块根紫金牛ISSR-PCR反应体系的建立与优化

采用正交和单因素试验设计方法,研究Mg2+、dNTP、引物、TaqDNA聚合酶、模板DNA、退火温度及循环次数对ISSR-PCR扩增效果的影响,建立块根紫金牛ISSR-PCR反应体系和扩增程序,即25 ul的体系中含Mg2+ 2.0 mmol/L,dNTP0.15 mmol/L,引物1.0μmol/L,TaqDNA聚合酶1.0U,模板DNA50ng,10×Buffer 2.5μl.适宜的扩增程序是94℃预变性5 min,94℃变性30s,56.6℃退火45s,70℃延伸2.0 min;45个循环;72℃延伸7min,4℃保存.采用正交和单因素试验可快速建立ISSR-PCR反应体系.该体系稳定、可靠,可用于块根紫金牛遗传多样性分析.     

广西特有植物小花异裂菊ISSR-PCR反应体系的建立

以小花异裂菊的叶片为材料,采用正交和单因素试验研究Mg2+、dNTP、TaqDNA聚合酶、引物、模板DNA浓度对ISSR-PCR扩增效果的影响。建立了适宜于小花异裂菊的ISSR-PCR反应体系,即25μL的体系中含Mg2+1.5 mmol/L,dNTP 0.20 mmol/L,TaqDNA聚合酶1.5 U,引物0.8μmol/L,模板DNA 50 ng,10×Buffer 2.5μL。该体系稳定、可靠,为利用ISSR分析小花异裂菊遗传多样性等工作奠定了良好的基础。     

毛竹ISSR-PCR反应体系的建立与优化

采用正交和单因素试验设计方法,研究Mg²⁺、dNTP、引物、TaqDNA聚合酶、模板DNA浓度及退火温度对毛竹ISSR-PCR扩增效果的影响。毛竹ISSR-PCR反应体系为:25μL的体系中含Mg²⁺1.5 mmol/L,dNTP 0.25 mmol/L,引物1.0μmol/L,TaqDNA聚合酶1.0 U,模板DNA 30 ng,10×Buffer2.5μL。扩增程序为94℃预变性5 min,94℃变性45 s,54℃退火60 s,72℃延伸1.5 min,40个循环;72℃延伸7 min,4℃保存。该体系稳定、可靠,可用于毛竹遗传多样性分析。     

黄枝油杉ISSR-PCR反应体系的建立与优化

先运用正交设计进行初步筛选,再用单因素设计逐一优化对ISSR-PCR扩增效果有影响的Mg2+、Taq DNA聚合酶、dNTP、引物、模板DNA、循环次数及退火温度.建立了黄枝油杉的最佳反应体系和程序,即25μL体系中含2.0mmol/L的Mg2+、1.5U Taq DNA聚合酶、0.10mmol/L的dNTP、1.0μmol/L的引物、30ng的模板DNA以及2.5μL10×PCR buffer,其余的用灭菌的ddH2O补够25μL.扩增程序:94℃预变性5min;94℃变性30s,48～56℃(不同的引物,其退火温度不同,根据具体引物而定)退火45s,72℃延伸90s,以上3个步骤循环50次;最后72℃延伸7min;扩增产物放在4℃冰箱中保存.该体系和程序稳定性良好,结果可靠,可用于黄枝油杉遗传多样性分析.     

枣ISSR扩增体系的建立

以冬枣DNA为研究对象,利用单因素试验,分析了Mg2+浓度、dNTP浓度、Taq酶含量、引物浓度、模板DNA含量以及退火温度对ISSR-PCR扩增的影响,经优化建立了枣属植物最适ISSR-PCR反应体系,25μL反应液中包含1×PCR Buffer、2.0 mmol/L Mg2+、0.25 mmol/L dNTP、1.5 UTaq酶、1.25μmol/L引物和50 ng模板,最适退火温度为58℃。PCR反应体系的建立为应用ISSR标记技术开展枣种群遗传变异分析和构建遗传图谱奠定基础。     

新疆贝母DNA提取及ISSR-PCR体系的建立与优化

目的:从新疆贝母干叶片中提取高质量的DNA,建立ISSR-PCR体系并进行优化。方法:通过改进的CTAB法提取基因组DNA;综合利用单因素实验和L9(34)正交实验2种方法,对镁离子、dNTP、模板DNA含量、TaqDNA聚合酶量、引物浓度等因素进行优化,并采用温度梯度筛选引物的退火温度。结果:新疆贝母20μL ISSR-PCR最适反应体系dNTP为0.3 mmol/L,Mg2+为2.0 mmol/L,模板浓度为40～100 ng/(20μL),引物为1.2 mmol/L,Taq DNA聚合酶为1.0 U/20μL,引物UBC 859的最适退火温度为50.7℃。结论:此提取方法得到的DNA纯度高,ISSR-PCR扩增效果显著,为新疆贝母种质资源鉴定、遗传多样性分析奠定研究基础。     

水葫芦苗ISSR-PCR体系的优化与引物筛选

以改良CTAB法提取水葫芦苗基因组DNA为模板,利用正交设计试验对影响ISSR-PCR反应体系的5个因素(TaqDNA聚合酶,dNTP,引物,模板及Mg~(2+))进行了优化筛选。经极差分析和方差分析,最终建立了水葫芦苗ISSR-PCR的最佳反应体系。结果表明,总体积20μL的反应体系中各因素浓度分别为:TaqDNA聚合酶0.025 U/μL、Mg~(2+)1.5 mmol/L、dNTP 1.0 mmol/L、引物0.5μmol/L、模板DNA 1.5 ng/μL。通过筛选得到10条扩增条带清晰的ISSR引物,并通过梯度PCR确定了各引物的最适退火温度。在此基础上对循环次数进行优化,最终确定最佳循环次数为40次。     

文冠果ISSR反应体系的建立及优化

为了建立文冠果ISSR-PCR最佳反应体系,从而为文冠果遗传多样性研究提供理论依据,从定西巉口林场文冠果人工林内选取健康嫩叶为试验材料,通过单因子试验研究了影响ISSR-PCR反应各因素的浓度,分析因子包括模板DNA用量、退火温度、dNTP浓度、Mg2+浓度、引物浓度及TaqDNA聚合酶用量等。通过研究,找出了各因子合适的条件,建立并优化了文冠果ISSR反应体系,即20μL总反应体系含模板DNA约30 ng,Mg²⁺2.5 mmol/L,引物0.2μmol/L,dNTP 0.20 mmol/L,Taq聚合酶1 U。试验结果表明,在此反应体系下可得到多样性好、条带清晰的图谱。     

柑桔SRAP和ISSR分子标记技术体系的建立与优化

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.     

酥瓜(Cucumis melo var. conomon Group)ISSR-PCR反应体系建立与优化

利用正交设计L_(16)(4~5),对酥瓜ISSR-PCR反应体系的5个影响因素(引物,d NTPs,Taq DNA聚合酶,Mg2+和模板DNA)在4个水平上进行优化试验,并在36℃~56℃范围内摸索退火温度,建立适合酥瓜ISSR-PCR反应体系,结果表明,在20μL反应体系中,含有引物0.2μmol/L、d NTPs 0.3 mmol/L、Taq DNA聚合酶1.2 U、Mg2+1.0 mmol/L、DNA模板70 ng、10×Buffer 2.0μL为最佳反应体系,ISSR-PCR扩增程序中最佳退火温度为52.5℃。该体系为酥瓜种质资源的遗传多样性分析评价提供了帮助。     

Location of a high-lysine gene and the DDT-resistance gene on barley chromosome 7

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.     

Simultaneous Determination of Four Phenolic Acids in Radix Salviae Miltiorrhizae Formula Granules by QAMS

[Objectives]This study aimed to establish a QAMS(quantitative analysis of multi-components by single-marker)method for simultaneous determination of four phenol...     

Pollen and pollination experiments. III. The viability of apple and pear pollen as affected by irradiation and storage

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.     

Evolution under domestication involving disturbance of genic balance

Summary An hypothesis is developed that the rapid change from wild plants into domesticated crops principally involves the selection of alleles with non-functional gene products which leads to reduced control of the highly integrated metabolism and morphogenesis previously accumulated by lengthy natural selection. Such disturbance of the genome produces altered physiological and morphological development which, although deleterious in nature, serves mankind better and has been selected.     

Genetic variation for disease and nematode resistances and forage quality in perennial diploid and tetraploid lucerne populations (Medicago sativa L.)

Summary Twenty-five lucerne populations of the Medicago sativa complex, which were either diploid or tetraploid and wild or cultivated, were analysed for their resistance to four different fungal diseases and to stem nematode. Forage quality, including stem digestibility and saponin content, was also tested.Populations varied in susceptibility to the diseases caused by Colletotrichum trifolii, Verticillium albo-atrum, Sclerotinia trifoliorum and Pseudopezizza medicaginis, and to the nematode Ditylenchus dipsaci. Except for Sclerotinia rot, sativa and falcata subspecies differed in susceptibility, but this grouping of populations did not account for the full range of variation among them. However, the resistance to P. medicaginis was much lower in the sativa than in the falcata populations.Populations also varied significantly in stem fiber content and digestibility. Stem digestibility was negatively correlated to forage yield. Wild sativa and falcata populations had lower fiber content and higher digestibility than cultivated sativa populations. The medicagenic acid was the sapogenin responsible for the anti-nutritional effect of the lucerne measured by the yellow mealworm larvae Tenebrio molitor. The medicagenic acid content was lowest for the pure sativa populations, highest for the pure falcata populations, and intermediate for the French sativa varieties that have some traits originating from falcata germplasm. Some populations could be used in breeding programs to improve disease and nematode resistance, and forage quality.     

Review of Effects of Active Components of Gastrodiae Rhizoma on Central Nervous System

In order to explore the pharmacological effects of active components of Gastrodiae Rhizoma on the central nervous system,through consulting related literatures,...     

Effects of Pb on Rhizosphere Soil Enzyme Activity and Chemical Constituents of Achyranthes bidentata Blume

[Objectives]The purpose of this study was to investigate the effects of Pb on rhizosphere soil enzyme activity and chemical constituents of Achyranthes bidentat...     

不同P-Zn配比对小麦幼苗微量元素营养的影响

采用了螯合-缓冲营养液培养方法对小麦进行了苗期培养试验,在3个P水平(0,0.6,3.0 mmol/L)和3个Zn水平(0,3,30 μmol/L)的完全组合下对小麦苗期生长及Zn、Fe、Cu、Mn营养进行了研究,旨在为小麦微肥施用提供理论依据.结果表明,P、Zn的正常供应促进了小麦生长,二者的缺乏与过量均会抑制小麦发育,且这种影响在冠部表现得更为明显.在小麦苗期,Zn与Cu的吸收存在明显的拮抗作用,但供Zn则促进了Zn和Cu的转运,而Mn转运则受到了抑制;过量供Zn时,大量Zn被转运到冠部,同时明显抑制了(Fe+Cu+Mn)的吸收总量;P的供应显著地抑制了Fe的吸收,但P的供应提高了Zn、Cu、Mn的转运率;P、Zn在对Zn与Fe、Cu、Mn间吸收竞争的影响中,Zn本身的影响要比P的影响更为明显,供Zn明显促进了小麦幼苗对Zn的吸收;在小麦幼苗冠部,Zn与Fe的竞争中,供P利于Zn的吸收,缺P则利于Fe的吸收;而Zn与Cu以及Zn与Mn间的竞争中,缺磷时利于Zn的吸收,供磷后则利于Cu和Mn的吸收.总之,小麦幼苗Zn、Fe、Cu、Mn营养中,P、Zn的不同配比会不同程度地改变Zn与Fe、Cu、Mn的协同或拮抗效应.     

Isozyme variation between diploid and tetraploid cytotypes of Glycine tabacina (Labill.) Benth.

Summary Glycine tabacina (Labill.) Benth. is a wild perennial species related to the cultivated soybean, G. max (L.) Merr. It is composed of diploid (2n=40) and tetraploid (2n=80) cytotypes. Currently, to differentiate the cytotypes, plants are grown out in the greenhouse and chromosome counts made on pollen mother cells. It is a laborious and time consuming process. The objective of this study was to determine whether electrophoretic techniques could be utilized to separate the cytotypes. Electrophoretic examination of seven isozyme systems from seed of 67 G. tabacina accessions revealed banding patterns that could be used to differentiate between diploid and tetraploid cytotypes in the species. Among the tetraploid accessions, the number of bands observed were always greater than the diploids. Some tetraploid banding patterns consisted of bands similar to the diploid tabacina and/or additional bands previously identified in other Glycine species. The patterns of isozyme multiplicity and variation in the tetraploid tabacinas suggests more than one mode of origin for the tetraploids.     

Some physiological and photosynthetic characters of rice × sorghum and rice × wheat hybrids

Summary Some photosynthetic characters as-leaf anatomy, leaf photosynthetic rate and CO₂ compensation pointdistinguishing C₃ and C₄ plants and physiological characters as leaf area ratio, nitrogen content, leaf stem ratio and total shoot-to-root and deep root-to-shoot ratios have been studied in rice × sorghum and rice × wheat hybrids. Rice × sorghum 1. has lower values of photosynthetic rate, leaf nitrogen, total root and deep root-to-shoot ratio and CO₂ compensation point as of rice parent where as, rice × sorghym 2. is superior in all these characters. Both hybrids lack kranz anatomy. Though both rice × sorghum hybrids show characters of C₃ rice plant but rice × sorghum 2. has improved drought tolerance and leaf characters in relation to yield. Rice × wheat hybrid have higher assimilatory area and higher total root-to-shoot ratio. Grains of rice × wheat hybrids are identical to rice grain. However, as grains of rice × wheat hybrid does not contain seed coat, it could be exploited as novel rice germplasm after improvement.Abbreviations T CO₂ compensation point - T21 CO₂ compensation point at 21% O₂ - T2 CO₂ compensation point at 2% O₂ - dR/sR deep Root-to-shoot Ratio - LAR Leaf Area Ratio - CER photosynthetic rate - TR/SR Total Root-to-Shoot Ratio