DNA barcode identification has wide applications in taxonomy, molecular phylogenetics, biodiversity science and species identification. This work developed a CRISPR-microfluidic array for rapid identification of DNA barcodes. This assay has successfully distinguish various types of Datura L. with single-copy sensitivity and rapidity. The detection system achieved 100% consistency with the morphological identification in detecting real-field seeds, holding great potential in the field of DNA barcoding and species recognition.

小麦花叶病毒(wheat mosaic virus,WMoV)是严重危害小麦、玉米等禾本科作物的种传病毒,在我国尚无发生,具有重要检疫意义.本研究利用引物WMoV-283F/R全国首次检测出玉米种子中的WMoV,成功扩增出了两个不同典型毒株NP序列,并分析其序列特征.根据WMoV-RNA3、WMoV-RNA3A和WMoV-RNA3B序列特点设计特异性引物,经RT-PCR扩增玉米种子样品2211-2和8044-10的RNA,扩增产物测序得到283 bp基因序列,分别与WMoV分离物GG1-RNA3B(GenBank登录号KT988872.1)、WMoV分离物K1-RNA 3(GenBank登录号KT988889.1)同源性为96.11％、96.47％.设计引物分别扩增玉米种子样品2211-2和8044-10的RNA核衣壳全长序列,得到WMoV-Germany RNA3、WMoV-Malaysia RNA3A和WMoV-Malaysia RNA3B全长序列.使用MEGA6软件采用Maximum likelihood法建立系统发育树,它们分别属于3个同源性差异明显的簇.通过DNAMAN序列比对分析三者的关系,RNA3序列在标尺碱基位点220～506区域序列差异性最大,而序列700～892、972～1102区段则相对保守.这为检测引物的设计提供了重要参考.WMoV的首次截获并鉴定为该病毒的检测鉴定和致病机理研究提供依据.

针对目前小麦不完善粒检测完全依靠人眼识别的低效性和误差较大的缺陷,构建了基于多元细粒度卷积神经网络的人工智能检测新方法.开展了人工智能检测新方法与传统人眼检测的对比验证,检测结果基本一致,符合国家标准中对不完善粒检测结果的允差要求,新方法模型对完善颗粒的识别准确率高达99.5％、不完善粒的识别准确率达91.7％,总体准确率高达99.1％.研究证明基于多元细粒度卷积神经网络的人工智能检测新方法具有准确、迅速、高效的特点,能够较好地适应小麦不完善粒检测需求,有望今后扩充到现有的标准体系中,逐步实现小麦不完善粒检测的人工智能化.

"Dalajiao" is a common variety of Phalaenopsis aphrodite. The individuals of the variety were cultured at a high temperature of 35℃, 14 h light/d with different durations. Throughout the treatment, their morphological changes were observed and the chlorophyll fluorescence parameters including the maximum quantum efficiency of photosystem II (PSⅡ) photochemistry(Fv/Fm), electron transfer rate under different light intensities were determined. Their corresponding explants were cultured in vitro to evaluate the potential for successful reproduction in vitro. We found that there existed a positive correlation between Fv/Fm of the stems and the successful rate of tissue culture in vitro with corresponding stems. When Fv/Fm of stems was less than or equal to 0.35, the stems were unable to be cultured in vitro because of lacking vitality. The Fv/Fm of the stems is better than that of the leaves in indicating their potential as explants for tissue culture in vitro.

采集了中国6个省份的铁皮石斛种质资源共22份,通过叶绿体基因matK和rbcL进行克隆测序,同时结合已报道的近缘物种matK序列33个和rbcL序列30个进行系统进化分析及分子鉴定.序列结果显示:matK基因序列平均长度为1708 bp,GC含量32％,保守位点768个,简约信息位点871个;rbcL基因序列平均长度1332 bp,GC含量42.9％,保守位点802个,简约信息位点24个.遗传多样性分析表明:matK序列单倍型数量(H)32个,单倍型多态性水平(Hd)0.842,核苷酸多态性(π)0.27016;rbcL序列H为10个,Hd为0.664,π为0.02611;序列分析及遗传多样性显示matK序列的分析结果具有更大的遗传多样性.遗传聚类结果显示不同产地的铁皮石斛与已有报道的石斛属各聚一支,相同产地铁皮石斛的聚类结果也与其地理分布基本一致.matK序列作为石斛属系统进化关系及亲缘关系分析具有更高的遗传多样性;中国6个省份铁皮石斛种质资源在进化上也具有一定的地域性特征.

[目的]为了解栽培丹参rDNA内转录间隔区(ITS)的遗传多样性、系统发育关系和寻找其单核苷酸多态性(SNP)指纹用以准确快速鉴别丹参栽培居群.[方法]采用PCR技术扩增了国内33个地区的42个栽培丹参居群的ITS序列并进行了比较分析.[结果]丹参rDNA的ITS1、5.8SrDNA和ITS2区的长度分别为203～229、164和226～229 bp;其ITS1区长度变化较大,而ITS2区核苷酸变异较大.其SNP变异位点存在于ITS1 和ITS2区,变异位点数分别为19和37个,变异率分别为8.30％和16.16％.[结论]基于ITS的SNP指纹只能鉴别3个栽培丹参居群;ITS系统发育树显示,丹参栽培居群之间的亲缘关系与地理来源关系不大.

为实现我国非重要的检疫性杂草——苍耳属Xanthium杂草快速高效的分子鉴定,利用Illu-mina二代测序平台对白苍耳X.albinum、意大利苍耳X.italicum、直刺苍耳X.ripicola、刺苍耳X.spinosum、柱果苍耳X.cylindricum、宾州苍耳X.pennsylvanicum、蝟实苍耳X.echinatum、苍耳X.si-biricum和北美苍耳X.strumarium共9种苍耳属杂草进行转录组测序,获得的unigene序列数分别为103362、93506、87452、83822、70994、66397、57598、57164和57134个.利用生物信息分析方法对9种苍耳属杂草转录组进行简单重复序列(simple sequence repeat,SSR)和物种间的特异性序列预测和筛选.经过PCR试验验证,筛选获得了可以有效鉴定上述9种杂草的6对SSR引物和13对特异性引物,提高了苍耳属杂草鉴定的准确率.

随着我国进出口贸易的扩大,杂草等有害生物传入我国的风险也大大增加,海关部门从进境货物中截获到大量杂草,其中以苍耳属植物的截获量最大.本文系统分析了2003-2017年我国海关部门截获苍耳种类的情况:全国进境货物中共截获苍耳属杂草28种,70 269种次,其中检疫性苍耳26种,56 888种次;美国、澳大利亚和巴西是截获苍耳种类最多的3个来源国,占截获总数的69％;截获苍耳种类最多的货物是大豆和羊毛.另外,本文归纳整理了苍耳属的常见分类学说和研究现状,为口岸提供检疫鉴定苍耳属植物的理论支撑.本文也对进境货物检疫查验和后续监管提出管理建议.

介绍冬虫夏草的寄主昆虫对药效的影响、生物学特性、人工培育、系统分类、种类、地理分布以及分子系统学等方面的研究,以期对冬虫夏草资源的合理利用及再生提供科学的方向.

小麦线条花叶病毒(wheat streak mosaic virus,WSMV)是禾本科粮食作物上重要的检疫性有害生物,能引起严重花叶、矮化和坏死.本文报道从美国进境的种用燕麦种子中检出WSMV,并研究了该病毒在种子内部的分布.挑选表面带有花纹的种子,利用两对不同的引物进行RT-PCR检测,结果均从样品中扩增出特异性条带.PCR产物测序分析结果表明为WSMV多聚蛋白部分序列.通过对幼苗及种子不同部位的检测,发现WSMV主要存在于种子稃皮及种皮内,而胚及胚乳中则未检出病毒,这一结果有助于提高WS-MV的检出效率.

本文采用ISSR分子标记方法分析了24份不同地区来源铁皮石斛种质资源的遗传多样性和亲缘关系.利用筛选出的7条ISSR引物对铁皮石斛基因组DNA进行PCR扩增,共获得92条DNA条带,其中多态性条带84条,多态性比率(PPB)为91.3％.利用POPGENE 32软件计算得出平均观察等位基因(Na)为1.913 0,平均有效等位基因数(Ne)为1.528 3,平均Nei's基因多样性指数(H)为0.310 0,平均Shannon's信息指数(I)为0.465 0.利用NTSYS-pc软件得出24份试验材料的遗传相似系数(GS)范围为0.478 3～0.913 0,聚类分析结果表明24份人工栽培居群的铁皮石斛分为3类,暗示铁皮石斛种质具有较为丰富的遗传多样性.该研究为全面掌握铁皮石斛种质资源遗传背景,选育、引种和推广优良铁皮石斛品种奠定了可靠的理论基础.

The DNA C-values of three Macromitrium species (including M.cavaleriei,M.japonicum and M.gymnostomum) were determined by flow cytometry by using Physcomitrella patens and Oryza sativa spp. japonica as standard samples,respectively.Thus the method of measuring DNA C-values of Macromitrium species by flow cytometry was established.The factors influencing the measuring result were also discussed.Finnally,variaiton patterns of bryophytes DNA C-values,their phylogenetic and taxonomic significances,and some interesting topics about DNA C-value research were remarked.

Previous studies on plant nuclear DNA content variation and its relationships with environmental gradients produced conflicting results. We speculated that the relationships between nuclear DNA content of a widely-distributed species and its environmental gradients might be non-linear if it was sampled in a large geographical gradient. Echinochloa crus-galli (L) P. Beauv. is a worldwide species, but without documents on its intraspecific variation of nuclear DNA content. Our objectives are: 1) to detect intraspecific variation scope of E. crus-galli in its nuclear DNA content, and 2) to testify whether nuclear DNA content of the species changes with environmental gradients following Gaussian models if its populations were sampled in a large geographical gradient. We collected seeds of 36 Chinese populations of E. crus-galli across a wide geographical gradient, and sowed them in a homogeneous field to get their offspring to determine their nuclear DNA content. We analyzed the relationships of nuclear DNA content of these populations with latitude, longitude, and nineteen bioclimatic variables by using Gaussian and linear models. (1) Nuclear DNA content varied from 2.113 to 2.410 pg among 36 Chinese populations of E. crus-galli, with a mean value of 2.256 pg. (2) Gaussian correlations of nuclear DNA content (y) with geographical gradients were detected, with latitude (x) following y = 2.2923*e (-(x-249360)2/2.6379452) (r = 0.546, P < 0.001), and with longitude (x) following y = 2.2933*e(-(x-116.1801)2/2.4474502) (r = 0.672, P < 0.001). (3) Among the nineteen bioclimatic variables, except temperature isothermality, precipitations of the wettest month, the wettest quarter and the warmest quarter, the others could be better fit with nuclear DNA content by using Gaussian models than by linear models. There exists intra-specific variation among 36 Chinese populations of E. crus-galli, Gaussian models could be applied to fit the correlations of its Nuclear DNA content with geographical and most bioclimatic gradients. (C) 2017 Elsevier Masson SAS. All rights reserved.

There are a variety of farm weeds in Aegilops species,of which Aegilops cylindrical and Aegilops tauschii are quarantine weeds and often intercepted in the import grain.Identification of quarantine Aegilops species is very difficult because they are very similar in morphology to their close species.In this paper,a PCR-RFLP molecular identification method was developed for the rapid and accurate separation of quarantine Aegilops species from non-quarantine species.

The global economy is developing forward to a more liberal and convenient direction,and the circumstance and rules of trade are making corresponding changes.China is taking the initiative to explore and adapt to the new free trade environment and rules through the establishment of free trade area.Pest risk assessment (PRA) is an important technical support to protect and promote the trade of agricultural products.This paper compared the development of pest risk assessment technology in China with that in Europe,USA and Australia.The results showed that there were some problems existed in the PRA of China,which were lacking of system planning,scattered,repetitive and slow in information construction,and lacking of basic and applied research.In order to better meet the requirements of the new situation of free trade in the future,it is suggested that China should clarify the important role and status of pest risk assessment,develop a framework system of pest risk assessment for the new trade situation,strengthen the investment in basic information construction,risk assessment talent training and basic theoretical research,and promote the practical application in the development of pest risk assessment standards and pest forecasting.

[目的]近年来,通过电子商务平台获取境外珍贵的多肉物种资源已成为一种重要渠道,大量濒危物种通过第三方物流方式非法流入我国.甄别濒危物种并梳理出多肉物种资源重点查验名单,能够为物种资源查验尤其是植物多肉类濒危物种查验提供参考.[方法]以跨境多肉物种资源交易的热门平台"多肉之家"为研究对象,基于网络爬虫技术平台,获取电商多肉植物926条种类数据,并对数据进行筛选和归类分析.[结果]"多肉之家"平台上交易的多肉植物共涉及23科878种,其中包含18种CITES附录Ⅰ以及120种CITES附录Ⅱ濒危物种,约占16%.进一步对濒危植物进行归类分析发现,濒危植物中仙人掌科和大戟科多肉植物种类最多,分别为66和36种,两者占总计濒危植物种类的74%.[结论]网络爬虫技术在获取电商类平台的交易植物的种类数据上具有较好的实用性.

This paper reviewed the five major new quarantine and management measures for animal and plant quarantine in Shanghai FTZ.This series of new measures emphasized the important role of pest risk analysis techniques,proposed to implement classified management measures based on risk classification,and eventually promoted the development of animals and plants trade and provided a large number of practical measures and experiences for the reform of animal and plant quarantine management in our country.With the further development of free trade,the objects of animal and plant quarantine management will shift to a larger number of medium-risk animal and plant products and will face the dual requirements for safety baseline and facilitation.Therefore,our reform should shift from the existing "external" reforms to "internal" reforms,that is,the focus of animal and plant quarantine management should be shifted to improving the technologies of animal and plant quarantine.The reform of administrative functions should be focused on creating the best environment for the improvement of animal and plant quarantine technology to achieve the organic combination of administrative functions and animal and plant quarantine technology and thus create a safe,convenient,effective and free trade environment for animals and plants in our country.

In recent years,Dickeya solani has established itself in a number of European countries.Investigation demonstrated that D.solani plays major role in heavy blackleg outbreaks.It has found to cause disease on flowering bulbs (eg hyacinth).It is necessary to strengthen the quarantine activity of imported hyacinth bulb for preventing the spread of bacteria from abroad.This paper will briefly describe the taxonomic status,host,distribution,symptoms of damage,biological characters and quarantine diagnosis of D.solani.

本文介绍了ISPM32号标准框架下基于有害生物风险的商品分类管理的总体思路和我国现行的植物检疫措施,比较了我国现行检疫措施与ISPM32号标准的区别,提出了基于检疫风险的植物产品动态分级管理模型,以期为建立适合我国国情的植物检疫分类管理体系,推进植物检疫措施规范化、标准化、国际化提供参考.