红火蚁与热带火蚁间的干扰竞争

【目的】为了探讨入侵火蚁在我国成功定殖及其之间的竞争机制。【方法】运用行为学方法研究红火蚁Solenopsis invicta(Buren)和热带火蚁Solenopsis geminata(Fabricius)在个体水平和群体水平上的攻击性、攻击手段及合作能力。【结果】一对一攻击试验中,红火蚁和热带火蚁之间攻击级别多集中在3级,两种入侵蚂蚁间以相互威胁为主;红火蚁大型工蚁与热带火蚁兵、工蚁间最为好斗,其攻击级别达到4级的比例最高,分别为33.04%、37.92%。热带火蚁兵蚁与各型红火蚁间攻击强度差异不显著;热带火蚁工蚁与红火蚁小型工蚁之间的攻击性最强,其攻击性(3.49)显著高于热带火蚁工蚁与红火蚁大、中型工蚁的攻击性(3.32和2.97)。在攻击手段上,3级打斗时各型红火蚁更倾向以物理攻击主动威胁热带火蚁,而热带火蚁兵、工蚁会采取多种方式主动攻击红火蚁,双方皆以躲避应对为主;4级打斗时两种火蚁主要以混合攻击为主动或应对手段。群体攻击试验显示,红火蚁群体间攻击强度和合作性会随着群体数量的增加而显著增加,热带火蚁合作性较差,其群体对抗红火蚁的优势仅仅是由于个体数量的增加。【结论】红火蚁比热带火蚁具有更强的竞争优势。研究结果为入侵蚂蚁间不对称竞争机制和长期群落替代的内在原因提供理论基础。     

红火蚁及其他两种蚂蚁毒腺细菌鉴定与多样性分析(英文)

【目的】本研究旨在分析红火蚁Solenopsis invicta毒腺细菌群落多样性,并与热带火蚁Solenopsis geminata和聚纹双刺猛蚁Diacamma rugosum比较毒腺细菌群落差异。【方法】采用Illumine Hiseq 2500测序平台对红火蚁(工蚁、有翅蚁和蚁后)、热带火蚁(工蚁)及聚纹双刺猛蚁(工蚁)毒腺细菌群落16S rRNA基因V3-V4区测序,基于测序数据进行生物信息学分析。【结果】变形菌门(Proteobacteria)在红火蚁工蚁、有翅蚁和蚁后以及热带火蚁工蚁毒腺中占优势,而厚壁菌门(Firmicutes)在聚纹双刺猛蚁工蚁毒腺中占优势。与红火蚁工蚁和有翅蚁相比,柔壁菌门(Tenericutes)在红火蚁蚁后毒腺中更丰富。广州红火蚁蚁后毒腺中假单胞杆菌Pseudomonas相对丰度显著高于其在有翅蚁及工蚁中的。螺原体Spiroplasma相对丰度在热带火蚁毒腺中显著高于在聚纹双刺猛蚁工蚁毒腺中的。毒腺中细菌多样性分析发现,芽孢杆菌属Bacillus和乳酸杆菌属Lactobacillus在广西红火蚁工蚁中的相对丰度显著高于在广州红火蚁工蚁中的。然而,乳酸杆菌属细菌在聚纹双刺猛蚁工蚁毒腺中的相对丰度显著高于广西的热带火蚁工蚁毒腺中的。【结论】毒腺细菌组成和多样性在3种不同种类蚂蚁工蚁和红火蚁不同品级中存在差异。     

Leber氏病的PCR基因诊断

木文报告了用PCR技术对线拉体DNA突变引起的人类Leber氏病的研究。此病惠者的 mtDNA第11778位由G突变成A,这种点突变使sfaNI酶识别位点消失,因此可通过PC1t扩增此特 异片段确定其酶切多态性。我们扩增了含sfaN1酶切识别位点在内的一段340bp的mtDNA, sfaNI酶 将正常人的mtDNA切成190师和150bp两片段,而患者的mtDNA则保留完整的340bp, 关键词：线粒体DNA, PCR, Leber氏肩     

基于基因组编辑技术的大片段克隆新策略

基因组大片段克隆技术是合成生物学研究领域关键的使能技术。传统的大片段克隆技术获取目的大片段的手段存在各种缺陷,比如随机建库克隆需要依靠高通量筛选;PCR难以扩增10 kb以上片段,从小片段拼装费时费力且突变率高;基于限制性酶切连接难以找到片段两端适宜的限制性内切酶酶切位点。最近全基因组合成等前沿研究创造了全新的高性能大片段克隆方法,比如CRISPR/cas9系统中cas9可识别并切割20 bp核酸序列解决了识别位点设计难题,可用来获取任意目的基因片段;组合Gibson或者酵母偶联重组技术组装技术,可高效克隆大片段基因。本文将分类介绍基因组大片段克隆技术,并提出适用不同尺度大小基因克隆的技术选择参考标准。     

两种火蚁与黑头酸臭蚁上颚形态及其超微结构观察

应用扫描电镜和光学显微镜观察了红火蚁Solenopsis invicta Buren、热带火蚁Solenopsis geminata Fabricius和黑头酸臭蚁Tapinoma melanocephalum Fabricius的上颚类型及形态特征。头宽和上颚长度测量结果表明:两种火蚁的头宽和上颚大小具有连续性特征,黑头酸臭蚁工蚁仅1个类型,3种蚂蚁中黑头酸臭蚁的上颚最小,其前端具4个锋利切齿叶,后部有一排小而钝的臼齿叶,上颚内缘较光滑。在形态上,红火蚁的大、中、小型工蚁与热带火蚁工蚁的上颚区别不大,表面纹理光滑,都具4个锋利小齿,上颚内缘也比较光滑;热带火蚁兵蚁上颚仅3个小齿且较钝,其上颚内缘凹面比两种火蚁工蚁的深且纹理粗糙,推测红火蚁工蚁不仅用于筑巢,还用于防御,而热带火蚁兵蚁的强壮上颚可能主要用于磨碎食物,而非保卫蚁巢。     

细胞质雄性不育高粱叶绿体 ndh D 基因的序列变异

片段SAAU－02 700特异地扩增自7种具可育细胞质的高粱材料的总DNA,含有叶绿体psa C(88bp)和ndh D(192bp)基因的部分序列。该片段与Eco Ri HindⅢ酶切的总DNA,线粒体DNA和叶绿体DNA杂交,在总DNA中获得了0.74kb的杂交带,而在叶绿体中获得0.74kb和0．45kb两条杂交带。与线粒体DNA无杂交;与经Hae Ⅲ酶切的总DNA杂交,在不育系中获得4．9kb的杂交带,而保持系的杂交带为4．45kb。参考GenBank中高粱的近缘物种玉米叶绿体基因组的序列,构建了ndh D基因区的酶切位点图谱,借此分析得出高粱不育系的叶绿体ndh D基因序列已发生改变。这种变异与高粱细胞质雄性不育反生的关系正在探讨中。     

鳞翅目基因组IIB型内切酶位点的统计分析

IIB型限制内切酶能够识别并切割特异酶切位点两端特定距离的DNA,形成粘性末端的30 bp左右的等长DNA片段。利用其特性与限制性酶切位点关联测序技术(RAD)相结合发展出2b-RAD简化基因组测序技术,应用于遗传图谱构建、种群遗传结构分析、性状定位以及细菌分型等多种研究领域。构建2b-RAD测序文库之前,需要对基因组中的IIB型限制内切酶位点进行预测与统计分析,制定有效的测序文库构建方案。本文利用Python语言构建分析基因组中IIB型限制内切酶位点的流程,预测并统计6个鳞翅目代表物种基因组含有的8个商业化IIB型限制内切酶的酶切位点,比较了各个基因组与IIB型限制内切酶之间含有的酶切位点总量、重复序列数量以及酶切间隔长度的关系,为在昆虫基因组中进一步试行2b-RAD研究提供了参考。     

幽门螺杆菌hpaA基因RFLP研究

用限制性片段长度多态性(RFLP)的方法评价幽门螺杆菌不同菌株鞭毛粘附素基因(hpaA)的变异性。PCR扩增9株幽门螺杆菌710bp的hpaA基因,用Hha Ⅰ、HaeⅢ限制性内切酶对该基因片段进行酶切分析。hpaA基因HaeⅢ单酶切可见4种带型,HhaⅠ单酶切出现5种带型。从临床分离的H.pylori菌株hpaA RFLP互有差异,且不同于国际标准菌株;临床分离株感染动物后分离得到的动物适应株其hpaA基因也发生了变异。不同H.pylori菌株间hpaA基因表现出明显的多态性,为开展H.pylori的分子流行病学调查提供了一种有效的方法。     

利用多重PCR反应同时筛选番茄Cf-9和Tm-1基因

利用同一PCR反应体系,对分别与番茄抗叶霉病的Cf-9基因和抗番茄烟草花叶病毒病的Tm-1基因紧密连锁的PCR标记进行了同时扩增筛选,扩增的特异性片段与单引物扩增片段吻合。其中与Cf-9基因紧密连锁的CAPs标记在抗感试材均可扩增出560bp的特异片段,且都存在TaqⅠ酶切位点,抗病基因型酶切后分别产生了450bp、330bp和290bp的不同特异性片段,而感病基因型试材酶切后产生450bp和290bp的特异性片段;与Tm-1基因紧密连锁的SCAR标记为显性标记,只有抗病试材产生750bp的特异片段,不能被TaqⅠ酶切。经反复验证,结果稳定准确,可用于在同一PCR反应体系中对两个抗病基因进行同时筛选鉴定。该体系的建立不仅省时、省工、节省费用,而且可用于苗期辅助选育,加快番茄抗病育种进程。     

动物遗传工程

951070利用PCR扩增的ZFX/ZFY位点的限制性片段多形性鉴别牛胚胎性别[英]/Pollevick,G.D.…,Bio/Technology.一1992,10(7).-805~'807[译自DBA,1992,11(17),92-09812] 克隆了雄z~ZFX和ZFY位点的PCR扩增片段并进行了测序。447bp扩增片段的检测显示若千多形性。检测出31个点突变,其中部分包括限僚5性内切酶位点。证实存在已介绍过的多形性Pstl位点(用于鉴别雌雄性),检测出2个更替性限制性片段长度多形性(RFLP)。FokI位点存在于ZFX位点中,而不存在于ZFY中,NsiI位点只存在于ZFY中。用牛ZFX和ZFY位点序列设计一套寡核苷酸,以提…     

Mechanisms of interspecific competition among an invasive and two native fire ants

The mechanisms of interspecific competition among an invasive and two native Solenopsis fire ant forms were investigated in a series of laboratory experiments. In separate trials each with a different food resource, the native S. geminata × xyloni retrieved the greatest amount of a protein- and lipid-rich artificial food resource and a high protein natural food resource, and the native S. geminata retrieved the greatest amount of a high carbohydrate food resource. In trials investigating aspects of interference competition at the colony level, the invasive S. invicta proved to be initially more aggressive than S. geminata , but less aggressive than S. geminata × xyloni . Solenopsis invicta eventually controlled more of the foraging arenas against both native forms when colonies were equivalent by worker biomass, but not when colonies were equivalent by worker number. When paired with S. invicta , S. geminata suffered a significantly greater proportional reduction in both workers and entire colonies when colonies were initially standardized by worker biomass, but not when colonies were standardized by worker number. When paired with S. invicta , a significantly greater proportional reduction of workers occurred in S. geminata × xyloni , regardless of how colonies were standardized. In pairwise trials at the individual level, majors always exhibited significantly less mortality than minors, regardless of the Solenopsis form. The majors of both native forms suffered significantly less mortality than those of S. invicta . Superiority in colony-level interference ability appears to be an important mechanism allowing S. invicta to displace native Solenopsis forms. The ability of S. invicta to reach high population densities, because of intrinsic biological characteristics or an escape from natural enemies, plays an important contributory role. Similar mechanisms may underlie the success of other invasive ant species.     

Phenology, distribution, and host specificity of Solenopsis invicta virus-1

Studies were conducted to examine the phenology, geographic distribution, and host specificity of the Solenopsis invicta virus-1 (SINV-1). Two genotypes examined, SINV-1 and -1A, exhibited similar seasonal prevalence patterns. Infection rates among colonies of S. invicta in Gainesville, Florida, were lowest from early winter (December) to early spring (April) increasing rapidly in late spring (May) and remaining high through August before declining again in the fall (September/October). Correlation analysis revealed a significant relationship between mean monthly temperature and SINV-1 (p<0.0005, r=0.82) and SINV-1A (p<0.0001, r=0.86) infection rates in S. invicta colonies. SINV-1 was widely distributed among S. invicta populations. The virus was detected in S. invicta from Argentina and from all U.S. states examined, with the exception of New Mexico. SINV-1 and -1A were also detected in other Solenopsis species. SINV-1 was detected in Solenopsis richteri and the S. invicta/richteri hybrid collected from northern Alabama and Solenopsis geminata from Florida. SINV-1A was detected in S. geminata and Solenopsis carolinensis in Florida and the S. invicta/richteri hybrid in Alabama. Of the 1989 arthropods collected from 6 pitfall trap experiments from Gainesville and Williston, Florida, none except S. invicta tested positive for SINV-1 or SINV-1A. SINV-1 did not appear to infect or replicate within Sf9 or Dm-2 cells in vitro. The number of SINV-1 genome copies did not significantly increase over the course of the experiment, nor were any cytopathic effects observed. Phylogenetic analyses of SINV-1/-1A nucleotide sequences indicated significant divergence between viruses collected from Argentina and the U.S.     

Detection of Thelohania solenopsae (Microsporidia: Thelohaniidae) in Solenopsis invicta (Hymenoptera: Formicidae) by multiplex PCR

Oligonucleotide primer pairs were designed to unique areas of the small subunit (16S) rRNA gene of Thelohania solenopsae and a region of the Gp-9 gene of Solenopsis invicta. Multiplex PCR resulted in sensitive and specific detection of T. solenopsae infection of S. invicta. The T. solenopsae-specific primer pair only amplified DNA from T. solenopsae and T. solenopsae-infected S. invicta. This primer pair did not produce any amplification products from DNA preparations from uninfected S. invicta, seven additional species of microsporidia (including Vairimorpha invictae), or Mattesia spp. The Gp-9-specific primers recognized and amplified DNA from Solenopsis xyloni, Solenopsis richteri, Solenopsis geminata, the invicta/richteri hybrid, and monogyne and polygyne S. invicta, but not from T. solenopsae, and, as such, served as a positive control verifying successful DNA preparation. Multiplex PCR detected T. solenopsae in worker fire ants infected with as few as 5000 spores. Furthermore, multiplex PCR detected T. solenopsae in all developmental stages of S. invicta. However, detection could be made more sensitive by using only the T. solenopsae-specific primer pair; ants infected with as few as 10 spores were able to be discerned. Multiplex PCR detection of T. solenopsae offers the advantages of a positive control, a single PCR amplification, detection of all developmental stages, and increased sensitivity and specificity compared with microscopy.     

Use of ribosomal DNA sequence data to characterize and detect a neogregarine pathogen of Solenopsis invicta (Hymenoptera: Formicidae)

A neogregarine parasite of the red imported fire ant, Solenopsis invicta, was discovered recently in Florida and tentatively placed in the Mattesia genus based on morphological characterization. S. invicta infected with this Mattesia species exhibited a characteristic yellowing of the cuticle which was designated Mattesia "yellow-head disease" (YHD). The 18S rRNA gene sequence from Mattesia YHD was elucidated and compared with the neogregarine pathogens, Mattesia geminata and Ophriocystis elektroscirrha. The sequence data support the previous conclusion that Mattesia YHD is a new species that infects S. invicta. Furthermore, high sequence identity between Mattesia YHD, M. geminata (95.7%), and O. elektroscirrha (86.2%) correctly place the YHD organism in the Mattesia genus and Neogregarinorida order. Oligonucleotide primer pairs were designed to unique areas of the 18S rRNA genes of Mattesia YHD and S. invicta. Multiplex PCR resulted in sensitive and specific detection of Mattesia YHD infection of S. invicta.     

Predaceous ants, beach replenishment, and nest placement by sea turtles

Ants known for attacking and killing hatchling birds and reptiles include the red imported fire ant (Solenopsis invicta Buren), tropical fire ant [Solenopsis geminata (Fabr.)], and little fire ant [Wasmannia auropunctata (Roger)]. We tested whether sea turtle nest placement influenced exposure to predaceous ants. In 2000 and 2001, we surveyed ants along a Florida beach where green turtles (Chelonia mydas L.), leatherbacks (Dermochelys coriacea Vandelli), and loggerheads (Caretta caretta L.) nest. Part of the beach was artificially replenished between our two surveys. As a result, mean beach width experienced by nesting turtles differed greatly between the two nesting seasons. We surveyed 1,548 sea turtle nests (2000: 909 nests; 2001: 639 nests) and found 22 ant species. S. invicta was by far the most common species (on 431 nests); S. geminata and W. auropunctata were uncommon (on 3 and 16 nests, respectively). In 2000, 62.5% of nests had ants present (35.9% with S. invicta), but in 2001, only 30.5% of the nests had ants present (16.4% with S. invicta). Turtle nests closer to dune vegetation had significantly greater exposure to ants. Differences in ant presence on turtle nests between years and among turtle species were closely related to differences in nest placement relative to dune vegetation. Beach replenishment significantly lowered exposure of nests to ants because on the wider beaches turtles nested farther from the dune vegetation. Selective pressures on nesting sea turtles are altered both by the presence of predaceous ants and the practice of beach replenishment.     

Colony genetic structure and queen mating frequency in fire ants of the subgenus Solenopsis (Hymenoptera: Formicidae)

Colony genetic structure was studied in natural populations of three fire ant taxa, Solenopsis richteri Forel, S. geminata (Fabr.), and hybrid S. invicta/richteri , using allozyme markers. All colonies studied exhibited arrays of female genotypes predicted under a model of monogyny (single functional queen) and monoandry (single insemination of queens). Males produced in the colonies appear to originate exclusively from the foundress queen, rather than from any virgin females present in the colonies. Thus these social insect colonies represent simple, albeit enormous, family groups. Single insemination and foundress parentage of males appear to be conserved reproductive traits in the subgenus Solenopsis , whereas another major determinant of colony genetic structure, the number of functional queens, is evolutionarily labile in this group.     

表皮碳氢化合物在3种火蚁属蚂蚁成虫鉴定中的应用

红火蚁Solenopsis invicta是火蚁属重要的入侵蚂蚁,与其近缘种黑火蚁S.richteri和杂交蚁S.invicta×S.richteri形态相似,难以区分。为了快速准确鉴定3种火蚁属近缘种,本研究利用气相色谱-质谱联用仪(GC-MS),解析3种火蚁的工蚁、有翅雌蚁、有翅雄蚁的表皮碳氢化合物种类和含量,并进行主成分分析、判别分析及聚类分析。结果表明：3种火蚁共检测到62种表皮碳氢化合物,主要包括一甲基烷烃、二甲基烷烃和正构烷烃等;红火蚁、黑火蚁及杂交蚁不同品级的表皮碳氢化合物种类及含量存在显著的种间差异,红火蚁不同地理种群的表皮碳氢化合物种类及含量相似度较高;建立3种火蚁相应品级的分类判别函数,可准确区分各品级下的3种火蚁。因此,表皮碳氢化合物组成分析可用于红火蚁及其近缘种的分类鉴定,为口岸火蚁属蚂蚁的快速检疫鉴定提供新技术。