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1.
无患子总DNA提取方法研究 总被引：3，自引：2，他引：1
针对无患子富含多酚、多糖、蛋白质及其他次生代谢物质的特点,以无患子叶片为材料,分别采用改良的CTAB法和CTAB与SDS相结合法提取不同种源无患子的总DNA。结果显示,改良的CTAB法提取的总DNA,得率较高,纯度好,可用于酶切、PCR扩增等后续试验,并能获得清晰、稳定的扩增产物。而采用CTAB与SDS相结合的方法得到的基因组DNA效果不稳定,无法检测出DNA。改良的CTAB法具有简单、经济的特点,可用于无患子总DNA的提取,成功地进行SRAP分子标记分析,为研究无患子遗传资源多样性奠定基础。 相似文献
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木材DNA条形码鉴定研究进展 总被引：1，自引：1，他引：0
木材种属鉴定具备显著的生物学与经济意义, 但传统的以显微特征观察为主的方法已不能适应高通量和精细化鉴别的需要。DNA条形码是根据特定基因片段的序列差异, 利用生物信息学技术对生物物种进行快速分类与鉴别的方法。近年来, DNA条形码技术已被陆续应用于木本植物及相应木材的种质鉴定, 在目标基因选择、木材DNA提取及生物信息学分析等方面均取得显著进展。在使用优化的微量DNA提取技术的前提下, 干燥木材中也可提取出满足扩增要求的DNA。经过生命条形码联盟等国际机构的长期努力, 确定了rbcL+ matK组合等通用植物条形码标记及ITS2等补充标记, 并建立了BOLD等数据库系统。传统的条形码序列分析主要通过BLAST比对、遗传距离分析及系统进化分析来实现, 近年来随着生物信息学的发展, DNA条码数据库不断完善, 新的数据分析方法和软件正不断涌现。文中在总结现有研究成果和实施规范的基础上, 综述国内外应用DNA条形码技术进行木材鉴别的新进展, 并着重阐述新型序列分析方法和相应的生物信息学工具。 相似文献
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金叶含笑叶片基因组DNA提取方法的比较 总被引：2，自引：0，他引：2
以金叶含笑叶片为实验材料,分别采用CTAB法、改良CTAB法、简易提取方法、高盐沉淀法和SDS法5种方法提取金叶含笑叶片的基因组DNA,并通过琼脂糖凝胶电泳、紫外分光光度计及ISSR扩增3种方法对所提取的DNA样品进行检测.DNA纯度、含量比较以及扩增结果表明:5种方法中简易提取法和SDS法提取的DNA纯度较高.高盐沉淀法得到的DNA产量较高.5种方法提取的DNA用于ISSR扩增,其扩增结果基本一致.简易提取方法操作简单.省时省力,是用于ISSR扩增的金叶含笑基因组DNA提取的最佳选择. 相似文献
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芒果属植物叶绿体DNA提取方法的研究 总被引：1，自引：0，他引：1
为了探究不同提取方法对芒果属植物叶绿体DNA提取质量的影响,以芒果和扁桃的成熟叶片为材料,比较植物叶绿体DNAout试剂盒法和高盐-低p H法分离叶绿体及提取叶绿体DNA效果。结果表明:叶绿体DNAout试剂盒法能够较好地去除蛋白质、酚类、多糖等代谢物质,OD260/OD230大于2.000,OD260/OD280在1.800~1.900之间。高盐-低p H法提取叶绿体DNA产率高达75.8 ng/μL,远大于叶绿体DNAout试剂盒法提取叶绿体DNA最高产率42.8 ng/μL,2种方法所得模板电泳图和SSR标记图谱谱带完整性皆好。显微镜下观察叶绿体,2种方法都可以得到杂质少、背景清晰的叶绿体显微成像效果,但高盐-低p H法提取的叶绿体细胞器密度更高。2种方法获得的叶绿体DNA均可满足后续研究工作的需要。 相似文献
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[目的]探讨适合舞毒蛾标本的保存和DNA提取方法,分析舞毒蛾标本的保存时间和保存方式对基因序列扩增的影响。[方法]利用SDS法、磁珠法以及E.Z.N.A.TM昆虫DNA提取试剂盒3种方法对舞毒蛾新鲜样本、干燥成虫标本和福尔马幼虫林标本进行DNA提取,并对3种方法提取DNA的浓度和纯度进行分析比较。利用1956年至1996年收集的舞毒蛾干燥成虫标本和福尔马林幼虫标本分析并比较保存时间和保存方式对15对COI基因(cytochrome oxidase subunit I)引物(目的片段长度为216 bp^977 bp)扩增成功率的影响。[结果]表明与SDS法和磁珠法相比,E.Z.N.A.TM昆虫DNA提取试剂盒对舞毒蛾3种标本的DNA提取效果最好,提取过程中未对标本DNA产生额外损伤。标本保存时间越久扩增长序列目的片段的成功率越低,福尔马林幼虫标本总体扩增成功率比干燥成虫标本的略高。[结论]E.Z.N.A.TM昆虫DNA提取试剂盒更适合舞毒蛾新鲜样本、干燥成虫标本和福尔马林幼虫标本的DNA提取。舞毒蛾标本的保存时间和保存方式均会对基因序列扩增产生影响。 相似文献
7.
土壤微生物总 DNA 提取及其 PCR 优化 总被引：1，自引：0，他引：1
采用3种不同土壤微生物总 DNA 提取方法对广东省乐昌杨东山十二度水自然保护区的样地土壤进行比较研究,进一步通过 PCR 扩增,研究牛血清白蛋白(BSA)对整个 PCR 扩增过程的影响。结果表明：试剂盒提取法相对于其他两种提取方法更加方便有效,当土壤样品量比较大时,直接提取法是最经济有效的方法;而在土壤微生物的 PCR 扩增实验中,加入2μl 的 BSA,可以有效地抑制腐殖酸等对后续 PCR 扩增的影响。 相似文献
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Y. Q. Hu X. C. Kong X. R. Wang T. K. Zhong X. W. Zhu M. M. Mota L. L. Ren S. Liu C. Ma 《Forest Pathology》2011,41(2):165-168
Pine wilt disease (PWD), caused by the pine wood nematode (PWN) Bursaphelenchus xylophilus, leads to serious losses to pine forestry around the world. Pinus massoniana, which is vulnerable to be attacked by the PWN, is the dominant species used in pine forestry in China. The objective of this study is to develop a direct PCR‐based method for detecting B. xylophilus in the wood of P. massoniana without a separate nematode extraction step. A simple procedure was first developed for isolating B. xylophilus DNA in 5 mg pine wood tissue samples harbouring PWN for detection by PCR amplification. A B. xylophilus‐specific amplicon of 403 bp (DQ855275) was generated by PCR from the infested wood tissue. The entire procedure can be completed within 5 h with one pair of primers. This assay can serve as a rapid, cheap and environmentally friendly method to detect B. xylophilus in samples of P. massoniana. 相似文献
10.
CTAB-silica Method for DNA Extraction and Purification from Castanea mollissima and Ginkgo biloba 总被引：4，自引：0，他引：4
ShenYongbao ShiJisen 《中国林学(英文版)》2003,5(3):10-12
A new method CTAB-silica for DNA extraction and purification from the leaves and buds of Castanea mollissima and Ginkgo biloba was tested. The method is based on the silica-based purification protocol developed by Boom et al. (1990). By modifying the protocol, plant genome DNA could be extracted easily from dormant buds, mature leaves, and other parts of plant. Our results showed that the purified DNA was of high purity and could be analyzed by PCR. Furthermore, this CTAB-silica method took much less time for a successful DNA purification process compared to the traditional methods (CTAB and SDS). By our method,the suitable DNA can be extracted and purified from over 10 plant samples by one person in an hour. 相似文献
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The selection of wood species and the styles of sculpture play key roles in the characterization of Buddhist statues. After Jianzhen, a Chinese Buddhist monk, visited Japan in the mid-eighth century, wood of the genus Torreya had been frequently used to produce single-bole statues. Establishing measures for the accurate identification of wood in the genus Torreya is effective for investigating the drastic change in the production of statues during this period. Analyzing the plastid deoxyribonucleic acid (DNA) fragments extracted from wood is considered helpful in the identification of species in the same genus. This study analyzed the sequences and residual amounts of plastid DNA fragments in the wood of Torreya nucifera. Nucleotide substitutions in the plastid DNA were clearly identified between T. nucifera and the species distributed in China, indicating that the wood of Torreya sp. can be discriminated based on the plastid DNA sequences. DNA polymorphism analyses revealed sequence diversity for the intergenic spacers on the T. nucifera plastid DNA. A series of polymerase chain reaction (PCR) analyses demonstrated that the plastid DNA fragments with a length of approximately 100 bp could be amplified from the residual DNA extracted from the T. nucifera sapwood with longer elapsed years after cutting. Therefore, an identification of wood species in the genus Torreya based on their plastid DNA is considered to be one of the most effective measures taken in the study regarding the historical changes of Buddhist statues. 相似文献
12.
We have extracted total DNA from different fractions of fresh wood as well as from cold-stored and air-dried wood harvested from transgenic aspen grown in the field. The highest amounts of DNA were obtained from bark/cambium tissue; the DNA quality, however, was poor. Best results in PCR and Southern blot analyses were obtained from DNA extracted from early wood. Using appropriate primer pairs, amplification products were obtained from both the foreign gene (transgene) and aspen genomic sequences. In Southern blot analyses transgene-specific hybridisation signals were obtained. This is the first report on the detection of foreign genes in wood sampled from genetically modified trees. 相似文献
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为建立思茅松大配子体ISSR分子标记的反应体系,以思茅松种子样本作为供试材料,比较不同思茅松胚乳DNA提取方法的效果,通过紫外分光度计、DNA琼脂糖电泳和PCR扩增分别予以检测并比较。结果表明SDS法优于CTAB法。同时,通过单因子分析法对ISSR-PCR反应体系中的模板DNA用量,Taq DNA聚合酶用量,Mg2＋浓度,dNTP浓度、引物浓度等主要影响因子进行优化,确立适用于思茅松的ISSR分子标记反应体系。对该体系进行稳定性检验,结果表明该反应体系适合思茅松ISSR-PCR扩增。 相似文献
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Lee Hong Tnah Soon Leong Lee Kevin K.S. Ng Naoki Tani Subha Bhassu Rofina Yasmin Othman 《Forest Ecology and Management》2009
The inbuilt unique properties of DNA within the timber could serve as tracking and monitoring tools to verify the legality of a suspected timber in the context of illegal logging, forest certification and chain of custody certification. By using Neobalanocarpus heimii (Dipterocarpaceae) as an example, a population identification database and haplotype distribution map in Peninsular Malaysia were generated for authenticity testing based on four chloroplast DNA markers (trnL intron, trnG intron, trnK intron and psbK-trnS spacer). Twenty one haplotypes were identified from 10 significant intraspecific variable sites. The results clearly revealed that only northern and southern regions of Peninsular Malaysia were distinguishable. Thus, this database could only be used to determine the wood lot of unknown origin at the regional level. Statistical procedure based on the composition of wood lot was used to test whether a suspected timber conforms to a given regional origin. Overall, the observed type I and II errors of the database showed good concordance with the predicted 5% threshold, which might indicate that the database is useful to reveal provenance and establish conformity of wood lot from northern and southern regions of Peninsular Malaysia. Applications of this database for timber tracking are discussed. 相似文献
15.
枣叶片DNA的提取以及AFLP反应体系的建立 总被引：5，自引：0，他引：5
扩增片断长度多态性(AFLP)实验过程中,DNA提取质量和适合的PCR扩增反应体系是2个关键因素.利用CTAB法对枣树叶片DNA的提取方法进行探讨.结果表明:改良CTAB法抽提到了较高质量的枣基因组DNA,可用于AFLP实验分析;通过对酶切-连接体系和PCR扩增体系中的关键因素实验分析,建立了适合枣AFLP荧光反应体系,并得到清晰的DNA指纹图谱. 相似文献
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Suma Arun Dev E. M. Muralidharan P. Sujanapal M. Balasundaran 《Annals of Forest Science》2014,71(4):517-522
Context
East Indian sandalwood (Santalum album L.) in commercial markets is highly prone to adulteration. A number of cases were registered with regard to the adulteration of East Indian sandalwood, but the lack of technical tools for the precise species identification of the source wood stalled most of the court cases.Aims
The standard DNA barcode regions, the rbcL, matK and trnH-psbA chloroplast genomic sequences recommended by the Consortium of Barcode of Life (COBOL) were analysed to distinguish wood adulterants of East Indian sandalwood.Methods
Standard polymerase chain reactions with COBOL recommended primers were performed for all three barcode loci. The PCR products after gel elution were sequenced and alignments were performed using CLUSTALX.Results
Single nucleotide polymorphisms (SNPs) identified with rbcL and trnH-psbA sequences of Erythroxylum monogynum Roxb. as well as with matK sequences of Osyris wightiana Wall ex. Wight could be efficiently utilized for the detection/monitoring of East Indian sandalwood adulterants. Among the two common adulterants O. wightiana and E. monogynum, the former was more similar to S. album and grouped together in the dendrogram.Conclusion
The study recommends the exploitation of DNA barcoding technique using standard barcodes to trace sandalwood timber adulterants. 相似文献18.
《Scandinavian Journal of Forest Research》2012,27(6):559-567
Abstract Methodology to assess the potential for energy wood procurement in Russia is described in this article and applied to the Leningrad region. Wood from thinnings, logging residues, non-industrial roundwood and residues from sawmilling are considered as sources for energy production. Energy wood available in the region, based on the 2004 actual cut, is approximately 4 million m3. Nearly 86% of this is non-industrial roundwood and felling residues, and 14% is by-products from sawmilling. Almost two-thirds of the non-industrial roundwood and felling residues are in cutting areas and one-third is in central processing yards. Deciduous tree species (birch and aspen) dominate in energy wood, representing about 65% of the total amount of felling residues and non-industrial wood. It is possible to intensify utilization of forest resources and thereby also to increase the use of wood in energy production. The total amount could be 54% higher if the allowable cut was fully utilized and 124% higher if thinnings were also utilized completely. There are, however, significant intraregional differences, as the current rate of utilization of forest resources varies in the region. 相似文献
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Zhang Qian Lin Shanzhi Lin Yuanzhen Zhang Zhiyi Liu Haijun Zou Yening Wang Zeliang College of Biological Sciences Biotechnology Beijing Forestry University Beijing P. R. China College of Resources Environment Beijing Forestry University Beijing P. R. China 《中国林学(英文版)》2004,6(3)
The putative transgenic hybrid triploid poplars [(P. tomentosa P. bolleana) P. tomentosa] with CpTI gene have been outplanted in test field for 2 years. Although the authors previous studies have proved that they are highly resistant to 3 species of poplar-threatening insect pests and contain high content of CpTI protein in foliage, incorporation status of foreign CpTI gene in poplar genome is uncertain. In this present study, the incorporation of foreign CpTI gene in genome of 5 transgenic poplars was confirmed by PCR and Southern blotting analysis. DNA amplification showed that there were clear DNA bands of about 450bp specific to CpTI gene in transgenic lanes, while no corresponding band in non-transgenic lane was observed. Correspondingly, clear DNA hybridization signals and no signal were exhibited on film for DNA Southern blotting analysis in transgenic lanes and non-transgenic lane, respectively, which further confirmed the stable integration of foreign CpTI gene in genome of 2-year-old transgenic poplar. 相似文献
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Alexandro B. Leverkus Carolina Puerta-Pi?ero José Ramón Guzmán-álvarez Javier Navarro Jorge Castro 《New Forests》2012,43(5-6):601-613
Post-fire salvage logging (i.e. felling and removing burnt trees, often eliminating the remaining woody debris) is a practice routinely performed by forest managers worldwide. In Mediterranean-type ecosystems, salvage logging is considered a measure to reduce future reforestation costs, but this assumption remains largely untested. We made a cost analysis of different management schemes, addressing the immediate post-fire burnt-wood management as well as the costs and success of subsequent reforestation efforts. Two experimental 25-ha plots were established in a burnt pine reforestation of SE Spain, in which three replicates of three post-fire treatments were applied: non-intervention (NI), partial cut plus lopping (PCL; felling and lopping off the branches from most of the trees, leaving all biomass in situ), and salvage logging (SL). After 4?years, a mechanised reforestation was undertaken, and seedling mortality was monitored for 2?years. The cost of all management operations was recorded in situ, and the cost of re-planting the dead seedlings was estimated according to the expenses of previous reforestation. Initial cost of wood management was greatest in SL and zero in NI. Reforestation cost was highest in NI and lowest in SL, and seedling-mortality rates proved lowest in PCL (43?% vs. 51?% and 52?% in SL and NI, respectively). Considering all the post-fire management operations, salvage logging did not provide particular economic advantages for forest restoration, and had an overall cost of 3,436?±?340?€/ha. By contrast, NI and PCL reduced total restoration costs by 50 and 35?%, respectively, and PCL indeed promoted restoration success. We suggest that the full cost of management operations needs to be considered when evaluating the economic implications of post-fire salvage logging. 相似文献