冷诱导下毛百合甘油-3-磷酸酰基转移酶活性变化及其基因保守区的克隆   总被引：1，自引：0，他引：1  

陈丽静  葛菱  李浩戈  郭志富  张丽  党晓璇  陶承光  李天来 《华北农学报》2011,(5):34-39

探讨了野生毛百合甘油-3-磷酸酰基转移酶活性在冷胁迫下的变化,用简并引物扩增了甘油-3-磷酸酰基转移酶基因保守区序列的结果表明,该保守区段长744 bp,推测编码247个氨基酸.在GPAT氨基酸序列中存在1个高度保守的区域(WIAPSGGRDRP),在NCBI上经过BLASTp比对分析,发现这段保守区序列为LPLAT基...  相似文献   

陆地棉SSR核心引物筛选及95份骨干种质的遗传多样性分析   总被引：6，自引：3，他引：3  

张小娟  何团结  陆徐忠  路曦结  倪金龙  马琳  杨剑波 《棉花学报》2011,23(6):529-536

从391对棉花SSR引物中筛选出均匀分布于棉花26条染色体的52对核心引物,对92份陆地棉和3份亚洲棉骨干种质进行多样性分析,结果表明,52对引物的等位基因数范围为2～13,平均5.7692个,多态信息含量(Polymorphism Information Contents,PIC)在0.3457～0.8800间,均值...  相似文献   

土壤微生物DNA木聚糖酶基因多样性的研究   总被引：2，自引：0，他引：2       下载免费PDF全文

张桂敏  庄永红  刘婷  马立新 《土壤学报》2006,43(2):295-299

采用国产硅胶Gk254（60型）代替进口Glass bead的改良方法抽提土壤微生物DNA，然后设计了一对扩增木聚糖酶基因片段的新简并引物，对抽提的土壤微生物DNA进行PCR扩增。扩增片段连接pMD18T载体，转化大肠杆菌，重组片段通过酶切进行RFLP分析后，测序分析得到10个木聚糖酶基因片段。对所得片段翻译的氨基酸序列进行BLAST分析表明有8个片段与来自放线菌的木聚糖酶具有较高的同源性，2个与假单胞菌的木聚糖酶具有较高的同源性。所得10个木聚糖酶片段的氨基酸序列同源性比较显示，第27个氨基酸均为天冬酰胺（N），暗示这些来自土壤微生物DNA的基因片段编码耐碱的木聚糖酶。通过构建系统进化树，发现扩增的木聚糖酶片段之间的相似性均在70％以上。  相似文献   

Diversity of laccase genes from basidiomycetes in a forest soil     

Patricia Luis  Grit Walther  Francis Martin 《Soil biology & biochemistry》2004,36(7):1025-1036

Fungal oxidative exo-enzymes lacking substrate specificity play a central role in the cycling of soil organic matter. Due to their broad ecological impact and available knowledge of their gene structure, laccases appeared to be appropriate markers to monitor fungi with this kind of oxidative potential in soils. A degenerate PCR-primer pair Cu1F/Cu2R, specific for basidiomycetes, was designed to assess directly the diversity of laccase genes in soils. PCR amplification of mycelial cultures and fruit-bodies of a wide spectrum of basidiomycetes, covering all functional groups (saprophytes, symbionts, and pathogens), produced multiple DNA fragments around 200 bp. A neighbor-joining tree analysis of the PCR-amplified laccase sequences showed a clear species-specificity, but also revealed that most fungal taxa possess several laccase genes showing a large sequence divergence. This sequence diversity precluded the systematic attribution of amplified laccase of unknown origin to specific taxa. Amplification of laccase sequences from DNA, extracted from a brown (moder) forest soil, showed a specific distribution of laccase genes and of the corresponding fungal species in the various soil horizons (O_h, A_h, B_v). The most organic O_h-horizon displayed the highest gene diversity. Saprophytic fungi appeared to be less widespread through the soil horizons and displayed a higher diversity of laccase genes than the mycorrhizal ones.  相似文献   

Applying the USLE Family of Models at the Sparacia (South Italy) Experimental Site     

《Land Degradation \u0026amp; Development》2017,28(3):994-1004

Soil erosion is a key process to understand the land degradation, and modelling of soil erosion will help to understand the process and to foresee its impacts. The applicability of the Universal Soil Loss Equation (USLE) at event scale is affected by the fact that USLE rainfall erosivity factor does not take into account runoff explicitly. USLE‐M and USLE‐MM, including the effect of runoff in the event rainfall–runoff erosivity factor, are characterized by a better capacity to predict event soil loss. The specific objectives of this paper were (i) to determine the suitable parameterization of USLE, USLE‐M and USLE‐MM by using the dataseries of Sparacia experimental site and (ii) to evaluate their performances at both event and annual scale. The measurements allowed to establish the relationships for calculating the factors of USLE, USLE‐M and USLE‐MM usable at the Sparacia experimental area. At first, for slope‐length values greater than 33 m, the calibration of USLE model at event scale pointed out that sediment delivery processes, that is processes involving deposition of the transported eroded soil particles, occur. The analysis showed that USLE and USLE‐M tend to overestimate low event soil losses, while for USLE‐MM, this tendency is less pronounced. However, the USLE‐MM performed better than USLE and USLE‐M and was able to reproduce better than other two models the highest soil loss values that are the most interesting from a practical point of view. The results obtained at annual scale were generally consistent with those obtained at event scale. Copyright © 2016 John Wiley & Sons, Ltd.  相似文献   

Molecular Variability in Different Indian Isolates of Equine Herpesvirus-1     

Gupta AK  Kaur D  Rattan B  Yadav MP 《Veterinary research communications》2005,29(8):721-734

Three abortigenic Indian isolates of equine herpesvirus-1 (EHV-1) (Tohana, Hisar and Bikaner), along with two exotic abortigenic isolates (AB4 and V592) and another EHV-1 isolate (Jind) obtained from a case of perinatal foal mortality, were studied for variability. For this purpose, PCR and restriction endonuclease (RE) digestion techniques were used simultaneously as a DNA fingerprinting system. Nine different regions of EHV-1 virus were amplified by PCR using primer pairs specific for the regions and the products obtained from these regions were subsequently subjected to various restriction endonucleases to further assess the variability in the number of RE sites as well as in their positions. No difference was observed in all the four abortigenic isolates in terms of the size of different PCR products amplified by all the nine primer pairs, except for primer pairs ‘E’ and ‘C’. PCR products obtained with primer pair E revealed that Tohana and Bikaner isolates were most similar while Hisar isolate was like V592 isolate. However, the PCR product obtained from Jind isolate had a size between the PCR products of Hisar and Tohan/Bikaner isolates. The primer pair ‘C’ used to amplify the region between 1151 to 3679 in ‘Gene 1,2,3’ clearly differentiated the EHV-1 isolate obtained from a case of perinatal foal mortality from isolates obtained from abortion cases. This primer pair needs to be exploited more extensively for use as a potential marker for differentiating the EHV-1 isolates, mainly the abortion cases from perinatal foal mortality ones. Restriction endonuclease studies done with PCR product of all the isolates with various primer pairs did not reveal any changes in the position or number of RE sites present in the products amplified, indicating no variation in different RE sites within the amplified PCR products. However, this study clarified that all the Indian isolates belonged to the IP group of EHV-1.  相似文献   

中国鲎微卫星引物扩增圆尾鲎DNA的PCR反应体系优化     

莫然  谢仰杰  肖志群  翁朝红 《厦门水产学院学报》2012,(2):89-95

采用正交设计法,从dNTPs浓度、引物浓度、Mg2＋浓度、Taq DNA聚合酶用量4个因素3个水平出发,优化设计圆尾鲎DNA的PCR反应体系（引物为中国鲎微卫星引物）.并采用直观分析方法分析正交试验结果,最终建立了圆尾鲎SSR-PCR最佳反应体系：总体积20μL,Taq DNA聚合酶1.5 U、dNTPs 0.16 mmol/L、引物0.2μmol/L、Mg2＋2.0 mmol/L;并通过PCR梯度实验进一步优化模板DNA质量浓度、退火温度及退火时间,获得最佳反应条件：模板DNA质量浓度为30 ng/μL,退火温度为48℃,退火时间为20～25 s.对最佳反应体系和反应条件进行了检验,结果显示该反应体系稳定性高、重复性好.  相似文献   

基于毛细管电泳技术的牡丹SRAP-PCR反应体系优化及引物筛选     

史倩倩  王雁  周琳  黄国伟 《江西农业大学学报》2012,(1):158-164

以牡丹叶片DNA为模板,对SRAP-PCR反应程序进行研究,确定适合的反应程序,即94℃预变性5 min;94℃变性1 min,33℃退火1 min,72℃延伸1 min,共5个循环;随后94℃变性1 min,52℃退火1 min,72℃延伸1 min,共35个循环;最后72℃延伸10 min。基于毛细管电泳技术,采用正交设计L18(37)对牡丹SRAP-PCR反应体系的5因素(Taq酶,Mg2+,模板DNA,dNTP,引物)3个水平进行了优化,构建了牡丹SRAP最佳反应体系:模板DNA 50 ng,dNTP 0.25 mmol/L,Mg2+浓度2.5 mmol/L,引物浓度0.4μmol/L,TaqDNA聚合酶0.5 U,总体积为25μL。各因素对扩增结果影响程度均不同:dNTPs>引物>DNA模板>Mg2+>Taq酶。运用该体系从756个SRAP引物组合中筛选出多态性好、条带清晰的26个引物组合,并证明了该体系稳定可靠。该体系的建立以及引物组合的确定为今后利用SRAP分子技术进行牡丹的相关研究奠定了科学基础。  相似文献   

本氏针茅SRAP PCR反应体系的建立及引物筛选     

井赵斌  俞靓  魏琳  程积民 《草业科学》2012,29(2):219-228

以本氏针茅（Stipa bungeana）幼嫩叶片为材料,建立适合本氏针茅基因组DNA提取的改良CTAB法,在此基础上采用正交试验设计和单因素分析相结合的方法,对本氏针茅SRAP PCR反应体系中的5个主要因素（DNA、Taq酶、dNTPs、Mg2+和引物）进行优化,旨在建立适合本氏针茅SRAP分析的反应体系。结果表明,在20 μL总的反应体系中各组分的加入量分别为：DNA（20 ng·μL-1）3 μL、Taq DNA酶(5 U·μL-1)0.2 μL、dNTPs（2.5 mmol·L-1)1.4 μL、引物（10 μmol·L-1）1.0 μL、Mg2+ (25 mmol·L-1)2.0 μL、10×Buffer 2.5 μL、ddH2O 8.9 μL。体系验证和引物筛选试验表明,该体系适于本氏针茅遗传多样性分析,该体系的建立可为本氏针茅种质资源遗传多样性研究和黄土高原植被恢复及生态建设提供理论基础。  相似文献   

正交设计优化草地早熟禾SRAP-PCR反应体系及引物筛选     

任小巍  王瑜  袁庆华 《草业科学》2012,29(3):411-416

采用L9(34)正交试验设计方法,对草地早熟禾(Poa pratensis)基因组DNA SRAP PCR反应体系中的Taq DNA聚合酶、Mg2+、引物及dNTP四因素的用量进行优化,并比较不同模板DNA用量对扩增的影响,建立草地早熟禾SRAP PCR最佳反应体系,同时,利用该体系对SRAP引物进行筛选。结果表明,草地早熟禾SRAP PCR最佳反应体系为Taq DNA聚合酶1.0 U、Mg2+ 1.75 mmol·L-1、引物0.25 μmol·L-1、dNTP 220 μmol·L-1、40 ng模板DNA、2 μL 10×PCR buffer,总体积20 μL。运用该体系从100对SRAP引物中筛选出43对引物能够产生清晰稳定的扩增条带且多态性丰富。优化体系的建立及引物的筛选可为今后利用SRAP标记技术对草地早熟禾进行遗传多样性分析、图谱构建、种质资源鉴定奠定技术基础。  相似文献