Biological control of alligator weed: Unsuccessful attempts to control terrestrial growth using the flea beetleDisonycha argentinensis [Col.: Chrysomelidae]

Biological control of alligator weed growing in aquatic habitats in Australia is successful but the agents, a flea beetleAgasicles hygrophila and a mothVogtia malloi, do not control terrestrial growth. Consequently another flea beetleDisonycha argentinensis was introduced into Australia specifically to control the terrestrial growth. Progeny of adults collected in Brazil from areas similar in climate and habitat to areas infested with alligator weed in Australia, were released but failed to become established. Eggs were laid by females released into a large field cage and some completed development, but the new adults failed to reproduce. Tentative conclusions are that microclimate or predation may have prevented establishment ofD. argentinensis but the results should not preclude attempts to establish this insect in North America, China or elsewhere.      

空心莲子草叶甲室内大量繁殖研究

为了大量繁殖供环境释放的空心莲子草叶甲Agasicles hygrophila以实现空心莲子草区域减灾, 我们探索出室内大量饲养与繁殖空心莲子草叶甲的方法与流程, 包括用叶片法或苗水培法孵化卵粒、用盒养法饲养各龄幼虫与成虫并供成虫产卵、用栽培活苗笼养法化蛹羽化。在室内成虫终日均能取食、交配、产卵, 产卵前期约4～5 d, 产卵高峰期在羽化后第7～24 天, 每雌平均产卵21.08块, 约570粒。盒养法叶片平均可着卵4.28块, 叶背与叶面着卵量相近; 笼养法叶片平均着卵为1.46块, 卵主要产于叶背。盒养法与笼养法得到的卵孵化率分别为94.02%与92.50%。空心莲子草叶甲除化蛹需在栽培活苗上完成外, 各龄幼虫与成虫均可用离体新鲜苗盒养法密集饲养。初孵1龄幼虫转株（叶）期、3龄老熟幼虫转化蛹苗期是室内大量饲养与繁殖空心莲子草叶甲的关键时期, 高密度成功饲养与繁殖空心莲子草叶甲的最适化蛹接虫量是每株8头, 产卵期雌虫的最适密度是每株5头。     

空心莲子草叶甲对越冬保护的响应与控害效能

空心莲子草叶甲Agasicles hygrophila是入侵杂草空心莲子草Alternan thera philoxeroide的重要天敌,冬春季低温冰冻是影响其安全越冬的关键胁迫因子。为了探明采取保护措施是否能提高越冬后的种群基数问题,在低温冰冻天气来临前,用塑料膜覆盖网室构建空心莲子草与空心莲子草叶甲保护生境(PH),分低温期(2009年1月21日至4月9日)、升温期(4月15日至5月1日)与自然生境(NH)中叶甲成虫初现期(从6月1日至29日)3个阶段,调查比较PH与NH两类生境中空心莲子草叶甲种群数量、结构、空间生态位分布特征及控害效能。结果表明:在保护生境中,低温期空心莲子草叶甲不休眠,且能安全越冬,当晴天温度较高时,成虫主要在直立茎或嫩枝上活动取食、补充营养,而在阴雨低温天气则停留在匍匐草层或土壤表层、缝隙中,少活动。随着气温回升,成虫转移到直立茎生态位上活动取食,并于4月上旬开始产卵繁殖,种群数量迅速增加,对空心莲子草的地上部分有较好的控制效果。在自然生境中,直到6月初才见少量成虫活动,此时草生长茂密,较低的空心莲子草叶甲种群难以发挥生防因子的作用。比较两种生境中6月1日与6月15日的控草效果显示,空心莲子草株高增长率分别为:PH-13.91%,NH-2.94%,NH中空心莲子草株高极显著高于PH,且单株生物量的控制率为PH(47.56%)极显著高于NH(4.89%)。在6月15日与29日的调查表明,单株叶片控制效果PH极显著优于NH,PH中的空心莲子草茎上均无叶,叶片或被取食殆尽、或被害后干枯脱落。茎被害率、茎蛀孔数与蛀茎率均为PH极显著高于NH。结果说明覆膜保护能显著提高叶甲越冬种群基数,使其提前进入繁殖期,从而能更有效地控制生境中的空心莲子草。     

Con‐specific neighbours may enhance compensation capacity in an invasive plant

Facilitation, both by inter‐ and intra‐specific neighbours, is known to be an important process in structuring plant communities. However, only a small number of experiments have been reported on facilitation in plant invasions, especially between invasive con‐specific individuals. Here, we focus on how con‐specific neighbours of the invasive alien plant alligator weed affect the tolerance of alligator weed to herbivory by the introduced biological control agent, Agasicles hygrophila. We conducted greenhouse and garden experiments in which invasive plant density and herbivory intensity (artificial clipping and real herbivory) were manipulated. In the greenhouse experiment, artificial clipping significantly reduced plant biomass when plants were grown individually, but when con‐specific neighbours were present in the same pot, biomass was not significantly different from control plants. Similarly, when compared to control plants, plants that were subjected to herbivory by A. hygrophila produced more biomass when grown with two con‐specific neighbours than when grown alone. Real herbivory also resulted in an increased number of vegetative buds, and again when two con‐specific neighbours were present this effect was increased (a 55.3% increase in buds when there was no neighbour, but a 111.6% increase in buds when two con‐specific neighbours were present). In the garden experiment, in which plants were grown at high density (6 plants per pot), alligator weed fully recovered from defoliation caused by insects at levels from 20–30% to 100%. Our results indicate that the con‐specific association may increase the compensatory ability to cope with intense damage in this invasive plant.     

除草剂胁迫对空心莲子草叶甲种群的影响及应对策略

为探讨除草剂胁迫对空心莲子草叶甲种群的影响,寻求合理施用除草剂,有效保护叶甲,提高其控草效果的应对策略,利用叶片残留法测试了在草甘膦、农达、水花生净与35%苄·丁可湿粉等除草剂胁迫下空心莲子草叶甲卵孵化率,幼虫、成虫存活率以及蛹羽化率。结果表明被测试的4种除草剂均不同程度地降低空心莲子草叶甲卵孵化率,幼虫与成虫存活率及蛹羽化率,特别是水花生净28.0、35.0 g/L两种浓度处理,35%苄·丁可湿粉4.0 g/L浓度处理显著降低卵孵化率;农达13.4 g/L浓度处理显著降低1龄前期幼虫存活率,农达40.1 g/L与 53.4 g/L两种浓度处理显著降低2龄前期幼虫存活率;水花生净28.0 g/L浓度处理显著降低3龄前期幼虫存活率;草甘膦97.6 g/L与 122.0 g/L浓度处理、农达26.7 g/L与66.8 g/L浓度处理、35%苄·丁可湿粉2.0 g/L浓度处理显著降低后期蛹的羽化率;35%苄·丁可湿粉1.0 g/L浓度处理显著降低羽化后3 d雌虫的存活率。除草剂除直接杀死空心莲子草叶甲,降低其种群数量外,使叶甲失去食物与避护所,快速地引起叶甲种群崩溃。建议合理施药,并在施药时,随机留若干小斑块不施药,保护叶甲,可望提高其田间控草效果。     

类雄激素受体在尖唇散白蚁繁殖蚁和工蚁卵子发生中的免疫细胞化学表达

为探讨类雄激素受体 (androgen receptor-like,AR-like)在白蚁卵子发生过程中的作用,采用免疫细胞化学方法对尖唇散白蚁Reticulitermes aculabialis雌性繁殖蚁和工蚁卵子发生中的类雄激素受体进行了定位检测。结果发现：繁殖蚁和工蚁卵巢中均有类雄激素受体免疫阳性反应。类雄激素受体在末龄若虫和成虫卵母细胞的分化期和生长期定位于卵母细胞质,而在成虫的卵黄形成期定位于滤泡细胞;在工蚁卵巢中,类雄激素受体定位于分化期和生长期的卵母细胞质,无卵黄形成期。结果提示类雄激素受体在白蚁的卵子发生过程中有重要作用： 工蚁的卵巢发育受抑制,其卵子发生相当于末龄若虫水平,但根据群体的变化又可以发育为补充繁殖蚁,有发育的潜能。     

捕食者对空心莲子草叶甲种群的生物胁迫

广食性捕食者广泛捕食植食性昆虫,常被用于有害生物的生物防治,也因此影响植食性昆虫对杂草的生物效果。空心莲子草叶甲(Agasicles hygrophila)(鞘翅目:叶甲科Chrysomelidae)作为入侵恶性杂草空心莲子草(Alternanthera philoxeroides)(苋科:莲子草属Alternanthera)的专性天敌,从美国的弗罗里达州引入中国,在释放地防治空心莲子草取得了较好的防治效果。虽然空心莲子草叶甲在引入地均已建立田间种群并有一定程度的自然扩散,但丰富的食物资源,并未使空心莲子草叶甲的自然种群数量变得繁荣,因此其未能有效抑制空心莲子草的扩散蔓延。在野外调查时发现空心莲子草生境中存在大量广食性捕食者。这些广食性捕食者是抑制空心莲子草叶甲种群数量扩张的生物胁迫因子吗?为此,选择捕食性昆虫龟纹瓢虫(Propylaea japonica)(鞘翅目:瓢虫科Coccinellidae)、蜘蛛类捕食者拟水狼蛛(Pirata subpiraticus)(蜘蛛目:狼蛛科Lycosidae)与斜纹猫蛛(Oxyopes sertatus)(蜘蛛目:猫蛛科Oxyopidae)为捕食者,分别以空心莲子草叶甲各虫态为猎物,构建简单的捕食者-猎物系统,在室内检测了上述3种捕食者对空心莲子草叶甲各虫态在不同密度下的日捕食量,以期了解捕食者对空心莲子草叶甲的捕食作用,客观评估空心莲子草叶甲的生物防治效能。研究结果表明:捕食者龟纹瓢虫、斜纹猫蛛与拟水狼蛛均捕食空心莲子草叶甲的卵粒及1龄、2龄幼虫,斜纹猫蛛与拟水狼蛛捕食3龄幼虫,捕食者的捕食量均随着猎物密度的升高而增加,寻找效应降低。三者均不捕食成虫。除拟水狼蛛对3龄幼虫的捕食用Holling II模型拟合不呈显著相关关系外,其余捕食反应均拟合Holling II模型并显著相关。通过拟合方程得出捕食者对空心莲子草叶甲卵粒的理论日最大捕食量为:斜纹猫蛛10.9粒,拟水狼蛛为6.2粒,龟纹瓢虫为5.6粒;对1龄幼虫的理论日最大捕食量为:斜纹猫蛛为17.1头;拟水狼蛛为35.8头,龟纹瓢虫为10.4头;对2龄幼虫的理论日最大捕食量为:斜纹猫蛛为6.6头,拟水狼蛛为11.2头,龟纹瓢虫为2.9头;对3龄幼虫的理论日最大捕食量为:斜纹猫蛛捕食12.3头,拟水狼蛛为1.1头。研究结果证实了捕食者可通过捕食作用降低空心莲子草叶甲种群密度,削弱空心莲子草叶甲对空心莲子草的控害效能,是空心莲子草叶甲种群存活的生物胁迫因子。建议在提高空心莲子草叶甲田间种群数量,达到对空心莲子有效的持续控制效果方面开展进一步研究。     

Disturbance-mediated competition: the interacting roles of inundation regime and mechanical and herbicidal control in determining native and invasive plant abundance

Disturbance is a key component of many successful plant invasions. However, interactions among natural and anthropogenic disturbances and effects of these interacting disturbances on invasive plants and desired vegetation are rarely examined. We investigated the effect of anthropogenic disturbance (herbicidal and mechanical) along a natural inundation gradient (20–282 days) on the biomass and resource allocation of the invasive wetland plant, alligator weed (Alternanthera philoxeroides), and two co-occurring competitor plants, the introduced grass, kikuyu (Pennisetum clandestinum), and the native grass, couch (Cynodon dactylon), over a 2-year period. In the absence of additional disturbance, kikuyu biomass was negatively affected, alligator weed biomass was positively affected, and couch biomass was not affected by inundation disturbance. In addition, kikuyu was not affected by the selective removal of alligator weed, while couch increased in wetter habitats where kikuyu was absent due to inundation stress. This suggests that kikuyu is a superior competitor in drier habitats and inundation facilitates the invasion of alligator weed, while couch is an inferior competitor to both kikuyu and alligator weed and is therefore suppressed across its entire niche by these two introduced species. Mowing alone had no effect on the biomass of the species, suggesting the plants are equally tolerant of shoot removal. Selective herbicide reduced alligator weed biomass by 97.5% and the combination of selective herbicide and mowing reduced the biomass of alligator weed significantly more than herbicide alone, by 98.6% compared with un-manipulated controls. To predict community change and prevent sequential exotic plant invasions after weed removal, it is necessary to consider the interacting effects of disturbance and the niche space of invasive species in the local propagule pool.     

Heat sensitivity of eggs attributes to the reduction in Agasicles hygrophila population

Agasicles hygrophila has been introduced worldwide as a control agent for the invasive weed Alternanthera philoxeroides.However,global warming has potential impact on its controlling efficacy.The aim of this research was to explore the primary factors responsible for the greatly reduced A.hygrophila population in hot summers.To imitate the temperature conditions in summers,different developmental stages of hygrophila were treated with high temperatures from 32.5℃ to 45℃ for 1-5 h.Based on the survival rate,the heat tolerance of each developmental stage was ranked from lowest to highest as follows:egg,1st,2nd,3rd instar larva,adult and pupa.Eggs showed the lowest heat tolerance with 37.5℃ as the critical temperature affecting larval hatching.Heat treatment of the A.hygrophila eggs at 37.5℃ for 1 h decreased the hatch rate to 24%.Our results indicated that when compared with the control at 25℃,1 h treatment at 37.5℃ prolonged the duration of the egg stage,shortened the duration of oviposition and total longevity,and changed the reproductive pattern of A.hygrophila.The net reproductive rate,intrinsic rate and finite rate were all significantly reduced.The results suggest that low heat tolerance of the eggs was the major factor responsible for the reduction of A.hygrophila populations,and the key temperature was 37.5°C.Therefore,appropriate measures should be taken to protect eggs in order to maintain the efficacy of A.hygrophila in the biological control of A.philoxeroides in hot summers.     

Genetic diversity of alligator weed ecotypes is not the reason for their different responses to biological control

Biological control of alligator weed Alternanthera philoxeroides (Mart.) Griseb. using Agasicles hygrophila, a Chrysomelid beetle, has been successful in limiting growth in water, but not on land. In order to understand a possible genetic basis of this difference, technique using inter-simple sequence repeats (ISSR) markers was applied to analyse genetic diversity of this invasive weed. No genetic variation was detected not only within or between populations growing in the same habitats, but also between land- and water-grown populations. Thus we consider that the genetic variation is not the baseline factor resulting in the biological control difference in China. The differential success of pupation by the beetle may be related to the phenotypic plasticity of the plant stem diameter, rather than to genotypic factors.     

Investigation of cis-acting sequences regulating expression of the gene encoding yolk protein 3 in Drosophila melanogaster.

Summary The regulatory sequences leading to the ovarian and fat body expression of yolk proteins 1 and 2 (YP1 and 2) of Drosophila melanogaster have been characterised in some detail. These genes (yp1 and yp2) share many enhancer elements, and some important regulatory sequences lie within the coding regions. We have begun to investigate the cis-regulation of the gene encoding yolk protein 3 (yp3). We describe a system for P element transformation using the complete and unaltered yp3 gene rather than reporter genes and describe sequences conferring correct expression in the ovary and carcass.     

Notch pathway regulates female germ cell meiosis progression and early oogenesis events in fetal mouse

A critical process of early oogenesis is the entry of mitotic oogonia into meiosis, a cell cycle switch regulated by a complex gene regulatory network. Although Notch pathway is involved in numerous important aspects of oogenesis in invertebrate species, whether it plays roles in early oogenesis events in mammals is unknown. Therefore, the rationale of the present study was to investigate the roles of Notch signaling in crucial processes of early oogenesis, such as meiosis entry and early oocyte growth. Notch receptors and ligands were localized in mouse embryonic female gonads and 2 Notch inhibitors, namely DAPT and L-685,458, were used to attenuate its signaling in an in vitro culture system of ovarian tissues from 12.5 days post coitum (dpc) fetus. The results demonstrated that the expression of Stra8, a master gene for germ cell meiosis, and its stimulation by retinoic acid (RA) were reduced after suppression of Notch signaling, and the other meiotic genes, Dazl, Dmc1, and Rec8, were abolished or markedly decreased. Furthermore, RNAi of Notch1 also markedly inhibited the expression of Stra8 and SCP3 in cultured female germ cells. The increased methylation status of CpG islands within the Stra8 promoter of the oocytes was observed in the presence of DAPT, indicating that Notch signaling is probably necessary for maintaining the epigenetic state of this gene in a way suitable for RA stimulation. Furthermore, in the presence of Notch inhibitors, progression of oocytes through meiosis I was markedly delayed. At later culture periods, the rate of oocyte growth was decreased, which impaired subsequent primordial follicle assembly in cultured ovarian tissues. Taken together, these results suggested new roles of the Notch signaling pathway in female germ cell meiosis progression and early oogenesis events in mammals.     

Mating affects reproductive investment into eggs,but not the timing of oogenesis in the flesh fly <Emphasis Type="Italic">Sarcophaga crassipalpis</Emphasis>

We examined the effects of mating on reproductive investment and the timing of oogenesis in the flesh fly Sarcophaga crassipalpis by exposing females to males or not. All females exposed to males were mated within a few days and we found that mating affected reproductive investment. Virgin females not exposed to males produced a large clutch of eggs (∼91), but females exposed to males and mated produced 10% more. There was no effect of mating on egg length or mass. There was also no effect of mating on the timing of oogenesis. Females in both treatments provisioned their eggs at the same rate with yolk first becoming visible in the oocytes on day three of adulthood and complete provisioning of eggs occurring by the seventh day of adulthood. We examined the biochemical basis of egg provisioning by identifying the yolk proteins and quantifying their blood titer during the oogenic period in both, females exposed to males and mated and those not exposed to males. There was no difference in the timing of the first appearance, peak titer, or disappearance of yolk proteins in the blood between the two treatments. However, consistent with our observation of greater egg production in mated females, these females contained a greater peak yolk protein titer.     

Multiple vitellogenins from the Haemaphysalis longicornis tick are crucial for ovarian development

Ovarian development and egg maturation are crucial processes for the success of reproduction in ticks. Three full-length cDNAs encoding the precursor of major yolk protein, vitellogenin, were obtained from cDNA libraries of the Haemaphysalis longicornis tick and designated as HlVg-1, HlVg-2 and HlVg-3. The HlVg mRNAs were found in fed females with major expression sites in the midgut, fat body and ovary. Native PAGE and Western blot demonstrated that HlVgs in the hemolymph, fat body and ovary of fed females consisted of four major polypeptides. RNAi results showed that HlVg dsRNA-injected ticks obtained lower body weight, egg weight and showed higher mortality of engorged females after blood sucking than control groups. Our results indicate that all HlVgs are essential for egg development and oviposition.     

酸雨和采食模拟胁迫下克隆整合对空心莲子草生长的影响

研究表明克隆整合可以显著提升异质环境中克隆植物的生长,然而当克隆植物遭受均质环境压力时,整合对植物生长影响的研究相对较少。本文以典型入侵克隆植物空心莲子草（Alternanthera philoxeroides）为例,研究均质环境压力酸雨和采食模拟胁迫对空心莲子草生长的影响,以及克隆整合在空心莲子草适应不利环境过程中所起的作用。酸雨设3种浓度梯度：pH值3.5 、pH值4.5和 pH值6.5（对照）;采食设3种水平：不去叶、去叶50%和去叶90%;整合水平：匍匐茎切断和连接。结果表明：无论保持或切断匍匐茎的连接,酸雨处理都不影响空心莲子草生物量。当保持匍匐茎连接时,pH值4.5酸雨处理增加了空心莲子草匍匐茎长度和分株数目,因此,低度酸雨可能对空心莲子草生长有一定的促进作用。同样,无论匍匐茎是否被切断,采食处理都显著降低了空心莲子草克隆片段生物量,而显著增加了叶片数目。当切断匍匐茎连接时,采食处理使空心莲子草分株数目显著增加。本文得出的结论是：空心莲子草能较好地适应酸雨和采食的环境压力,当空心莲子草全部克隆分株遭受均质环境胁迫时,克隆整合并不能显著改善它的生长。     

A Critical Role of the Nuclear Receptor HR3 in Regulation of Gonadotrophic Cycles of the Mosquito Aedes aegypti

The orphan nuclear receptor HR3 is essential for developmental switches during insect development and metamorphosis regulated by 20-hydroxyecdysone (20E). Reproduction of female mosquitoes of the major vector of Dengue fever, Aedes aegypti, is cyclic because of its dependence on blood feeding. 20E is an important hormone regulating vitellogenic events in this mosquito; however, any role for HR3 in 20E-driven reproductive events has not been known. Using RNA interference (RNAi) approach, we demonstrated that Aedes HR3 plays a critical role in a timely termination of expression of the vitellogenin (Vg) gene encoding the major yolk protein precursor. It is also important for downregulation of the Target-of-Rapamycin pathway and activation of programmed autophagy in the Aedes fat body at the end of vitellogenesis. HR3 is critical in activating betaFTZ-F1, EcRB and USPA, the expressions of which are highly elevated at the end of vitellogenesis. RNAi depletion of HR3 (iHR3) prior to the first gonadotrophic cycle affects a normal progression of the second gonadotrophic cycle. Most of ovaries 24 h post second blood meal from iHR3 females in the second cycle were small with follicles that were only slightly different in length from of those of resting stage. In addition, these iHR3 females laid a significantly reduced number of eggs per mosquito as compared to those of iMal and the wild type. Our results indicate an important role of HR3 in regulation of 20E-regulated developmental switches during reproductive cycles of A. aegypti females.