Seasonal and geographical variation in diapause and cold hardiness of the Asian corn borer,Ostrinia furnacalis

Asian corn borer, Ostrinia furnacalis (Guenée), is a key corn pest in the Asian‐Western Pacific countries. It overwinters as full‐grown larvae in plant stalks or in a spun‐silk covering located in the plant debris in the temperate regions of China. Supercooling point (SCP) and survival rate after low sub‐zero temperature treatment were assessed for field‐collected populations in the laboratory using a cool bath with a 1°C/min cooling rate until ?40°C. Mean SCPs were varied among geographical populations, with a significant decline from ?22.7°C of Haikou, the multivoltine tropical population in the south, to ?28.5°C of Gongzhuling, the univoltine temperate population in the northeast of China. In addition, there was more than 1°C difference in SCP between Gongzhuling univoltine and bivoltine populations that were from the same geographic origin. Mean SCPs of the Guangzhou population fluctuated over the year, with significantly lower SCPs in winter than in other seasons, which correlated with a significantly higher proportion of diapausing larvae in winter than in other seasons. Over 41% of overwintering larvae from the northeast population could withstand to be supercooled for a few minutes to the low sub‐zero temperature of ?40°C, but only 6.7% of their southern counterparts did so. The findings from this study suggest that O. furnacalis mostly takes advantage of freeze avoidance as diapausing larvae for overwintering in the southern region, whereas it exhibits freeze tolerance in diapause in the northeastern region.     

Inheritance of photoperiodic induction of larval diapause in the Asian corn borer Ostrinia furnacalis

The Asian corn borer Ostrinia furnacalis (Guenée) enters facultative diapause as fully‐developed larvae in response to short‐day conditions. As a consequence of geographical variation in photoperiodic response, moths from Nanchang (28°46′N, 115°50′E) enter diapause in response to short day‐lengths (D strain), even at the high temperatures whereas moths from Ledong (18°47′N, 108°89′E) exhibit almost no diapause under the same conditions (N strain). In the present study, crosses between the two strains are used to evaluate the inheritance of diapause under different photoperiods at temperatures of 22, 25 and 28 °C. The moths, both reciprocal crosses and backcrosses, show a clear long‐day response, similar to that of the D strain, suggesting that the photoperiodic response controlling diapause in this moth is heritable. However, the critical day‐length for induction of diapause is shorter in hybrids than in the D strain. The N strain also shows a short‐day photoperiodic response at the lower temperature of 22 °C, indicating that the N strain still has the capacity to enter a photoperiodically‐induced diapause, depending on the rearing temperature. The incidence of diapause in all crosses is highest with D strain fathers or grandfathers and lowest with N strain fathers or grandfathers, indicating that the male parent has significantly more influence on the incidence of diapause of subsequent progeny than the female. The results obtained from all crosses under LD 12 : 12 h or LD 13 : 11 h photocycles at 25 °C show that inheritance of diapause in O. furnacalis does not fit an additive hypothesis and that the capacity for diapause is transmitted genetically in the manner of incomplete dominance.     

亚洲玉米螟滞育诱导外源性因子研究

研究了食料、温度、光周期等外源性因子对亚洲螟滞育的影响,试验证明,食料性质对其滞育诱导影响显著,取食玉米雌的幼虫以滞育诱导光周反应的敏感性强于取食棉铃的幼虫,在一定范围内,食料中可溶性糖含量增加可促进幼虫的滞育诱导,而粗蛋白含量的增加则具有抑制滞育诱导的作用,两者的互作效应不显著。高温抑制滞育诱导,你 促进滞育诱导。短光周对幼虫的滞育诱导无特定的敏感龄期,但以３龄与２龄对短光周刺激敏感性较强,包光     

Age‐dependent changes in tolerance to cold and accumulation of cryoprotectants in overwintering and non‐overwintering larvae of European corn borer Ostrinia nubilalis

Abstract The age‐dependent cold hardiness profile of Ostrinia nubilalis is compared between nondiapausing and diapausing larvae, as well as with field‐collected larvae. The results suggest that both cold tolerance and accumulation of cryoprotectants depends upon the age of O. nubilalis larva. Late fifth‐instar nondiapausing larvae are more cold tolerant than younger fifth‐instars because they show enhanced ability to withstand sub‐zero temperatures. No appreciable difference is observed between the experimental groups of diapausing larvae as far as their supercooling ability and tolerance at sub‐zero temperatures above the supercooling point. In general, both field‐collected and diapausing larvae are more cold tolerant than nondiapausing larvae, indicating a direct link between diapause and cold hardiness. The age of diapausing larvae affects the ability to accumulate glycerol. Glycerol levels of 45‐day‐old diapausing larvae are significantly higher (2.7‐fold) compared with 90‐day‐old diapausing larvae. Moreover, diapausing larvae display a five‐ to 13‐fold higher glycerol content compared with nondiapausing larvae. There is a trend for an age‐dependent cold hardiness profile in O. nubilalis and further tests that could demonstrate a causal relationship between age and cold tolerance are needed.     

Transgenic ice nucleation-active Enterobacter cloacae reduces cold hardiness of corn borer and cotton bollworm larvae

The ice nucleation (IN) gene iceA of Erwinia ananas 110 was integrated into the chromosomes of two Enterobacter cloacae strains (Enc1.2022 and Enc1.181). These two newly derived transgenic strains, designated Enc2022-I and Enc181-I, respectively, possessed ice nucleation activity at -2.5 degrees C, significantly higher than their parent strains (active at approx -10 degrees C or lower). After ingesting these transgenic bacteria, the mean supercooling points (SCPs) of corn borer and cotton bollworm larvae were -3 to -4 degrees C, significantly higher than those of untreated controls. The SCPs remained significantly elevated over the 9-day period after ingestion, which matched well with the efficient gut colonization of the bacteria during this period. All treated larvae froze and eventually died after exposure for 6 h to a temperature of -7 degrees C, and more than 95% died after 12 h at -5 degrees C. In contrast, few or none of the untreated control larvae froze and died under the same conditions. Furthermore, the growth ability of these transgenic ice nucleation-active (INA) En. cloacae strains on corn leaves was reduced, compared to that of wild-type epiphytic E. ananas, as revealed by pot tests conducted in both greenhouse and outdoor conditions. The stable colonization in insect guts and their lower affinity to plants would make these transgenic INA bacteria useful as a novel tool for biological control of insect pests in agricultural fields.     

亚洲玉米螟对玉米果穗挥发物的定向行为反应

为了明确玉米果穗挥发物对亚洲玉米螟Ostrinia furnacalis(Guenée)定向行为的影响,本研究采用气相色谱-质谱联用(GC-MS)、气相色谱-触角电位(GC-EAD)技术以及行为实验测定了亚洲玉米螟对玉米果穗挥发物的电生理及行为反应。结果表明:玉米果穗气味对其未交配雄蛾、雌蛾均具有引诱作用;从中分离出35种化合物,其中(+)-环苜蓿烯含量最高,达到(2391.60±161.34)ng;6种化合物能引起亚洲玉米螟未交配成虫的GC-EAD反应,其中顺-3-己烯-1-醇、(+)-环苜蓿烯、β-石竹烯对未交配雄蛾、雌蛾定向行为具有引诱作用。     

Cold hardiness of larvae of the southwestern corn borer, Diatraea grandiosella

The cold-hardening capacity of field-collected larvae from southeast Missouri and laboratory-reared larvae of the southwestern corn borer, Diatraea grandiosella Dyar, was examined. Supercooling points of non-diapause and diapause larvae collected from maize plants grown in Missouri (36°30 N lat.) were ca.-7.0°C. The hemolymph melting points of diapause field larvae (-0.8°C) were significantly lower than those of non-diapause larvae collected in July (-0.5°C). The supercooling points of hemolymph from non-diapause and diapause field larvae ranged randomly from-10° to-18°C. Supercooling points of non-diapause laboratory larvae increased from-13° to-10°C prior to pupation, whereas those of diapause larvae increased similarly before the onset of diapause, but then decreased during diapause to ca.-17°C. No change in supercooling points or capacity to survive in the presence of ice was observed in diapause laboratory larvae acclimated at 4°C for 63 days. Laboratory and field larvae began to freeze at ca.-1.5°C in the presence of ice, but survived to several degrees below their melting points. The high supercooling points of field larvae appeared to be due to the presence of an environmental ice-nucleator. Although data for laboratory larvae indicate sufficiently low supercooling points to permit winter survival in southeastern Missouri, considerable larval mortality occurs in the field due to inoculative freezing and the presence of an ice-nucleator.     

亚洲玉米螟幼虫体内保幼激素二醇激酶cDNA片段克隆与序列分析

为研究亚洲玉米螟Ostrinia furnacalis(Guenée)幼虫体内保幼激素二醇激酶(JHDK)表达调控的分子机理,根据不同昆虫保幼激素二醇激酶基因序列的保守区域,设计合成简并引物,采用RT-PCR技术从亚洲玉米螟5龄幼虫中扩增出一段cDNA片段,大小为189bp,编码63个氨基酸,预测分子量为6.78ku,理论等电点pI值为4.57。该基因序列中含有保守的GTP结合蛋白特征指纹基序∑3和∑1。BlastP分析结果表明:该片段氨基酸序列与烟草天蛾JHDK氨基酸序列的一致性最高,为69%;与家蚕和小菜蛾JHDK氨基酸序列的一致性分别为55%和52%。构建系统发育树分析了3种鳞翅目昆虫JHDK进化关系,结果显示:亚洲玉米螟cDNA片段氨基酸序列与家蚕JHDK的亲缘关系最近,与小菜蛾JHDK的亲缘关系最远。半定量PCR结果表明:JHDK基因在中肠中表达量最高,随着5龄幼虫的发育,JHDK基因在血细胞、脂肪体和体壁组织中表达量有下降趋势,但在中肠组织中表达量明显增强。     

Study on the physiology of diapause, cold hardiness and supercooling point of overwintering pupae of the pistachio fruit hull borer, Arimania comaroffi

The pistachio fruit hull borer, Arimania comaroffi (Ragonot) (Lepidoptera: Pyralidae), is a key pest of pistachio orchards in Iran. This pest passes the winter as diapausing pupae. In this study, some physiological changes in relation to environmental temperature were investigated in field collected pupae. The relationship between supercooling point, cold hardiness and physiological changes of a wild population of this pest was also investigated. The glycogen content decreased with decrease in environmental temperature. Decrease in glycogen content was proportional to increase in total body sugar, trehalose, myo-inositol and sorbitol contents. In January with mean ambient temperature of 5.4°C, glycogen (5 mg/g fresh body weight) content was at the lowest level whereas total body sugar (10.3 mg/g fresh body weight), trehalose (8.6 mg/g fresh body weight), myo-inositol (5.3 mg/g fresh body weight) and sorbitol (2.6 mg/g fresh body weight) were at the highest levels. Total body sugar, trehalose, myo-inositol and sorbitol contents increased as mean temperature decreased from 22.7°C in October to 5.4°C in January. Total body lipid decreased during overwintering and reached to the lowest level at the end of March. Supercooling points were decreased from October to January and reached to the lowest level (-16°C) in January with minimum ambient temperature of -10°C. Survival at low temperature after 24 h was also greatest in January with 72% survival at -10°C, 39% survival at -15°C and 0% survival at -20°C. Increase in temperature from February onward, was proportional with increase in supercooling points and decrease in survival rate. Regardless of sampling date, all pupae died after 24 h at -20°C, whereas none pupae died after 24 h at -5°C. This indicates that this insect is freeze-intolerant.     

亚洲玉米螟对丁硫克百威的抗性筛选及交互抗性

采用点滴法测定丁硫克百威对亚洲玉米螟Ostrinia furnacalis(Guenée)幼虫的致死中量,并采用亚致死剂量筛选亚洲玉米螟10代。结果表明,丁硫克百威处理亚洲玉米螟5代后,抗性即显著上升,抗性倍数为3.23倍,到10代时,抗性达到了5.43倍。对高效氯氟氰菊酯、辛硫磷和丁硫克百威3种玉米螟抗性品系进行交互抗性测定,发现丁硫克百威与其它2种农药存在交互抗性。建议在害虫防治谨慎使用丁硫克百威。     

一种快速鉴别亚洲玉米螟蛹雌雄的方法

报道了一种采用体视镜和三维显微镜对亚洲玉米螟Ostrinia furnacalis(Guenée)蛹雌雄进行外部形态学观察,快速准确通过外部形态特征区分玉米螟蛹雌雄的方法。结果表明,雌蛹生殖孔与产卵孔连接成1条短裂缝且都位于第8腹节,生殖孔与第7腹节后缘相连;雄蛹生殖孔位于第9腹节,距离第7腹节后缘约390μm。根据羽化后成虫鉴定结果,此方法鉴定准确率为100%。     

Regulation of larval diapause by juvenile hormone in the European corn borer,Ostrinia nubilalis

The effects of juvenile hormone (JH) on the larval diapause of the European corn borer, Ostrinia nubilalis, were investigated. The larval period was prolonged and the development of gonads was retarded if non-diapause larvae were treated with JH. Pupation of diapausing larvae was accelerated by allatectomy. It was concluded that the diapause of this borer is largely regulated by JH.     

Volatiles induced by the larvae of the Asian corn borer (Ostrinia furnacalis) in maize plants affect behavior of conspecific larvae and female adults

Abstract  Effects of maize ( Zea mays L.) volatiles induced by larvae of the Asian corn borer, Ostrinia furnacalis (Guenée), on the orientation behaviors of Asian corn borer larvae and oviposition of the females were investigated. Nineteen volatile chemicals, with terpenes being the major components, were identified from maize plants attacked by third instar Asian corn borer larvae. Coupled gas chromatographic-electroantennographic detection (GC-EAD) analyses revealed some electroantennographic differences between female and male Asian corn borer antennae in response to larvae-induced maize volatiles; female responded to ( E )-2-hexenal, nonanal, ( Z )-3-hexen-1-ol and three unknown compounds while the male only responded to ( E )-2-hexenal, nonanal and one unknown compound. In laboratory orientation bioassays, Asian corn borer neonate larvae were attracted to extracts collected from Asian corn borer-damaged plants as well as to synthetic farnesene, but were repelled by ( Z )-3-hexen-1-ol. In laboratory oviposition bioassays, gravid females laid fewer eggs on plants damaged by larvae than on mechanically damaged plants or undamaged plants. Adult Asian corn borer females deposited fewer eggs on wax paper treated with ( E )-2-hexenal or ( Z )-3-hexen-1-ol than on wax paper treated with hexane (control). The results suggest that Asian corn borer can affect the behaviors of conspecific larvae and adults by changing host plant volatiles.     

亚洲玉米螟幼虫酚氧化酶原基因序列的生物信息学分析

酚氧化酶原PPO是昆虫免疫的关键酶, 本文从生物信息学角度对亚洲玉米螟Ostrinia furnacalis Guenée幼虫PPO进行分析, 为进一步研究其高级结构与功能的关系提供理论依据。利用我们已提交到GenBank的数据, 采用在线分析及MEGA4和RasMol软件对亚洲玉米螟酚氧化酶原（Of-PPO）的核苷酸和氨基酸序列、系统发生关系和蛋白质三级结构进行分析。结果表明: Of-PPO全长cDNA序列有2 686 bp, 包含一个2 079 bp的开放阅读框, 其推导的693个氨基酸序列中包含6个组氨酸残基构成的2个铜离子结合位点, 以及保守的硫羟酸酯区域。Of-PPO属于PPO2类群, 其N端不含信号肽, 无跨膜结构域区域, 无糖基化位点, 44个磷酸化位点均匀分布于整个多肽链中, 有2段序列可能形成卷曲螺旋, 有5个区域的氨基酸具较强疏水性, 其二级结构中α-螺旋占22.54%, 随机卷曲占56.79%。同源建模显示其三级结构为“α/β型”中的“滚筒结构”, 存在一个明显的空位, 可能与该酶催化活性有关。本文可为Of-PPO的实验研究和应用开发提供有价值的信息。     

苯甲酰脲类杀虫剂对亚洲玉米螟生长发育和繁殖的效用

为全面评价苯甲酰脲类(BPUs)杀虫剂对亚洲玉米螟Ostrinia furnacalis(Guenée)的持续控制作用,采用叶片浸渍法,测定氟啶脲、除虫脲、氟铃脲和虱螨脲对亚洲玉米螟生长发育和繁殖的影响。结果表明,经4种BPUs处理后,亚洲玉米螟存活幼虫的历期延长了2.4～4.3d,平均单头雌、雄蛹重分别减轻了10.3～13.2mg和0～7.7mg,幼虫化蛹率和成虫交配成功率分别下降了14.1%～43.3%和5.6%～25.6%,产卵量降低了25.5%～53.3%,但卵孵化率没有明显改变。亚洲玉米螟雌、雄蛾吸食蔗糖水药液后,成虫寿命增加了1.3～4.3d,产卵量下降40.3%～58.2%,卵孵化率降低了32.9%～64.3%,F1代幼虫死亡率是对照组的3.5～8.5倍,表明苯甲酰脲类杀虫剂显著抑制了亚洲玉米螟幼虫生长发育和成虫的繁殖力。     

Landscape genetic analyses reveal host association of mitochondrial haplotypes in the Asian corn borer,Ostrinia furnacalis

Crop expansion often leads to high pest pressure. These pests may have fitness trade-offs related to host use, and some host-associated genotypes may benefit and increase in frequency. However, evidence concerning the effect of host availability on spatial distribution and frequency of mitochondrial haplotypes is scarce. We studied genetic variation of the Asian corn borer, Ostrinia furnacalis (Guenée), across a large area during 2 years (2016 and 2017). Mitochondrial sequence data were obtained from 530 individuals collected from 79 locations in Shandong Province, China. In total, 155 haplotypes were found based on the combined cytochrome oxidase subunit 1 (COI) and COII genes. Three haplotypes (H2, H12, and H23) were dominant, whereas most of the other haplotypes occurred in low frequency. A haplotype network showed that the 155 haplotypes can be grouped into three clusters. Haplotype clusters seemed to be randomly distributed. The frequency of H12 (in Cluster 1) was positively correlated with maize crop proportion, but negatively correlated with other crops (primarily vegetables, oilseed crops, and cotton) at all spatial scales (1-, 3-, and 5-km radius). Cluster 2 had haplotype H23, and this cluster was negatively correlated with semi-natural habitats. Cluster 3 had no dominant haplotype and was not affected by landscape factors. We conclude that H12 may be a maize-associated haplotype. Further study is needed to verify the possibility that the carriers of this haplotype may possess some fitness trade-offs. Our study highlights the importance of host availability in O. furnacalis haplotype distribution and frequency.     

亚洲玉米螟性体型二型性及幼虫发育历期与蛹重的关系

【目的】探明亚洲玉米螟不同地理种群性体型二型性及幼虫发育历期与蛹重的关系。【方法】在26℃,L︰D=16︰8条件下,详细记录了来自热带地区的海南省三亚种群,来自亚热带地区的广东省广州和江西省永修种群,及来自温带地区的河北廊坊种群的雌雄幼虫发育历期、蛹重、成虫重,并计算幼虫发育速率。【结果】不同地理种群的雌性个体均显著大于雄性个体。三亚种群雌雄个体的发育历期没有差异,但雌性的生长速率显著大于雄性个体;广州种群、永修种群及廊坊种群,雌雄的生长速率没有差异,但雌性幼虫的发育历期显著长于雄性幼虫。各地理种群蛹重随幼虫发育历期的延长呈下降趋势。【结论】亚洲玉米螟存在明显的性体型二型性,雌雄个体发育历期和生长速率存在地理差异。     

Factors influencing parasitism of Trichogramma dendrolimi on eggs of the Asian corn borer, Ostrinia furnacalis

Laboratory studies were made to determine the capacity of Trichogramma dendrolimi to parasitize eggs of Ostrinia furnacalis, as affected by the rearing host species, substrate of host eggs, host age, original locality of host populations, and cold storage of host eggs. Wasps reared from eggs of Antheraea pernyi showed parasitic capacity on eggs of O. furnacalis on average twice as high as that of the wasps reared from eggs of Corcyra cephalonica. When the age of O. furnacalis eggs at 26 °C increased from 0–6 h to 18–24 h, the proportion of wasps that successfully parasitized host eggs, the number of host eggs parasitized, and the rate of parasitization all decreased by >50%. The number of O. furnacalis eggs parasitized per female T. dendrolimi increased with the number of host eggs available, and reached 22.9 in a 24 h period. However, the parasitic capacity of female T. dendrolimi on eggs of O. furnacalis laid on plant leaves was similar to that of O. furnacalis eggs laid on wax paper. Levels of parasitism of O. furnacalis eggs from two widely separated localities, i.e. Changchun (43.50° N, 125.20° E) and Hangzhou (30.18° N, 120.07° E), were similar. Cold storage of O. furnacalis eggs at 4 °C for 5 days did not affect parasitization. Results obtained in this study indicate that although O. furnacalis is a less preferred and less suitable host than many other hosts, such as Dendrolimus punctatus, Actias selene ningpoane, Philosamia cynthia, A. pernyi, C. cephalonica, within the host-species range of T. dendrolimi, the parasitoid has the potential to achieve 50–60% or even higher rates of parasitization of O. furnacalis eggs in corn fields under suitable conditions, and could be used in the biological control of the pest.     

The role of juvenile hormone in the larval diapause of the European corn borer,Ostrinia nubilalis

The possible role of juvenile hormone (JH) in the induction and termination of larval diapause in the European corn borer, Ostrinia nubilalis, was investigated using topical applications of both JH I and a JH mimic as well as by monitoring JH titers with the Galleria bioassay. Neither JH nor the JH mimic ZR515 was capable of influencing diapause termination when administered topically. The Galleria bioassay revealed little or no JH in the hemolymph of mid diapause (>30 days) insects, indicating no demonstrable role for JH in diapause maintenance. When ZR515 was administered to nondiapause, newly ecdysed fifth instar larvae the pupal molting cycle was delayed. By use of photoperiodic regimes we were able to show that the molting delay was not equivalent to diapause induction. The Galleria bioassay showed differences in JH titer profiles between diapause and nondiapause animals during the final larval stadium. The nondiapause insects showed titers that decline rapidly to trace amounts following the molt to fifth instar then rose prior to pupation. The diapause insects had generally higher titers and exhibited a more gradual decline after the molt. No evidence was obtained to support the hypothesis that JH plays a key role in the induction, maintenance, or termination of larval diapause.