus phenotype characterizing a commercial strain Cr02 of the superior type could potentially be used to distinguish this strain from the other 22 studied commercial strains. To our knowledge, this study is the first to describe the development of a multiplex PCR technique based on SCAR markers for detecting the molecular phenotypes among commercial strains of L. edodes in China.