广东省主要红树林植物DNA条形码评价     

武锋  裴男才  廖宝文  管伟  姜仲茂  李玫 《东北林业大学学报》2020,(4):42-49

为评估DNA条形码对鉴定红树林植物的通用性和有效性,在广东红树林分布区域共计采集红树林植物16科22属23种,共144个样品,并进行DNA条形码测序。结果表明:选择的rbcL、matK和trnH-psbA 3个DNA片段的PCR扩增成功率分别为100%、80.29%±8.49%、99.38%±1.25%。测序成果率最高为rbcL 100%,trnH-psbA次之94.57%±5.06%,matK最低75.04%±6.26%。表明rbcL和trnH-psbA片段在红树林群落中都具有较好的通用性。应用BLAST和NJ Tree两种方法计算红树植物的物种识别率。BLAST结果表明,单片段中trnH-psbA的物种识别率最高,为84.48%±12.09%,rbcL次之,matK最低。NJ Tree分析显示单片段中rbcL的物种识别率最高,为66.65%±17.35%;trnH-psbA片段次之,matK片段最低。两张分析方法都显示多个片段组合使用时,rbcL或trnH-psbA是提高物种平均识别率的主要片段。利用单片段rbcL即可获得平均节点支持率最高的红树植物系统发育树,且能准确区分不同树种。trnH-psbA片段可以识别rbcL片段不能识别的物种,可以作为补充片段。综合比较,推荐rbcL、trnH-psbA作为红树林植物DNA条形码片段。  相似文献   

转基因玉米TC1507质粒DNA标准物质的研制     

瞿展  杨立桃 《浙江农业学报》2021,33(3):390

标准物质具有特定量值、均匀性和稳定性三大典型特征,也是其作为测量标尺的依据。转基因生物标准物质是我国转基因产品标识制度顺利实施的关键技术支撑之一。文章以转基因玉米TC1507为对象,制备了转化体特异性的新型质粒DNA标准物质pTC1507,并对其均匀性、稳定性、量值进行了评价和测定。测试结果表明,制备的质粒DNA标准物质具有良好的瓶间和瓶内均匀性,pTC1507稳定性可靠,可以在-20 ℃稳定放置6个月以上。经过测序和实时荧光定量PCR定值以及不确定度评估,pTC1507标准物质的量值是1.01±0.053。均匀性、稳定性和量值结果表明,研制的转基因玉米TC1507质粒分子标准物质符合标准物质的典型要求,可以代替传统的基体标准物质应用于转基因玉米检测,解决传统标准物质获取困难、制备复杂、成本高等不足。  相似文献   

DNA条形码技术在中药领域的应用     

吕小旭  关鉴茹  樊敏  赵小荣 《中兽医医药杂志》2021,(1):42-45

DNA条形码技术可对物种进行快速自动鉴定,具有鉴定准确、结果稳定、操作简便、适用广泛等特点,目前在中药领域应用较多。从DNA条形码技术在中药材鉴定、种植、流通、市场监管、中成药鉴定和药用植物种质资源调查等中药领域的多个方面进行综述,并探讨DNA条形码技术在中药领域的优势和不足,以期为DNA条形码技术在中药领域的研究提供新的思路。  相似文献   

Immunotherapy in Veterinary Oncology     

Philip J. Bergman 《Veterinary Clinics of North America: Small Animal Practice》2014

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Characterization of Genomic Integration and Transgene Organization in Six Transgenic Rapeseed Events     

WU Yu-hua  ;ZHANG Li  ;WU Gang  ;NIE Shu-jing  ;LU Chang-ming 《中国农业科学(英文版)》2014,(9):1865-1876

To characterize the DNA rearrangement of both the T-DNA region and the genomic insertion site during T-DNA insertion, the Genomewalker strategy was used to isolate the junctions between the inserted DNA and the plant genomic DNA in six rapeseed events as well as the genomic DNA at the sites before integration. During transformation in each of the six events, portions of both the right border（RB） and left border（LB） regions of the T-DNA were deleted, ranging from a 7 nucleotide deletion of the LB repeats in event RF1 to a 207 bp deletion of the LB region in event RF2. For the six events, T-DNA integration resulted in a deletion at the target site spanning less than 100 bp. Sequence analysis indicated that the T-DNA was integrated into the coding region of various native rapeseed genes in events RF1 and RF2. Duplications of the genomic DNA target site were observed in events RF2, RF3 and Topas 19/2. And multimerization of transgenes was found in event Topas 19/2, in which, the T-DNA was integrated as a head-to-head（RB-to-RB） concatemer into the recipient genome. In event MS1, chromosomal translocation or a large target-site deletion may have occurred during T-DNA integration, which was identified due to a failure to amplify the presumptive insertion site based on the flanking rapeseed DNA sequences. Our results provide comprehensive data concerning transgene organization and the genomic context of the T-DNA in six rapeseed events, which can aid in the developing of insert fingerprinting and the monitoring of long-term genetic stability and potential unintended effects of transgenic events.  相似文献   

Controlling sprouting in potato tubers using ultraviolet-C irradiance     

《Postharvest Biology and Technology》2014

Legislation limiting the use of chlorpropham (CIPC), the major potato sprout suppressant, has led to a need for new technologies to extend storage life of tubers. Ultra violet C (UV-C) has been used postharvest to reduce disease incidence on many crops, yet its use and efficacy as a sprout suppressant has not been investigated. The aim of this project was to identify the optimum dose and treatment timing of UV-C treatment on potato tubers as an alternative method of sprout suppression to reduce the dependence on chemical sprout suppressants. Up to six potato cultivars over two seasons were treated with varying doses of UV-C ranging from 0 to 30 kJ m⁻² either at harvest or at first indication of dormancy break. The tubers were stored at 9 °C and sprout growth and incidence assessed. Treatment with moderate UV-C doses (5–20 kJ m⁻²) suppressed sprout length and sprout incidence in a range of cultivars. Periderm DNA damage and programmed cell death were not detected in response to any of the UV-C doses. The inactive ABA metabolite, ABA-GE, increased in response to 10 or 20 kJ m⁻² within 72 h of treatment. Multivariate analysis showed a negative relationship between ABA metabolites and sprout growth/incidence during storage. This study found that UV-C reduced sprout growth in potato with no deleterious effects on tuber quality. This suggests potential for further development as an alternative or supplement to conventional sprout suppressant technologies.  相似文献   

不同发育时期菊芋块茎DNA提取效果比较研究     

宋向阳  赵孟良  王丽慧  李莉 《种子》2017,(7)

为筛选菊芋块茎DNA提取的适宜生育时期,以“青芋1号”菊芋品种为试材,用改良CTAB法对不同发育时期菊芋块茎DNA进行提取,经琼脂糖凝胶电泳及全波长分光光度计检测总DNA的纯度、浓度及质量,选用4条ISSR引物进行PCR验证.结果表明:不同发育时期提取菊芋块茎DNA效果较好,DNA浓度呈现单峰曲线变化,峰值出现在第9周,与琼脂糖凝胶电泳检测呈现的亮度结果一致,PCR验证结果显示,提取的DNA能够满足后续相关分子生物学的要求.  相似文献   

Study on Liver-protective Effect of Chinese Herbal Compound Probiotics on Acute Liver Injury Layers     

YU Jia-min  XIE Quan-xi  ZHANG Jian-mei  GU Wei 《中国畜牧兽医》2016,43(7):1904-1909

This experiment was conducted to investigate the liver-protective effect of Chinese herbal compound probiotics (CHCP) on acute liver injury layers.One hundred and eight 1 day old hens were divided into 4 groups with 3 replicates in each group and 9 layers per replicate in trial 1.The layers in model groups Ⅰ to Ⅲ were gavaged with 10% (V/V) soybean oil solution of carbon tetrachloride (SCCl₄) according to 1,2 and 4 mL/(kg·BW) at 14,28 and 35 d,respectively.The layers in control group were gavaged with 2 mL/(kg·BW) soybean oil.In trial 2,sixty 1 d layers were divided into 5 groups:Control group (soybean oil),model control group(SCCl₄)and low-dose,middle-dose and high-dose CHCP group (SCCl₄+1‰ CHCP,SCCl₄+2‰ CHCP and SCCl₄+4‰ CHCP respectively).CHCP were used by drinking water since 7 days.SCCl₄ were gavaged according to 2 mL/(kg·BW) at 14 and 28 d.The results showed as follows:The model of layers liver damage could be built by intragastric administration of 2 mL/(kg·BW) 10%(V/V) SCCl₄ at 14,28 d respectively,with the signs of hepatic steatosis,severe vacuolar degeneration,nuclear condensation and necrosis.Compared to the model control group,the serum AST levels in low,medium and high dose CHCP groups were decreased by 4.35% (P > 0.05),7.57% (P > 0.05) and 9.79% (P < 0.05),the serum ALT levels in medium and high dose CHCP groups were decreased by 34.92% (P < 0.01),36.51% (P < 0.01),the serum total bilirubin content in medium and high dose CHCP groups were decreased by 25.49% (P < 0.01),27.45% (P < 0.01).The liver cell congestion was reduced to varying degrees in different dose CHCP groups,and the liver cell had no vacuoles,arranged in neat rows,abundant cytoplasm and uniform in different dose CHCP groups.In conclusion,2‰,4‰ CHCP could reduce hepatocyte necrosis,decrease the serum activities of ALT,AST and total bilirubin levels,and had protective effect on hepatic injury induced by SCCl₄.  相似文献   

高含固率木质纤维素厌氧发酵物总DNA的4种提取方法比较     

马旭光  刘素君  江滔  常佳丽  唐琼 《中国沼气》2020,(2):28-35

不同的样本特性和提取方法对获得微生物总DNA的质量有重要影响。文章基于高含固率木质纤维素厌氧发酵物腐殖酸、酚类物质含量高、质地均一性差、微生物浓度低的特点,研究了4种方法提取不同高含固率粪秸厌氧发酵物中微生物总DNA的效果。结果表明,常规的十二烷基磺酸钠法(sodium dodecyl sulfate,SDS)、十二烷基磺酸钠和溴化十六烷基三甲铵结合法(sodium dodecyl sulfate and cetyltrimethyl ammonium bromide,SDS-CTAB)和商业的粪便试剂盒法提取的DNA质量均较差,SDS法和试剂盒法未能获得聚合酶链式反应(polymerase chain reaction,PCR)扩增目的条带,SDS-CTAB法得到的条带较模糊;改进SDS-CTAB法获得的DNA杂质少、纯度高,具有较好的稳定性,A260/A280和A260/A230值分别为1.74~1.86和1.65~1.86,每克样品的DNA浓度在50 ng·μL^-1以上,电泳条带单一齐整、清晰明亮,PCR扩增的目的条带清晰度高,适宜后续分子生物学技术的分析。林格氏液洗脱、聚乙烯吡咯烷酮-40(Polyvinyl Pyrrolidone-40,PVP-40)洗涤液除杂以及裂解液和多种酶联合破壁是改进SDS-CTAB法获得该类专一性样本高质量微生物总DNA的关键步骤。  相似文献   

入侵性食叶害虫椰子织蛾的单倍型多样性分析     

李优佳  杨帆  吕宝乾  何荣晓  蔡波 《华南农业大学学报》2020,41(4):76-81

【目的】分析危害棕榈科Palmae植物的一种重要入侵食叶害虫椰子织蛾Opisina arenosella单倍型在原产地和入侵地的分布特点,揭示椰子织蛾入侵我国的虫源信息。【方法】利用线粒体COI基因分析16个地理种群共计172个样本,比较椰子织蛾印度种群和入侵地(中国、马来西亚和泰国)种群的遗传关系。【结果】片段长度为625 bp的172条序列共鉴定出12个单倍型,包含15个变异位点,构成2个明显的单倍型分支,其中一个分支由11个单倍型(IN1～IN11)组成,均来自印度种群,单倍型IN1是6个印度种群的共享单倍型,IN2～IN11为独享单倍型;另一个分支为单倍型HAP,由来自中国、马来西亚和泰国的种群共享;HAP与11个来自印度的单倍型IN1～IN11均存在4个变异位点。【结论】入侵地区的椰子织蛾种群来自同一基因型或者具有相同的入侵源;椰子织蛾种群入侵后受环境选择压力,在新栖息地产生新的突变或杂交。  相似文献