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1.
以舟山群岛的普陀樟(Cinnamomum japonicum var.chenii)新鲜叶片为实验材料,采用L16(45)正交试验设计,对影响普陀樟SRAP-PCR反应的Mg2+浓度、dNTPS、Taq DNA聚合酶、引物和模板DNA用量5因素组合进行了筛选。结果表明:适于普陀樟的SRAP-PCR反应体系(20μL)为2.5 mmol/L Mg2+、0.2 mmol/L dNTPs、1.5 U Taq DNA聚合酶、0.4μmol/L引物、10 ng模板DNA、2μL 10×PCR buffer,该体系位点清晰,扩增稳定;利用该反应体系从100对SRAP引物组合中筛选出多态性好的引物24对。  相似文献   

2.
采用单因素试验对白桦SRAP-PCR反应体系中的5个因素(Mg2+、d NTP、引物、Taq DNA聚合酶和模板DNA)进行优化,结果表明:在25μL反应体系中Mg2+浓度2.0 mmol·L-1、Taq DNA聚合酶1.5 U、d NTPs 0.15 mmol·L-1、引物浓度0.4μmol·L-1、模板DNA 30 ng,该体系能够很好地满足白桦SRAP扩增的要求。优化体系的建立将为今后利用SRAP标记技术对白桦进行分子遗传育种研究奠定基础。  相似文献   

3.
为确立按树SRAP-PCR反应体系,并对桉树品种进行遗传多样性分析,以巨尾桉GL -9号嫩叶提取的DNA为模板,进行Mg2+、dNTP、引物和Taq DNA聚合酶4个因素3个水平L9(34)正交试验,并比较了不同浓度模板DNA对PCR扩增效果的影响.结果显示:桉树的SRAP-PCR最佳反应体系为Mg2+2.5 mmol/L、dNTP 0.20mmol/L、引物0.4μ mol/L、Taq DNA聚合酶1.5U,DNA模板最佳浓度为10ng.利用最佳反应体系对桉树品种进行引物组合多态性筛选,从40个引物组合中筛选出多态性引物组合17个.挑选12个多态较高的引物组合对11个按树品种进行遗传多样性分析,通过PCR扩增,得到109个谱带.其中多态性条带95条,平均每个引物组合产生7.92个多态性条带,显示了相对较高的多态性.表明SRAP标记可应用于按树分子生物学研究.  相似文献   

4.
以刨花润楠(Machilus pauhoi)1.5 a生小苗幼嫩叶片为试材,对影响刨花润楠SRAP-PCR扩增的模板DNA量、引物、dNTP和Mg2+体积摩尔浓度、Taq DNA聚合酶、退火温度6个主要因素进行优化.结果表明,SRAP-PCR的最佳反应体系为:25μL的SRAP-PCR反应体系中,2.5μL 10×PCR buffer、模板DNA量60 ng、Mg2+2.0 mmol/L、dNTP 0.225 mmol/L、引物0.3μmol/L和Taq DNA聚合酶1.25 U.对优化的反应体系和扩增程序的验证结果表明,优化的刨花润楠SRAP-PCR反应体系和扩增程序是稳定可行的.  相似文献   

5.
以西伯利亚杏为试验材料,进行基因组DNA的提取、扩增及电泳试验。通过L16(45)正交试验,确定西伯利亚杏SRAP-PCR优化反应体系(反应体系总体积20μL):Mg2+2.0 mmol·L-1、dNTPs0.25 mmol·L-1、引物浓度1.2μmol·L-1、Taq聚合酶1.0 U、DNA模板20 ng。采用SRAP引物组合ME5/EM10,对优化的SRAP-PCR反应体系进行初步验证,24个西伯利亚杏无性系SRAP分子标记试验共扩增出谱带431条,引物扩增出23条谱带,其中338条为多态性谱带,多态百分率为78.42%。  相似文献   

6.
利用正交试验L16(45),结合单因素试验对厚朴相关序列扩增多态性(Sequence-related amplified polymorphism,SRAP)标记反应体系的5个因素(Mg2+,dNTPs,引物,Taq酶和模板DNA)进行优化试验,结果表明:各因素水平变化对PCR反应的影响从大到小依次为:dNTPsTaq酶模板DNAMg2+引物;筛选出各反应因素的最佳水平,建立厚朴SRAP-PCR反应的最佳体系(25μL)为:Taq酶1.5 U,Mg2+1.8 mmol.L-1,模板DNA100 ng,dNTP 0.24 mmol.L-1,引物0.40μL。试验表明,该体系重复性好、稳定性强。  相似文献   

7.
为了建立锥栗SRAP-PCR反应体系,从而为今后的锥栗分子生物学研究提供理论参考,以锥栗叶片基因组DNA为材料,采用单因素试验和正交试验相结合的研究方法,对影响锥栗SRAP-PCR反应体系的模板用量、Mg2+浓度、dNTP浓度、引物浓度和Taq DNA聚合酶浓度这5个主要因素进行了优化试验。试验确立的适合锥栗的SRAP-PCR反应体系为:20μL体系中,模板DNA用量为40 ng,Mg2+浓度为1.8 mmol/L,dNTP浓度为280μmol/L,Taq DNA聚合酶浓度为2.0 U,引物浓度为0.6μmol/L。  相似文献   

8.
红松SRAP-PCR反应体系的优化   总被引:1,自引:0,他引:1  
通过单因素试验,对红松SRAP—PCR反应体系的主要影响因子(Taq DNA聚合酶、Mg^2+、模板DNA、dNTPs、引物浓度)进行了优化筛选,得出红松SRAP—PCR反应的最佳体系:20μL反应体系中,模板90ng,Mg^2+1.0mmol/L,dNTPs0.15mmol/L,Taq DNA聚合酶1.5 U,引物0.3~0.4μmol/L。  相似文献   

9.
以广西南宁产的金樱子叶片为材料,采用L16(45)正交试验设计,对SRAP-PCR反应体系中的Mg2+、dNTPs、TaqDNA聚合酶、引物和模板DNA用量5因素进行了优化。结果表明:适用于金樱子SRAP-PCR的最佳反应体系为:反应总体积10μL,包含2.0 mmol/L Mg2+、0.4 mmol/L dNTPs、0.5 UTaqDNA聚合酶、3μmol/L引物、25ng DNA及10×PCR Buffer;各因素对PCR反应影响由大到小依次为:Mg2+、DNA、dNTPs、引物、TaqDNA聚合酶。用7份不同来源的金樱子材料对优化的SRAP-PCR反应体系进行验证,均获得了条带清晰、多态性丰富的扩增图谱,证实了该体系的稳定可靠。  相似文献   

10.
油茶SRAP标记的PCR体系建立与优化   总被引:2,自引:1,他引:1  
油茶是我国主要的木本食用油料树种,拟建立油茶新型SRAP标记(序列相关扩增多态性)的优化PCR体系。以油茶优良品种的总DNA为材料,分析了影响油茶SRAP-PCR的主要参数,包括模板DNA量、引物浓度、dNTPs、Mg2+浓度、TaqDNA聚合酶用量、退火温度6个因素的影响,建立了适合油茶SRAP-PCR的优化体系为:20μL的PCR反应体系中,2.0μL 10×PCR buffer,2.0 mmol/L Mg2+,225μmol/L dNTPs,0.6μmol/L引物,30 ng模板DNA和0.75 U Taq DNA聚合酶;最佳PCR循环条件为:94℃预变性5 min;94℃变性1 min,35℃复性1 min,72℃延伸1 min,5循环;94℃变性1 min,52℃复性1 min,72℃延伸1 min,35循环;最后72℃延伸10 min;4℃保存。本研究结果可为今后利用SRAP这一新型标记技术进行油茶分子遗传学与标记辅助选择育种研究打下基础。  相似文献   

11.

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.

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.

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.

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.
Contour hedgerows of multipurpose tree species in the sloping tea lands of Sri Lanka are expected to reduce soil erosion and also add significant amounts of plant nutrients to the soil via periodic prunings. The objective of this experiment was to characterize the biomass decomposition pattern and quantify the amount of nutrients added through prunings of six tree species (Calliandra calothyrsus, Senna spectabilis, Euphatorium innulifolium, Flemingia congesta, Gliricidia sepium and Tithonia diversifolia) currently being used in hedgerows associated with tea. Withered leaf and stem prunings (50 g) were enclosed in 2-mm litter bags, placed at 5-cm depth and retrieved after one, three, six, nine and 12 weeks. Loss of initial dry weight, N, P and K was measured. Single exponential decay function adequately described both dry weight and nutrient loss. Tree species differed significantly in their rate of breakdown with decomposition constants (k) varying from 0.0299 to 0.2006 week−1 for leaves and from 0.0225 to 0.0633 week−1 for stems. Gliricidia showed the highest k for leaves with the rest in the following descending order: Senna > TithoniaEuphatorium > Calliandra > Flemingia. A similar pattern was observed for loss of all nutrients with Calliandra and Flemingia always having lower k values than the rest. Although N immobilization was not observed, immobilization of P and K was observed during the first week of incubation in some species, particularly in stem prunings. Annual biomass of prunings differed significantly between tree species in the following descending order: Calliandra > Senna > Flemingia > Tithonia > Gliricidia > Euphatorium. Calliandra added the greatest amount of nutrients annually to the soil with Euphatorium adding the least. Calliandra prunings provided the annual total K requirement and 49% of the N requirement of mature tea. However, none of the species provided more than 5% of the P requirement. It is concluded that among the tree species tested, Calliandra and Flemingia are the most suitable for contour hedgerows in tea plantations of this agroclimatic region because of their higher soil nutrient enrichment capacity and slower decomposition rates which would minimize leaching losses. This revised version was published online in June 2006 with corrections to the Cover Date.  相似文献   

16.
Heterobasidion parviporum and Heterobasidion annosum are widely distributed root‐rot fungi that infect conifers throughout Europe. Infection of conifer stumps by spores of these pathogens can be controlled by treating fresh stumps with a competing non‐pathogenic fungus, Phlebiopsis gigantea. In this study, growth of three Latvian strains of P. gigantea and the biological control agent ‘Rotstop’ strain was evaluated in stem pieces of Norway spruce, Scots pine, lodgepole pine, Douglas‐fir, Weymouth pine, Siberian larch and Sitka spruce. The growth rates of one H. parviporum and one H. annosum isolate were also measured in the same stem pieces. The growth rate of P. gigantea varied greatly in wood of different conifer species. It was higher in the three pine species, lower in Norway spruce and lowest in Sitka spruce and Siberian larch, and in Douglas‐fir, this fungus did not grow. The largest area of wood occupied by P. gigantea was in lodgepole pine. Growth of Latvian isolates of P. gigantea in the wood of Pinus and Picea species was comparable to that of the Rotstop isolate. Consequently, stump treatment with local P. gigantea isolates should be recommended. However, our results suggest that Douglas‐fir stump treatment against Heterobasidion by P. gigantea may be ineffective and other stump treatment methods should be considered.  相似文献   

17.
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   

18.

The effects of seed weight and seed type on seedling growth of Pinus sylvestris (L.) were studied by seeding individually weighed orchard and stand seed in different mixtures under harsh (direct seeding in field) and optimal (seeding in nursery) conditions. In the nursery experiment an increase in the seed weight from 3 to 7 mg increased the seedling height by 10-27% and total weight by 27-113%, and decreased the height/diameter ratio by 5-6% after 2 yrs. With elimination of seed weight effects, orchard seedlings were 2% taller than stand seedlings in year 2. Without elimination of seed weight effects, orchard seedlings were 7-13% taller. In the field experiment an increase in the seed weight from 3 to 7 mg increased seedling height by 18-65%, stem volume by 81-274% and the number of top-buds by 23-34% in year 5. After elimination of seed weight effects, orchard seedlings were 7-13% taller than stand seedlings and without elimination of seed weight effects 20-21% taller after 5 yrs. Even after elimination of both seed weight and genetic effects orchard seedlings were 3-9% larger than stand seedlings in the field experiment. In conclusion, the influence of seed weight and seed type on growth traits and slenderness is highly significant and the influence seems to be greater in harsh conditions.  相似文献   

19.
Baobab leaves form an important part of the local diet in Sahel countries and elsewhere in Africa. Existing leaf nutritional data and agroforestry performance information are based solely on Adansonia digitata L., the baobab of continental Africa. The introduction potential of Adansonia species from the center of diversity in Madagascar and from Australia remains untapped. To assess this potential, the mineral contents and B1 and B2 vitamin levels of dried baobab leaves were determined for five-year old trees of A. digitata, A. gibbosa (A. Cunn.) Guymer ex D. Baum, A. rubrostipa Jum. & H. Perrier (syn. A. fony Baill.), A. perrieri Capuron and A. za Baill. grown in an introduction trial in Mali. Nutritional data were evaluated against survival and vigor to identify promising germplasm. Leaf vitamin and crude protein contents were highest in the Madagascar species, especially A. rubrostipa (B1 88 mg 100 g−1, B2 187 mg 100 g−1, protein 20.7% dry weight). However, the local species far outperformed the introductions in survival, tree height, basal diameter and resistance to termites. We suggest grafting as a way of harnessing the vigor of well-adapted local baobab varieties to the superior nutritional profiles of A. rubrostipa and others. Cross-species grafting tests in Adansonia were successful, thus creating new agroforestry possibilities with different scion/rootstock combinations.  相似文献   

20.

Mean age, mean and top heights and yield were studied in 20 mixed stands of birch ( Betula pubescens Ehrh. and B. pendula Roth) and Picea abies (L.) Karst. and nine mixed stands of birch and Pinus sylvestris L. in south-eastern Norway. Each mixed stand and the adjacent pure coniferous stand (control) were growing under the same site conditions and had not been commercially thinned. There were no significant differences in mean age at breast height or in top heights between birch and conifers in the mixed stands, while mean height was significantly higher for birch than for spruce. A growth index was calculated based on total volume and age at breast height. For the spruce sites the growth in young mixed stands (birch < 17 m) was superior to that of pure spruce, while the difference was insignificant in older stands. The growth index correlated positively with the ratio between generatively and vegetatively regenerated birch trees, and negatively with the age of the oldest species in the mixture and with site quality. For the pine sites there was no significant difference in the growth index between mixed birch-pine and pure pine stands. A mixture effect of birch on the volume yield of spruce or pine could not be demonstrated.  相似文献   

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