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。     

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。     

中国松树枯梢病菌遗传多态性的RAPD分析

运用随机增增多态ＤＮＡ（ＲＡＰＤ）技术对发生于我国１３省区１６种松树和其它２种针叶树上的松枯梢病菌（Ｓｐｈａｅｒｏｐｓｉｓ ｓａｐｉｎｅａ）的５５个菌株进行基因组ＤＮＡ多态性分析。用１７个随机引物经ＰＣＲ扩增共得到２００个ＲＡＰＤ标记,其中９８．５％具有多态性。ＵＰＧＭＡ聚类分析确定了供试菌株间的亲缘关系,将５５个菌株分为３个类群。各菌株间的差异与其寄主种类无明显关系,与其地理来源在某些类群间有一定联系,但在大多数菌株间相关趋势不明显。     

西种竹子的RAPD指纹图谱的初步研究

本实验以ＲＡＰＤ技术对考顺竹，凤尾竹，绿竹，白绿竹四个品种用２０种已知序列的１０ｎｔ引物进行ＰＣＲ反应扩增，其中有１７种可扩增出ＤＮＡ条带，构成ＲＡＰＤ指纹图谱，各中引物ＰＣＲ扩增的ＤＮＡ条带数目在０－６条之间，大小在０．３－２．０ｋｂ之间，各种竹子的１７种引物扩增的ＤＮＡ条带总数在２３－４４条之间，四种竹子两两之间的相似系数在３６－７３％之间。     

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了比较研究。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,由于能获得清晰话带的同功酶种类有限,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。     

板栗主要栽培品种的分子鉴别

应用ＲＡＰＤ分子标记手段，研究板栗品种的遗传多样性，建立了板栗品种ＲＡＰＤ标记的标准程度，以及板栗品种种的ＤＮＡ指纹数据库，并为板栗品种鉴别提供了准确、可靠的标准方法，４６个板栗品种的样品的ＤＮＡ，用１６个引物进行扩增反应的位点总数为１１９个，产生６９个多态位点，建立了４６个板栗品种的ＤＮＡ指纹数据库，并分析出这些板栗品种分子鉴别的最优化引物组合。对特定的引物组合，计算品种间的遗传距离矩阵，从而确定引物组合对这些品种鉴别的特异性。     

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。     

杉木幼林根圈土壤磷酸酶活性、磷组分及其相互关系

通过对杉木人工林根圈土壤两种磷酸酶活性、８种磷组分及其相互关系研究，结果表明：（１）根圈酸性、中性磷酸酶活性分别比根圈外高１．４４、０．６６酚ｍｇ／ｇ（３７℃，１２ｈ），Ｒ／Ｓ值分别为１．９１和２．０１；（２）根圈全Ｐ、ＤＡ－Ｐ、Ａｌ－Ｐ、Ｆｅ－Ｐ、Ｈ２ＳＯ４－Ｐ分别比根圈外高２０３．４５，２．２１，３．０５，１０．９３，１０．３３ｍｇ／ｋｇ，Ｒ／Ｓ值分别为１．６１，４．６８，３．６１，１．９７，２．４６，Ｃａ－Ｐ比根圈外低２．３４，Ｒ／Ｓ值为０．７２，Ｉ－Ｐ和Ｏ－Ｐ无显著差异；（３）根圈酸性磷酸酶活性与ＤＡ－Ｐ和Ｈ２ＳＯ４－Ｐ呈显著正相关，ｒ值分别为０．５８４和０．５７９，中性磷酸酶活性与全Ｐ、Ｉ－Ｐ和Ｏ－Ｐ呈显著或极显著正相关，ｒ值分别为０．５９４，０．７７３和０．６８６。     

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．     

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。     

DNA fingerprinting of Populus trichocarpa clones using RAPD markers

Nine trees from a single, natural population of black cottonwood (Populus trichocarpa Torr. et Gray) in Alaska were screened for randomly amplified polymorphic DNA (RAPD) markers with ten different 10-base random oligonucleotide primers in order to evaluate the use of RAPD analysis for distinguishing black cottonwood clones. Nine primers amplified the genomic DNA targets; two primers were able to differentiate all clones. Eight clones were distinguished among the nine tree samples assayed. Two trees showed identical banding patterns with all primers used; therefore it is suggested that these trees are from the same clone. The RAPD fingerprinting method is simple and powerful-one primer can distinguish different clones, while the use of multiple primers reduces fingerprint similarity and resolves discrepancies.     

Differentiation of poplar and willow clones using RAPD fingerprints

The technique of random amplified polymorphic DNA (RAPD) fingerprinting was used to differentiate species and identify clones of 15 poplar and 15 willow clones (ramets of three poplar clones were also included to verify the stability of the results). Four random DNA primers (Deca-11, Chl-1, Chl-4 and Chl-10) and an M13 universal primer were used to determine DNA polymorphism among the clones. Based on the DNA banding pattern obtained with the four DNA primers by polymerase chain reaction (PCR) amplification we conclude that RAPD fingerprinting with properly selected primers can be used for clonal and species differentiation of poplar and willow. The technique is simple, accurate and inexpensive.     

Multiallelic and multilocus simple sequence repeats(SSRs) to assess the genetic diversity of a Salix spp. germplasm collection

Salix L.(willow) is the largest genus of the family Salicaceae and plays an important role in riparian habitats,wetlands and in shrub tundra.Due to the different implications for the species belonging to this family,it is fundamental to identify molecular tools characterizing relevant clones.A set of six multilocus and multiallelic simple sequence repeat(SSRs) markers are presented,leading to390 polymorphic fragments considered as single dominant markers and able to discriminate successfully 92 S.alba L.from 24 Salix spp.The polymorphic fragments have been used to perform genetic diversity studies,and to investigate population structures and cluster analysis in a germplasm collection.The results highlight the capability of the six SSRs to be powerful genetic resources in applied forestry research,both to distinguish S.alba clones from Salix spp.and to perform genetic population studies for breeding programs.     

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。     

常山胡柚优良无性系的RAPD分析

采用改良的CTAB法成功提取到常山胡柚叶片DNA，并对常山胡柚的12个优良无性系及7个优株之间进行了RAPD分析。从初选的0peron公司的60个随机引物中又筛选出12个扩增效果较好的引物，共产生随机扩增位点103个，平均每个引物产生8．6个识别位点，但都没有发现多态性位点。由此表明，目前所选的常山胡柚优良无性系及优株之间在分子水平上并无什么差异，其经济性状方面的差异可能主要受生长发育阶段、生态环境条件及经营管理水平等因素的影响。     

8个柳树无性系冠形、主干分枝结构及生长比较研究

柳树无性系,变异大,形态多样,根据平原地区用材、防护、观赏的需要,对比分析柳树优良品种苏柳172、苏柳795、金丝垂柳1011和新选出的苏柳549、苏柳797、苏柳335、苏柳736、苏柳1034等新无性系的生长、冠形和枝干结构的差异。     

四种竹子的RAPD指纹图谱的初步研究

本实验以RAPD技术对考顺竹、凤尾竹、绿竹、白绿竹四个品种用20种已知序列的10nt引物进行PCR反应扩增。其中有17种可扩增出DNA条带,构成RAPD指纹图谱。各种引物PCR扩增的DNA条带数目在0—6条之间,大小在0．3—2．0kb之间。各种竹子的17种引物扩增的DNA条带总数在23—44条之间。四种竹子两两之间的相似系数在36—73％之间。     

运用层次分析法优选临夏北塬农田防护林树种

选取对农田防护林体系有重要影响的树种适应性、树种防护效果和社会经济状况3大类13个指标,利用层次分析法,对所选10个树种(或无性系)进行优选评价.结果表明,在影响优选评价因子中,树种的保存率贡献率最大,说明选择适生树种是营造农田防护林成功的前提;防护效果评价因子,尤其是树种的防风效果,是农田防护林能否得到充分发挥综合防护效益的关键,对优选分析评价具有重要影响;同时考虑农村社会经济方面因素,才能得出客观、准确的优选评价结果.10个树种(或无性系)优选评价排序为:苏柳J172＞苏柳J369＞苏柳J194＞青刚柳＞准噶尔柳＞二白杨＞新疆杨＞河北杨＞三倍体毛白杨BT85＞三倍体毛白杨B1008.     

Development of AFLP and RAPD markers linked to a locus associated with twisted growth in corkscrew willow (Salix matsudana 'Tortuosa')

Salix matsudana Koidz. cultivar 'Tortuosa' (corkscrew willow) is characterized by extensive stem bending and curling of leaves. To investigate the genetic basis of this trait, controlled crosses were made between a corkscrew female (S. matsudana 'Tortuosa') and a straight-stemmed, wild-type male (Salix alba L. Clone 99010). Seventy-seven seedlings from this family (ID 99270) were grown in the field for phenotypic observation. Among the progeny, 39 had straight stems and leaves and 38 had bent stems and curled leaves, suggesting that a dominant allele at a single locus controls this phenotype. As a first step in characterizing the locus, we searched for amplified fragment length polymorphism (AFLP) and randomly amplified polymorphic DNA (RAPD) markers linked to the tortuosa allele using bulked segregant analysis. Samples of DNA from 10 corkscrew individuals were combined to produce a corkscrew pool, and DNA from 10 straight progeny was combined to make a wild-type pool. Sixty-four AFLP primer combinations and 640 RAPD primers were screened to identify marker bands amplified from the corkscrew parent and progeny pool, but not from the wild-type parent or progeny pool. An AFLP marker and a RAPD marker linked to and flanking the tortuosa locus were placed on a preliminary linkage map constructed based on segregation among the 77 progeny. Sectioning and analysis of shoot tips revealed that the corkscrew phenotype is associated with vascular cell collapse, smaller cell size in regions near the cambium and less developed phloem fibers than in wild-type progeny. Identification of a gene associated with this trait could lead to greater understanding of the control of normal stem development in woody plants.     

紫胶虫主要生产种的RAPD分子标记分析

用RAPD技术研究了紫胶虫7种主要生产种12个群体之间的亲缘关系,研究中每个群体使用了24个标本,共有288个个体用于研究.在试用的45种随机引物中,筛选出9种引物用于随机引物扩增,共得到121个清晰稳定的位点,其中118个为多态性位点,多态百分率为97.52%,每条引物检测到的位点数为11～16个,长度为150～3 000 bp.根据扩增结果,用UPGMA法对Nei's遗传距离进行聚类分析,构建分子系统树.7种紫胶虫聚类结果显示:紫胶虫各群体间的遗传距离为0.044 3～0.791 7,其中,种间的遗传平均距离为0.443 0,种内的遗传平均距离为0.170 7.根据聚类分析,讨论了紫胶虫主要生产种的亲缘关系.