利用SSR和RAPD标记分析中国部分地区晚疫病菌群体遗传多样性

利用SSR和RAPD对来自福建、黑龙江、河北和内蒙古4个地理群体的80个马铃薯晚疫病菌（phytophthora infestans）株进行了遗传多样性分析。13对SSR引物共扩增出76条谱带,多态性条带比率78.9%,相似系数变化范围0.00～0.42之间;筛选出的14条RAPD引物共扩增出189条谱带,多态性条带比率95.2%,相似系数变化范围0.04～0.66之间。遗传多样性分析表明,在4个群体中,福建群体的多样性更为丰富。遗传相似性分析显示,黑龙江和内蒙古两个群体间的遗传相似性最高,而福建和河北两个群体间的遗传相似性最低。聚类分析显示,来自南方福建的菌株与来自北方黑龙江、河北和内蒙古的菌株亲缘关系较远,且福建群体分布于更多的聚类组,显示出更高的遗传变异度。     

黑木耳单核杂交菌株遗传分析

为实现黑木耳菌株快速鉴定和栽培性状定向育种,研究通过荧光核染色筛选,酯酶同工酶酶谱比较及RAPD分子鉴定,筛选出了黑木耳杂交单核体的交配核型及杂交新种,并对杂交新种进行了遗传分析和栽培比较.结果显示,2个亲本筛选出4种不同类型原生质体单核体,8808亲本单核体为A1、A2型,981亲本单核体为B1、B2型.单核体单-单杂交获得核型为A1B2的杂交菌株,命名为Z1.酯酶同工酶谱及RAPD图谱结果显示Z1菌株含有来源单核的互补型条带和亲本条带.聚类分析将被试菌株分为3类,Z1菌株与981相似度高于8808菌株.栽培试验显示Z1与981栽培性状更近似而又集合了双亲的性状优势.研究的开展可为将来黑木耳菌株鉴定及杂交子遗传分析等提供快速、可靠检测方法.     

广西金福菇菌株筛选及配套栽培技术研究与示范

引进多个金福菇菌株,通过试验筛选出适合广西地区栽培的金福菇菌株,研究总结出一套成熟的栽培技术。试验成功筛选出适合广西本地气候和原料栽培的品种两个,分别为桂菌Tg505和金福菇K,并研究出适合本地的金福菇栽培技术。经产业化示范,南宁市青秀区刘圩镇2018—2019年的金福菇菌株总产值达到116万元,新增种植收入68万元。     

基于ISSR指纹的甘薯食用品种的遗传多样性分析

运用ISSR分子指纹技术检测甘薯基因组DNA的多态性,是鉴定甘薯遗传多样性和亲缘关系有效的方法之一。本研究选用6个ISSR标记对17份甘薯品种进行了DNA指纹扩增,共扩增出55条谱带,其中多态性谱带50条,多态性水平为90.91%;UBC825引物扩增出带型最多,有16种;根据Nei’s遗传距离计算获得的聚类树状图表明,在GS为0.625时,参试的17份甘薯品种分为4个组群;遗传相似性分析表明,顶芽菜用品种间遗传相似系数最高0.877,叶柄食用品种和烤薯型品种之内的遗传相似系数分别为0.778和0.740。结果表明甘薯主要食用间具有较好的遗传多样性,但同类型品种间却又有较高的遗传相似性,这类品种需要引进或创制新资源。     

基于ISSR分析28份香蕉种质的基因组DNA多样性

利用ISSR标记技术分析了28份香蕉种质的遗传多态性。从100个ISSR引物中筛选出8个多态性引物,共扩增出55条DNA带,其中46条为多态性带,占83.6%,平均每个引物扩增的DNA带数为6.88条。依相似系数0.73的水平,将香蕉28个品种划分为6大类。其中云南BB（BB）和东莞高把大蕉（ABB）在相似系数为0.94时,二者的亲缘关系较近。Pisang Ceylan（AAB）和FHIA-18（AAAB）相似系数水平接近为1,表明二者亲缘关系最近。本研究为香蕉遗传关系的建立及品种鉴定提供理论基础。     

杂交稻同核异质不育系的同工酶与遗传鉴定

研究分析了7个同核异质不育系和保持系花药中的酯酶同工酶和过氧化物酶同工酶,结果表明:在这两种同工酶的酶谱中,有些谱带是不育系与保持系共有的,不受雄配子的育性和胞质差异的影响。有些谱带则伴随雄配子的败育而出现(或缺失),其余谱带组分,则因胞质背景的不同而存在差异。据此认为,可以通过同工酶酶谱分析,对细胞质进行遗传鉴定。     

马氏珠母贝杂交与自繁后代及企鹅珍珠贝间同工酶差异分析

利用垂直板状聚丙烯酰胺梯度凝胶电泳分离技术,对马氏珠母贝（PinctadamartensiiD.）和企鹅珍珠贝（Pteria penguinR.）的鳃、后闭壳肌2种组织的5种同工酶进行检测,以比较珍珠贝及马氏珠母贝不同养殖群体酶谱差异.分析表明,不同种间的个体酶谱表型有稳定的差异,筛选出的鳃组织过氧化物歧化酶（SOD）、酯酶（EST）及闭壳肌组织的苹果酸脱氢酶（MDH）、苹果酸酶（ME）、葡萄糖-6-磷酸脱氢酶（G6PDH）等同工酶,具有种特异性,这些酶带清晰稳定,可作为区别种的分子标记.马氏珠母贝群间杂交与群内自繁群体中,不同个体的酶谱表型比较相似,但自繁后代群体酶谱的相似程度更高,位点Sod-2杂合度仅为0.05、近交系数高达0.857,而杂交后代位点杂合度为0.55、F=-0.173,在不同种组织中,两群体间SOD同工酶均存在稳定差异,这种与亲缘关系相关联的酶谱表型,作为蛋白分子标记,可望应用于珠母贝品种改良及品系鉴定上.     

23种野生百合遗传关系的SRAP分析

为探讨百合属属下类群间亲缘关系,本研究选用19对SRAP (sequence-related amplified polymorphism)标记引物对百合属(Lilium) 23个野生种基因组DNA进行遗传多样性及遗传关系分析;NTSYS-pc2.1和POPGEN1.32数据处理软件分析数据.共得到清晰条带236条,多态性条带233条,多态性条带比率为98.72％,平均每对引物产生12.3条多态性条带.相似系数变化范围为0.3750～0.7679,遗传距离变化范围为0.2642～0.9808,UPGMA聚类结果将23个供试材料分成4类.岷江百合与宜昌百合亲缘关系最近,野百合与朝鲜百合亲缘关系最远,来源相同、形态相似的材料大部分聚在一起,钟花组与卷瓣组存在基因交流.聚类分析结果与形态分类结果大致吻合.SRAP标记适合于百合属植物遗传多样性检验与遗传关系分析.     

甘蔗及其近缘属植物同工酶特点的比较研究

用聚丙烯酰胺凝胶电泳法,对我院近年收集的甘蔗及其近缘属植物中,一些代表性材料的过氧化物酶、酯酶同工酶进行分析。通过对植株不同组织、不同叶位叶片间,甘蔗属内不同种间,甘蔗各近缘属间,以及有密切亲缘关系的若干甘蔗栽培品种的同工酶谱特点的比较,对所研究材料的亲缘关系,在进化和分类上的地位,以及同工酶分析在品种鉴定上的意义等问题进行了讨论。     

铁观音和黄棪杂交品种遗传多样性分析

为研究福建省选育的与铁观音或黄棪有亲缘关系品种的遗传多样性,本研究以12个有亲缘关系品种和15个对照种为试验材料,利用SSR分子标记技术分析参试品种的遗传多样性。结果表明,铁观音与黄棪杂交选育的4个品种的遗传相似性与生物学形态和制茶品质相互印证;12个有亲缘关系品种的聚类图能清晰地呈现出品种间的遗传多样性,且大部分品种遗传背景更靠近黄棪,表明在育种阶段选择遗传相似系数更接近黄棪的株系,出现优良品种的可能性更高;各品种与其对应亲本的相似性分析表明,紫玫瑰与铁观音亲缘关系最近,瑞香与黄棪亲缘关系最近;对所有参试品种的相似系数分析发现,有亲缘关系品种的相似系数介于0.64~0.92之间,其中0.80以上的占19.70%,而对照种的相似系数介于0.63~0.81之间,大于0.80的仅占5.71%,二者的聚类图对比分析也表明,对照种的遗传多样性高于有亲缘关系品种。本研究结果为了解与铁观音或黄棪有亲缘关系品种的遗传多样性提供了数据支持,为茶叶生产中的选种和引种提供了遗传多样性指导,同时为茶树的杂交育种与遗传改良提供了理论参考。     

Isozyme analysis of indigenous cassava germplasm for identification of duplicates

Isozyme analysis was utilized for identification of duplicates in indigenous collection of cassava germplasm. 786 indigenous accessions of cassava were screened for 11 key morphological characters and the morphological duplicates were identified. Two hundred and eighteen accessions of cassava consisting of ninety one sets of morphological duplicates were analysed for esterase isozyme polymorphism. Thirty seven polymorphic bands were obtained, showing high polymorphism for this enzyme. Thirty seven double sets and 17 multiple sets were found to be similar at isozyme level. These isozyme duplicates will be further analysed for DNA similarity before eliminating them from the field germplasm. The dissimilar sets showed variation in minor morphological characters, thus confirming the relation between isozyme polymorphism and phenotypic variation.     

35个山东主栽平菇菌株的ISSR遗传差异分析

平菇是我国北方栽培的主要食用菌品种,其菌株多,遗传背景复杂。本研究采用ISSR标记技术对35个山东省主要栽培的平菇菌株进行DNA指纹分析,并利用NTSYS-PC分析软件对这些供试菌株的遗传差异性进行聚类分析。研究结果表明,从107个随机引物中共筛选到16个备用引物,通过利用16个引物对这35个平菇菌株的DNA进行ISSR-PCR,共扩增出259条清晰的DNA片段,大小介于0.3～4.2kb。聚类分析发现,相似水平在66.5%左右时,35个菌株分为3个组群,即黑色、灰黑色和白色子实体3个组群。本研究为山东省平菇遗传信息库的建立和平菇遗传育种工作奠定了一定的基础。     

辐照诱导的新雄性不育系过氧化物酶和脂酶同工酶分析

采用聚丙烯酰胺凝胶垂直板电泳法,对甘蓝型油菜细胞质雄性不育系Polima、Shan2A、Ogu和辐照处理的新雄性不育系Xin1和Xin2及其保持系Shan2B的种子进行了脂酶和过氧化物酶分析,结果表明:5个细胞质雄性不育系比保持系Shan2B多1条迁移率为0.45的脂酶(EST)谱带;Xin1、Shan2A和Polima各有4条过氧化物酶(POD)谱带,保持系缺少1条迁移率为0.61的POD谱带,推测这一同工酶的表达可能与雄性不育相关。     

The Genetic Diversity and Similarities Among <Emphasis Type="Italic">Kengyilia</Emphasis> Species Based on Random Amplified Microsatellite Polymorphism (RAMP)

The genetic diversity and similarities among 32 Kengyilia accessions, distributed to 14 species and one variety were analyzed by using random amplified microsatellite polymorphism (RAMP) markers. Of the 160 RAMP primer combinations tested, 40 (25%) produced polymorphic and clear bands. A total of 264 bands were produced by 40 primer combinations, among which 231 out of 264 bands (87.5%) were polymorphic. Two to 11 polymorphic bands could be amplified from each primer combination, with an average of 5.8 bands. The data of 264 bands were used for RAMP assay. By NTSYS-pc program, genetic similarity coefficients were generated and dendrogram was constructed using UPGMA. The genetic similarity coefficients ranged from 0.477 to 0.965 with the mean of 0.714. The results showed as follows: (1) distinct genetic differences were present among the different species; (2) the different accessions in a species were clustered together, respectively, which had larger genetic similarities and closer relations; (3) the species with similar morphological characters and the species from the same areas or neighboring geographical regions were clustered together; (4) the lowest genetic similarity was found between K. hirsuta (PI531618) and K. laxiflora (PI531631), while the highest genetic similarity was observed between K. hirsuta (Y2364) and K. hirsuta (Y2368); (5) RAMP results are basically comparable with those obtained from studies on morphology and cytology. It is a useful method for analysis of the genetic diversity and similarities in Kengyilia.     

The Genetic Diversity Among Leymus Species Based on Random Amplified Microsatellite Polymorphism (RAMP)

The genetic diversity and similarities among 40 accessions of Leymus Hochst., distributed in 19 species and 1 subspecies, were analyzed by using random amplified microsatellite polymorphism (RAMP) markers. Of the 120 RAMP primer combinations tested, 24 (20%) produced polymorphic and clear bands. A total of 192 bands were amplified by 24 primer combinations, among which 179 (93.23%) bands were found to be polymorphic. Three to thirteen polymorphic bands were amplified by each primer combination, with an average of 7.64 bands. The data of 192 RAMP bands were used to generate Jaccard's similarity coefficients and to construct a dendrogram by means of UPGMA in the NTSYS-pc computer program. The genetic similarity coefficients ranged from 0.10 to 0.73 with the mean of 0.34. The results showed as follows: (1) Distinct genetic differences were present among the different species; (2) The different accessions in a species were clustered together, respectively, which had larger genetic similarities and closer relations; (3) The species with similar morphological characters and the species from the same areas or neighboring geographical regions were clustered together; (4) RAMP results are basically comparable with those obtained from studies on morphology. It is a useful method for analysis of the genetic diversity and similarities in Leymus.     

The genetic characterization of Turkish watermelon (<Emphasis Type="Italic">Citrullus lanatus</Emphasis>) accessions using RAPD markers

Genetic diversity of the Turkish watermelon genetic resources was evaluated using different Citrullus species, wild relatives, foreign landraces, open pollinated (OP) and commercial hybrid cultivars by RAPD markers. The germplasm was consisted of 303 accessions collected from various geographical regions. Twenty-two of 35 RAPD primers generated a total of 241 reproducible bands, 146 (60.6%) of which were polymorphic. Based on the RAPD data the genetic similarity coefficients were calculated and the dendrogram was constructed using UPGMA (Unweighted pair-group method with arithmetic average). Cluster analysis of the 303 accessions employing RAPD data resulted in a multi-branched dendrogram indicating that most of the Turkish accessions belonging to var. lanatus of Citrullus lanatus (Thunb.) Matsum et Nakai were grouped together. Accessions of different Citrullus species and Praecitrullus fistulosus (Stocks) Pangalo formed distant clusters from C. lanatus var. lanatus. Among 303 accessions, a subset of 56 accessions was selected representing different groups and a second dendrogram was constructed. The genetic similarity coefficients (GS) within the Turkish accessions were ranged from 0.76 to 1.00 with 0.94 average indicating that they are closely related. Taken together, our results indicated that low genetic variability exist among the watermelon genetic resources collected from Turkey contrary to their remarkable phenotypic diversity.     

生物表面活性剂产生菌的筛选及其发酵条件的初步优化

以石油为 C 源,经过富集培养从石油污染土壤中筛选出 15 株具有较强产生物表面活性剂能力的菌株.其中菌株 BS-5 产生物表面活性剂的能力最强,该菌发酵液的表面张力可达 27.3 mN/m(空白发酵液表面张力为 54.5 mN/m).通过红外光谱分析发现该菌产生的生物表面活性剂为糖脂类物质.另外,通过正交试验对该菌的培养基条件进行初步优化,结果以植物油 10 g/L、MgSO4 0.2 g/L、K2HPO4 1.0 g/L、KH2PO4 1.0 g/L、蛋白胨 1.0 g/L、FeSO4 0.05 g/L、CaCl2 0.02 g/L、初始 pH 值 7.5 为最佳.通过16S rDNA 测序结果表明该菌为铜绿假单胞菌 (Pseudomonas aeruginosa).     

绿僵菌航天诱变菌株的DNA变异性分析

昆虫病原真菌绿僵菌M2189菌株经航天搭载诱变后,分离得到200多个单孢菌株。以9个随机引物对47个诱变菌株进行PCR扩增,获得了38个位点的共1381条DNA片段。统计分析表明,47个诱变菌株在DNA水平上发生了不同程度的变异,变异系数在0.062~0.406之间,平均为0.176,>0.3的有3个菌株。由聚类分析可将47个诱变菌株划分为14个组群,组间变异率小于0.2,与原始菌株相比变异系数接近的诱变菌株无规律地分散在不同组群中,说明航天诱变导致DNA变异的位点和频率是随机的、分散的,并无特定位点。航天诱变为选育生物防治优良菌株提供了新途径。     

The assessment of genetic variation in Spanish primitive cultivars of barley, Hordeum vulgare L., by a combination of isozymes and hordeins

Five isozyme and endosperm reserve protein systems were analysed using electrophoretic techniques in order to investigate the genetic diversity of 222 accessions of Spanish, local varieties of barley, Hordeum vulgare L., maintained at the Centro de Conservación de Recursos Fitogeneéticos of the I.N.I.A. (Alcalá de Henares, Spain). The esterase (EST) isozymes provided more information than did the other systems analysed, showing a total of 14 variable markers. The cathodic peroxidases (CPX) and acid phosphatases (ACPH) were also polymorphic. Malate dehydrogenase (MDH) and glutamate oxaloacetate transaminase (GOT) were monomorphic. The hordeins showed patterns of up to 15 bands, the majority of which were very useful in distinguishing genotypes. 17.2% of accessions showed a uniform genotype, 29.8% showed practically identical genotypes and 53% showed mixtures of different genotypes. It is noteworthy that the use of only two systems (EST and hordeins) and the analysis of only six loci (Est-1, Est-2, Est-4, Hor-1, Hor-2 and Hor-3) is sufficient to reveal the genetic diversity of the collection.