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以杜鹃花为试验材料,采用正交实验方法,研究模板DNA浓度、ISSR引物浓度、dNTPs浓度、Taq DNA聚合酶浓度、Mg2+浓度等5个因素对ISSR-PCR反应体系的影响,以建立适合杜鹃花的ISSR-PCR最佳扩增体系。结果表明:杜鹃花ISSR反应体系的最佳条件为模板DNA用量为60ng/20μL,ISSR引物浓度0.60μmol/L,dNTPs浓度0.50μmol/L,Taq DNA聚合酶浓度30U/mL,Mg2+浓度为0.6mmol/L。采用该反应体系可以从10份杜鹃花(R.simsii)基因组内扩增出稳定性高、重复性好ISSR-PCR产物。该研究为杜鹃花的遗传多样性分析、ISSR指纹图谱构建、亲缘关系鉴定等研究奠定了基础。 相似文献
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以30个茶花品种为试材,采用5因素4水平正交实验以及单因素优化试验方法,研究Mg2+浓度、dNTP浓度、引物浓度、Taq DNA聚合酶浓度和模板DNA浓度对ISSR-PCR指纹图谱条带清晰度的影响,以进行茶花品种ISSR-PCR反应体系优化及引物筛选。结果表明:茶花品种ISSR-PCR最适扩增条件为25μL反应体系中,Mg2+3.0mmol/L、dNTPs 0.2mmol/L、引物0.3mmol/L、Taq DNA聚合酶0.5U、模板DNA 80ng以及52.1℃退火温度;试验从100个ISSR引物中筛选出12个适用引物;并对12个ISSR引物的多态性和稳定性进行了检验。 相似文献
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采用改良CTAB法提取古樟叶片基因组DNA.利用单因素试验设计,对反应体系中退火温度、模板DNA含量、Mg2+浓度、dNTP浓度、引物浓度、Taq DNA聚合酶用量、去离子甲酰胺等7种因素不同梯度对扩增结果的影响进行了比较分析.结果表明:古樟ISSR-PCR最佳反应体系为,在20 μL PCR反应体积中,含50 ng DNA模板,2.5 mmol/L MgCl2,0.20 mmol/LdNTPS,1 U Taq DNA聚合酶,0.5μmol/L引物浓度,2％去离子甲酰胺.该反应体系的建立为今后利用ISSR分子技术对古樟的遗传多样性分析提供了技术支撑. 相似文献
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平欧杂种榛ISSR反应体系的优化与验证 总被引：1，自引：0，他引：1
以5-’(AG)8(AGC)C-3’为引物对育种代号为82-6的平欧杂种榛品系的ISSR反应体系中各个主要影响因子进行优化筛选。结果表明:在20μL ISSR反应体系中各组分的最适浓度分别是,DNA模板为2.5 ng/μL,Mg2+浓度为1.875 mmol/L,1×buffer(不含MgCl2),引物的浓度为0.3μmol/L,Taq DNA聚合酶1 U,dNTP浓度为200μmol/L。并利用该优化体系对平欧杂种榛的15个品种(系)进行了ISSR扩增,证实该体系稳定可靠。 相似文献
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《中国南方果树》2017,(4)
为更好地将ISSR标记应用于番石榴种质研究,以"维邦2号"番石榴DNA为筛选体系试验材料,采用单因子试验对ISSR-PCR反应中Mg~(2+)浓度、dNTPs浓度、引物浓度、Taq酶浓度、DNA浓度进行了优化;选用引物UBC810对9份番石榴种质进行PCR反应,选取"南宁本地番石榴实生2号"DNA作为模板对8条ISSR引物进行PCR反应,对优化扩增体系进行验证。结果表明,番石榴20μL最适ISSR-PCR反应体系为1.0mmol/L Mg~(2+),0.25mmol/L dNTPs,0.8μmol/L引物,0.2U Taq酶,10ng的DNA(2.0μL 10×Buffer,加入适量ddH2O至20μL)。验证试验得到的扩增产物条带多态性丰富,且特异性强、重复性好。试验确立的最适ISSR-PCR反应体系适用于番石榴的ISSR分子标记。 相似文献
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利用正交设计L16(45)对金线莲ISSR-PCR反应体系的5因素(模板DNA、Taq酶、引物、Mg2+和dNTP)在4个水平上进行优化试验,采用直观分析法获得影响因素最佳反应水平。结果表明:最终建立的金线莲ISSR-PCR的最佳反应体系为,在25μL的反应体系中,DNA模板为40ng、Taq酶为1.60U、引物浓度为0.80mmol/L、dNTP浓度为0.96mmol/L、Mg2+浓度为2.40mmol/L。该反应体系的建立为金线莲种质资源分类、遗传多样性分析提供了更客观可靠的方法。 相似文献
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以大白菜为试材,采用新型植物基因组DNA提取试剂盒提取大白菜基因组DNA,采用正交实验设计方法,对dNTPs、Mg2+、Taq DNA聚合酶、引物、及模板DNA五因素四水平进行优化,筛选并建立了适合大白菜的ISSR-PCR反应体系。结果表明:25μL的反应体系中含有1.5mmol/L Mg2+、200μmol/L dNTP、0.5UTaq DNA聚合酶、0.7μmol/L引物、30ng模板DNA。在此基础上探讨了最佳循环次数,应用该优化反应体系,用3个不同循环数对资源DNA进行ISSR-PCR扩增,结果显示优化的反应体系适宜的循环次数是30。 相似文献
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Zusammenfassung Die Leistungsprüfungen wurden im Zeitraum 1997 bis 2003 mit den Unterlagen Gisela 4 und 5, den Klonnummern 195/20 und 497/8 aus der Gisela-Serie sowie Weiroot 10, 13, 53, 72 und 158 durchgeführt. Dabei dienten Sämlinge von P1 (bulgarische Selektion aus Prunus mahaleb) als Kontrolle. Alle Unterlagen waren mit der Sorte Stella veredelt und im Dezember 1996 in der Versuchsanlage der Agraruniversität in Plovdiv, Bulgarien, im Abstand von 6 m×4,5 m gepflanzt worden. Dabei erfolgte ein Pflanzschnitt. Nach Abschluss der natürlichen Kronenentwicklung wurde jedes Jahr ein Winterschnitt vorgenommen. Der Boden wurde durch mechanische Bearbeitung offen gehalten und nach dem 4. Standjahr wurden die Baumstreifen mit Herbiziden behandelt. Die Wasserversorgung erfolgte durch eine dem natürlichen Gefälle folgende Überflutung, allerdings nicht immer zum optimalen Zeitpunkt, da keine eigene Wasserquelle zur Verfügung stand.Basierend auf den Ergebnissen bis zum Anfang des 7. Standjahres können die untersuchten Unterlagen in zwei Gruppen differenziert werden: starkwüchsig—Weiroot 10, P1 und Weiroot 13; mittelstarkwachsend bis schwachwüchsig—Gi 497/8, Gisela 4, Weiroot 53, Weiroot 158, Gi 195/20, Weiroot 72 und Gisela 5. Letztere zeichnete sich durch besondere Schwachwüchsigkeit aus. Die meisten Wurzelschosser bildeten Gisela 4, Weiroot 10 und Weiroot 13. Weiroot 53, Weiroot 72 und Weiroot 158 entwickelten deutlich weniger und P1, Gisela 5, Gi 195/20 sowie Gi 497/8 keine Wurzelschosser. Den frühesten Blühbeginn induzierte Gisela 4. Die anderen Unterlagen führten, in Abhängigkeit von den Temperaturbedingungen des jeweiligen Jahres, zu einer Verspätung der Blüte: P1 und Weiroot 10 um 1–2 Tage; Gi 497/8, Weiroot 13 und Weiroot 158 um 2–4 Tage; Weiroot 72 um 2–7 Tage; Gi 195/20 um 3–6 Tage; Weiroot 53 um 3–8 Tage und Gisela 5 um 3–10 Tage. Die Reifezeit der Früchte war bei den Bäumen auf Gisela 5 im Vergleich zu den anderen Varianten um 2–3 Tage verspätet. Gisela 5, Weiroot 72 und Gisela 4 induzierten bei der aufveredelten Sorte die höchsten Ertragsleistungen, P1 die geringsten. Bei den Bäumen auf Gisela 5 war die Fruchtgröße geringer als bei den anderen Unterlagen. Bäume auf Gisela 5 brauchen intensive Pflege. Nur wenn alle Produktionsfaktoren und kulturtechnischen Maßnahmen optimiert werden, kann das hohe Ertragspotenzial dieser Unterlage ausgeschöpft werden. 相似文献
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AIM: To examine the autoantibody against α1-adrenoceptor and its biologic activities during the development of renal hypertension. METHODS: Renal hypertension of rat was achieved by clipped renal artery, the titre of autoantibody to α1-adrenoceptor was detected using ELISA immunoassay. Furthermore, the biological offects of these autoantibodies on cultured cardiomyocytes were also examined. RESULTS: After two weeks of clipping renal arteries, both the frequency of occurrence and the titre of autoantibodies to cardiac α1-adrenergic receptor were significantly increased as compared with the control of pre-treatment. The increased autoantibodies lasted for several weeks and then automatically decreased gradually to the pre-clipping level at 12 weeks. The biological effects of these autoantibodies displayed an "agonistic-like" activities on the beating frequency of cultured neonatal cardiomyocytes. CONCLUSION: Autoantibodies against α1-adrenoceptor may play a role in the elevation of peripheral vascular resistance and in the development of cardiac hypertrophy in rats with renal hypertension. 相似文献
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AIM: Although endovascular radiotherapy inhibits neointimal hyperplasia, the exact alterations induced by β-particles irradiation remain to be elucidated. The objective of this study was to investigate the ability and the cellular mechanism of local β-particles emission from 188Re to inhibit vascular smooth muscle cells (SMCs). METHODS: The SMCs in vitro were irradiated by 188Re with single doses of 2.6 Gy-25.8 Gy. The effects of β-particles on SMCs, such as effective irradiate doses, the period of inhibition for SMCs proliferation, the changes of cell proliferation rate and DNA synthesis rate, cell cycle progression and related gene expression, were investigated by cell count, [3H]-TdR incorporation, cell cycle progression analysis, cell viability and immunocytochemistry, respectivecy. RESULTS: β-particles irradiation with dose of 5.2 Gy could inhibit significantly SMCs proliferation. At dose of 20.6 Gy DNA synthesis inhibitory rate was 92%, SMCs proliferation rate was only 3%. Renoval of 188Re did not abolish the inhibitory effects of β-particles on SMCs proliferation. The expression of P53 was up regulation and PCNA was down regulation after irradiation. CONCLUSION: β-particles from 188 Re was significantly effective and permanent in inhibiting SMCs proliferation, and inhibitory effect was in dose-dependet manner ED50was 5 Gy, the best dose to inhibit SMCs proliferation was 20 Gy. β-particles irradiation induced SMCs to occur G0/G1 arrest, damaged the ability of SMCs reproliferation and led to cell clonogenic death. P53 and PCNA had regulatiory effects on SMCs proliferation after β-particles irradiation. 相似文献
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《International Journal of Fruit Science》2013,13(1):139-156
Abstract This review is based partly on complete articles and partly on abstracts. Three of the 60 articles deal with the total uptake of elements in strawberry plant organs in two different strawberry production systems, both considered as optimal concerning amount and balance of elements. The effect on fruit quality may be dramatic if the level of a particular element is outside this range, but there may also be effects initiated by differences within the optimal range of elements. Most articles refer to product oriented quality, but some focus on consumer oriented quality, as discussed by Shewfelt (1999). The discussion here is on a general basis, so one should keep in mind that there are cultivar differences and that specification of nutrition ideally should mirror the needs of a single cultivar, or a group of cultivars with similar requirements. Also, to get a complete understanding of the subject future reviews should embrace a broader access of information including the effect on plant development of individual elements, such as the role of calcium in fruit firmness and its importance in cell wall structure. However, the intention here is to narrow the information to results that suggest a direct connection between nutrient uptake and fruit quality. 相似文献
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AIM:To study the effect of L-Arg on plasma content of endothelin (ET) and the expression of proto-oncogene c-fos mRNA in the left ventricle of rats with renovascular hypertensive hypertrophy. METHODS: The level of c-fos mRNA were measured by in situ hybridization. The ET in plasma were measured by radioimmunoassay. RESULTS:After eight weeks of treatment with L-Arg, the expression of c-fos decreased markedly (P＜0.01). The ET content in plasma also decreased significantly by L-Arg(P＜0.01).CONCLUSION: Plasma ET content and the expression of c-fos in the left ventricle of rats with renovascular hypertensive hypertrophy could be decreased by L-Arg administration. 相似文献
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GONG Yong-sheng LI Ji-wu YANG Peng-lin FAN Xiao-fang HU Liang-gang ZHENG Lü-zhen JIANG Zhong-sun 《园艺学报》2003,19(9):1261-1264
AIM:To investigate the effect of puerarin on pulmonary vessel collagen metabolism in pulmonary hypertension rats induced by chronic hypoxia and hypercapnia.METHODS:Collagen Ⅰ, Ⅲ and their mRNA were observed in pulmonary arterioles by the technique of immunohistochemistry and in situ hybridization.RESULTS:① Light microscopy showed media thickness of pulmonary arterioles was much higher in HH(hypoxic-hypercapnia) group than that of NC(normal control) group, and, vessel cavity turned more straiter in HH group than that of NC group.However, the damage of pulmonary arterioles in HP(hypoxic-pueratin) group was much slighter than that of HH group. ② The levels of plasma ET-1 and lung homogenates Hyr were much higher in HH group than those of NC group(P＜0.01), and lower in HP group than HH groups(P＜0.01).Plasma NO content in group HH was lower than that of group NC(P＜0.01), it was higher in group HP than that of group HH(P＜0.01).③Expression of collagen Ⅰ and collagen Ⅰ mRNA in pulmonary arterioles were significantly higher in HH groups than those of NC group (P＜0.01), and they were lower in HP group than those of HH group (P＜0.01).Expression of collagen Ⅲ and collagen Ⅲ mRNA showed no difference among three groups(P＞0.05).CONCLUSION:Puerarin inhibited the deposition of collagen and improved pulmonary vessel remodeling. 相似文献
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Fulé Peter Z. Crouse Joseph E. Heinlein Thomas A. Moore Margaret M. Covington W. Wallace Verkamp Greg 《Landscape Ecology》2003,18(5):465-486
Fire regime characteristics of high-elevation forests on the North Rim of the Grand Canyon, Arizona, were reconstructed from fire scar analysis, remote sensing, tree age, and forest structure measurements, a first attempt at detailed reconstruction of the transition from surface to stand-replacing fire patterns in the Southwest. Tree densities and fire-/non-fire-initiated groups were highly mixed over the landscape, so distinct fire-created stands could not be delineated from satellite imagery or the oldest available aerial photos. Surface fires were common from 1700 to 1879 in the 4,400 ha site, especially on S and W aspects. Fire dates frequently coincided with fire dates measured at study sites at lower elevation, suggesting that pre-1880 fire sizes may have been very large. Large fires, those scarring 25% or more of the sample trees, were relatively infrequent, averaging 31 years between burns. Four of the five major regional fire years occurred in the 1700s, followed by a 94-year gap until 1879. Fires typically occurred in significantly dry years (Palmer Drought Stress Index), with severe drought in major regional fire years. Currently the forest is predominantly spruce-fir, mixed conifer, and aspen. In contrast, dendroecological reconstruction of past forest structure showed that the forest in 1880 was very open, corresponding closely with historical (1910) accounts of severe fires leaving partially denuded landscapes. Age structure and species composition were used to classify sampling points into fire-initiated and non-fire-initiated groups. Tree groups on nearly 60% of the plots were fire-initiated; the oldest such groups appeared to have originated after severe fires in 1782 or 1785. In 1880, all fire-initiated groups were less than 100 years old and nearly 25% of the groups were less than 20 years old. Non-fire-initiated groups were significantly older (oldest 262 years in 1880), dominated by ponderosa pine, Douglas-fir, or white fir, and occurred preferentially on S and W slopes. The mixed-severity fire regime, transitioning from lower-elevation surface fires to mixed surface and stand-replacing fire at higher elevations, appeared not to have been stable over the temporal and spatial scales of this study. Information about historical fire regime and forest structure is valuable for managers but the information is probably less specific and stable for high-elevation forests than for low-elevation ponderosa pine forests.This revised version was published online in May 2005 with corrections to the Cover Date. 相似文献
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《International Journal of Fruit Science》2013,13(3):101-120
Abstract Saskatoon berry (Amelanchier alnifolia Nutt., Rosaceae) and blueberry (Vaccinium corymbosum L., Ericaceae) are substantially equivalent in all characteristics that are important to the consumer, including fruit color, shape, size, nutrition, texture, and uses. In addition, both fruits are native to North America and they have practically identical historical uses and known health benefits. Their composition, processing, nutritional value and metabolism, intended uses, and levels of undesirable substances are compared. 相似文献
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The objective of this study was to establish a cryopreservation protocol for hawthorn shoot apices (Crataegus pinnatifida Bge.). Cryopreservation was carried out via encapsulation–dehydration, vitrification, and encapsulation–vitrification on shoot apices excised from in vitro cultures. We began by showing that cold-acclimation enhanced the regrowth of cryopreserved apices from 10.0 to 65.5% in encapsulation–dehydration. We then decided that the encapsulation–dehydration method was an optimal cryopreservation method for hawthorn shoot apices in terms of its high recovery after cryopreservation as well as its ease of use compared with vitrification and encapsulation–vitrification. In encapsulation–dehydration, the protocol leading to optimal regrowth was as follows: after cold-acclimation at 5 °C in the dark for 2 weeks, excised shoot tips were pretreated for 24 h at 25 °C on hormone-free Murashige and Skoog [Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol. Plant. 15, 473–497] (MS) basal medium with 0.4 mol/L sucrose, then encapsulated and precultured in liquid MS medium with 0.8 mol/L sucrose for 16 h at 25 °C. Precultured beads were dehydrated for 6 h at 25 °C in the dessicator containing 50 g silica gel to a moisture content of 15.3% (fresh-weight basis) before cryostorage for 1 h. In addition, we examined the effect of adding glycerol to both the alginate beads and loading solution to enhance regrowth after cryopreservation in encapsulation–dehydration. In the present study, it was shown that adding 0.5 mol/L glycerol resulted in high regrowth percentages (82.5–90.0%) in four Crataegus species. 相似文献
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Habitat for wide-ranging species should be addressed at multiple scales to fully understand factors that limit populations.
The marbled murrelet (Brachyramphus marmoratus), a threatened seabird, forages on the ocean and nests inland in large trees. We developed statistical relationships between
murrelet use (occupancy and abundance) and habitat variables quantified across many spatial scales (statewide to local) and
two time periods in California and southern Oregon, USA. We also addressed (1) if old-growth forest fragmentation was negatively
associated with murrelet use, and (2) if some nesting areas are more important than others due to their proximity to high
quality marine habitat. Most landscapes used for nesting were restricted to low elevation areas with frequent fog. Birds were
most abundant in unfragmented old-growth forests located within a matrix of mature second-growth forest. Murrelets were less
likely to occupy old-growth habitat if it was isolated (> 5 km) from other nesting murrelets. We found a time lag in response
to fragmentation, where at least a few years were required before birds abandoned fragmented forests. Compared to landscapes
with little tono murrelet use, landscapes with many murrelets were closer to the ocean's bays, river mouths, sandy shores,
submarine canyons, and marine waters with consistently high primary productivity. Within local landscapes (≤ 800ha), inland
factors limited bird abundance, but at the broadest landscape scale studied (3200 ha), proximity to marine habitat was most
limiting. Management should focus on protecting or creating large, contiguous old-growth forest stands, especially in low-elevation
areas near productive marine habitat.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献