红火蚁的社会型及其相关基因Gp-9的研究综述

红火蚁Solenopsis invicta Buren的蚁巢具有2种基本的社会组织形态:单后型和多后型。蚁巢中蚁后的数量是由工蚁和蚁后的基因型共同决定的。红火蚁的Gp-9基因编码一种气味结合蛋白并控制这种社会多态性的表达。Gp-9基因的发现首次证明单个基因在昆虫复杂的社会行为中起关键的作用。从红火蚁的社会型特征、Gp-9基因与不同社会型的关系以及Gp-9基因的相关研究对该领域的研究进展进行综述。     

不同方法对红火蚁社会型鉴定效果的比较

红火蚁Solenopsis invicta Buren的社会型——单蚁后(Monogyne)和多蚁后(Polygyne)型的鉴定方法众多,对于鉴定方法之间的比较较少。为了探究鉴定红火蚁社会型的有效实验方案,本研究对采集自中国南方的红火蚁种群样品进行了生物学观察,并采用了几种常用的方法对样品社会型进行了测定,此外还对比了4种不同DNA聚合酶在实验中的效果。多重PCR技术(Multiplex Polymerase Chain Reaction)、Gp-9 b等位基因扩增两种方法成功率高,能准确鉴定红火蚁社会型。4对同时扩增B、b等位基因的引物成功率不高。实验结果表明,将多重PCR技术、Gp-9 b等位基因扩增,与生物学观察相结合,能更准确地对红火蚁社会型进行鉴定。在PCR实验中选用具有适度保真性和热启动效果的DNA聚合酶能提高红火蚁社会型鉴定的准确性。     

红火蚁雌蚁生殖发育的研究

通过解剖多后型红火蚁Solenopsis invicta Buren雌蛹、有翅雌蚁和脱翅雌蚁,观察了雌蚁内生殖系统的结构和发育进度。结果表明,多后型雌蚁内生殖系统形成于蛹期,其结构与单后型雌蚁存在一定差异。从发育进度来看,虽然雌蚁羽化后其卵巢大小和卵巢管数量仍有一定增长,但蛹期是卵巢大小、卵巢管数量快速增长的阶段;卵子发生始见于羽化后第4d,且卵量随个体发育而快速增加,直至脱翅后第10d达到高峰,随后开始下降;生殖系统的其它结构的变化较小。雌蛹、有翅雌蚁和脱翅雌蚁发育后期的卵巢平均大小分别为1149．23×712．42、1594．88×834．26和1975．60×1303．57μm;而卵巢管数量则分别为62．67、81．33和86．33条。至雌蚁脱翅后10d,卵巢中成熟卵和总卵量分别为20．00±4．62粒和34．00±2．31粒。     

入侵我国红火蚁的三种单倍型

红火蚁Solenopsis invicta Buren是重要的经济害虫,特别是在农业和生态系统危害方面。2004年底在广东和香港的部分地区发现红火蚁入侵。本文用线粒体细胞色素氧化酶Ⅰ（CO Ⅰ）基因对境内的13个种群进行了入侵蚁巢是否独立传入进行了分析。通过对共计28个蚁巢56个体的包含904个碱基的CO Ⅰ基因分析,发现入侵中国的红火蚁中存在3种单倍型。基于未修正的配对序列变异分析表明,单倍型Ⅱ与单倍型Ⅲ之间的变异最小。而香港种群同时具有这三种单倍型,是入侵中国的红火蚁种群中多样性最丰富的地区。这三种单倍型都分别在阿根廷红火蚁种群中有记录。同源性分析显示,中国的红火蚁可能起源于阿根廷或南美洲,而香港可能是所研究种群的第一入侵地点。     

红火蚁对我国一些生物潜在影响的初步分析

通过分析国际上对红火蚁影响生境中其他生物的研究成果,认为红火蚁主要通过以下3种方式危害其他生物:(1)攻击、捕食刚孵化的地栖性卵生动物个体,或者以群体力量捕食昆虫幼虫、成虫等;(2)竞争有限的食物资源,导致其他物种因为缺乏足够食物供给而种群数量减少甚至灭绝;(3)通过叮咬而使得某些动物存活率降低,改变生境,甚至弃巢外逃,或者因为受攻击活动量加大而增加被捕食的几率。同时初步分析在《国家重点保护野生动物名录》中列举的379种野生动物名录中,有22种鸟类(占9.6%)、1种两栖类(占14%)、所有的18种爬行类(占100%)可能因为红火蚁的入侵导致种群数量下降甚至灭绝。     

香菇单核体交配型及杂交后代的等位基因特异性PCR鉴定

【背景】 食用菌交配型和单单杂交杂合子的鉴定通常采用显微镜检测观察是否具有锁状联合的方式进行,存在耗时长、工作量大且易出现误检等问题。【目的】 建立一种鉴定香菇单核体交配型和单单杂交后代的分子辅助育种技术,为提高育种效率提供技术支撑。【方法】 利用交配型因子保守序列单核苷酸多态性(single-nucleotide polymorphism, SNP)位点设计分型引物,建立等位基因特异性PCR (allele-specific PCR, AS-PCR)技术,鉴定香菇L808和YX7的单孢分离株交配型及其杂交后代。【结果】 AS-PCR鉴定结果表明,L808的38个单孢分离株中,交配型为A1B1、A2B2、A1B2和A2B1的单核体分别有6、13、8和11个;YX7的45个单孢分离株中,交配型为A3B3、A4B4、A3B4和A4B3的单核体分别有15、8、8和12个,交配型为A3A4B3B4的异核体2个;12个单单杂交菌株中,10个为真正的杂合子,2个为非杂合子。传统方法与AS-PCR分子鉴定结果完全一致,但前者容易将异核体误判为单核体。【结论】 基于SNP位点的AS-PCR技术能有效鉴别香菇单核体交配型和单单杂交后代,区分单核体与异核体,具有精准、高效的特点,是一种香菇分子辅助育种的理想工具。     

红火蚁工蚁对绿僵菌侵染蛹的防御行为

研究了红火蚁工蚁感染绿僵菌后在蛹室的行为变化,以及健康工蚁对侵染蛹的行为保护机制.结果表明: 工蚁被绿僵菌侵染后,在蛹室的活动时间逐渐减少,由第1天的103.4 s降至第3天的38.5 s;而且育幼时间占蛹室活动时间的比例也下降,由第1天的13.6%降至第3天的3.5%.当蛹被绿僵菌侵染后,工蚁对侵染蛹的梳理总时间为对照组的5.3倍,每次梳理的平均持续时间为对照组的5.2倍.梳理行为能显著减少侵染蛹的体表分生孢子数量,在无工蚁、2只工蚁和10只工蚁存在条件下,蛹体表平均孢子数分别为103.1、51.6和31.3个.工蚁的存在能抑制蛹体表孢子的萌发,处理20 h后,无工蚁、2只工蚁和10只工蚁存在条件下,蛹体表孢子萌发率分别为95.1%、80.4%和59.9%.蛹的羽化率随着工蚁数量增加显著升高.红火蚁工蚁通过社会行为防御病原真菌侵染蛹的策略为种群的延续和发展提供了保障.     

红火蚁迁巢时间动态及与生境类型和蚁群社会型的关系

红火蚁Solenopsis invicta是我国具有重大生态和经济影响的入侵物种,明确其蚁群迁移(迁巢)发生情况对理解其生态适应性及科学制定防控策略具有重要意义,迄今虽已有这方面的一些报道,但有关其迁巢发生程度、季节特点及不同生境中是否存在差异等缺乏认识。研究对浙江中部一草地中两种社会型红火蚁迁巢情况进行了观察,同时在浙江、江西、广东调查了31个红火蚁发生区块中(以绿化带和荒草地为主)的空巢比例。结果显示,在单蚁后型红火蚁活动季节,几乎各个时期均有蚁巢被废弃,以2-7月废弃比例较高(最高接近12%),8月下旬后则明显较低;废弃巢穴和新巢形成之间无明显数量关系。废弃蚁巢被重新利用的现象也在各月均有发生,且与废巢总数量存在显著正相关。多蚁后型弃巢比例总体要低于单蚁后型。在所调查的各地31个区块中,有27个(87.1%)见到空巢;空巢比例在不同社会型、生境和区块间存在一定差异。其中绿化带中80%所查区块的空巢比例为10%-35%,在荒草地中则接近70%的区块空巢比例不到10%,但这两类生境中均有个别区块空巢比例高于35%。表明红火蚁迁巢是一种普遍现象并存在明显的季节特征,迁巢后所形成的空巢比例因地而异,且与生境类型、蚁群社会型等存在一定关系。     

基于Y型驱避测定尺的红火蚁驱避剂筛选新方法

筛选测定方法是开发红火蚁Solenopsis invicta Buren驱避剂的重要环节,目前尚缺乏针对攻击水平较高的红火蚁驱避活性的测定方法。本研究利用自主研发的一种Y型驱避测定尺监测了红火蚁工蚁的活动规律,对比了裸型和绷带型Y型驱避测定尺对攀爬红火蚁工蚁数量的影响,并测定了避蚊胺对红火蚁工蚁的驱避活性。结果表明：Y型驱避测定尺的侧臂长度与红火蚁工蚁的密度呈负相关,选择侧臂长度11 cm的Y型驱避测定尺用于驱避活性测定。室内红火蚁工蚁受惊扰后75 s内数量涨幅较大,而田间红火蚁工蚁受扰动后30 s时数量可达到峰值,裸型和绷带型Y型驱避测定尺的工蚁数量在不同时间点上均无显著性差异（P> 0.05）。避蚊胺对红火蚁的驱避效果显著,随着处理浓度上升,驱避率和有效驱避时间也随之提高,裸型和绷带型Y型驱避测定尺分别在5.0%和10.0%浓度时的驱避率达到100%,当避蚊胺浓度为5.0%时,裸型尺和绷带型尺的有效驱避时间分别可达198.16 min、228.14 min。本研究设计并优化的Y型驱避测定尺可直接测定化合物对惊扰状态下红火蚁的驱避活性,为红火蚁驱避剂的筛选与评价提供了一种新方法,同时也为红火蚁个体防护剂的研发提供了技术支撑。     

等位基因特异PCR技术的研究与应用

生物的单核苷酸多态性(Single-nucleotide polymorphism,SNP)具有数量多、分布广、易于分型、稳定性强等优点,很适合于用做分子标记.等位基因特异PCR(Allele-specific PCR,AS-PCR)是根据SNP位点设计3'末端与SNP位点碱基互补或错配的特异PCR引物,通过凝胶电泳等方法检测PCR扩增产物的有或无,从而检测基因型中SNP的一种技术.经过不断地改进与完善,基于SNP的等位基因特异PCR标记已逐渐成为一种快速、简便、低成本、可靠、高通量的检测基因型SNP的方法.本文应用等位基因特异PCR技术,根据小麦TaDREB1基因在旱选10和鲁麦14的120(C→A)SNP成功地开发了一个SNP分子标记,证明了该方法的有效性和可行性.     

Major gene effects on phenotype and fitness: the relative roles of Pgm-3 and Gp-9 in introduced populations of the fire ant Solenopsis invicta

The objective of this study was to disentangle the relative effects of Pgm-3 and Gp-9 and/or other closely linked genes on the phenotypes and reproductive success of queens in introduced (USA) populations of S. invicta. Gp-9 or a closely linked gene(s) was found to have major effects on queen weight, the likelihood that queens shed their wings (a behaviour associated with the onset of reproduction), and the probability that queens are accepted in polygyne (multiple-queen) colonies. Our analyses show that once the effect of Gp-9 genotype is taken into account, Pgm-3 genotype no longer is significantly associated with differences in queen phenotype or the probability of queens being accepted in polygyne colonies. This suggests that the associations of Pgm-3 genotype with weight, wing shedding rate and probability of acceptance by polygyne colonies previously reported in studies that did not control for the effects of Gp-9 are due to the strong linkage disequilibrium that exists between Pgm-3 and Gp-9, or to linkage disequilibria between these and other genes affecting queen phenotype and fitness. Several lines of evidence, including data from the native South American range, suggest that additional cryptic alleles at Gp-9, or additional genes in the same linkage group as Gp-9, must be involved in controlling queen phenotype and the large suite of traits important in determining social organization of S. invicta colonies.     

Unusual Behavior of Polygyne Fire Ant Queens on Nuptial Flights

This study reports previously undescribed behavior of fire ant queens (Solenopsis invicta) on their nuptial flights. We captured large numbers of alate (winged) queens flying at low altitudes in dense swarms that were virtually devoid of males. We assayed the genotypes of these alate queens at the locus Gp-9, which exhibits strong genotype frequency differences between monogyne (single-queen) and polygyne (multiple-queen) populations, and found that almost all of these low-flying queens originated from polygyne colonies. Comparisons of mtDNA haplotype distributions of these queens to those of alates leaving polygyne nests suggest that the flying queens had not dispersed more than a few hundred meters. Moreover, the proportion of flying queens that were mated did not differ significantly from the proportion of reproductive queens that were mated within the same sites. Thus the flight behavior appears to occur subsequent to mating. We suggest that the flying queens are sampling the local environment in order to select a suitable landing site. Such a site would contain established polygyne nests into which the queens may be adopted as new reproductives.     

红火蚁多蚁后型种群有效蚁巢局域分布的空间相关性

应用地统计学的方法分析了红火蚁多蚁后型种群有效蚁巢的空间变异规律,建立了多个间隔距离与半方差值的球状模型.对吴川市调查样地的研究表明, 多个样地不同方向上红火蚁有效蚁巢不同间隔距离上的半方差值呈明显规律性变化,表现为随着间隔距离的增大,半方差值增大,具有空间依赖性.其5个球状模型的变程分别为9.1、7.6、23.5、7.5和14.5 m,平均为12.4 m.在这些距离内的抽样空间上任何两点红火蚁有效蚁巢数量间存在相关关系.所建模型块金常数占基台的比例,即空间不连续性较大,分别为0.7034、0.9247、0.4398、1.1196和0.4624,表明红火蚁有效蚁巢分布存在一定的空间随机性和结构性.深圳市的5块样地10个方向上有7个方向半方差值随间隔距离呈明显规律性变化,表现为空间依赖性,变程分别为14.5、11.2、10.8、17.6、11.3、9.9和12.8 m,平均为12.6 m.     

Parasitoid–host relationships when host size varies: the case of Pseudacteon flies and Solenopsis fire ants

Abstract. 1. Phorid flies in the genus Pseudacteon are parasitoids of ants. Variation in host size preferences of four South American and two North American Pseudacteon species on monogyne and polygyne forms of their host Solenopsis species [ S. invicta Buren and S. geminata (F.), respectively] was documented.
2. Monogyne Solenopsis workers were, on average, significantly larger than polygyne workers, and the average size monogyne worker attacked was significantly larger than the average size polygyne worker attacked in four of the six Pseudacteon species.
3. Three South American Pseudacteon species attacked larger than average size workers, whereas one attacked smaller than average size workers, in both monogyne and polygyne forms. Both North American Pseudacteon species attacked larger than average size polygyne workers and smaller than average size monogyne workers.
4. Three Pseudacteon species were reared from eggs to adults in infected ants in the laboratory. The size of the emergent phorid fly was related positively to the size of the host worker ant, with females emerging from larger hosts. Similar patterns were documented for both monogyne and polygyne forms.
5. The mean size of worker host from which phorids emerged did not differ significantly between the monogyne and polygyne forms in the subsample of phorids reared to adults.
6. The observed patterns elucidate factors that may cause variation in Pseudacteon sex ratios, and have implications for biological control efforts of pest Solenopsis species.     

Molecular evolutionary analyses of the odorant-binding protein gene Gp-9 in fire ants and other Solenopsis species

The fire ant Solenopsis invicta exists in two social forms, one with colonies headed by a single reproductive queen (monogyne form) and the other with colonies containing multiple queens (polygyne form). This variation in social organization is associated with variation at the gene Gp-9, with monogyne colonies harboring only the B allelic variant and polygyne colonies containing b-like variants as well. We generated new Gp-9 sequences from 15 Solenopsis species and combined these with previously published sequences to conduct a comprehensive, phylogenetically based study of the molecular evolution of this important gene. The exon/intron structure and the respective lengths of the five exons of Gp-9 are identical across all species examined, and we detected no evidence for intragenic recombination. These data conform to a previous suggestion that Gp-9 lies in a genomic region with low recombination, and they indicate that evolution of the coding region in Solenopsis has involved point substitutions only. Our results confirm a link between the presence of b-like alleles and the expression of polygyny in all South American fire ant species known to possess colonies of both social forms. Moreover, phylogenetic analyses show that b-like alleles comprise a derived clade of Gp-9 sequences within the socially polymorphic species, lending further support to the hypothesis that monogyny preceded polygyny in this group of fire ants. Site-specific maximum likelihood tests identified several amino acids that have experienced positive selection, two of which are adjacent to the inferred binding-pocket residues in the GP-9 protein. Four other binding-pocket residues are variable among fire ant species, although selection is not implicated in this variation. Branch-specific tests revealed strong positive selection on the stem lineage of the b-like allele clade, as expected if selection drove the amino acid replacements crucial to the expression of polygyne social organization. Such selection may have operated via the ligand-binding properties of GP-9, as one of the two amino acids uniquely shared by all b-like alleles is predicted to be a binding-pocket residue.