利用ＲＣＲ—ＲＡＰＤ法和同功酶分析法日本柳杉无性系的识别方法研究

以日本福岛县林业试验场日本柳杉种子园的２４个无性系为材料,利用ＰＣＲ－ＲＡＰＤ法和同功酶分析法对日本柳杉无性系的识别方法进行,结果显示：同功酶法和由７个标记构成的ＲＡＰＤ分析法不能完全识别２４个无性系,用１７个和２７个标记构成的ＲＡＰＤ分析法识别这２４个无性系是可能的;同功酶标记是一种基因型标记,具有稳定性高的特点,适合于分析遗传的构造的相似程度,特别适合于群体遗传分析,但是,由于能获得清晰谱带的同工酶种类有限,在用于个体识别上比较困难;ＲＡＰＤ标记是一种显性标记,稳定性较差,但只要反应体系和条件稳定,利用ＲＡＰＤ标记正确进行无性系识别是完全可能的。     

板栗优良品种（无性系）苗木分子标记鉴别研究）

利用ＲＡＰＤ标记对浙江省林木良种审定委员会审定（认定）和通过省级鉴定的８个板栗品种及无性系进行指纹分析，鉴别出各板栗品种（无性系），实验过程中对６８４个随机核苷酸引物进行了重复筛选，经筛选共获得９个稳定的ＲＡＰＤ标记，构建了板栗的ＮＤＡ指纹图谱，经过大量的实验，确定了鉴别板栗优良品种（无性系）的较为理想的程序，获得了ＲＡＰＤ良好的重复性，建立了较完整的板栗ＲＡＰＤ分析的技术体系，用大田苗进行步验证，结果可靠。     

运用RAPD技术进行杉木无性系识别研究

以湖北省的４０个杉木优树为研究材料,初步开展了运用ＰＣＲ—ＲＡＰＤ技术对杉木优树进行识别的研究。研究表明,运用 ８个引物的 １７个分子标记可以对其中的３８个无性系进行识别,只有２个无性系相互不能识别。说明湖北省杉木遗传资源变异幅度较大,可以运用 ＤＮＡ技术对优树进行品系管理。     

用RAPD技术估测柳树种及无性系的变异

利用１２个引物对柳树２种１６个无性系基因组分别进行了ＲＡＰＤ检测，从电泳图谱可直观看出，引物ＣＨＬ－１可对柳树两种识别：引物Ｄｅｃａ－４可对１６个无性系基因组扩增产物经电泳产生的图谱中ＤＮＡ条的统计，利用ＮＴ－ＳＹＳ对其进行分析建立系统树，结果表明，利用ＲＡＰＤ获得的两种柳树种的分类结果与经典方法得到的结果一致。     

银杏主要栽培品种的分子鉴别

以１５个银杏主要栽培品种的种子胚乳为材料,利用筛选出来的１２种具多态型的引物进行ＲＡＰＤ分析．结果表明,１５个品种之间在分子水平上存在很大差异,并确定了这些品种的ＲＡＰＤ标记基因型．以标记基因型为依据,可以利用单倍体种子胚乳和二倍体叶片组织在ＤＮＡ水平上对这些品种进行有效的鉴别．依据各个品种的标记基因型对这些品种进行分子聚类的结果与传统的三大类分类法非常吻合．     

筛选与杨树抗云斑天牛基因连锁的RAPD标记

筛选与杨树抗云斑天牛基因连锁的ＲＡＰＤ标记王京兆，王斌，卞学瑜，韩一凡（中国科学院遗传研究所北京１００１０１）（中国林业科学研究院林业研究所北京１０００９１）关键词杨树，云斑天牛，ＲＡＰＤ，多态性产物植物抗虫基因工程是当前比较活跃的研究领域，但真正能...     

日本柳杉无性系的引种保存与扦插繁殖

为了引种抗雪压能力强的日本柳杉优良繁殖材料,从日本冬季降雪多的地区引进了52个柳杉无性系.通过就地栽植和假植两种方式进行保存,并开展扦插繁殖、促萌增穗试验.结果表明:提前整地和遮阴保湿是成功保存日本柳杉无性系的关键措施;不同无性系类别间的扦插成活率有差异,用黄心土作基质扦插的效果好于石英砂;截枝法、截干法均能促进枝条萌发,截枝法略优于截干法.     

美洲黑杨(Populus deltoides Marsh)×青杨(P.cathayana Rehd.)分子连锁图谱的构建

利用随机扩增ＤＮＡ多态性ＲＡＰＤ方法，在美洲黑杨（Ｐ．ｄｅｌｔｏｉｄｅｓＭａｒｓｈ）×青杨（Ｐ．ｃａｔｈａｙａｎａＲｅｈｄ．）３代谱系中分析分子标记，构建出第１张美洲黑杨×青杨分子连锁图谱。共从３００个１０ｍｅｒ随机引物中筛选出７９个适合引物，检测出可供构图的分离标记１８０个。该图谱由２０个连锁群，１１０个ＲＡＰＤ标记组成。总图距为覆盖基因组总长度的７０３５％，标记间的平均间距为１７２７ｃＭ．连锁群长度在３７１～１８９８，相应标记分别在３～１０。本图谱为杨树抗病、虫和其它性状基因定位提供了框架结构，为实现杨树分子遗传育种迈进了最重要的一步。     

杨树抗云斑天牛纤维材无性系选育*

以较抗天牛的Ｉ－６９杨为母本，以Ｉ－６３杨、欧洲黑杨和晚花杨２７２为父本，于１９８３年春进行人工控制授粉，组合编号分别为３４、３２和３７。经苗期选择选出优良单株营造比较林，经５年生无性系比较林和９年生无性系比较林测定，采用主成分分析、主成分遗传距离聚类分析和选择指数，选出速生、抗虫和材质较好的纤维材无性系３４－３０１和３４－２８６。这两个新选出的无性系（３４－３０１和３４－２８６），在９年生时与对照Ｉ－６９杨相比材积分别提高２４．９％和１６．６％，抗虫性明显提高（单株幼虫数分别为０．４和０．５，而Ｉ－６９杨为５．３８）；与３４－３１４相比材积分别提高１６．７％和８．９％。化学物质测定表明，抗虫性与树皮中酚类含量和丹宁含量呈正相关。分子生物学测定表明，杨树对云斑天牛的抗性受基因控制，找到了与抗虫基因连锁的ＲＡＰＤ分子标记。     

美洲黑杨基因资源收存及其遗传评价的研究

从１７个国家引进３３１个黑杨派无性系（其中５２个美洲黑杨无性系）,在山东省长清县营建我国第一个黑杨派无性系基因库。连续１０年对基因库内美洲黑杨无性系进行了多性状系统研究,结果表明各无性系间在物候期、生长、生根、抗寒和抗病虫等方面均存在着显著的差异,遗传变异丰富;系间的主要性状与无性系起源纬度及各性状间存在一定相关性。主要材料性状变异的研究结果表明,３８个８年生美洲黑杨无性系在木材基本密度、纤维长度无性系间变异达到极显著水平;木材密度和纤维长度与树高和胸径呈一定相关性;木材密度和纤维长度这两个性状在遗传上相互独立,受不同遗传机制控制。利用分子遗传标记ＲＡＰＤ技术研究库内美洲黑杨无性系ＤＮＡ多态性,结果表明本库美洲黑杨ＤＮＡ多态率为８６％,再次证明本库美洲黑杨遗传多样性高,可为我国杨树改良提供丰富的育种材料。     

Molecular identification of sex in <Emphasis Type="Italic">Hippophae rhamnoides</Emphasis> L. using isozyme and RAPD markers

In many dioecious plants, gender affects economic value, breeding schemes and opportunities for commercial harvests. Hippophae rhamnoides L. is a dioecious plant species in which female genotypes are commercially preferred over male genotypes. Its berries have rich medicinal, nutritional and pharmaceutical properties because of their large amounts of vitamins, essential oils, proteins, fatty acids, free amino acids and flavanoids. Primary limitation for breeding H. rhamnoides L. is its dioecious nature, since gender cannot be identified by traditional methods. Therefore, some reliable and quick methods need to be developed. This communication deals with the development of isozyme and RAPD markers for early sex identification in this dioecious tree. The isozyme analysis was conducted with four enzyme systems, viz. peroxidase, esterase, malate dehydrogenase and catalase. The peroxidase enzyme system produced a female specific sex marker, which successfully differentiated between the staminate and pistillate genotypes of H. rhamnoides L. Thirty five random decamer primers were used in our study and one male sex linked marker was identified. OPD-20 (5′-ACTTCGCCAC-3′) displayed a band at 911 bp that expressed polymorphism between male and female genotypes. The staminate and pistillate genotypes could be distinguished using RAPD marker OPD-20911. These results revealed the immense potential of peroxidase isozyme patterns and RAPD as genetic markers for sex identification in H. rhamnoides L.     

美国山核桃主要栽培品种的RAPD鉴定

以叶片为材料,采用改良CTAB法和16种Operon引物对从原产地美国引进的30个美国山核桃主要栽培品种进行RAPD分子分析。结果表明：参试品种问存在明显的基因带型差异,多态型比率高达78．1％;根据RAPD特异分离谱带确定了各品种的标记基因型．以此可实现对美国山核桃无性系品种DNA水平上的鉴别;以标记基因型对30个美国山核桃品种进行系统聚类,结果与传统形态学及Marquard等人的同工酶分类基本吻合。     

分子标记在林业辅助选择育种中的应用

本文主要阐述分子标记在林木辅助选择育种中的应用。利用多种分子标记(RAPD,RFLP,AFLP,STS,SSR,STR等),可以在林木早期生长阶段对一些性状进行鉴别, 构建单种分子标记遗传连锁图谱或几种分子标记共存的混合连锁图谱和对控制数量性状的基因进行定位, 对林木群体遗传结构、遗传变异、遗传分化和基因流动进行研究, 在基因工程中, 能够追踪目的基因行为和对控制质量性状的基因进行鉴别, 对单株进行指纹图谱, 对种子质量进行监测和对品系、品种和无性系进行鉴定。     

白桦种群间木材纤维性状的表型变异与分子标记的遗传多态性

以东北地区的5个白桦天然种群为研究材料,在木材纤维形态性状测定的基础上,采用ISSR、RAPD分子标记研究了5个种群的遗传变异以及种群间纤维形态性状与DNA分子标记的相关性.结果表明:白桦天然林成熟材(18～31年生)的木材纤维长度在种群间差异不显著,纤维宽度和长宽比差异显著,帽儿山种群的木材纤维形态性状最好,汪清最差;利用ISSR和RAPD 2种标记检测5个白桦种群的遗传多样性,所得的研究结果基本一致,多态位点比率最高的为辽宁新宾种群,其次是帽儿山和汪清,而塔河和金山屯最低;ISSR标记的AMOVA分析表明白桦种群间的遗传变异占总遗传变异的10.82%;RAPD标记的AMOVA分析表明白桦种群间的遗传变异占总遗传变异的12.94%,研究结果均显示白桦的遗传变异主要发生在种群内部.利用2种标记技术的种群聚类结果基本一致,这一结果与基于木材纤维性状的表型聚类基本一致, 而且种群间遗传距离和地理距离之间存在一定的相关性.     

Adaptive genetic diversity and population structure of the “algarrobo” [<Emphasis Type="Italic">Prosopis chilensis</Emphasis> (Molina) Stuntz] analysed by RAPD and isozyme markers

The “algarrobo” [Prosopis chilensis (Molina) Stuntz] is a tree species that represents an important natural resource in arid and semi-arid regions of Argentina. In this paper, we analysed and compared the variability of 46 RAPD (Random Amplified Polymorphic DNA) loci with previous estimates obtained from 12 isozyme markers in nine Argentinean populations of P. chilensis representative of the whole range of this species in Argentina. We evaluated the population structure and the existence of genetic variants associated with environmental variables. Expected heterozygosity (H _e) estimated from RAPD varied significantly among populations and regions. Hierarchical analysis of genetic variability (AMOVA) showed that most (88.1%) of the total diversity occurs within populations, the component among populations within regions (9.3%) was intermediate, while the between-region component was the lowest (2.6%). All three variance components were highly significant. The MDS plot from pair-wise Φ _ST matrix was consistent with the highly significant among-region differentiation indicated by the AMOVA. All 12 variable isozyme loci and 26 out of 46 RAPD loci showed significant or highly significant association with at least one geographic/climatic variable according to the stepwise multiple regression analysis. These results imply that the genetic differentiation among populations is better explained by environmental or biogeographical grounds than by geographical distances, suggesting gametic disequilibrium with loci responsible for the adaptation to particular environmental conditions. The information from RAPD markers would provide a relevant criterion to preserve genetic diversity in programmes for conservation and rationale use of this species.     

利用RAPD标记进行银杏雄株无性系的识别

利用RAPD分子标记技术对来自10个省份的银杏雄株10个无性系的研究表明,12对引物共扩增出了96条带,多态性条带41条,占总带数的42%,扩增片段长度为300~2 000 bp,10个银杏雄株无性系间表现出较丰富的多态性;利用12对引物的41个分子标记完全可以对10个不同银杏雄株无性系进行识别,谱带清晰,因此利用RAPD技术进行银杏雄株无性系识别是可行的。     

Molecular characterization and marker based chemotaxonomic studies of Podophyllum hexandrum Royle

Detailed chemical studies and RAPD analysis were done in different populations of Podophyllum hexandrum collected from high altitude regions of North Western Himalayas. Random amplified polymorphic DNA (RAPD) analysis revealed a high degree of genetic diversity among the 12 collected accessions, attributed to their geographical and climatic conditions. HPLC analysis also revealed variation in the concentration of two major marker compounds which lead to the identification of a chemotype. The study demonstrated that RAPD and chemical markers are very useful tools to compare the genetic relationship and pattern of variation among such prioritized and endangered medicinal plants.     

Use of DNA markers in forest tree improvement research

DNA markers are rapidly being developed for forest trees. The most important markers are restriction fragment length polymorphisms (RFLPs), polymerase chain reaction- (PCR) based markers such as random amplified polymorphic DNA (RAPD), and fingerprinting markers. DNA markers can supplement isozyme markers for monitoring tree improvement activities such as; estimating genetic diversity in breeding populations, germplasm identification, verifying controlled crosses, and estimating seed orchard efficiencies. Because the number of DNA markers is potentially limitless, it should be possible to map individual quantitative trait loci (QTL) by linkage analysis with high-density maps. Finally, if such associations can be found, it may also be possible to design marker-assisted breeding strategies for forest trees.     

Genetic diversity and phylogenetic relationships between and within wild Pistacia species populations and implications for its conservation

Although cultivation and utilization of Pistacia are fully exploited, the evolutionary history of the Pistacia genus and the relationships among the species and accessions is still not well understood. The aim of this study was to analyze random amplified polymorphic DNA(RAPD) in a total of 50 accessions of wild pistachio species, which included five populations Pistacia vera, Pistacia khinjuk,Pistacia atlantica, Pistacia mutica, and Pistacia eurycarpa.High levels of genetic diversity were detected within wild pistachio accessions, as revealed by using the unweighted pair-group method with arithmetic averaging and supported via analysis of molecular variance. The objectives of this investigation were to estimate marker indices, polymorphic information contents(PICs), and genetic similarities(GS)for RAPD markers; assess the genetic diversity of Pistacia species, using GS estimated from RAPD fingerprints and molecular characterization; and facilitate the use of markers in inter-specific introgression and cultivar improvement.Out of the 149 polymerase chain reaction fragments that were scored, 146(97.98 %) were polymorphic. Genetic similarities ranged from 0.3 to 0.86 %, marker indices ranged from 2.98 to 17.74 %, and PICs ranged from 0.80 to0.99 %. Our results provided great molecular identification of all assayed genotypes, which have shown that there is large quantity of genetic diversity among the pistachio accessions. This finding might render striking information in breeding management strategies for genetic conservation and cultivar development.