黄精属植物DNA分子鉴定技术应用研究进展

黄精属植物分布广泛、资源丰富、类群复杂、栽培历史悠久,具有重要的药用、经济、观赏与文化价值.在大健康的时代背景下,黄精作为中国传统中药材,其潜在市场和广泛用途不断受到人们的重视与发掘.随着分子生物学科研究的完善与深入,先后出现了多种基于DNA水平的分子鉴定技术,加速黄精属相关领域的研究并取得了较为丰硕的成果.本文对近年...     

分子遗传标记技术在蜻蜓目昆虫研究中的应用

分子生物学技术应用于昆虫系统学研究在近些年发展相当迅速。目前已出现了几十种分子遗传标记。该文简述了酯酶同工酶电泳、核酸序列分析以及AFLP、RAPD、核酸探针等分子生物学技术在蜻蜓目昆虫研究中的应用情况。     

六种食用蜻蜓稚虫营养成分分析和评价

以云南省和贵州省采集到的6种常见食用蜻蜓为材料,采用凯氏定氮法、索氏提取法、蒽酮比色法、氨基酸自动分析法、原子吸收法、电感耦合等离子质谱法(ICP-MS)和高效液相色谱法(HPLC)法分别测定了6种蜻蜓稚虫的基本营养组成、氨基酸、矿质元素和脂溶性维生素的含量,评价其营养价值。结果显示,6种蜻蜓稚虫平均水分含量为81.17%,干基蛋白质、脂肪、糖和灰分的平均含量分别为65.62%,9.80%、3.01%和5.47%,氨基酸组成合理,符合WHO/FAO提出的优质蛋白条件,主要限制性氨基酸为Met+Cys和Trp,与全鸡蛋蛋白质贴近度为0.59~0.66。富含各类人体必需矿质元素,尤其是Ca和Se,脂溶性维生素中分别测定出0.252~3.859mg·kg~(-1)的α-生育酚,具有高蛋白、低脂肪、氨基酸组成合理和富含矿质元素的特点,营养价值较高,具有开发利用前景,但需注意部分种类体内检测出重金属元素超标。     

滇重楼的psbA-trnH条形码分子鉴定研究北大核心CSCD

为探讨滇重楼药材鉴定新方法,本研究通过对重楼样品提取基因组DNA,PCR扩增叶绿体psb A-trn H序列并进行双向测序,所得序列经Codon Code Aligner拼接后,采用MEGA5.0软件进行序列比对,计算种内和种间遗传距离(K2P),构建邻接数(neighbor-joining tree)进行结果分析。研究表明,叶绿体psb A-trn H片段在滇重楼种内变异较小,平均K2P遗传距离为0.007,而滇重楼与其它重楼种间平均K2P遗传距离为0.025,滇重楼种间最大K2P遗传距离明显小于滇重楼与重楼属其它种内的最小K2P遗传距离。中药材DNA条形码鉴定系统比对和NJ树鉴定结果均表明psb A-trn H序列可区分滇重楼及其同属物种,且具有较好的稳定性和准确性,为保障临床安全用药提供了新的技术手段。     

一种新的DNA分子克隆方法

DNA分子克隆是基本的分子生物学实验技术,传统的分子克隆方法大多需经过酶切链接过程,但在某些情况下,没有合适的酶切位点往往会成为阻碍克隆进行的障碍.本文描述了一种新的分子克隆方法,称为不依赖酶切和链接的分子克隆（RLIC）.利用RLIC,将3种不同大小的DNA片段克隆到3种不同载体,证明了这种方法的有效性和可靠性.由于该方法不受限制性酶切序列限制,省去了酶切连接步骤,因此具有很大的灵活性和简便性,在分子生物学研究方面有广泛应用前景.     

基于ITS条形码序列的刺桐姬小蜂分子鉴定

【目的】刺桐姬小蜂Quadrastichus erythrinae Kim体型小,传统的形态学鉴定方法难以快速准确识别。【方法】本研究测定了刺桐姬小蜂的rDNA ITS1和ITS2序列,根据18S rDNA部分序列,利用MEGA的最大相似法(Maximum Likehood)构建系统发育树。根据刺桐姬小蜂ITS1和ITS2序列设计了特异引物,应用特异引物对单只刺桐姬小蜂进行PCR扩增,可稳定地扩增出明显的目的DNA条带。【结果】研究表明,基于ITS基因的DNA条形码技术可以用于刺桐姬小蜂的快速准确鉴定。【结论】因此,采用ITS1和ITS2区的特异性引物可对刺桐姬小蜂进行快速分子鉴定。     

果园三种天蛾的DNA条形码鉴定

DNA条形码(DNA Barcoding)可以利用一段通用基因短片段对物种(包括不同生活史阶段)进行区分。本文尝试应用该技术对深色白眉天蛾Hyles gallii(Rottemburg)、构月天蛾Parum colligata(Walker)、桃红六点天蛾Marumba gaschkewitschi(Bremer et Grey)等3种天蛾进行了鉴别。测定了3种天蛾的COⅠ基因序列,并在BOLD(Barcode of Life Data Systems)及GenBank中进行了检索。结果发现BOLD系统比GenBank更能帮助准确鉴定到物种,而在GenBank中只有1种能鉴定到种。将实验室获得序列与GenBank中下载的474条同源序列整合后,利用MEGA4.0的Kimura-2-Parameter模型进行了遗传距离分析,利用MEGA4.0及PAUP4.0进行了系统发育树的构建,结果发现待鉴定标本序列在各个系统发育树的位置基本一致,并能准确鉴定出一种天蛾。本研究表明,随着BOLD系统、GenBank等公共数据库的不断完善,DNA条形码将有助于更加有效地鉴定特定类群物种。     

一种改进的小型节肢动物无形态损伤的DNA提取方法

本文介绍了一种改进的小型节肢动物无形态损伤的DNA提取方法,并在双尾纲、原尾纲和弹尾纲中进行了实验验证。结果表明,该方法可以高效的提取三类小型节肢动物的DNA,并用于扩增目标基因序列,凭证标本的回收质量高,有助于进一步的分类鉴定。该方法有望对螨、蚜虫、介壳虫、蚤等其他小型节肢动物的分子鉴定提供方便。     

试论昆虫种类鉴定的准确性

昆虫分类学有３个主要的任务：（１）鉴定昆虫种类;（２）研究昆虫的区系;（３）探索昆虫的自然系统发育。这３项工作既相互联系,又有各自的侧重点。鉴定昆虫种类的核心问题是识别昆虫是哪一个种,它的正确学名（有效名）是什么。在许多类似的昆虫中区别它们是否为同一个种。１物种概念是昆虫鉴定的指导思想物种是什么？依据什么来确定一个物种？这是昆虫鉴定最基本的问题。这一概念的理解对鉴定工作有着最直接的影响。昆虫种类鉴定就是在这一概念指导下进行的。物种的定义有很多,花保祯博士曾在其博士论文中例举了最具代表性的一些定义…     

入侵种的DNA条形码鉴定

生物安全研究的范畴涉及任何由生物威胁所造成的风险。随着害虫优先考虑级别的不断变化,以及国家和部门间相互协作的不断加强,对DNA分子鉴定技术的标准化提出了更迫切的要求,而DNA条形码的出现为此类问题的解决提供了很好的机遇。我们以前人对毒蛾和果蝇的研究为例,比较了DNA条形码技术与PCR-RFLP等传统方法的鉴定效果,在此基础上提出了建立一个可对不同入侵种进行快速和准确鉴定的条形码技术平台的构想。     

云南食用蜜蜂种类调查及鉴定

在云南,人们有食用蜜蜂幼虫和蛹的习俗。市场上销售的蜜蜂幼虫和蛹多经热水烫过或煮过,因而缺少成虫形态,俗名易混用,食用种类长期未予明确。本文在市场调查的基础上,收集其中部分样品,通过形态学和mtDNA的COI基因部分序列初步鉴定其分类地位,以明确食用种类。结果表明,云南蜜蜂的食用集中在横断山脉南部地区,接受人群广泛,食用方式多样;云南食用本地分布的全部5种野生蜜蜂,包括东方蜜蜂(Apis cerana)、小蜜蜂(A.florea)、大蜜蜂(A.dorsata)、黑大蜜蜂(A.laboriosa)和黑小蜜蜂(A.andreniformis);基于COI序列构建的邻接法(NJ)和最大简约法(MP)系统发育树支持现有的蜜蜂属系统发育关系;现有的食用蜜蜂大多采集自野外,亟待开展遗传资源保护。本研究为食用蜜蜂的鉴定提供了数据支持,也为蜜蜂自然资源的利用和保护提供参考。     

Phylogenomic relationships and species identification of the olive genus Olea (Oleaceae)

The olive genus Olea includes c. 30–40 taxa in three subgenera (Olea, Tetrapilus, and Paniculatae) within the family Oleaceae. Historically, the Olea genus was classified into four groups that were overall well supported by reconstructed phylogenies, despite incomplete sampling of subgenus Tetrapilus and poor resolution within clades. These analyses also showed that the genus was not monophyletic. Reliable identification of Olea species is important for both their conservation and utilization of this economically important genus. In this study, we used phylogenomic data from genome skimming to resolve relationships within Olea and to identify molecular markers for species identification. We assembled the complete plastomes, and nrDNA of 26 individuals representing 13 species using next-generation sequencing and added 18 publicly available accessions of Olea. We also developed nuclear SNPs using the genome skimming data to infer the phylogenetic relationships of Olea. Large-scale phylogenomic analyses of 138 samples of tribe Oleeae supported the polyphyly of Olea, with Olea caudatilimba and Olea subgenus Tetrapilus not sharing their most recent common ancestor with the main Olea clade (subgenus Paniculatae and subgenus Olea). The interspecific phylogenetic resolution was poor owing to a possible rapid radiation. By comparing with the plastome data, we identified the markers ycf1b and psbE-petL as the best Olea-specific chloroplast DNA barcodes. Compared with universal barcodes, specific DNA barcodes and super-barcode exhibited higher discriminatory power. Our results demonstrated the power of phylogenomics to improve phylogenetic relationships of intricate groups and provided new insights into barcodes that allow for accurate identification of Olea species.     

中文蜻蜓常用名称考

在我国,蜻蜓有多种名称,目前仍在使用的有蜻蜓、蜻蛉、水虿、、豆娘等。它们当中,有些是源自蜻蜓类昆虫的体色,如“蜻、”;有些是由蜻蜓的身体形状而来,如“蜓、蛉、水虿”。而“豆娘”出现最晚,应用时间也最短。中文之“豆娘”与日文中的“豆娘”关系甚为复杂,目前尚无定论。     

Dragondrop: a novel passive mechanism for aerial righting in the dragonfly

Dragonflies perform dramatic aerial manoeuvres when chasing targets but glide for periods during cruising flights. This makes dragonflies a great system to explore the role of passive stabilizing mechanisms that do not compromise manoeuvrability. We challenged dragonflies by dropping them from selected inverted attitudes and collected 6-degrees-of-freedom aerial recovery kinematics via custom motion capture techniques. From these kinematic data, we performed rigid-body inverse dynamics to reconstruct the forces and torques involved in righting behaviour. We found that inverted dragonflies typically recover themselves with the shortest rotation from the initial body inclination. Additionally, they exhibited a strong tendency to pitch-up with their head leading out of the manoeuvre, despite the lower moment of inertia in the roll axis. Surprisingly, anaesthetized dragonflies could also complete aerial righting reliably. Such passive righting disappeared in recently dead dragonflies but could be partially recovered by waxing their wings to the anaesthetised posture. Our kinematics data, inverse dynamics model and wind-tunnel experiments suggest that the dragonfly''s long abdomen and wing posture generate a rotational tendency and passive attitude recovery mechanism during falling. This work demonstrates an aerodynamically stable body configuration in a flying insect and raises new questions in sensorimotor control for small flying systems.     

来自东亚的黑耳属一新种,亚东黑耳

“亚东黑耳”是西藏近几年备受关注的一种特色野生珍稀食用菌,但对其分类学地位尚无定论。本文基于分子系统学和形态学研究了东亚、欧洲和北美洲与亚东黑耳相近的黑耳属标本,发现碟形黑耳Exidia recisa是个物种复合群,已知包括4个种,欧洲有碟形黑耳和浅波黑耳E. repanda两个种,北美洲有钝齿黑耳E. crenata,东亚的为1新种,本文将其描述为亚东黑耳Exidia yadongensis。这4个种在系统发育关系中相近,形态相似,但亚东黑耳的子实体边缘基本是全缘,担孢子大小为12-16×3-4μm (Q=3.62-3.83),而欧洲的碟形黑耳E. recisa和北美洲的钝齿黑耳E. crenata子实体边缘通常锯齿状,担孢子分别为13-15.5×2.8-3.8μm (Q=4.51)和12-14×3-4μm (Q=4.11),浅波黑耳E. repanda具有垫状至波浪状折叠的子实体,区别于其他3个种的盘状至耳状的子实体,其担孢子为12.7-15.7×2.8-3.7μm (Q=4.33-4.57)。此外,亚东黑耳具有树状分枝和叉形的子实层侧丝,但其他3个种只有树状分枝的子实层侧丝。尽管亚东黑耳广泛分布于东亚地区,但由于亚东黑耳的汉语名称已经在中国普遍使用,所以我们提出了上述拉丁学名。     

一种褐飞虱病原真菌的分子生物学鉴定

从田间褐飞虱Nilaparvata lugens( St(a)l)罹病虫体分离得到一株绿僵菌,以该菌为供试菌体,采用氯化苄法、CTAB法以及裂解液法分别提取了该菌的基因组DNA;以ITS1和ITS4为绿僵菌通用引物,对供试菌株的rDNA ITS序列进行PCR扩增、琼脂糖凝胶电泳检测和序列分析,并在核酸序列数据库中进行同...     

Molecular identification of species in Prunus sect.Persica (Rosaceae), with emphasis on evaluation of candidate barcodes for plants

Species of Prunus L. sect. Persica are not only important fruit trees, but also popular ornamental and medicinal plants. Correct identification of seedlings, barks, or fruit kernels is sometimes required, but no reliable morphological characters are available. Nowadays, the technique of DNA barcoding has the potential to meet such requirements. In this study, we evaluated the suitability of 11 DNA loci (atpB-rbcL, trnH-psbA, trnL-F, trnS-G, atpF-H, rbcL, matK, rpoB, rpoC1, nad1, and internal transcribed spa...     

Rapid mitochondrial genome sequencing based on Oxford Nanopore Sequencing and a proxy for vertebrate species identification

Molecular information is crucial for species identification when facing challenging morphology‐based specimen identifications. The use of DNA barcodes partially solves this problem, but in some cases when PCR is not an option (i.e., primers are not available, problems in reaction standardization), amplification‐free approaches could be an optimal alternative. Recent advances in DNA sequencing, like the MinION device from Oxford Nanopore Technologies (ONT), allow to obtain genomic data with low laboratory and technical requirements, and at a relatively low cost. In this study, we explore ONT sequencing for molecular species identification from a total DNA sample obtained from a neotropical rodent and we also test the technology for complete mitochondrial genome reconstruction via genome skimming. We were able to obtain “de novo” the complete mitogenome of a specimen from the genus Melanomys (Cricetidae: Sigmodontinae) with average depth coverage of 78X using ONT‐only data and by combining multiple assembly routines. Our pipeline for an automated species identification was able to identify the sample using unassembled sequence data (raw) in a reasonable computing time, which was substantially reduced when a priori information related to the organism identity was known. Our findings suggest ONT sequencing as a suitable candidate to solve species identification problems in metazoan nonmodel organisms and generate complete mtDNA datasets.     

分子标记在食（药）用真菌遗传多样性研究中的应用

食（药）用真菌在经济和生态方面都具有重要意义,其遗传多样性研究是资源可持续利用和生物保护学研究的基础,有利于食（药）用真菌种质资源的收集、保存、评价和利用,也有助于其分类学、系统学及进化等的研究。遗传多样性的研究方法很多,分子标记是目前最常用最有效的方法之一。综合分析了分子标记在食（药）用真菌遗传多样性研究中的应用,比较了各种标记的应用范围、优缺点,探讨了分子标记用于食（药）用真菌遗传多样性评价的前景及问题。