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1.
风信子ISSR-PCR体系的优化及引物筛选 总被引：1，自引：0，他引：1
以风信子基因组DNA为ISSR-PCR扩增模板,采用单因素试验方法,对影响PCR扩增体系中Mg2+浓度、dNTPs、模板DNA及引物浓度、Taq酶的用量5个因素进行研究,建立了风信子ISSR-PCR扩增最佳反应体系,即:20μL反应体系中分别加入2μL10×Buffer、1.4μL Mg2+(25mmol/L)、1.5μL dNTPs(2.5mmol/L)、1.5μL引物(10pmol/μL),0.2μLTaq酶(5U/μL),1.2μL模板(30ng/μL),ddH2O补足体积。并以此体系对110条引物进行筛选,最终获得了多态性高,重复性好的引物12条。 相似文献
2.
《华北农学报》2017,(Z1)
为了对玉米弯孢病菌的鉴定、区域发生关系以及遗传多样性等进行研究,采用单因素试验优化玉米弯孢叶斑病菌ISSR-PCR反应体系中模板DNA、引物、dNTPs、Taq酶、Mg~(2+)的用量,并确定每条引物的最佳退火温度。结果表明,最佳反应体系为:模板DNA 1.6μL(25 ng/μL)、引物1.4μL(5μmol/L)、dNTPs 0.25μL(2.5 mmol/L)、Taq酶0.4μL(2.5 U/μL)、Mg~(2+)0.8μL(25 mmol/L)、10×Taq Buffer 2μL、dd H2O 13.55μL。在该体系下选用3个玉米弯孢病菌的全基因组DNA,分别对52条ISSR引物进行筛选,筛选出16条多态性高、扩增稳定的ISSR引物。玉米弯孢病菌ISSR-PCR反应体系的建立为利用ISSR分子标记技术对该病原菌进行遗传分析奠定了基础。 相似文献
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建立最适的PCR反应体系是成功进行ISSR-PCR的关键,且反应的扩增效果易受多种因素的影响,本研究采用正交试验和单因素筛选试验两种方法,选用凤仙花叶片DNA为实验材料,针对影响凤仙花ISSR-PCR反应较大的 Mg~(2+)、dNTPs、Taq酶、引物、模板DNA这5个因素进行优化并筛选,最终建立凤仙花ISSR-PCR最佳反应体系:25μL PCR反应体积中, Mg~(2+)浓度为2.0 mmol/L、引物浓度为0.4μmol/L、模板DNA浓度为150 ng、dNTPs浓度为0.15 mmol/L、Taq酶为0.5 U。利用该体系筛选出14条可用于凤仙花ISSR-PCR扩增的引物。通过建立凤仙花属最佳ISSR-PCR反应体系,为研究凤仙花属植物提供分子标记水平上的理论依据和技术方法。 相似文献
5.
为建立适合新疆野生欧洲李的ISSR-PCR反应体系,本研究以野生欧洲李为供试材料,采用L16(45)正交试验设计和单因素试验设计相结合的方法,对影响野生欧洲李ISSR-PCR反应体系的5个因素(DNA模板,Taq酶,dNTPs,引物,Mg2+)进行筛选与优化,对16条引物的退火温度进行筛选。结果表明,Taq酶对PCR扩增反应的影响最大,野生欧洲李ISSR-PCR最佳反应体系为:总体积20μL,5 U Taq酶0.10μL,10 mmol/L引物1.00μL,2.5 mmol/L dNTPs 1.50μL,10×Buffer(含Mg2+)2.00μL,50 ng/μL DNA模板1.00μL,双蒸水14.40μL。建立的ISSR-PCR反应体系经过22份野生欧洲李样品验证,表明反应体系稳定可靠,可用于后续遗传多样性研究,为野生欧洲李种质资源的保护和利用提供理论参考。 相似文献
6.
柑桔SRAP和ISSR分子标记技术体系的建立与优化 总被引：16，自引：0，他引：16
通过对PCR反应程序、反应体系(DNA模板量、PCR反应体积、Mg2 浓度、dNTP浓度、Taq酶用量、引物量)、电泳检测方法的系统优化,建立了柑桔SRAP-PCR和ISSR-PCR体系;以此进行大规模引物筛选,从而建立了柑桔SRAP和ISSR分子标记技术体系.SRAP-PCR:25μL体系,模板DNA25ng,Tris-HCl10 mmol/L,KCl50 mmol/L,Mg2 1.2 mmol/L,dNTP 120 μmol/L,Taq酶1.5U,引物0.4μmol/L,反应程序为94℃预变性5min,35个循环(94℃ 30s,47℃ 1min,72℃ 1min),72℃延伸10min;ISSR-PCR:25μL体系,模板DNA25ng,Tris-HCl10mmol/L,KCl50mmol/L,Mg2 1.6 mmol/L,dNTP200μmol/L,Taq酶1 U,引物0.8μmol/L.筛选出稳定性好、多态性高的24对SRAP引物和13条ISSR引物. 相似文献
7.
通过研究月季杂交品系的遗传多样性,解决其在月季品种中的分类问题.本试验利用正交设计,从Mg2+、dNTPs、引物、Taq聚合酶和模板DNA 5种因素和4个浓度水平来优化月季杂交品系ISSR-PCR反应体系,其建立的最佳反应体系为:在25μL反应体系中分别加入Mg2+ 2.0 mmol/L、dNTPs 2.0 mmol/L、引物10μ mol/L、Taq酶0.4 U、模板DNA 20 ng/25μL,并含2.5μL的10×PCR Buffer(Mg2+ free)加dd H2O至25μL.此ISSR-PCR反应优化体系为进一步利用ISSR分子标记技术进行月季遗传多样性分析奠定了良好的技术条件和试验基础. 相似文献
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《分子植物育种》2020,(15)
以藜芦为试验材料,通过L_(16)(4~5)正交试验与单因素实验两种方法对藜芦ISSR-PCR反应体系中的5个主要因素(模板DNA,引物, Taq DNA聚合酶, dNTPs, Mg~(2+))进行筛选与优化,建立藜芦ISSR-PCR最佳反应体系;在此优化体系下,从100条引物中筛选适宜的ISSR引物,设置温度梯度检测各引物最佳退火温度。结果表明,藜芦20μL ISSR-PCR反应体系包括:0.30μmol/L引物、24 ng/20μL模板DNA、2 U/20μL Taq DNA聚合酶、0.20 mmol/L dNTP、1.00 mmol/L Mg2+;从100条引物中筛选出16条扩增稳定、多态性丰富的ISSR引物。利用14份藜芦样品验证该优化体系的稳定性,证明该体系稳定可靠,可为后续藜芦的遗传多样性分析研究提供帮助。 相似文献
10.
《分子植物育种》2020,(6)
为建立适合蓝花丹的ISSR-PCR反应体系,本研究以蓝花丹为供试材料,结合L_(16)(4~5)正交试验和单因素试验设计对影响蓝花丹ISSR-PCR反应体系的5个因素(dNTPs, Mg~(2+),模板, Taq酶,引物)进行优化,并在最优反应体系的基础之上进行了引物及其最佳退火温度的筛选。结果表明,5个影响因素中,Mg~(2+)对扩增反应的影响最大;不同退火温度对扩增效果具有显著的影响,但将退火温度设置为55℃时,大部分的引物都能扩增出清晰的条带。最终建立的蓝花丹ISSR-PCR反应体系为:总体积25μL,其中含Taq酶1 U,模板DNA 100 ng,dNTPs 0.15 mmol/L,引物0.4μmol/L,Mg~(2+)0.5 mmol/L,10×Buffer 2μL。研究结果为蓝花丹及其近缘种的遗传多样性研究以及遗传图谱的构建提供分子标记水平上的依据。 相似文献
11.
Biological Activity and Quantification of Suspected Allelochemicals from Alfalfa Plant Parts 总被引：7，自引：0，他引：7
Autotoxicity restricts reseeding of alfalfa (Medicago sativa L.) after alfalfa until autotoxic chemical(s) breaks down or is dispersed into external environments. A series of aqueous extracts from leaves, stems, roots and seeds of alfalfa ‘Vernal’ were bioassayed against alfalfa seedlings of the same cultivar to determine their autotoxicity. The highest inhibition was found in the extracts from the leaves. Extracts at 40 g dry tissue l?1 from alfalfa leaves were 15.4, 17.5 and 28.7 times more toxic to alfalfa root growth than were those from roots, stems and seeds, respectively. A high‐performance liquid chromatography (HPLC) analysis with nine standard compounds showed that the concentrations and compositions of allelopathic compounds depended on the plant parts. In leaf extracts that showed the most inhibitory effect on root growth, the highest amounts of allelochemicals were detected. Among nine phenolic compounds assayed for their phytotoxicity on root growth of alfalfa, coumarin, trans‐cinnamic acid and o‐coumaric acid at 10?3 m were most inhibitory. The type and amount of causative allelochemicals found in alfalfa plant parts were highly correlated with the results of the bioassay, indicating that the autotoxic effects of alfalfa plant parts significantly differed. 相似文献
12.
C. J. Spurr D. A. Fulton P. H. Brown R. J. Clark 《Journal of Agronomy and Crop Science》2002,188(4):275-280
Development of onion (Allium cepa L., cv. ‘Early Cream Gold’) seed under cool climate conditions in Tasmania, Australia occurred over a longer duration than previously reported, but similar patterns of change in yield components were recorded. In contrast to previous studies, umbel moisture content declined from 85 to 67 % over 57 days while seed moisture content decreased from 85 to 31 %. Seed yield continued to increase over the duration of crop development, with increasing seed weight compensating for seed loss resulting from capsule dehiscence in the later stages of maturation. Germination percentage was high and did not vary significantly from 53 to 77 days after full bloom (DAF), but mean germination time declined and uniformity of germination increased significantly over the same time period. The percentage abnormal seedlings declined with later harvest date, resulting in highest seed quality at 77 DAF. The results of this study suggest that the decision to harvest cool climate onion seed crops before capsule dehiscence will result in a loss of potential seed yield and quality. 相似文献
13.
Jens Jensen 《Euphytica》1979,28(1):47-56
Summary The high-lysine gene in Risø mutant 1508 conditions an increased lysine content in the endosperm via a changed protein composition, a decreased seed size, and several other characters of the seed. The designation lys3a, lys3b, and lys3c, is proposed for the allelic high-lysine genes in three Risø mutants, nos 1508, 18, and 19. Linkage studies with translocations locate the lys3 locus in the centromere region of chromosome 7. A linkage study involving the loci lys3 and ddt (resistance to DDT) together with the marker loci fs (fragile stem), s (short rachilla hairs), and r (smooth awn) show that the order of the five loci on chromosome 7 from the long to the short chromosome arm is r, s, fs, lys3, ddt. The distance from locus r to locus ddt is about 100 centimorgans. 相似文献
14.
Hongyan WEI Jian ZHOU Jinguang LU Jingzheng SONG Qiongxi YU Zhihong JIANG 《Medicinal Plant》2019,(6):27-29
[Objectives]This study aimed to establish a QAMS(quantitative analysis of multi-components by single-marker)method for simultaneous determination of four phenol... 相似文献
15.
Chunli TANG Fengxian ZHAO Hongxia CHEN Jiabao MA Jiangcun WEI Meiyan QIU Zhen XIE 《Medicinal Plant》2019,(6):60-65
[Objectives]To optimize the water extraction process of Chinese Herbal Compound Man Gan Ning and establish a method for its extraction and content determination... 相似文献
16.
D. A. Bond G. J. Jellis G. G. Rowland J. Le Guen L. D. Robertson S. A. Khalil L. Li-Juan 《Euphytica》1993,73(1-2):151-166
Progress is being made, mainly by ICARDA but also elsewhere, in breeding for resistance to Botrytis, AScochyta, Uromyces, and Orobanche; and some lines have resistance to more than one pathogen. The strategy is to extend multiple resistance but also to seek new and durable forms of resistance. Internationally coordinated programs are needed to maintain the momentum of this work.Tolerance of abiotic stresses leads to types suited to dry or cold environments rather than broad adaptability, but in this cross-pollinated species, the more hybrid vigor expressed by a cultivar, the more it is likely to tolerate various stresses. 相似文献
17.
Summary Apple and pear pollen was irradiated with doses of 0, 50, 100, 250 and 500 krad (gamma rays) and stored at 4°C and 0–10% r.h. From the in-vitro germination percentages an average LD 50 dose of about 220 krad was estimated. For both irradiated and untreated pollen a close and corresponding lineair relationship existed between germination percentage and pollen tube growth.Irradiated pollen was much more sensitive to dry storage conditions than untreated pollen, resulting in less germination and more bursting. Apparently, irradiation caused the pollen cell membrane to lose its flexibility faster than normal. Rehydration of dry-stored, irradiated pollen in water-saturated air restored germination percentages up to their initial levels. The importance of this procedure in germination trials is stressed. 相似文献
18.
[Objectives] To determine the optimum extraction technology for total phenols of leaves in Acanthopanax giraldii Harms.[Methods]The single factor test and ortho... 相似文献
19.
E. Keep 《Euphytica》1986,35(3):843-855
Summary Cytoplasmic male sterility (cms) is described in the F1 hybrids Ribes × carrierei (R. glutinosum albidum × R. nigrum) and R. sanguineum × R. nigrum. In backcrosses to R. nigrum, progenies with R. glutinosum cytoplasm were either all male sterile, or segregated for full male fertility (F) and complete (S) and partial (I) male sterility. Ratios of F:I+S suggested that two linked genes controlled cms, F plants being dominant for one (Rf
1) and recessive for the other (Rf
2).Segregation for cms in relation to three linded genes, Ce (resistance to the gall mite, Cecidophyopsis ribes), Sph
3(resistance to American gooseberry mildew, Sphaerotheca mors-uvae) and Lf
1(one of two dominant additive genes controlling early season leafing out) indicated that Rf
1and Rf
2were in this linkage group. The gene order and approximate crossover values appeared to be: % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXafv3ySLgzGmvETj2BSbqef0uAJj3BZ9Mz0bYu% H52CGmvzYLMzaerbd9wDYLwzYbItLDharqqr1ngBPrgifHhDYfgasa% acOqpw0xe9v8qqaqFD0xXdHaVhbbf9v8qqaqFr0xc9pk0xbba9q8Wq% Ffea0-yr0RYxir-Jbba9q8aq0-yq-He9q8qqQ8frFve9Fve9Ff0dme% aabaqaciGacaGaamqadaabaeaafaaakeaacaWGdbGaamyzamaamaaa% baGaaiiiaiaacccacaGGWaGaaiOlaiaacgdacaGG0aGaaiiiaiaacc% caaaGaaiiiaiaacccacaGGGaGaamOuaiaadAgaliaaigdakmaamaaa% baGaaiiiaiaacccacaGGGaGaaiiiaiaaccdacaGGUaGaaiOmaiaacs% dacaGGGaGaaiiiaiaacccacaGGGaGaaiiiaaaacaWGsbGaamOzaSGa% aGOmaOWaaWaaaeaacaGGGaGaaiiiaiaacccacaGGGaGaaiiiaiaacc% cacaGGGaGaaiiiaiaacccaaaGaamitaiaadAgaliaaigdakmaamaaa% baGaaiiiaiaacccacaGGGaGaaiiiaiaacccacaGGGaGaaiiiaiaacc% cacaGGGaGaaiiiaiaacccacaGGGaaaaiaadofacaWGWbGaamiAaSGa% aG4maaaa!6E4D!\[Ce\underline { 0.14 } Rf1\underline { 0.24 } Rf2\underline { } Lf1\underline { } Sph3\]. Crossover values of 0.36 for Ce-Lf
1, and 0.15 for Lf
1-Sph
3were estimated from the relative mean differences in season of leafing out between seedlings dominant and recessive for Ce and Sph
3.It is suggested that competitive disadvantage of lf
1-carrying gametes and/or zygotes at low temperatures may be implicated in the almost invariable deficit of plants dominant for the closely linked mildew resistance allele Sph
3. Poor performance of lf
1- (and possibly lf
2-) carrying gametes and young zygotes during periods of low temperature at flowering might also account for the liability of some late season cultivars and selections to premature fruit drop (running off). 相似文献
20.
Parasitic angiosperms cause great losses in many important crops under different climatic conditions and soil types. The most widespread and important parasitic angiosperms belong to the genera Orobanche, Striga, and Cuscuta. The most important economical hosts belong to the Poaceae, Asteraceae, Solanaceae, Cucurbitaceae, and Fabaceae. Although some resistant cultivars have been identified in several crops, great gaps exist in our knowledge of the parasites and the genetic basis of the resistance, as well as the availability of in vitro screening techniques. Screening techniques are based on reactions of the host root or foliage. In vitro or greenhouse screening methods based on the reaction of root and/or foliar tissues are usually superior to field screenings and can be used with many species. To utilize them in plant breeding, it is necessary to demonstrate a strong correlation between in vitro and field data. The correlation should be calculated for every environment in which selection is practiced. Using biochemical analysis as a screening technique has had limited success. The reason seems to be the complex host-parasite interactions which lead to germination, rhizotropism, infection, and growth of the parasite. Germination results from chemicals produced by the host. Resistance is only available in a small group of crops. Resistance has been found in cultivated, primitive and wild forms, depending on the specific host-parasite system. An additional problem is the existence of pathotypes in the parasites. Inheritance of host resistance is usually polygenic and its transfer is slow and tedious. Molecular techniques have yet to be used to locate resistance to parasitic angiosperms. While intensifying the search for genes that control resistance to specific parasitic angiosperms, the best strategy to screen for resistance is to improve the already existing in vitro or greenhouse screening techniques. 相似文献