瘿蚊科3种昆虫核糖体DNA的克隆及序列分析

瘿蚊科是双翅目昆虫中的重要类群?本研究对瘿蚊科3种昆虫—麦红吸浆虫?菊花瘿蚊和食蚜瘿蚊的核糖体DNA进行PCR扩增?克隆?测序及序列分析, 并对ITS-1在麦红吸浆虫4个地理种群中的遗传变异情况进行分析?结果表明, 从3种昆虫中获得的核糖体DNA序列包括:部分的18S rDNA(44 bp)?28S rDNA(37 bp), 全部的ITS-1(487~535 bp), 5.8S rDNA(121 bp)及ITS-2(336~352 bp)序列?3种昆虫ITS序列的碱基差异百分比在17.21%~29.59%之间, 共含有206个变异位点?ITS-1序列在麦红吸浆虫4个地理种群中比较保守, 只有4个变异位点, 单倍型多样性为0.311 7~0.796 5, 核苷酸多样性为0.000 6~0.002 2?本研究为今后瘿蚊科昆虫的分类鉴定?系统发育和遗传进化等相关研究提供了基础?     

欧洲纵坑切梢小蠹的检疫鉴定研究

云浮口岸从土耳其进境木质包装中截获多头活小蠹虫,本研究对该小蠹虫的形态特征进行描述,同时采用小蠹科通用引物CI-J-2183和TL2-N-3014进行PCR扩增该虫的COⅠ基因序列。结果获得758 bp的基因序列,通过与Gen Bank数据库比对分析显示,该序列与Gen Bank数据库上的3条欧洲纵坑切梢小蠹COⅠ序列相似性高达99%;采用邻接法(NJ)构建的系统发育树也显示该虫与欧洲纵坑切梢小蠹亲缘关系极近且在同一进化分支中。结合形态特征和DNA条形码序列分析,可最终判定所截获昆虫为欧洲纵坑切梢小蠹。     

亚洲玉米螟6-磷酸葡萄糖异构酶基因(Pgi)全序测定及分析

为进一步从核酸水平上研究亚洲玉米螟Pgi基因相关特性,采用反转录PCR及RACE等技术对该基因编码序列及DNA全序列进行了测定,与Gen Bank中其它昆虫Pgi基因相关信息进行比较分析,并构建了系统发育树。结果表明:亚洲玉米螟Pgi基因编码区序列长为1 671 bp,共编码556个氨基酸;其DNA序列全长为10 078 bp(短序列为9 311 bp),由12个外显子与11个内含子镶嵌而成;各外显子长度与鳞翅目大部分昆虫相同,介于95~188 bp之间,内含子序列总长度为8 407 bp(短序列为7 640 bp),各内含子长度介于418~1 547 bp之间,且在内含子3、4、5、11上均发现杂合现象。系统发育结果显示,大部分昆虫Pgi基因c DNA严格按物种聚类,除了双翅目的家蝇Musca domestica与黑森瘿蚊Mayetiola destructor出现一定的交叉现象外,其余没有出现交叉现象。     

介绍两种窃蠹科害虫——浓毛窃蠹,灵芝窃蠹

介绍两种窃蠹科害虫——浓毛窃蠹、灵芝窃蠹林其铭（福州动植物检疫局３５００１３）陈启宗（郑州粮食学院）窃蠹科昆虫全世界目前已记述有２０００多种，分隶于８个亚科、１６０多个属。但其形态特征差异不大，种乃至属的正确鉴定存在一定困难。我国对该科昆虫的研究尚浅...     

三种重要皮蠹科害虫的幼虫形态与分子鉴定

为寻求白腹皮蠹和赤毛皮蠹幼虫简便科学的鉴定方法,利用形态学结合分子生物学对其做了鉴定研究,首先对其触角、上内唇、臀突和胸足胫节的特征进行了形态描述;再分别提取白腹皮蠹、赤毛皮蠹和外群黑毛皮蠹幼虫的虫体DNA进行PCR扩增,通过测序比对构建了系统进化树。结果表明:白腹皮蠹幼虫上内唇的中间2根缘刚毛呈蘑菇状,粗短,而赤毛皮蠹幼虫的则呈矛状,细长,二者上内唇的形态特征差异非常明显,可用于二者形态上的鉴定;对3种皮蠹幼虫的序列进行比对,白腹皮蠹与赤毛皮蠹的碱基相似率为89.97%,黑毛皮蠹与白腹皮蠹和赤毛皮蠹的碱基相似率分别为77.54%和77.05%,差异均明显,可用于3种皮蠹幼虫种间鉴定的线粒体COⅠ基因标记序列。     

进境木质包装疫情的国别(地区)分析

对近年来我国口岸截获进境木质包装疫情进行了统计,通过聚类分析方法可以将主要疫情来源国家(地区)按照疫情截获情况分为4类。长蠹科、长小蠹科、小蠹科、鼻白蚁科、天牛科、蚁科(仅含红火蚁一种)等6科昆虫及线虫(主要为松材线虫)为进境木质包装中的主要检疫性有害生物。因子分析表明,中国台湾、印度、东南亚地区及美国、俄罗斯、中国香港、德国、日本、韩国为木质包装疫情最为严重的地区,不同国家(地区)主要检疫性有害生物类别特征明显。此外,给出了木质包装中主要检疫性有害生物名录及检疫监管对策。     

6种木材钻蛀性昆虫的声学特征与比较

为了在口岸检疫中应用木材钻蛀害虫声检测,测试了鳞毛粉蠹(Minthea rugicollis Waller)、双钩异翅长蠹(Heterobostrychus aequalis(Waterhouse))、棕异翅长蠹(Heterobostrychus brunneus(Murray))、黑双棘长蠹(Sinoxylon conigerum Gerstaecker)、刺角楝天牛(Cordylomera spinicornis(Fabricius))和宽斑脊虎天牛(Xylotrechus colonus Fabricius)等6种昆虫幼虫蛀食声的声学特征。6个种的蛀食声均为不连续的单个声脉冲,波形包络前大后小,逐渐收敛;这些蛀食声均属于离散谱,且频带较宽,主要能量集中在10kHz以下,一般都至少有一个明显的高峰(主频)。单个脉冲的持续时间和主频在不同种间有差异,并存在这些昆虫科间主频差异的可能性。     

利用近等基因系构建黏虫黑化型AFLP-SCAR标记

意义与目的迁飞昆虫种群内存在丰富的遗传变异,如外翅类昆虫的翅型分化和内翅类昆虫的迁飞型分化现象。黏虫迁飞型的分化很大程度上受环境因素调控,限制了从分子水平鉴定迁飞型分化。黏虫黑化型是种内发生变异后形成的一种新基因型,体色差异主要表现在成虫,而其他发育阶段均与正常型无明显差别。黑化性状是受常染色体上单对等位基因中隐性基因调控,符合孟德尔完全隐性遗传规律。其行为、生理特征及生活史对策符合滞留型标准。为揭示黏虫黑化型变异的分子基础,通过构建黑化型与正常型近等基因系,应用AFLP技术筛选与黑化连锁的DNA标记,并将…     

从印尼胶合板上截获一种双棘长蠹

从印尼胶合板上截获一种双棘长蠹１９９３年１０月１３日，汕头动植物检疫局植检科对“布米”轮运载进口的印尼胶合板实施检疫时发现一长蠹科害虫，经鉴定该虫为ＳｉｎｏｘｙｌｏｎｃｏｎｉｇｅｒｕｍＧｅｒｓｔａｃｋｅｒ（长蠹科、大长蠹亚科、长蠹族、双棘长蠹属）。为...     

基于DNA条形码的红翅大小蠹和黄杉大小蠹分子鉴定

外来有害生物的快速准确鉴定有助于提高口岸检疫效率。红翅大小蠹(Dendroctonus rufipennis)和黄杉大小蠹(D. pseudotsugae)作为大小蠹属的非中国种是我国口岸原木检疫中频繁截获的检疫性有害昆虫。本研究以mtDNA COI基因5′端和3′端序列为标记对红翅大小蠹与黄杉大小蠹的11个样本开展分子鉴定有效性评价。结果显示,当以COI基因5′端和3′端为靶标时,两种大小蠹间的平均遗传距离分别为0.181和0.144,是种内遗传距离的60.3和48倍,且种内、种间遗传距离间无重叠现象,在系统发育树上都能聚为独立的分支,表明mtDNA COI基因5′端和3′端序列均可有效区分红翅大小蠹与黄杉大小蠹。     

Occurrence and Identification of Phytophthora spp. Pathogenic to Pear Fruit in Irrigation Water in the Wenatchee River Valley of Washington State

ABSTRACT Seven hundred forty-nine isolates of Phytophthora spp. were obtained from irrigation canals in eastern Washington State during the 1992 to 1995 and 1999 growing seasons. Isolates were retrieved using pear baiting techniques. All isolates were pathogenic to pear and were present in irrigation water beginning early in fruit development. Over the course of the 5 year study, 10 and 5% of isolates were identified as P. cactorum and P. citricola, respectively, using morphological criteria. The remaining isolates could not be identified using morphological criteria. Colony morphology of these isolates was characterized during all years of the study. In 1999, more detailed studies utilizing polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis of entire internal transcribed spacer (ITS) regions (ITS1, 5.8S, and ITS2) of ribosomal DNA for 180 isolates, and sequence analysis of ITS2 for 50 isolates, were used to investigate genetic variation and phylogenetic relationships among isolates. Isolates were divided into 12 groups based on their growth type on corn meal agar. Restriction digestion of the entire ITS region with three enzymes revealed 11 restriction digestion patterns among 180 isolates. PCR-RFLP and sequence data were obtained for 12 reference Phytophthora spp. (two species in each of Waterhouse's six morphological groups). Phylogenetic analysis of ITS2 regions revealed nine clades, each with strong bootstrap support. Molecular analyses revealed 23 isolates that were in the P. gonapodyides clade, 9 in the P. parasitica clade, 1 in the P. cactorum clade, 7 in the P. citricola/capsici clade, and 4 in the P. cambivora/pseudotsugae clade. The three isolates comprising clade 5 were significantly distinct from all other Phytophthora spp. in the databases and may represent a new Phytophthora sp. Colony morphology was not consistently correlated to PCR-RFLP pattern or ITS2 phylogeny, suggesting that the former criterion is insufficient for species identification. The results of this study indicate that at least nine phylogenetically distinct taxa of Phytophthora pathogenic to pear are present in irrigation water in North Central Washington.     

Whole-genome QTL analysis of Stagonospora nodorum blotch resistance and validation of the SnTox4-Snn4 interaction in hexaploid wheat

Necrotrophic effectors (also known as host-selective toxins) are important determinants of disease in the wheat-Stagonospora nodorum pathosystem. To date, five necrotrophic effector-host gene interactions have been identified in this system. Most of these interactions have additive effects while some are epistatic. The Snn4-SnTox4 interaction was originally identified in a recombinant-inbred population derived from a cross between the Swiss winter wheat cultivars 'Arina' and 'Forno' using the S. nodorum isolate Sn99CH 1A7a. Here, we used a recombinant-inbred population consisting of 121 lines developed from a cross between the hexaploid land race Salamouni and the hexaploid wheat 'Katepwa' (SK population). The SK population was used for the construction of linkage maps and quantitative trait loci (QTL) detection using the Swiss S. nodorum isolate Sn99CH 1A7a. The linkage maps developed in the SK population spanned 3,228 centimorgans (cM) and consisted of 441 simple-sequence repeats, 9 restriction fragment length polymorphisms, 29 expressed sequence tag sequence-tagged site markers, and 5 phenotypic markers. The average marker density was 6.7 cM/marker. Two QTL, designated QSnb.fcu-1A and QSnb.fcu-7A on chromosome arms 1AS and 7AS, respectively, were associated with disease caused by the S. nodorum isolate Sn99CH 1A7a. The effects of QSnb.fcu-1A were determined by the Snn4-SnTox4 interaction and accounted for 23.5% of the phenotypic variation in this population, whereas QSnb.fcu-7A accounted for 16.4% of the phenotypic variation for disease but was not associated with any known effector sensitivity locus. The effects of both QTL were largely additive and collectively accounted for 35.7% of the total phenotypic variation. The results of this research validate the effects of a compatible Snn4-SnTox4 interaction in a different genetic background, and it provides knowledge regarding genomic regions and molecular markers that can be used to improve Stagonospora nodorum blotch resistance in wheat germplasm.     

内蒙古白刺属植物遗传多样性的RAPD分析

利用随机扩增DNA多态性 (RAPD)技术对4种不同地理种群的白刺属(Nitraria L.)植物进行了遗传多样性分析.15个引物扩增出118个位点,其中多态性位点95个(80.5%),各引物的Nei's指数和Shannon's 多样性指数差别较大,利用UPGMA方法构建分子系统树.结果表明:白刺属植物的遗传分化程度和基因流因种而异,除球果白刺以外,其余3种白刺的种群间有很大的基因流,其遗传一致度在0.8561～0.9694之间,而种间遗传分化程度较低,故4种白刺有着相近的祖先,这与形态学和细胞学的分析结果一致,说明RAPD分析方法可从分子生物学角度为白刺属植物的系统学分类提供更为有利和可靠的证据.     

Molecular identification of weedy glyphosate-resistant Lolium (Poaceae) in California

The origins of invasive weed populations can involve multiple species, introductions and genotypes. In California, USA, self-incompatible (SI) Lolium species are highly successful agricultural weeds. During the last decade, resistance to glyphosate has evolved in California populations of Lolium . However, the species identity of glyphosate-resistant populations is unclear. Occurrence of three SI species, L. multiflorum , L. rigidum and L. perenne , and the lack of reproductive barriers between them, suggests that any or all species may be contributing to resistant populations. To elucidate the identity and evolutionary history of resistant Lolium , we assessed genetic variation of known species and unknown California individuals using five simple sequence repeat loci, a nuclear DNA sequence and a chloroplast DNA sequence. Employing two analytical approaches, we identified three major genetic groups within the accessions of SI species that were similar, but not identical, to the taxonomic species identities. California glyphosate-resistant and susceptible plants were most closely related to the L. multiflorum group. However, a few glyphosate-resistant plants and a third of the glyphosate-susceptible plants also identified with L. rigidum and L. perenne . While L. rigidum and L. perenne contribute to invasive populations within California, the majority of glyphosate-resistant individuals tested were L. multiflorum .     

河南商丘地区棉花黄萎病菌分离鉴定和致病力分析

为探讨河南商丘地区棉花黄萎病菌的致病型群体变异,对该地区棉花上分离的8株单孢菌株的菌落形态、显微结构、致病力、ITS序列、系统进化及菌体蛋白等方面进行了研究。结果表明:这些菌株均属于棉花黄萎病菌Verticilliumdahliae;系统进化树显示8株菌株并没有聚在同一进化枝上;8株黄萎菌菌株存在致病力差异,SQ4菌株致病力最强,属于落叶型,而其它致病力较弱的7个菌株属于非落叶型;不同致病力的菌株间蛋白谱带存在差异。     

法国向日葵种子中向日葵黑茎病菌的首次截获与检测

从法国进境向日葵种子中分离到3株疑似向日葵黑茎病的菌株,对所有菌株进行形态学、致病性测定和分子序列比对分析。分离菌菌落乳白色或象牙色至灰白色,有大量黑褐色分生孢子器产生,分生孢子器球形至扁球形,内含无色单胞、卵圆形分生孢子,有明显或不明显油球;针刺接种4片真叶向日葵幼苗的下胚轴,7～9d后茎部产生典型黑茎病黑色椭圆形病斑,病斑上着生黑色分生孢子器;菌丝DNA用ActF1/R1和ITS1/ITS4扩增,序列与NCBI基因库中P.macdonaldii序列相似性为98%～100%。形态学、分子生物学及致病性检测结果显示,截获的3株菌均为向日葵黑茎病菌。     

Emergence of a New Cucurbit-Infecting Begomovirus Species Capable of Forming Viable Reassortants with Related Viruses in the Squash leaf curl virus Cluster

ABSTRACT Cucurbit leaf curl virus (CuLCV), a whitefly-transmitted geminivirus previously partially characterized from the southwestern United States and northern Mexico, was identified as a distinct bipartite begomovirus species. This virus has near sequence identity with the previously partially characterized Cucurbit leaf crumple virus from California. Experimental and natural host range studies indicated that CuLCV has a relatively broad host range within the family Cucurbitaceae and also infects bean and tobacco. The genome of an Arizona isolate, designated CuLCV-AZ, was cloned and completely sequenced. Cloned CuLCV-AZ DNA A and B components were infectious by biolistic inoculation to pumpkin and progeny virus was transmissible by the whitefly vector, Bemisia tabaci, thereby completing Koch's postulates. CuLCV-AZ DNA A shared highest nucleotide sequence identity with Squash leaf curl virus-R (SLCV-R), SLCV-E, and Bean calico mosaic virus (BCaMV) at 84, 83, and 80%, respectively. The CuLCV DNA B component shared highest nucleotide sequence identity with BCaMV, SLCV-R, and SLCV-E at 71, 70, and 68%, respectively. The cis-acting begomovirus replication specificity element, GGTGTCCTGGTG, in the CuLCV-AZ origin of replication is identical to that of SLCV-R, SLCV-E, and BCaMV, suggesting that reassortants among components of CuLCV-AZ and these begomoviruses may be possible. Reassortment experiments in pumpkin demonstrated that both reassortants of CuLCV-AZ and SLCV-E A and B components were viable. However, for CuLCV-AZ and SLCV-R, only one reassortant (SLCV-R DNA A/CuLCV-AZ DNA B) was viable on pumpkin, even though the cognate component pairs of both viruses infect pumpkin. These results demonstrate that reassortment among sympatric begomovirus species infecting cucurbits are possible, and that, if generated in nature, could result in begomoviruses bearing distinct biological properties.     

对小麦黄花叶病毒存在抗性差异的2种小麦品种中eIF4E序列分析

正小麦黄花叶病于1927年在日本首次报道~([1]),其病原包括小麦黄花叶病毒(Wheat yellow mosaic virus,WYMV)和中国小麦花叶病毒(Chinese wheat mosaic virus,CWMV)。我国各省麦区在1980年以后陆续报道了其发生与危害,对小麦生长发育构成严重影响。WYMV属于马铃薯Y病毒科(Potyviridae)大