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1.
利用正交试验对Taq酶浓度、引物浓度、dNTPs浓度、Mg2+浓度和模板DNA浓度5个因素进行优化,从而建立了一套适宜构树的ISSR-PCR反应体系.在20 μL的反应体系中,各反应物的最适宜含量为:0.07 U·μL-1 TaqDNA聚合酶、0.35 μmol·L-1ISSR引物、0.3 mmol·L-dNTPs,2.0 mmol·L-Mg2+、1.2 ng· μL-1模板DNA.PCR扩增程序为:94℃预变性5min,94℃变性30 s,46℃退火45 s,72℃延伸2 min,40个循环. 相似文献
2.
鹅掌楸ISSR-PCR反应体系的建立及优化 总被引：2，自引：0，他引：2
为建立鹅掌楸简单序列重复区间扩增ISSR的PCR优化反应体系,以鹅掌楸叶片基因组DNA为材料,系统地测试了模板DNA、引物、dNTPs、Mg2+浓度、TaqDNA聚合酶用量及退火温度对ISSR-PCR反应体系的影响。结果表明:优化的PCR反应体系为:20μL总体系中,含30 ng模板DNA,0.3μmol.L-1随机引物,0.2 mmol.L-1dNTPs,1.4 mmol.L-1Mg2+,0.8 UTaqDNA聚合酶;最佳退火温度为60℃;PCR反应程序为:94℃预变性5 min;94℃变性1 min,60℃退火45 s,72℃延伸2 min;45个循环;72℃再延伸7 min。 相似文献
3.
油茶随机扩增多态DNA条件的研究 总被引：10，自引：0，他引：10
以改进的CTAB法提取油茶嫩叶总DNA,进行随机扩增多态DNA(RAPD)分析,分别测试了Mg2+浓度、引物浓度、dNTP浓度、模板DNA浓度和TaqDNA聚合酶用量对反应结果的影响,确定了油茶RAPD分析的最适反应体系:在25μlPCR反应体积中,含20ng模板DNA,2 5mmol·L-1MgCl2,0 2mmol·L-1dNTP,0 3μmol·L-1引物,1UTaqDNA聚合酶。扩增程序为:94℃预变性180s;94℃变性60s,37℃退火90s,72℃延伸120s,反应40个循环;最后在72℃延伸5min。 相似文献
4.
以红花石蒜叶片基因组DNA为模板,分析了模板DNA、引物、dNTPs、Mg2 的浓度及Taq DNA聚合酶的用量对ISSR-PCR扩增结果的影响.建立了石蒜ISSR分析最优化的反应体系及应用程序:即25 μL反应体系中,有20 ng模板DNA、0.5 μmol·L-1随机引物、150 μmol·L-1 dNTPs、2.0 mmol·L-1 Mg2 、1.0 U Taq DNA聚合酶.反应程序为:94 ℃预变性5 min;然后45个循环:每个循环94 ℃变性45 s,55 ℃退火60 s,72 ℃延伸2 min;循环结束后72 ℃延伸7 min. 相似文献
5.
狗牙根ISSR—PCR反应体系的建立与优化 总被引：2，自引：0，他引：2
以改良的CTAB法提取的狗牙根叶片DNA为模板,采用单因素多水平方法对狗牙根ISSR反应体系进行构建和优化,系统分析Taq酶、Mg2+、模板DNA、dNTPs、引物和buffer这6种1SSR反应成分的浓度对扩增结果的影响,结果表明:20.0μL PCR反应体积中,20.0 ng模板DNA,0.4 μmol/L引物,0.5mmol/L Mg2+,0.05mmol/L dNTPs,0.2U TaqDNA聚合酶,2.0 μL 10×buffer.反应程序为;94℃预变性5 min;94℃变性45 s,55 C退火45 s,72℃延伸1.5 min,35个循环;再72℃延伸7 min,4℃保温. 相似文献
6.
模板DNA、引物、dNTPs、Mg^2＋的浓度,Taq DNA聚合酶的用量以及退火温度是影响简单序列重复区间扩增（ISSR-PCR）结果的主要因素.以桉树叶片基因组DNA为试材,系统地测试了这6个因素对桉树ISSR反应结果的影响.结果表明：最优化的反应体系即20μL反应体系中,含20 ng模板DNA、0.4μmol.L^-1随机引物、0.15 mmol.L^-1dNTPs、2.0 mmol.L^-1Mg^2＋、1.25 U TaqDNA聚合酶.最佳退火温度为55℃;PCR反应程序为94℃预变性5 min;94℃变性45 s,55℃退火45 s,72℃延伸1.5 min,35个循环;72℃再延伸7 min. 相似文献
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8.
以改进的CTAB高盐法提取油茶嫩叶总DNA,进行简单重复序列间扩增(ISSR)分析,分别测试了退火温度、模板DNA浓度、M g2+浓度、dNTP s浓度、T aqDNA聚合酶用量和是否加入去离子甲酰胺对反应结果的影响.油茶ISSR分析的最适反应体系为:在20μL PCR反应体积中,含20 ng模板DNA,2.5 mm o l.L-1M gC l2,0.2 mm o l.L-1dNTP s,1UT aqDNA聚合酶,0.5pm o l.μL-1引物,1%去离子甲基酰胺.扩增程序为:94℃预变性2 m in;94℃变性30 s,52℃退火30 s,72℃延伸90 s,反应38个循环;最后在72℃延伸7 m in. 相似文献
9.
应用L9(34)正交表建立了适合于樟树ISSR-PCR的优化反应体系,即20μL ISSR反应体系中含有1×PCR buffer、dNTP0.25 mmol/L、引物0.3μmol/L、Mg2 2.0 mmol/L、Taq酶1 U和模板DNA50 ng.适宜的扩增条件为:94℃预变性4 min;94℃变性40 s,58℃退火1 min,72℃延伸2 min,42个循环;72℃延伸10 min;4℃保存. 相似文献
10.
杨树基因组SRAP扩增体系的建立与优化 总被引：4，自引：0，他引：4
以响叶杨和银白杨及F1代为实验材料,利用正交设计直观分析法和单因素随机试验对杨树SRAP反应体系中各组分(TaqDNA聚合酶、dNTP、引物和Mg2+)的浓度进行优化;通过实验确定10μL反应体系为:引物0.35μmol/L,TaqDNA聚合酶0.5U,dNTP0.2 mmol/L,Mg2+2.0 mmol/L,10×PCRbuffer 1μL,模板DNA 1μl(20 ng/μL).同时利用最佳反应体系以及正交设计直观分析法对PCR扩增程序进行筛选,并通过梯度PCR试验,确定引物最佳退火温度.结果表明最佳反应程序为:开始94℃变性5min,反应前5个循环在94℃1 min,37℃30 s,72℃ 1min条件下运行;之后35个循环在94℃1 min,54℃ 1 min,72℃ 1 min条件下运行,最后延伸7 min;12℃保存.并利用两树种的杂种F1代对优化的反应体系和程序的可行性进行了验证. 相似文献
11.
《Scandinavian Journal of Forest Research》2012,27(5):427-435
In Scandinavia, moose (Alces alces L.) sometimes cause severe browsing damage to economically-important pine. Moose-vehicle accidents have spurred construction of fences along roads, and these may interfere with moose migration between summer and winter ranges, or the road alone may be a barrier. If this happens and moose build up along roads, landowners may suffer economically. Therefore, this study investigated whether roads, fences or other factors influence the use of young pine stands by moose. Eighty stands along roads in northern Sweden were evaluated in which individually-browsed branches were counted on 9972 pines. Moose browsing was not significantly related to birch (Betula pendula Roth, B. pubescens Ehrh.) density, nor did it differ between pines (Pinus contorta Douglas or P. sylvestris L.). However, increased pine density, site productivity and proximity to a highway were associated with increased browsing. Further large-scale studies are needed to understand moose habitat selection and the effects of roads. 相似文献
12.
《Scandinavian Journal of Forest Research》2012,27(3):206-217
The root systems of 2-yr-old Picea glauca, Picea mariana and Pinus banksiana seedlings were submitted to various frost temperatures during an artificial frost to induce different levels of root damage. Frost-damaged and control seedlings were placed in a greenhouse under high and low soil moisture regimes. Seedling growth and physiology were evaluated periodically. Seedling survival was reduced when root damage reached levels of 60-80%. Root systems of all three species showed partial to total recovery by the end of the experiment. In general, root freezing damage caused reductions in seedling growth, with these reductions becoming less significant over time. Root damage had little to no effect on black spruce and jack pine seedling physiology, while white spruce CO 2 uptake decreased with increasing root damage. Shoot nitrogen content of all three species decreased slightly with increasing root damage. 相似文献
13.
《Scandinavian Journal of Forest Research》2012,27(1):53-61
This study investigated the stand structure in pine, spruce and deciduous forests in the border district of Finland and Russia. A total of 46 mature forest stands was selected as pairs, the members of each pair being as similar as possible with respect to their forest site type, age, moisture and topography. The stands were then compared between the two countries by means of basal areas and number of stems. The proportions of dominating tree species were 2-12% lower, and correspondingly the proportions of secondary tree species higher, in Russian forests. The density of the forest stock was also higher in each forest type in Russia. The forests in the two countries differed most radically in terms of the abundance of dead trees. The amount was two to four times higher in Russian deciduous and spruce forests, and in pine forests the difference was 10-fold. The stand structures indicated that Russian coniferous stands, in particular, were more heterogeneous than intensively managed pine and spruce stands in Finland. 相似文献
14.
《Scandinavian Journal of Forest Research》2012,27(1):15-27
The root collar diameter and the height:diameter ratio are of particular importance in container-grown seedlings where a high density in the containers may produce spindly seedlings. Temperature regimes and light quality are known to affect plant growth. The aim of this study was to identify responses in Picea abies (L.) Karst. seedlings grown with light providing different red:far-red ratios and under temperature regimes with alternating day (DT) and night temperature (NT) from negative (DT < NT) to positive (DT > NT) difference (DIF) between DT and NT. Experiments were conducted in controlled environment chambers and in a daylight phytotron. Only limited thermoperiodic responses appeared in P . abies seedlings with respect to seedling height and dry weight accumulation. The formation of terminal buds, however, was clearly delayed in seedlings grown at negative DIF. The results indicate a requirement for day extension light that is high in far-red, to prevent terminal bud formation under natural short-day conditions. An extended study should be conducted to clarify the minimum level of light intensity and the optimal light quality needed to prevent terminal bud formation under natural short-day conditions. 相似文献
15.
本文分析了CAD在设计中引起的正面和负面影响,并进行了系统的阐述,从而使设计者在应用中能够保持客观的态度。 相似文献
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17.
《Scandinavian Journal of Forest Research》2012,27(5):472-479
Based on an enquiry, risk perception among non-industrial private forest owners is described in relation to climate change and forestry hazards. Of the respondents, 11% took action to remedy the effects of climate change. Out of a given set, hazards were ranked according to each respondent's experience of recent substantial financial loss to the estate and in relation to his or her willingness to make investments aimed at risk reduction. For each hazard, the respondent assessed the risk in four classes ranging from very high to negligible risk. Six hazards were considered most problematic in all three aspects: browsing damage, falling timber prices, damage by wind, spruce bark beetle, root rot and pine weevil. A majority of the respondents claimed to take action to reduce the risk associated with at least one hazard, while 35% did not know whether they did. Excluding climate change, the need for decision support was the largest in relation to damage by wind owing to a combination of perceived high risk and a high level of ignorance in relation to whether risk-reducing measures were taken. 相似文献
18.
Srisuda Thippayarugs Banyong Toomsan Patma Vityakon Viriya Limpinuntana Aran Patanothai Georg Cadisch 《Agroforestry Systems》2008,72(2):137-148
In situ produced plant residues contain a mixture of different plant components of varying quality. To assess synergistic
or antagonistic interactions occurring during the decomposition and mineralization of such mixtures, individual plant parts
(stems, leaves, leaf litter and roots) or the mixture of stems, leaves and leaf litter of the agroforestry species pigeonpea
(Cajanus cajan) or of crop residues of peanut (Arachis hypogaea) or of the weed hairy indigo (Indigofera hirsuta) were incubated in pots for 19 weeks. Periodically, remaining plant residues were sieved out (>2 mm), weighed and N content
as well as soil mineral N determined. Above- and below-ground residues of peanut decomposed fastest and showed the largest
N release in agreement with their high N concentration and low-acid detergent fibre (ADF) : N ratio. Hairy indigo was hypothesized
to be of lower quality than pigeonpea because of its high-polyphenol content. However, it decomposed faster than pigeonpea,
largely because of the higher N and lower lignin concentration of its components. Ranking of individual plant components for
N mineralization resulted in the following pattern, leaves > leaf litter > roots > stems. In mixtures of the different plant
components a similar species order in decomposition was obtained, e.g. peanut > hairy indigo > pigeonpea. The amount of N
released from the mixture was dominated by stem material that comprised 46–61% of the mixture. The interactions in mixtures
were relatively small for peanut (generally high-quality components) as well as for pigeonpea (low proportion of high-quality
components, i.e. N rich leaf material). However, a positive interaction occurred during later stages of N mineralization in
the mixture of hairy indigo as it had a significant proportion of N rich components and absence of highly reactive polyphenols.
Thus, for plants with low to intermediate chemical quality attributes, manipulating plant composition (e.g. by varying harvest
age, affecting stem and leaf proportions) will be important to obtain significant interactions during decomposition when its
components are mixed. 相似文献
19.
A. Rigueiro-Rodríguez M. R. Mosquera-Losada E. Gatica-Trabanini 《Agroforestry Systems》2000,48(3):245-256
The use of organic waste materials such as milk sewage as an organic fertilizer could have the dual advantages of organic-waste
disposal and reduced dependence on inorganic fertilizers. The effects of fertilization with (1) conventional mineral fertilization,
(2) milk sewage sludge at 40 kg N ha−1 target rate and (3) no fertilization on pasture production and tree growth were examined in an experiment consisting of two
pasture mixtures under a one-year-old Pinus radiata plantation with a density of 2500 trees ha−1. The two pasture mixtures were: (1) Dactylis glomerata L. var. saborto (25 kg ha−1) + Trifolium repens L. group Ladino (4 kg ha−1) + Trifolium pratense L. var. Marino (1 kg ha−1); (2) Lolium perenne L. var. Tove (25 kg ha−1) + Trifolium repens L. group Ladino (4 kg ha−1) + Trifolium pratense L. var. Marino (1 kg ha−1). The experiment began in the spring of 1995 using a randomized block design with three replicates in Castro Riberas de Lea
(Lugo, Galicia, north-western Spain). Plot size was 12 × 8 m2, with a 1 m buffer strip between plots. Two-year data showed that fertilization with either material had a positive effect
on pasture production, with no significant difference between the two fertilization treatments. Tree growth in the milk sewage
sludge plot was significantly higher than in the control plots. Inorganic fertilization increased pasture production, but
affected tree growth negatively. The results show that milk sewage sludge could be used as a fertilizer in silvo-pastoral
systems.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
20.
《Scandinavian Journal of Forest Research》2012,27(2):166-178
This study investigated the forest history, structural characteristics, the presence of fungal species suggested to indicate ecological continuity in old-growth forest, and the species composition of saproxylic beetles in 30 old forest ''woodland key habitats'' (WKHs), and compared them with production forest in the same age-class in south-eastern Norway. No statistically significant differences in forest characteristics, community of saproxylic beetles or number of red-listed beetles were found between the WKHs and the production forest, probably owing to a combination of profound long-term logging and a lack of will to delineate sufficiently large WKHs in the one area with significantly less forestry impact. The study indicates the advantage of including forest history information in WKH selection where such data exist, and the importance of restoring habitats in heavily exploited forest landscapes. 相似文献