杜鹃花属植物基因组DNA RAPD-PCR反应体系的优化

试验通过系统比较研究,确立了杜鹃属植物RAPD-PCR的最佳反应体系,即20μL反应体系中:模板DNA(10 g.L-1)1.5μL;10×PCR buffer 2μL;Taq酶(0.5 U.μL-1)0.3μL;引物(0.8μmol.L-1)4μL;MgCl2(25 mmol.L-1)1.8μL;dNTPs(dATP、dCTP、dTTP、dGTP各含1.0 mmol.L-1)0.6μL。经试验验证,该反应体系重复性高,扩增条带亮度均匀,弥散现象少。     

平菇RAPD—PCR反应体系优化研究

以平菇黑丰268为试材,对RAPD—PCR反应体系的各项影响因素进行了优化,建立了平菇RAPD—PCR反应体系。结果表明,平菇RAPD—PCR最优体系为：20μl PCR反应体系中,Mg^2＋浓度为1．4mmol／L,模板DNA为50ng,引物浓度为0．6—0．8μmol／L,dNTPs浓度为250μmol／L,Taq酶为1．2U,10×Buffer2．0μl。     

荷包猪RAPD-PCR反应条件研究

[目的]建立适合荷包猪RAPD-PCR反应的最佳反应体系。[方法]以荷包猪为试验材料,以MgCl2浓度、引物浓度、dNTP浓度、模板DNA用量、TaqDNA聚合酶用量及退火温度为影响因子,在保持其他影响因子一致的条件下,变化单一因子,筛选最优参数,研究各影响因子对RAPD-PCR反应的影响。[结果]最佳反应体系的总体积为20.0μl,MgCl2浓度为2.5μmol/L、引物浓度2.0μmol/L、dNTP浓度400.0μmol/L、模板DNA为100 ng/μl、TaqDNA聚合酶为1.0 U。PCR反应程序:94℃预变性2 min(94℃变性1 min,36℃退火1 min,72℃延伸1 min,循环40次),72℃延伸5 min。此反应体系所扩增出来的结果比较稳定,带型清晰且亮度适中。[结论]该研究为应用RAPD技术对荷包猪作进一步的遗传分析奠定了基础。     

Patch Size and Patch Quality of Gall-inducing Aphids in a Mosaic Landscape in Israel

For weak flying insects feeding on two different host plants during their life cycle, such as gall-inducing aphids, patch and matrix characteristics may play a critical role in patch occupancy and population size in occupied patches. The aims of the present study were to define the basic patch size of Baizongia pistaciae (L) (Aphididae, Fordini), an aphid inducing galls on Pistacia palaestina Boiss (Anacardiaceae) using a genetic approach, and to estimate the impact of landscape structure and patch quality on patch occupancy and gall density on occupied trees of this aphid and four other closely related species. Using 42 genetic markers detected by RAPD-PCR in 117 clones of the galling aphid Baizongia pistaciae, we calculated Wright's F statistics and estimated the number of winged migrants between demes. We found that host trees at least 150 m apart supported genetically differentiated demes of B. pistaciae, and formed distinct patches. Since the annual cycle of this aphid involves alternation between two different hosts, P. palaestina trees and Poaceae roots, patch – the smallest area that sustains a deme – is a relatively small area that must be composed of at least a single P. palaestina tree and nearby secondary hosts. To assess the impact of landscape structure and patch quality on patch occupancy and gall abundance in occupied patches, two field surveys of P. palaestina trees in natural Mediterranean maquis were performed. Among the five species of gall-inducing aphids found, B. pistaciae was the most abundant of those surveyed. Host trees were occupied more often in the ecotone, the transition zone between Mediterranean closed maquis and open bata, than in the maquis. Mature and old trees were more often occupied than young ones, and shrubs more often than tree-like plants. There was no difference in the proportion of occupied trees between isolated host trees or those growing in groups. Species richness showed similar trends. We also found no significant differences in gall abundance in occupied trees among tree quality categories, except that trees growing in the ecotone tended to carry more galls than those growing in the maquis. In conclusion, the best patch of gall-inducing aphids seems to be a small area, composed of an old shrub of P. palaestina standing in an open landscape with nearby secondary hosts, grass roots, available for colonization by winged migrants.     

蘑菇分离物的DNA指纹鉴别

按照植物理学原理，蘑菇分离物应依据柯赫法则（Koch's postulate)回接，在一定条件下，人工诱用蘑菇）难以人工诱发子实体，即使能形成子实体也费时费力，本文介绍DNA指纹鉴别技术，即运用RAPD－PCR技术制成分离物及对应子实体指纹图谱，分析它们间DNA同源性及遗传相似度以鉴别分离物真的，探求快速，简便，有效的鉴别方法。     

Molecular systematics in Chrysanthemum × grandiflorum (Ramat.) Kitamura

An attempt was made to understand the molecular systematics and genetic differences between 10 original chrysanthemum cultivars and 11 mutants. The similarity among the cultivars and mutants varied from 0.17 to 0.90 using RAPD analyses, a simple but efficient method to distinguish cultivars and to assess parentage. Two distinct groups were found. Two cultivars were present as a separate group showing differences from all other cultivars. Mutants with different flower colour could be identified at the molecular level using RAPD technique holding promise to identify unique genes as SCAR markers. A high genetic distance among the different chrysanthemums showed that there exists a possibility of introgressing new and novel genes from the chrysanthemum gene pool.     

正交试验设计在建立杜鹃花RAPD-PCR反应体系中的应用

利用正交试验对影响RAPD-PCR反应体系的各种因素进行研究,从而建立适合杜鹃花RAPD反应的最佳反应体系,最终选择杜鹃基因组RAPD-PCR的最佳扩增反应体系,即模板DNA 80ng、随机引物0.5μmol/L、dNTPs 1.1mmol/L、MgCl2 2.5mmol/L,1.2μmol/LTag-DNA聚合酶,buffer 2μL,总反应体系20μL.试验设计的6个因子、5个水平中,Mg2 、dNTPs和引物的浓度是主要因子,其中Mg2 的浓度对试验的影响最大,dNTPs和引物浓度在较低的水平就可满足需要,高浓度并没有显著提高试验效果.     

Molecular analysis of an interspecific hybrid ornamental eucalypt for parental identification

The parentage of ‘Urrbrae Gem’, an ornamental hybrid Eucalyptus, was investigated using RAPD-PCR. The female parent was known to be E. erythronema var. erythronema but previous opinion, based on adult morphological characters, placed the male parent as either E. stricklandii or E. gomphocephala. Samples of DNA, extracted from leaves of different individuals within each species, were amplified with six different 10-mer primers to produce RAPD fingerprints. These were used to generate a UPGMA dendrogram based on genetic similarities, and an ordination plot, derived by multi-dimensional scaling (MDS), and minimum spanning tree (MST) to show the relative dissimilarities between the individuals tested. The UPGMA dendrogram divided the samples into three clusters, with the hybrid slightly closer to E. stricklandii than E. gomphocephala. The MDS ordination and MST placed the hybrid between E. erythronema var. erythronema and E. stricklandii, supporting E. stricklandii as the male parent. This revised version was published online in August 2006 with corrections to the Cover Date.     

洋葱(Allium cepa L.)RAPD-PCR反应体系及扩增程序的优化

为建立多态性高、稳定性好的洋葱RAPD-PCR反应体系,采用正交设计,研究了Taq酶、Mg2 、引物和dNTP 4种RAPD-PCR反应组分浓度变化对扩增结果的影响,在此基础上对模板DNA用量、扩增程序中退火温度和反应循环次数进行了筛选。试验结果表明,洋葱20μl RAPD-PCR优化反应体系为1×Buffer、2.0 mmo1/L Mg2 、1.0 UTaqDNA聚合酶、200μmo1/L dNTP、0.6μmo1/L引物、2%甘油和15 ng DNA模板;PCR扩增程序为94℃预变性4 m in;94℃变性30 s,35℃退火40 s,72℃延长1.5 m in,45个循环;72℃保温延伸7 m in。     

长白山区中华蜜蜂与西方蜜蜂寄生瓦螨比较分析

采用形态测定和RAPD-PCR相结合的方法对寄生于长白山区的中华蜜蜂和西方蜜蜂的瓦螨进行比较分析.结果表明,中、西蜂寄生瓦螨无论在形态水平还是分子水平上都是具有差别的.在选定的8个形态指标中,寄生于中华蜜蜂的瓦螨个体的指标有7个大于寄生于西蜂的瓦螨,其中有6个指标的均数差异达到显著(0.01相似文献