香菇菌株限制性片段长度多态性

香菇菌株的ＤＮＡ片段通过隆转移到质粒载体Ｂｌｕｅｓｃｒｉｐｔ Ｍ１３－（ｐＢＳ）以获得部分基因文库。文库三个随机克隆和一个密环菌中１ＤＮＡ克隆被用作限制性片段长度多态性探针。其中三个探针可检测到多态性，提示了３２个野生和栽培香菇菌株的遗传分离。菌株间的相似性用平均链锁聚类方式（ＵＰＧＭＡ）聚类，相似性超过６０％的分为一组。２３个菌株被分成４组。９个菌株与其它菌株相似性均低于６０％而独立存在。从我们     

猕猴桃的组织培养和快速繁殖

１ 植物名称 金魁猕猴桃（Ａｃｔｉｎｉｄｉａ ｄｅｌｉｃｉｏｓａｖａｒ．Ｊｉｎ Ｋｕｉ）２ 材料类别 嫩茎３ 培养条件 培养基：①ＭＳ＋ＺＴＷ．０ｍｓ／Ｌ；②ＭＳ＋ＺＴ１．０ｍｐ／Ｌ；＠ ＭＳ＋６ＢＡ２．０ｍｇ／Ｌ＋ＮＡＡ ０．１ｍｇ／Ｌ；④１／２ＭＳ＋ＩＢＡ０．５～０．７ｍｇ／Ｌ。 蔗糖（市售白砂糖）浓度①～③培养基为３．０％，④培养基为 ２．０％；琼脂浓度为 ０．６５％～０．７０％，ｐＨ５．８～６．０，培养温度 ２５～２８℃，每天光照 １３ｈ，光照强度 １５００～２０００ １Ｘ。４ 生长与分化情况４．１ 愈伤…     

猕猴桃ACC氧化酶cDNA克隆及全序列测定

从‘秦美’猕猴桃（ＡｃｔｉｎｉｄｉａｄｅｌｉｃｉｏｓａＣ．Ｆ．ＬｉａｎｇｅｔＡ．Ｒ．Ｆｅｒｇｕｓｏｎ．ｃｖ．Ｑｉｎｍｅｉ）呼吸高峰跃变初期的果实中提取总ＲＮＡ，然后纯化得ｍＲＮＡ，用Ｏｌｉｇｏ（ｄＴ）作为引物反转录合成ｃＤＮＡ第一链，以此为模板，用人工合成的寡聚核苷酸作为引物进行扩增，得到大小约０．９５Ｋｂ基因片段，并将其克隆到ｐＧＥＭ－５ｚｆ（＋）载体上。经限制性内切酶图谱分析，组建亚克隆并进行了全序列测定，在其开放读码框架内，由９５７个ｂｐ编码３１９个氨基酸组成，该ｃＤＮＡ与海沃德ＡＣＣ氧化酶ｃＤＮＡ的核苷酸和氨基酸残基同源率分别达９５．０１％和９５．６１％，证明己克隆到完整的秦美猕猴桃ＡＣＣ氧化酶基因编码区。     

运用RAPD对核桃属种间亲缘关系的研究

用随机扩增多态性ＤＮＡ（ＲＡＰＤ）技术对核桃属内９个种及近缘属的２个种进行基因组ＤＮＡ多态性分析,共选用２０个１０ｂｐ随机引物扩增出３１１个ＤＮＡ片段,其中铁核桃Ｊ．ｓｉｇｉｌｌａｔａ,甘肃枫杨Ｐ．ｍａｃｒｏｐｔｅｒａ显示出自身特征性标记,利用这些片段进行遗传关系分析,根据ＵＰＧＭＡ方法构建聚类树状图。研究结果表明：铁核桃为核桃属中一个独立种;核桃属内组间和种间亲缘关系与经典分类学的结果完全一致;     

提高苹果基因转化效率的研究

采用根瘤农杆菌（Ａｇｒｏｂａｃｔｅｒｉｕｍ　ｔｕｍｅｆａｃｉｅｎｓ）强株系ＥＨＡ１０５（ｐ３５ｓＧＵＳ－ｉｎｔｒｏｎ）研究了影响‘皇家嘎拉’苹果（Ｍａｉｎｓ ｄｏｍｅｓｔｉｃａ Ｂｏｒｋｈ．）外植体的β-葡萄糖醛酸酶（ＧＵＳ）基因的瞬时、稳定表达水平和转基因植株的再生。证明在培养基生长素（ＩＢＡ、ＮＡＡ）存在的条件下,外植体的ＧＵＳ基因的瞬时表达水平提高了３－４倍,而共培养两周后稳定表达水平提高２倍以上,产生９．８个ＧＵＳ愈伤组织表达区域。白化处理促进外植体的基因转化,白化处理的新梢顶端第一节间外植体ＧＵＳ表达区域比常用的叶片外植体高４倍。在生长素存在的条件下 ２％外植体获得了转基因植株。Ｓｏｕｔｈｅｒｎ　ＢｌｏｔＤＮＡ杂交和组织化学染色分析证明ＧＵＳ基因已整合到苹果的染色体上,并得以表达。     

埃斯(Ace)苹果的离体培养及繁殖

埃斯（Ａｃｅ）苹果的离体培养及繁殖１植物名称苹果品种埃斯。２材料类别多年生树茎尖生长点。３培养条件（１）起始培养基：ＭＳ＋６－ＢＡ３．０ｍｇ／Ｌ＋ＮＡＡ０．０５ｍｇ／Ｌ＋ＣＨ３００ｍｇ／Ｌ＋３％食用绵白糖；（２）增殖培养基：ＭＳ＋６－ＢＡ１．５ｍｇ／...     

籽用西瓜品种(系)间亲缘关系的RAPD分析

用随机扩增多态DNA（RAPD）标记方法对籽用西瓜的8个品种（系）和西瓜种内其它变种的4个品种（系）进行了遗传多样性的检测。结果表明：筛选出的14个随机引物都能扩增出多态性DNA片段,反映出了不同品种（系）的遗传差异;计算了这12个西瓜种品种（系）间的Net’s相似系数,建立了UPGMA系统树,得出了它们之间的亲缘关系。     

大白菜品种同功酶及水溶蛋白的遗传多样性分析

用电泳技术分析了大白菜（ＢｒａｓｉｃａｃａｍｐｅｓｔｒｉｓＬ．ｓｐ．ｐｅｋｉｎｅｓｉｓ）４９个品种苗的葡萄糖磷酸变位酶（ＰＧＭ）、过氧化物酶（ＰＯＸ）、脂酶（ＥＳＴ）和６－磷酸葡萄糖酸脱氢酶（６－ＰＧＤ）的同功酶谱，以及种子水溶蛋白（ＰＲＯＴ）的图谱。从中分析了１３个表现出品种之间多样性的位点。根据这些位点上带型的差异，可以区分供试的所有大白菜品种。图谱上品种之间的差异位点，可以用于杂交种Ｆ１代的纯度鉴定     

桃果实ACC合酶cDNA的克隆

根据其它植物ＡＣＣ合酶氨基酸保守区设计两组简并引物,用套式ＰＣＲ技术从桃成熟果实ｃＤＮＡ中扩增出１．１ｋｂ大小特异性片段,将其克隆至ｐＧＥＭ－Ｔ载体上,对重组克隆ｐＰＡＣＳＩ进行全序列测定表明,插入片段全长１１００个碱基,编码３６６个氨基酸,该基因与番茄、笋瓜、小西葫芦、康乃馨、苹果ＡＣＣ合酶ｃＤＮＡ氨基酸序列同源性分别为７２．７％,７１．３％,７１．１％,６９．１％,６５．４％。ＲＮＡ点杂交表明ｐＰＡＣＳＩ基因随着桃果实成熟表达活性增强,乙烯处理能诱导桃果实该基因的表达。     

蜂斗叶的组织培养与快速繁殖

通过组织培养，蜂斗叶（ＰｅｔａｓｉｔｅｓｊａｐｏｎｉｃｕｓＭｉｑ．）的叶柄和地下茎切段在ＭＳ＋ＢＡ２ｍｇ／Ｌ的分化培养基上诱导出愈伤组织，并进一步分化出芽丛，切下２～３ｃｍ高的芽，移入１／２ＭＳ＋ＮＡＡ０．１ｍｇ／Ｌ的生根培养基中，生根率达１００％。以再生苗的叶片或叶柄为外植体，在分化培养基上很快便可再形成愈伤组织并分化出芽。     

河南17 个桂花品种的RAPD 分析

 采用改良CTAB 法提取河南17 个桂花品种的基因组DNA , 利用RAPD 技术对其进行鉴定和分类研究。从100 个10 bp 的随机引物种筛选出15 个扩增效果较好的引物进行扩增, 共产生121 条带, 其中87 条为多态性带。根据扩增结果进行聚类分析, 得出反映各品种间亲缘关系的树状图。各个品种群内的不同品种间的亲缘关系接近, 而不同品种群间亲缘关系较远。RAPD 对基因组的分析结果与传统分类学的结果基本相符。     

Characterization of white sapote (Casimiroa edulis Llave & Lex.) germplasm using floral morphology,RAPD and AFLP markers

Floral morphology, random amplified polymorphic DNA (RAPD), and amplified fragment length polymorphism (AFLP) were used to characterize and verify genetic diversity within a white sapote cultivar collection and to develop molecular markers for germplasm identification. On the basis of floral morphology, the cultivars were classified into three types: type I included 23 cultivars with large ovaries and small anthers; type II included 13 cultivars with small ovaries and large anthers; and type III included one cultivar, named ‘Maltby’, with a large ovary and large anthers. DNA was isolated from 39 cultivars of white sapote and subjected to RAPD and AFLP analysis using 24 and 7 primers, respectively. One hundred and sixty-eight RAPD and 286 AFLP bands were used to assess genetic characterization among white sapote. Sixty percent of the RAPD and 77% of the AFLP amplification products were polymorphic among accessions. RAPD or AFLP markers differentiated all white sapote cultivars effectively. Moreover, each flower type was characterized as specially associated with two RAPD bands. UPGMA dendrograms based on RAPD and AFLP data, showed the majority of the cultivars from flower type I and flower type II clustering together. Finally 101 RAPD markers and 220 AFLP markers were used to construct a neighbor-joining dendrogram. This showed that the 37 cultivars could be classified into six distinct clusters, between which the similarity coefficient was as low as 0.00–0.55, even though the cultivars were morphologically very similar. The remaining two cultivars namely ‘Smathers’ and ‘Maltby’ were found genetically very distant from the other cultivars in RAPD, AFLP or combined RAPD and AFLP based dendrograms. The results suggested that the level of genetic variation among white sapote cultivars is diverse and the morphological and molecular data may lead to representation of the cultivar relationships as well as flower type discrimination.     

应用RAPD分析新疆主要梨品种的遗传关系

利用RAPD分析了新疆18个梨品种间的亲缘关系。从160个随机十聚体引物中筛选出12条RAPD引物。18份材料共扩增133条带,平均每条引物扩增约11条带,其中14条带为所有材料的共有带,有119个多态性位点,占总带数的89.47%,Nei相似系数为0.5～0.923。用PHYLIP构建MP树,认为(1)对选用的形态学经典分类已较明确的新疆梨、白梨、砂梨、西洋梨、秋子梨,RAPD分析数据完全与其相符;(2)几个芽变品种间高度相似;(3)杂种新梨1号与砀山梨、杂种新梨6号与香梨、杂种早酥梨与苹果梨分别并类;(4)苹果梨独立于供试的5个种之外;(5)库尔勒香梨归属于新疆梨。     

不同山楂品种亲缘关系的RAPD分析

为探讨山楂品种间的亲缘关系,采用RAPD技术对20个不同品种的山楂材料进行了基因组DNA多态性分析。从120个引物中筛选出15个10bp的随机引物对所选山楂品种的DNA样品进行PCR扩增,共得到216条谱带,177条表现多态性,多态性比率达81.9%,其中包含27条特异性谱带,揭示了山楂植物丰富的遗传多样性。且利用NTSYS软件和UPGMA法对扩增结果进行了品种间相似系数的计算及聚类分析,结果表明相似系数在0.71～0.87,实生楂与其他山楂品种亲缘关系较远。     

利用RAPD和SCAR标记鉴定草莓品种

 利用RAPD和SCAR标记对32份草莓材料进行了鉴定, 结果表明: 8个RAPD引物共扩增出85个标记, 其中71个为多态性标记, 多态性比率为83。5%。25份草莓材料的RAPD图谱差异较大, 易于区分。利用具有多态性的RAPD标记, 对28份草莓试材的亲缘关系进行分析, 初步鉴定出同名异物和同物异名品种。两个RAPD标记被转化为片段长度分别为378 bp和214 bp的显性SCAR标记, 其多态性与相应RAPD标记一致。利用这两个SCAR标记对草莓品种进行了初步鉴定。     

Analysis of genetic diversity among Indian short-day onion (Allium cepa L.) cultivars using RAPD markers.

Summary

Randomly amplified polymorphic DNA (RAPD) markers were used to estimate genetic diversity among 24 cultivars of short-day onions. Total genomic DNA was extracted and subjected to RAPD analysis using 90 arbitrary 10-mer primers. Of these, 15 primers were selected which yielded 137 bands, 91.24% of which were polymorphic. None of the primers produced a unique banding pattern for each cultivar. RAPD data were used to calculate a Squared-Euclidian Distance matrix which revealed a minimum genetic distance between cultivars ‘AFLR-722’ and ‘PBR-140’, and a maximum genetic distance between cultivars ‘PBR-139’ and ‘A.Kalyan’, and ‘MS-48’ and ‘A.Kalyan’. Based on the distance matrix, cluster analysis was done using a minimum variance algorithm.The dendrogram thus generated, based on Ward’s method, grouped the 24 onion cultivars into two major clusters. The first cluster consisted of cultivars from the northern region, and the second of cultivars from the southern region of India. The present study shows that there is high diversity among the onion cultivars selected and indicates the potential of RAPD markers for identification and maintenance of onion germplasm for crop improvement purposes.     

15个樱桃品种的RAPD分析

利用RAPD技术,用104条随机引物对甜樱桃的14个品种和中国樱桃的1个品种进行遗传多样性分析,其中有14条引物的多态性较好。用任意一条能出现扩增带的引物,能明显区分开中国樱桃和甜樱桃,RAPD标记能够准确地进行种间的区分;而用一个引物或两个引物的组合只能鉴定出甜樱桃的一个品种。聚类结果显示,中国樱桃和甜樱桃的遗传距离最远;黄色果肉的养老和其他红色果肉的品种遗传距离较远。RAPD分析基本能够反映甜樱桃品种间的遗传多样性,但效果不理想,鉴定品种较为困难。     

Molecular assessment of polymorphism among local Jordanian genotypes of the common fig (Ficus carica L.)

This research was conducted to assess polymorphism among local genotypes of common fig available in Jordan (one of countries of origin). Leaf samples were collected in spring for DNA isolation from 20 different local genotypes (5 cultivars and 15 landraces). Two more wild types and one foreign cultivar were included. The genotypes were assessed using the randomly amplified polymorphic DNA (RAPD) technique. Six of the 19 screened primers showed reproducible polymeric profiles. Out of 62 amplified bands, 48 were polymorphic (77%). They generated 1104 data entries (591 for present and 513 for absent bands). After determining Jaccard similarity index, some genotypes showed high genetic similarity (90% between F20 and F22), while other were less similar (3–18% between F11 and all other genotypes). Moreover, the primers were evaluated for their discriminating power, where primer RAPD06 showed the weakest power (0.431), while highest values of 0.989 and 0.996 were achieved for primers RAPD02 and RAPD13, respectively.     

Characterization of progenies derived from pollination of pomegranate cv. Malase-Tourshe-Saveh using fruit traits and RAPD molecular marker

Fruit traits and molecular markers (RAPD) were used to survey genetic similarities and inheritance pattern of offspring, derived from self- and open pollination of pomegranate cv. Malase-Tourshe-Saveh (MTS) as well as progenies derived from cross between ‘MTS’ and ‘Narm-Daneh’ (ND) a soft-seed cultivar in the present study. Clustering based on fruit traits, divided the accessions firstly into two major groups, sweets and sour or sweet–sours. Members of sub-clusters were different in fruit characters, and seed softness also was affecting the grouping of individuals. Offspring of ‘MTS’ × ’ND’ showed somehow seed softness with varying degrees compared to other individuals. In RAPD analysis, by using 26 polymorphic primers, 325 fragments were produced from 39 individuals, among which 70 reproducible, polymorphic bands were scored for data analysis. NTsys and Bootstrap software were used for RAPD data analysis and respective dendrogram depicted by Jaccard's similarity coefficients using UPGMA method. The similarity coefficients were ranging from 0.2 to 0.94 and the lowest was obtained between pollen parent (ND cv.) and a genotype from open pollination of MTS cultivar. The highest similarity coefficient (0.94) was observed between two offspring both from self-pollination of MTS cultivar. In corresponding dendrogram, pollen parent was separated from other genotypes and laid individually in a sub-cluster, while the mother plant clustered with its progenies. In addition, some marker–trait associations were observed, especially for seed softness.     

利用RAPD标记对桃品种特殊种质分析

采用RAPD技术，用从200 个随机引物筛选的22 个10碱基随机引物对148个桃品种进行DNA扩增。根据各品种扩增的RAPD带和聚类结果，分析具特殊位点，并在聚类图中聚在外围、相似系数较低的品种有玉露蟠桃、秋蜜、爱保太、佛尔蒂尼莫蒂尼和金皇后等特殊品种种质。从分子生物学角度，为种质保存和育种提供理论依据。