Effects of elevated CO2 on the development and physiological metabolic activities of Nilaparvata lugens in response to the infection of Trichoderma asperellum

Trichoderma spp. are widely used in agriculture as biofungicides. In this study, effects of elevated CO₂ on the growth and enzyme activities of Nilaparvata lugens infected by T. asperellum were studied. The results showed that fecundity of N. lugens adversely decreased under elevated CO₂ relative to ambient CO₂ irrespective of infection with T. asperellum. The contents of protein and alkaline phosphatase significantly decreased in N. lugens nymphs with T. asperellum infection under elevated CO₂ relative to ambient CO_2. Significantly higher activities of carboxylesterase and acetylcholinesterase were observed in N. lugens infested with T. asperellum under elevated CO₂ compared with ambient CO₂. While significantly lower catalase and glutathione S-transferase were observed in N. lugens nymphs infested by T. asperellum under elevated CO₂ relative to ambient CO₂. Measuring the antioxidases and detoxification enzymes in N. lugens infected with T. asperellum can partly explain the response of fitness parameters of N. lugens under elevated CO₂.     

Evaluation on the fitness and population projection of Nilaparvata lugens in response to elevated CO2 using two-sex life table

Abstract

The life history and fitness of Nilaparvata lugens being reared under ambient level (current CO₂ concentration), medially elevated (550?µL/L) and highly elevated (750?µL/L) CO₂ concentration for long-term generation were compared using two-sex life table instead of traditional age-specific life table. The results showed that significantly longer larval duration and lower fecundity of N. lugens were observed in 750?µL/L relative to 550?µL/L treatment (P?<?0.05). Accordingly, 550?µL/L CO₂ significantly enhanced population parameters of N. lugens, including significantly higher intrinsic rate of increase (r), finite rate (λ) and net reproductive rate (R_O), but not for 750?µL/L CO₂. Taken together, N. lugens performs enhanced development rate, fecundity and survival in individual life history and higher potential in population multiplication under 550?µL/L CO₂ level, while only indicates the less enhanced development rate and survival without significant increased capacity of population expansion under 750?µL/L CO₂ level. These results should facilitate predicting the fitness and potential population damage of N. lugens, which is valuable for the integrated control of N. lugens in the future increasing CO₂ concentration.     

广西兴安地区褐飞虱发生动态及迁飞轨迹分析

为明确广西兴安地区单双季稻混作模式下褐飞虱发生规律,采用灯光诱捕和田间系统调查的方法研究了该地区早、中、晚稻田褐飞虱发生动态,并运用大气质点轨迹分析平台HYSPLIT 4.8对2011年褐飞虱迁飞高峰进行了迁飞轨迹分析。广西兴安地区褐飞虱种群数量随季节变化明显,年发生6代,迁飞活动主要集中在7月上旬至9月上旬。褐飞虱发生高峰期与单季中稻生长期吻合,在单季中稻田的发生量最大。2011年广西兴安褐飞虱5—6月迁入虫源主要来自海南稻区以及泰国、越南和老挝中部稻区;7—8月迁入虫源来自广西南部、东南部和广东西部稻区;回迁虫源来自湖南西南部和江西西部;10月从本地迁出的虫群主要迁向广西中部和贵州南部。研究表明,偏南方向的低空急流为褐飞虱的大规模迁入提供了运载气流,降雨是迫使稻飞虱降落的主要原因。     

Effects of different treatment methods of the fungicide jinggangmycin on reproduction and vitellogenin gene (Nlvg) expression in the brown planthopper Nilaparvata lugens Stål (Hemiptera:Delphacidae)

Jinggangmycin is an antibiotic fungicide against the rice sheath blight, Rhizoctonia solani, in China. Previous investigations have shown that jinggangmycin leaf spray stimulated the fecundity of Nilaparvata lugens Stål (Hemiptera:Delphacidae), but the effects of different application methods and concentrations of jinggangmycin on the reproduction of N. lugens have not been investigated. Here, we investigated three treatment methods (foliar and stem sprays and topical treatment) and four concentrations (100, 200, 400 and 800 ppm) of jinggangmycin on the reproduction of adult female N. lugens. The results showed that leaf spray significantly stimulated the fecundity of N. lugens and increased the vitellin content in female ovaries and the gene expression level of vitellogenin (Nlvg), but there was no significant effect on reproduction found for stem spray. In four concentrations of jinggangmycin, leaf sprays at concentrations of 100, 200 and 400 ppm and topical treatments of 100 and 200 ppm significantly increased the number of eggs laid, the vitellin content and the gene expression level of Nlvg compared to the control. Therefore, we suggest that stem spray at high concentration should be applied (if possible) when jinggangmycin is used against rice sheath blight to facilitate harmonious control of rice pests.     

稻飞虱及黑肩绿盲蝽在探照灯下的扑灯节律

为明确稻飞虱及其天敌黑肩绿盲蝽在探照灯下的扑灯节律,2012和2013年在广西兴安县应用探照灯诱虫器对稻飞虱及黑肩绿盲蝽进行了分时段监测。结果表明：稻飞虱和黑肩绿盲蝽年度间扑灯量有所不同,夜晚扑灯类型属于整夜扑灯型,晚上一般有1~2个扑灯高峰。2012年稻飞虱和黑肩绿盲蝽、2013年白背飞虱在迁入高峰期扑灯高峰不固定,2013年褐飞虱扑灯高峰主要集中在19：30～21：00时段,黑肩绿盲蝽2013年扑灯高峰主要集中在00：00～01：30和01：30～03：00时段。稻飞虱和黑肩绿盲蝽迁出高峰期及黑肩绿盲蝽本地活动期扑灯高峰主要集中在前半夜,稻飞虱本地活动期扑灯高峰主要集中在前半夜和04：30～06：00时段。     

杀虫剂对转Bt水稻靶标害虫二化螟和非靶标害虫稻飞虱种群数量的影响

为明确杀虫剂对转Bt水稻品种华恢1号(Huahui 1,HH1)靶标害虫二化螟Chilo suppressalis及其非靶标害虫白背飞虱Sogatella furcifera和褐飞虱Nilaparvata lugens种群数量的影响,于2014—2015年在江西省南昌市南昌县莲塘镇的广福试验基地进行田间试验,种植转Bt水稻和对照亲本水稻,分别对其进行施用杀虫剂和未施用杀虫剂处理,然后对经2种处理后转Bt水稻和对照亲本水稻田中靶标害虫二化螟幼虫数量及非靶标害虫白背飞虱、褐飞虱的若虫数量、成虫数量、总虫量(若虫数量和成虫数量之和)进行调查,并对这3种害虫田间种群数量的年份、水稻品种、杀虫剂影响因子进行方差分析。结果显示,转Bt水稻种植对二化螟幼虫数量影响显著,而施用杀虫剂处理未显著影响二化螟幼虫数量。转Bt水稻种植对白背飞虱和褐飞虱的若虫数量、成虫数量和总虫量无显著影响;而施用杀虫剂处理显著降低转Bt水稻及其对照亲本水稻稻田内白背飞虱和褐飞虱的若虫数量及总虫量,并显著降低白背飞虱成虫数量。2014年和2015年,施用杀虫剂和未施用杀虫剂转Bt水稻田内二化螟幼虫数量极低或未调查到;未施用杀虫剂转Bt水稻上二化螟幼虫数量仅在2014年显著低于未施用杀虫剂对照亲本水稻。2014年,未施用杀虫剂转Bt水稻和对照亲本水稻上白背飞虱总虫量最大值分别为1 037、1 376头/百丛,施用杀虫剂后分别降低至233、253头/百丛;未施用杀虫剂转Bt水稻上和对照亲本水稻上褐飞虱总虫量最大值分别为4 120、2 413头/百丛,施用杀虫剂后降低至367、410头/百丛;2015年白背飞虱和褐飞虱的发生规律与2014年相同。表明转Bt水稻可显著降低二化螟幼虫数量,但褐飞虱及白背飞虱数量没有降低,而施用杀虫剂可有效降低褐飞虱及白背飞虱数量。     

Mating pair combinations of insecticide-treated male and female Nilaparvata lugens Stål (Hemiptera: Delphacidae) planthoppers influence protein content in the male accessory glands (MAGs) and vitellin content in both fat bodies and ovaries of adult females

The brown planthopper, Nilaparvata lugens (Stål) (Hemiptera: Delphacidae), is an insect pest in which offspring are produced by the mating of adult males with adult females. This species is a classic case in which pest resurgence is induced by insecticides. In the past, studies of resurgence mechanisms have focused on insecticide-induced stimulation of reproduction in adult females. To date, however, the role that males play in the resurgence mechanisms of N. lugens has not been investigated. The aim of the present study is to examine changes in protein levels in male accessory glands (MAGs) induced by the insecticides triazophos and deltamethrin and to determine their relationship with vitellin content in the fat bodies and ovaries of adult females in the context of mating pairs. Our results show that protein content in MAGs is significantly affected by male mating status, insecticide type, and insecticide concentration. Insecticide application induced increased protein levels in MAGs. A greater quantity of MAG products was transferred to females via mating. Thus, protein levels in MAGs significantly decreased after mating. Experimental matings indicate that vitellin content in both fat bodies and ovaries of adult females in mating pairs consisting of a treated male and an untreated female (♂_t × ♀_ck) is significantly greater than that of females in pairs consisting of an untreated male and an untreated female (♂_ck × ♀_ck). Under various concentrations of the two insecticides, vitellin levels are highest in mating pairs consisting of a treated male and a treated female (♂_t × ♀_t), followed by mating pairs consisting of an untreated male with a treated female (♂_ck × ♀_t). These findings demonstrate that (1) insecticides have an effect on males; (2) insecticide effect can be transferred to females; and (3) the reproductive effect of insecticides is strongest in mating pairs in which both the males and females are treated compared to pairs in which only one individual is treated. These findings provide valuable information about the role of males in pesticide-induced resurgence of N. lugens.     

贵州三都一季中稻区褐飞虱主害代的发生规律

为明确贵州三都喀斯特褐飞虱主害代的发生规律,2009-2011年采用灯光诱测和田间系统调查方法对该地区一季中稻区的褐飞虱种群消长动态进行了系统研究.结果表明,各迁入代灯下诱捕虫量和田间虫量在不同年份间存在差异,2009年单日最大诱虫量达6 688头,分别是2010年和2011年的15.3倍和2.5倍.2009、2010、2011年预测圃虫量高峰分别为10777、4 630、1 615头/百丛,但主害代均为第4代和第5代,始见日与主迁入峰日呈显著相关.不同年份、不同移栽期稻田第4代、5代褐飞虱各虫态发生程度不同,2010、2011年褐飞虱若虫高峰日集中在7月下旬至8月上旬,晚栽稻田成虫、若虫的发生高峰日均迟于早栽类型田;早、晚栽时间差异形成的水稻不同生育期并存,利于褐飞虱栖居、繁殖.天敌蜘蛛、黑肩绿盲蝽与褐飞虱的种群消长具有同步性,但其自然控制作用不足以控制褐飞虱危害.     

Triazophos resistance mechanisms in the rice stem borer (Chilo suppressalis Walker)

A field population of the rice stem borer (Chilo suppressalis Walker) with 203.3-fold resistance to triazophos was collected. After 8-generation of continuous selection with triazophos in laboratory, resistance increased to 787.2-fold, and at the same time, the resistance to isocarbophos and methamidophos was also enhanced by 1.9- and 1.4-fold, respectively, implying some cross-resistance between triazophos and these two organophosphate insecticides. Resistance to abamectin was slightly enhanced by triazophos selection, and fipronil and methomyl decreased. Synergism experiments in vivo with TPP, PBO, and DEM were performed to gain a potential indication of roles of detoxicating enzymes in triazophos resistance. The synergism results revealed that TPP (SR, 1.92) and PBO (SR 1.63) had significant synergistic effects on triazophos in resistant rice borers. While DEM (SR 0.83) showed no effects. Assays of enzyme activity in vitro demonstrated that the resistant strain had higher activity of esterase and microsomal O-demethylase than the susceptible strain (1.20- and 1.30-fold, respectively). For glutathione S-transferase activity, no difference was found between the resistant and the susceptible strain when DCNB was used as substrate. However, 1.28-fold higher activity was observed in the resistant strain when CDNB was used. These results showed that esterase and microsomal-O-demethylase play some roles in the resistance. Some iso-enzyme of glutathione S-transferase may involve in the resistance to other insecticides, for this resistant strain was selected from a field population with multiple resistance background. Acetylcholinesterase as the triazophos target was also compared. The results revealed significant differences between the resistant and susceptible strain. The V_max and K_m of the enzyme in resistant strain was only 32 and 65% that in the susceptible strain, respectively. Inhibition tests in vitro showed that I₅₀ of triazophos on AChE of the resistant strain was 2.52-fold higher. Therefore, insensitive AChE may also involved in triazophos resistance mechanism of rice stem borer.     

Molecular characterization of two acetylcholinesterase genes from the brown planthopper, Nilaparvata lugens (Hemiptera: Delphacidae)

Acetylcholinesterase (AChE), which is encoded by the ace gene, catalyzes the hydrolysis of the neurotransmitter acetylcholine to terminate nerve impulses at the postsynaptic membrane. AChE is a primary target of many insecticides including organophosphates (OP) and carbamates (CB). In this study, full-length cDNA sequences of two ace genes (Nlace1 and Nlace2) were sequenced from the brown planthopper (BPH) Nilaparvata lugens, the most destructive insect pest of rice crops. Nlace1 cDNA is 2842 nucleotides long and contains an ORF potentially encoding a 790 amino acid peptide. Nlace2 cDNA is 2852 bp in length and contains an ORF that potentially encodes a 672 amino acid peptide. NlAChE1 has an identity of 40% with NlAChE2 at the amino acid sequence level. Phylogenetic analysis of 59 AChEs from 32 animal species showed that NlAChE1 is most closely related to AChE1s from Blattella germanica and Nephotettix cincticeps, while NlAChE2 is most closely related to AChE2 from N. cincticeps. Quantitative RT-PCR analysis showed that Nlace1 is expressed at a much higher level than Nlace2 in all developmental stages and tissues, demonstrating that NlAChE1 may be the dominant AChE form of the two enzymes. This result will help reveal the resistance mechanism of N. lugens to organophosphorous and carbamate insecticides and promote development of more selective insecticides targeting the main NlAChE1.     

不同作业方式和施药模式下杀虫剂对褐飞虱的防治效果

为探讨植保无人机施药沉积利用率高但防治效果不稳定的问题,利用液相色谱串联质谱法测定植保无人机和背负式喷雾器喷施后药液在水稻植株上的农药沉积率,选择烯啶虫胺、吡蚜酮、呋虫胺和毒死蜱4种杀虫剂分别对分蘖期和拔节期水稻进行不同施药模式的室内喷施试验,以模拟分析水稻非靶标部位的农药沉积对褐飞虱防治效果的影响。结果表明,植保无人机喷施后水稻中上部的农药沉积率为34.83%,显著高于背负式喷雾器的农药沉积率(15.30%),但药后7 d的校正防治效果分别为61.4%和64.4%,两者之间差异不显著。烯啶虫胺、呋虫胺和吡蚜酮3种内吸性杀虫剂室内模拟喷施后1 d和4 d均为直接喷施处理的水稻基部农药含量最高,且该处理药后1、4和7 d的校正死亡率最高,分别为23.33%～70.00%、46.67%～80.42%和70.00%～87.50%;其次为遮挡茎部喷施处理,而水中加药和叶片受药处理的防治效果最差。对于兼有触杀和熏蒸毒性的有机磷类杀虫剂毒死蜱,水中加药处理和直接喷施处理药后1、2和4 d的校正死亡率均在98.33%以上,显著高于其他2种处理。表明不同杀虫剂的吸收传导特性不同,这直接影响其能否到达水...     

三唑磷与氟虫腈复配对粘虫的增效机理研究

以粘虫Mythimna separate 4龄幼虫为试虫,从表皮穿透、生物转化、作用靶标3个方面研究了三唑磷·氟虫腈复配剂的增效机理。结果表明,三唑磷与氟虫腈复配后,相互促进了对方对粘虫表皮的穿透作用;三唑磷对酸性磷酸酯酶(ACPase)的竞争性抑制与氟虫腈对ACPase的非竞争性抑制与反竞争性抑制的混合型抑制相结合,使复配剂对ACPase的抑制能力大幅度提高;对谷胱甘肽S-转移酶(GSTs)的联合抑制系数(JIC)达到425.87,表现出显著的增效作用;三唑磷和氟虫腈对乙酰胆碱酯酶(AChE)只有微弱的交互作用。表皮穿透率的提高及其对ACPase和GSTs抑制能力的增强可能是该复配剂对粘虫增效的重要机理。     

苏云金芽胞杆菌工程菌WG-001可湿性粉剂的增效化合物筛选

为了筛选出对苏云金芽胞杆菌具有增效作用的无机化合物,并确定其最佳使用浓度,于苏云金芽胞杆菌工程菌WG-001可湿性粉剂(Bt WG-001 WP)中添加MgCl2、CaCl2、NaHCO3、MgSO4、NaH2PO4、Na2CO3、Na2HPO4、KH2PO4等8种化合物,测定其对小菜蛾的毒力.结果显示,添加0.4％MgCl2、0.2％CaCl2、0.4％MgSO4、0.2％Na2CO3时,共毒因子分别为46.75、36.27、41.51、31.03,均大于20,而添加另外4种化合物,共毒因子均低于20;添加0.4％MgCl2、0.2％CaCl2、0.4％MgSO4、0.2％Na2CO3,增效比最大,分别为1.46、1.33、1.34、1.29.田间试验结果显示,Bt WG-001 WP中添加0.4％MgSO4稀释500倍施用时,药后7天对小菜蛾的田间平均防效最高,为80.51％,且与其它处理差异显著.表明添加0.4％MgSO4可显著提高Bt对小菜蛾的田间防效.     

长翅型与短翅型褐飞虱对杀虫剂的敏感性比较

采用稻苗浸渍法测定了长、短翅型褐飞虱对烯啶虫胺、环氧虫啶、呋虫胺、噻虫嗪、噻虫胺、吡虫啉、毒死蜱、敌敌畏、噻嗪酮、异丙威、吡蚜酮和醚菊酯的敏感性,并对其体内解毒酶活力进行了比较分析。结果表明:长翅型与短翅型褐飞虱若虫对新烟碱类杀虫剂呋虫胺、噻虫嗪、噻虫胺和吡虫啉的敏感性存在显著差异,长翅型比短翅型更敏感;相反,对于有机磷类杀虫剂毒死蜱,短翅型褐飞虱则更敏感;2种生物型对烯啶虫胺、环氧虫啶、敌敌畏、噻嗪酮、异丙威、吡蚜酮和醚菊酯的敏感性无显著差异。解毒酶相对比活力测定结果表明,长翅型褐飞虱若虫酯酶比活力显著高于短翅型,细胞色素P450单加氧酶比活力显著低于短翅型,而谷胱甘肽S-转移酶比活力无显著性差异。本研究结果可为褐飞虱的有效防控提供科学参考。     

水稻锯齿叶矮缩病毒对褐飞虱生长发育和生殖的影响

为明确水稻锯齿叶矮缩病毒（rice ragged stunt virus,RRSV）对褐飞虱Nilaparvata lugens生长发育和生殖的影响,分别比较带毒褐飞虱与未带毒褐飞虱的若虫发育历期、成虫寿命、性比、翅型及产卵量,并利用实时荧光定量PCR（real-time quantitative PCR,RT-qPCR）技术分析RRSV侵染后褐飞虱体内卵黄原蛋白（vitellogenin,Vg）与其受体（vitellogenin receptor,VgR）以及海藻糖代谢途径中相关基因表达量的情况。结果显示,带毒褐飞虱的若虫历期显著高于未带毒褐飞虱的若虫历期,分别为17.56 d和15.90 d;带毒褐飞虱的成虫寿命、雌虫比例和短翅比例均高于未带毒褐飞虱的,但两者间均无显著差异;带毒褐飞虱较未带毒褐飞虱单雌日产卵量显著提升,分别为11.26粒和6.45粒;带毒褐飞虱的Vg、VgR和海藻糖转运蛋白基因（trehalose transporter,TRET）表达量均显著高于未带毒褐飞虱的。表明RRSV侵染会显著延长褐飞虱若虫发育历期,同时可能通过上调Vg、VgR以及TRET的表达量以促进褐飞虱生殖。     

大气CO_2浓度增高和施氮量对抽穗期春小麦光合参数及水分利用效率的影响

在开顶式小型温室内模拟大气CO2浓度增高,并在N0,N100,N200计3个氮素水平〔施N量分别为纯N0、0.1、0.2 g/kg(土)〕下盆栽小麦,研究小麦光合作用和水分利用效率(WUE)对大气CO2升高的响应及其氮素调控机制。结果表明,760μmol/molCO2浓度×N素小麦叶片的净光合速率、叶片胞间CO2浓度、气孔限制值及叶片水分利用效率较对照呈明显上升趋势,叶片气孔导度显著降低,蒸腾速率则呈先降后升的趋势;在400μmol/molCO2浓度下,N0和N100处理的小麦光合速率明显下降,但N200处理较当前CO2浓度处理明显升高;气孔导度,胞间CO2浓度以及蒸腾速率呈现下降的趋势,小麦的气孔限制值Ls和WUE在N200条件下表现最高,较各个处理均达到极显著水平。两种大气CO2浓度下的小麦叶片光合速率和气孔导度均随氮素水平的升高而明显升高,胞间CO2浓度和蒸腾速率表现均不一致。因此,长期高CO2浓度使得小麦叶片WUE和Ls值显著升高,在低中氮处理下产生明显的光合下调现象,但在高氮处理下却不发生。     

Decrease of rice plant resistance and induction of hormesis and carboxylesterase titre in brown planthopper, Nilaparvata lugens (Stål) by xenobiotics

The brown planthopper (BPH), Nilaparvata lugens (Stål) is a serious threat to the rice production throughout Asia. The indiscriminate application of various xenobiotics in rice ecosystem is perceived as one of the factors for the frequent outbreak of BPH. The present study has critically analysed the secondary effects of some xenobiotics used in rice field on certain plant and insect parameters that subsequently favour BPH outbreak. Application of 2,4-D, carbendazim, deltamethrin and urea reduced the innate BPH resistance of PTB 33 rice variety due to favourable alterations in rice free amino acid and sucrose content. Similarly, these chemicals also induced hormesis and enhanced feeding in BPH. Alternatively, soil amendment with neem seed powder and Calotropis gigantea leaves improved plant innate resistance and showed no sign of hormesis or enhanced feeding in BPH. In addition, deltamethrin has the ability to stimulate BPH carboxylesterase titre. Native PAGE analysis of esterases from whole body homogenate of BPH revealed at least five esterase isozyme bands, prominent being E1 and E2. However, no difference in BPH esterase banding pattern was observed between different xenobiotic treatments. All these esterase bands are classified under carboxylesterase based on their inhibition by class specific esterase inhibitors.     

表达Harpin蛋白质Hpa1功能片段的转基因小麦对赤霉病的抗性

来自水稻黄单胞菌的harpin蛋白质Hpa1有促进植物生长、诱导植物抗病性的功能,Hpa1序列10~42氨基酸片段（Hpa1_10-42）的活性比全长分子高1.3~7.5倍。为研究Hpa1_10-42在转基因小麦体内表达以后对赤霉病的影响和评价转基因小麦抗病水平与应用潜力,对6个小麦转基因系进行了测定。结果显示,Hpa1_10-42在转基因系T₃~T₅代呈现稳定表达,用禾谷镰刀菌一个分离物进行接种以后,转基因系发生赤霉病的程度较非转基因小麦显著降低,且转基因系T₃~T₅代赤霉病降低的趋势一致。另外,转基因系小麦病害减轻的程度,与在非转基因小麦上使用杀菌剂的效果相当,表明Hpa1_10-42转基因表达对小麦赤霉病有防治作用。