RAPD-PCR

摘要： Populus alba is a large woody deciduous plant.The plant has been introduced to shooting,then multiplication of rooting on Murashige and Skoog(MS)medium.This work was designed to estimate the effect of two factors(low levels of 1-Naphthaleneacetic acid NAA and sucrose)on P.alba response resulting in 6 treatments compared to the control,with twelve measured responses.There was a significant difference in some measurements in morphology,like plantlets fresh-weight,shoot-,root-length,and leaf number.In the physiological measurements,there were significant differences in all the measured parameters.The low concentrations of sucrose and media composition/power(MS grams/L)led to starvation in plants;however,these conditions led to enhancement in some morphological and physiological parameters to overcome the starvation effect,compared to the control.The RAPD-PCR molecular marker(four decamers)was used to evaluate the new individuals’genetic variation(instability),resulting in a total polymorphism percentage of 50.83%.It was formerly known that the plantlets were identical to each other and to the mother plant.In this study,however,the use of distinct media power,hormonal and sucrose levels resulted in molecular variation reflected in P.alba’s morphological and physiological responses.

摘要： 通过RAPD标记法对5类喀斯特小生境中小蓬竹的遗传结构及其多样性进行了分析。RAPD随机引物8个扩增共得87个清晰度高且重复性好的条带,分子量200～3 000 bp,含多态68条,物种PPL=78.16%,Nei’s基因多度H=0.291 3,Shannon多样性指数I=0.431 0。土面(TM)生长的小蓬竹遗传多样性水平最高(PPL=47.13%,H=0.195 3,I=0.283 4),石缝(SF)中的小蓬竹遗传多样性水平最低(PPL=39.08%,H=0.147 4,I=0.219 2)。POPGENE 32分析5类小生境中小蓬竹的遗传分化系数G_(st)=0.415 1,基因流N_m=0.704 5。5类小生境的小蓬竹间遗传相似系数(I)变化范围0.777 3～0.999 4,平均为0.865 6;遗传距离(D)变化范围0.000 6～0.251 9,平均为0.148 3。各类小生境的遗传多样性水平PPL均值为43.91%,5类小生境间的遗传分化G_(st)=0.405 4。研究结果说明,小生境间的差异增加了小蓬竹的遗传多样性。

摘要： Hyalomma dromedarii ticks are important disease vectors to camels in the UAE and worldwide. Ticks can be identified using DNA-based techniques. In addition, such techniques could be utilized to study the intraspecific genetic diversity in tick populations. In this study, the genetic diversity of four H. dromedarii populations was investigated using mitochondrial cytochrome c oxidase subunit I (COI) gene and randomly amplified polymorphic DNA polymerase chain reaction (RAPD-PCR). The results showed that both of the aforementioned techniques produced similar grouping patterns. Moreover, they revealed that the four tick populations had high levels of genetic similarity. However, one population was slightly different from the three other populations. The current study demonstrated that H. dromedarii ticks in the UAE are very similar at the genetic level and that investigating more locations and screening larger numbers of ticks could reveal larger genetic differences.

摘要： 以兰州百合鳞片和叶片为试材,采用改良CTAB法获得了高质量的基因组DNA.为优化兰州百合RAPD-PCR反应体系,利用正交试验设计,对兰州百合RAPD-PCR反应体系中的5种主要因素进行4水平共计16个组合设计,并选用3种反应程序,通过琼脂糖凝胶电泳结果分析,最终获得最佳反应体系为:20 uL反应体系中含30 ng模板DNA、0.5 μmol/L随机引物、0.2 mmol/L dNTPs、2.25 mmol/L MgCl2,及1.5U Taq DNA聚合酶.最佳反应程序:94°C预变性4min;94°C变性1 min,36°C退火1.5 min;72°C延伸2 min;35个循环;最后72°C延伸7min.

摘要： 以兰州百合鳞片和叶片为试材,采用改良CTAB法获得了高质量的基因组DNA.为优化兰州百合RAPD-PCR反应体系,利用正交试验设计,对兰州百合RAPD-PCR反应体系中的5种主要因素进行4水平共计16个组合设计,并选用3种反应程序,通过琼脂糖凝胶电泳结果分析,最终获得最佳反应体系为:20 uL反应体系中含30 ng模板DNA、0.5μmol/L随机引物、0.2 mmol/L dNTPs、2.25 mmol/L MgCl2,及1.5U Taq DNA聚合酶.最佳反应程序:94°C预变性4 min;94°C变性1 min,36°C退火1.5 min;72°C延伸2 min;35个循环;最后72°C延伸7 min.

摘要： AIM To evaluate effect of treatment failure on cag A and vac A genotypes in Helicobacter pylori(H. pylori) isolates from Colombia.METHODS One hundred and seventy-six participants infected with H. pylori from Colombia were treated during 14 d with the triple-standard therapy. Six weeks later, eradication was evaluated by 13C-Urea breath test. Patients with treatment failure were subjected to endoscopy control; biopsies obtained were used for histopathology and culture. DNA from H. pylori isolates was amplified using primers specific for cag A and vac A genes. The phylogenetic relationships among isolates obtained before and after treatment were established by conglomerate analysis based on random amplified polymorphic DNA(RAPD) fingerprinting.RESULTS Treatment effectiveness was at 74.6%. Of the par-ticipants with treatment failure, 25 accepted subjected to a second endoscopy. Prevalence of posttreatment infection was 64%(16/25) and 40%(10/25) by histology and culture, respectively. Upon comparing the cag A and vac A genotypes found before and after therapy, multiple cag A genotypes(cag A-positive and cag A-negative) were found before treatment; in contrast, cag A-negative genotypes decreased after treatment. vac A s1m1 genotype was highly prevalent in patients before and after therapy. The 3'cag A region was successfully amplified in 95.5%(21/22) of the isolates obtained before and in 81.8%(18/22) of the isolates obtained after treatment. In the isolates obtained from patients with treatment failure, it was found that 72.7%(16/22) presented alterations in the number of EPIYA motifs, compared to isolates found before treatment.CONCLUSION Unsuccessful treatment limits colonization by lowvirulence strains resulting in partial and selective eradication in mixed infections, and acts on the cag A-positive strains inducing genetic rearrangements in cag A variable region that produces a loss or gain of EPIYA repetitions.

摘要： 为了应用RAPD分子标记技术分析马铃薯早疫病菌遗传多样性,采用单因素水平对影响RAPD反应的Mg2+,dNTPs、Taq DNA聚合酶、引物浓度及引物退火温度等条件进行了优化.结果表明:建立了适宜于早疫病菌的RAPD最佳反应体系,25μL的反应体系中,Mg2+2.5 mmo1·L-1,dNTPs 0.20 mmol·L-1,Taq DNA聚合酶1U,引物0.40 μmol· L-1.

摘要： 利用RAPD‐PCR及ISSR‐PCR两种分子标记技术，分析了七里海中华绒螯蟹第6代繁育群体的遗传多样性。用10个多态性高的RA PD引物对24个七里海中华绒螯蟹样本个体进行扩增，共检测出107个不同的扩增位点，扩增片段大小为300～2300 bp。其中多态位点总数为66个，平均多态位点比例为61．32％，群体内香农－维纳多样性值为0．1714。用8个多态性高的ISSR引物对24个个体共检测到106个位点，扩增片段为200～1500 bp。其中多态性位点79个，平均多态位点百分率为74．53％，群体内香农－维纳多样性值为0．1778。结果显示，该群体的多态位点比例和香农－维纳多样性值均较大，说明其有较丰富的遗传多样性，同时也说明RAPD和ISSR技术用于七里海中华绒螯蟹核基因组的遗传多样性分析，具有较高的检出率和灵敏度。%Two types of molecular markers ,namely ,Randomly Amplified Polymorphic DNA (RAPD) and Inter‐Simple Sequence Repeat (ISSR) were used to detect the genetic diversities at the intra sixth genera‐tion breeding population of Chinese mitten handed crab Eriocheir sinensis from Qilihai .Using 10 RAPD primers selected from 33 primers ,a total of 107 amplified loci ranging from 300 to 2300 bp were observed from 24 individuals ,and 66 of 107 loci detected were polymorphic with the percent of 61 .32% ,and the Shannon′s index of 18 .3419 .Eight ISSR primers of 50 primers used generated 106 reproducible and stable bands ,ranging from 200 to 1500 bp , 79 of 107 loci detected were polymorphic with the percent of 74 .53% ,and the Shannon′s index of 18 .8425 .These findings indicated that the sixth generation breeding population had high genetic diversity ,and that the proportion of polymorphic loci and Shannon′s index of Qilihai Chinese mitten handed crab were high ,indicating that they have abundant genetic diversity .

摘要： [目的]研究采自云南的柚喀木虱Cacopsylla citrisuga Yang&Li种群的遗传多样性.[方法]利用线粒体细胞色素C氧化酶亚基I基因(COI)条形码片段对采自云南瑞丽和石屏的柚喀木虱样本进行描述,并进行遗传距离和系统发育分析.[结果和结论]经鉴定共得到2种单倍型C1和C2,瑞丽种群的单倍型均为C1,而石屏的种群分化出了2种单倍型C1和C2.分析表明,柚喀木虱的最大序列分歧度特别低,仅为0.16％.此COI片段能很好地区分柚喀木虱与同属的其他几种木虱.

摘要： 为了建立小蓬竹（ Drepanostachyum luodianense） RAPD-PCR反应的最优体系，以小蓬竹基因组DNA为模板，对影响其RAPD扩增的dNTPs浓度、模板DNA浓度、Taq DNA聚合酶量、引物浓度、Mg2＋浓度等重要参数进行了单因子和正交试验。试验得出小蓬竹RAPD最优反应体系为：20μL反应体系，10×PCR buffer为1/10体积，dNTPs为100μmol· L-1，模板DNA量为30 ng，Taq DNA聚合酶为1.0 U，引物浓度为0.2μmol· L-1，Mg2＋浓度为1.5 mmol· L-1。优化后的RAPD-PCR反应程序为：94°C预变性5 min，然后进入35个循环，即94°C变性1 min，35°C退火30 s，72°C延伸90 s，循环完毕后于72°C延伸7 min，最后在4°C保持。%The Drepanostachyum luodianense RAPD-PCR reaction system was optimized for the analysis of genetic variation and structure of D.luodianense.The genomic DNA of D.luodianense was extracted by improved CTAB method.Then, both single factor test and orthogonal design were applied to study the effects of main factors on the RAPD-PCR system for D.luodianense, in which the main factors included the concentration of dNTPs , primers and Mg2+.The content of template DNA and Taq DNA polymerase and an optimal 20μL RAPD-PCR reaction system for D.luodianense was established, including 1/10 volume 10 ×PCR buffer, 100 μmol· L-1 dNTPs, 30 ng template DNA, 1.0 U Taq DNA polymerase, 0.2 μmol· L-1 primers and 1.50 mmol· L-1 Mg2+.The optimized reaction program was initially at 94°Cfor 5 min, followed by 35 cycles at 94°Cfor 1 min, at 35°Cfor 30 s, at 72°Cfor 90 s, and then held at 72°Cfor 7 min, and finally kept at 4°C.

摘要： 本文研究目的:从不同蜜蜂种类、品系的DNA基因组水平寻找其优良性状的分子标记,并对优良性状的分子标记进行测序和探针的制备,为了应用于全国800万群中优良蜂种的普查及筛选。研究方法:为分离纯化工蜂蜂蛹中基因组DNA,通过RAPD-PCR技术,制备基因组DNA多态性图谱寻找优良性状的分子标记,对分子标记进行核酸序列测定,使用地高辛将序列标记成核酸探针,最后利用基因组多态性图谱或探针可筛选出优良性状的蜂种。研究结果:(1)找到了采蜜高,产浆量多以及易感白垩病等3种分子标记,它们分别是K700bp、W316bp、P380bp;(2)对3种分子标记进行核酸测序;(3)将3种分子标记用地高辛标记成探针;(4)使用3种分子标记的DNA多态性图谱及核酸探针可以快速(48h)的应用于各蜂场进行优良蜂种的普查及筛选。

摘要： 本文研究目的:从不同蜜蜂种类、品系的DNA基因组水平寻找其优良性状的分子标记,并对优良性状的分子标记进行测序和探针的制备,为了应用于全国800万群中优良蜂种的普查及筛选。研究方法:为分离纯化工蜂蜂蛹中基因组DNA,通过RAPD-PCR技术,制备基因组DNA多态性图谱寻找优良性状的分子标记,对分子标记进行核酸序列测定,使用地高辛将序列标记成核酸探针,最后利用基因组多态性图谱或探针可筛选出优良性状的蜂种。研究结果:(1)找到了采蜜高,产浆量多以及易感白垩病等3种分子标记,它们分别是K700bp、W316bp、P380bp;(2)对3种分子标记进行核酸测序;(3)将3种分子标记用地高辛标记成探针;(4)使用3种分子标记的DNA多态性图谱及核酸探针可以快速(48h)的应用于各蜂场进行优良蜂种的普查及筛选。

摘要： 本文研究目的:从不同蜜蜂种类、品系的DNA基因组水平寻找其优良性状的分子标记,并对优良性状的分子标记进行测序和探针的制备,为了应用于全国800万群中优良蜂种的普查及筛选。研究方法:为分离纯化工蜂蜂蛹中基因组DNA,通过RAPD-PCR技术,制备基因组DNA多态性图谱寻找优良性状的分子标记,对分子标记进行核酸序列测定,使用地高辛将序列标记成核酸探针,最后利用基因组多态性图谱或探针可筛选出优良性状的蜂种。研究结果:(1)找到了采蜜高,产浆量多以及易感白垩病等3种分子标记,它们分别是K700bp、W316bp、P380bp;(2)对3种分子标记进行核酸测序;(3)将3种分子标记用地高辛标记成探针;(4)使用3种分子标记的DNA多态性图谱及核酸探针可以快速(48h)的应用于各蜂场进行优良蜂种的普查及筛选。

摘要： 本文研究目的:从不同蜜蜂种类、品系的DNA基因组水平寻找其优良性状的分子标记,并对优良性状的分子标记进行测序和探针的制备,为了应用于全国800万群中优良蜂种的普查及筛选。研究方法:为分离纯化工蜂蜂蛹中基因组DNA,通过RAPD-PCR技术,制备基因组DNA多态性图谱寻找优良性状的分子标记,对分子标记进行核酸序列测定,使用地高辛将序列标记成核酸探针,最后利用基因组多态性图谱或探针可筛选出优良性状的蜂种。研究结果:(1)找到了采蜜高,产浆量多以及易感白垩病等3种分子标记,它们分别是K700bp、W316bp、P380bp;(2)对3种分子标记进行核酸测序;(3)将3种分子标记用地高辛标记成探针;(4)使用3种分子标记的DNA多态性图谱及核酸探针可以快速(48h)的应用于各蜂场进行优良蜂种的普查及筛选。

摘要： 本文研究目的:从不同蜜蜂种类、品系的DNA基因组水平寻找其优良性状的分子标记,并对优良性状的分子标记进行测序和探针的制备,为了应用于全国800万群中优良蜂种的普查及筛选。研究方法:为分离纯化工蜂蜂蛹中基因组DNA,通过RAPD-PCR技术,制备基因组DNA多态性图谱寻找优良性状的分子标记,对分子标记进行核酸序列测定,使用地高辛将序列标记成核酸探针,最后利用基因组多态性图谱或探针可筛选出优良性状的蜂种。研究结果:(1)找到了采蜜高,产浆量多以及易感白垩病等3种分子标记,它们分别是K700bp、W316bp、P380bp;(2)对3种分子标记进行核酸测序;(3)将3种分子标记用地高辛标记成探针;(4)使用3种分子标记的DNA多态性图谱及核酸探针可以快速(48h)的应用于各蜂场进行优良蜂种的普查及筛选。

摘要： 按照植物病理学原则,蘑菇分离物应依据柯赫法则(KOCh'S postulate)回接.在一定条件下,人工诱发子实体得以鉴别.但是一些蘑菇种类(如某些共生食用蘑菇)难以人工诱发子实体,即使能形成实体也费时费力.本文介绍DNA指纹鉴别技术,即运用RAPD-PCR技术制成分离物及对应子实体指纹图谱,分析它们间DNA同源性及遗传相似度以鉴别分离物真伪,探求快速;简便;有效的鉴别方法.

摘要： 按照植物病理学原则,蘑菇分离物应依据柯赫法则(KOCh'S postulate)回接.在一定条件下,人工诱发子实体得以鉴别.但是一些蘑菇种类(如某些共生食用蘑菇)难以人工诱发子实体,即使能形成实体也费时费力.本文介绍DNA指纹鉴别技术,即运用RAPD-PCR技术制成分离物及对应子实体指纹图谱,分析它们间DNA同源性及遗传相似度以鉴别分离物真伪,探求快速;简便;有效的鉴别方法.

摘要： 按照植物病理学原则,蘑菇分离物应依据柯赫法则(KOCh'S postulate)回接.在一定条件下,人工诱发子实体得以鉴别.但是一些蘑菇种类(如某些共生食用蘑菇)难以人工诱发子实体,即使能形成实体也费时费力.本文介绍DNA指纹鉴别技术,即运用RAPD-PCR技术制成分离物及对应子实体指纹图谱,分析它们间DNA同源性及遗传相似度以鉴别分离物真伪,探求快速;简便;有效的鉴别方法.

摘要： 按照植物病理学原则,蘑菇分离物应依据柯赫法则(KOCh'S postulate)回接.在一定条件下,人工诱发子实体得以鉴别.但是一些蘑菇种类(如某些共生食用蘑菇)难以人工诱发子实体,即使能形成实体也费时费力.本文介绍DNA指纹鉴别技术,即运用RAPD-PCR技术制成分离物及对应子实体指纹图谱,分析它们间DNA同源性及遗传相似度以鉴别分离物真伪,探求快速;简便;有效的鉴别方法.

摘要： 按照植物病理学原则,蘑菇分离物应依据柯赫法则(KOCh'S postulate)回接.在一定条件下,人工诱发子实体得以鉴别.但是一些蘑菇种类(如某些共生食用蘑菇)难以人工诱发子实体,即使能形成实体也费时费力.本文介绍DNA指纹鉴别技术,即运用RAPD-PCR技术制成分离物及对应子实体指纹图谱,分析它们间DNA同源性及遗传相似度以鉴别分离物真伪,探求快速;简便;有效的鉴别方法.