Seasonal changes in glycerol content and cold hardiness in two ecotypes of the rice stem borer, Chilo suppressalis, exposed to the environment in the Shonai district, Japan

The rice stem borer, Chilo suppressalis, is divided into at least two ecotypes in Japan, the Shonai ecotype (SN) which is distributed in the northern part of Japan, and the Saigoku ecotype (SG) which is distributed in the southwestern region. Cold hardiness is positively correlated with the level of glycerol in both ecotypes. To investigate whether ecological distribution affects glycerol accumulation and cold hardiness development in these two ecotypes, overwintering larvae of the SN and SG ecotypes were concurrently exposed to the Shonai district. Obvious differences in the progress of glycerol accumulation and cold hardiness development in SN and SG larvae were found in early winter in the Shonai district. The levels of glycerol content and cold hardiness were low in October and high in January in both ecotypes, but those levels were different within this period (November and December) between ecotypes; the levels in SN larvae quickly reached their maximum, whereas, in SG larvae levels increased slowly. Under controlled conditions, the effect of the period of acclimation at 10 degrees C and subsequent low-temperature (5 degrees C) exposure on glycerol accumulation was investigated. These results indicated that glycerol accumulation in SN was stimulated by the progression of diapause termination, whereas a higher cumulative effect on glycerol production in SG was found when diapause was in a deep state.     

Seasonal changes of phospholipids in last instar larvae of rice stem borer Chilo suppressalis Walker (Lepidoptera: Pyralidae)

Phospholipids of the cell membrane have been studied from the viewpoint of how overwintering insects inhabiting temperate zones adapt to low temperature. The transition of cell membrane phospholipids from a liquid crystalline phase to a gel phase is a crucial cause of cold or freezing injuries. We determined the qualitative and quantitative changes of phospholipids in the last instar larvae of the rice stem borer in summer and winter. We found that the total amount of their phospholipids did not change significantly between summer and winter and that the sum of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) comprised about 85% of their total phospholipids. In summer, the ratio of their PE to PC was almost one, while from autumn to mid‐winter it increased and reached three in February. The fatty acid compositions of PC hardly changed, and the percentage of unsaturated fatty acids did not exceed 50%. In contrast, the percentage of unsaturated fatty acids of PE in overwintering larvae increased up to 80% as ambient temperatures fell and oleic acid mainly contributed to the high percentage of unsaturation. In the present study, the relationship between the quantitative and qualitative changes of phospholipids and adaptation of the rice stem borer to cold winter climate are discussed.     

CO_2倍增和转Bt水稻对二化螟幼虫的生理影响

通过开顶式气室研究了CO2浓度倍增(750vs.375μL/L)对取食转Bt水稻及其对照亲本秀水11的二化螟Chilo suppressalis(Walker)幼虫体内代谢酶乙酰胆碱酯酶(AChE)和保护酶(超氧化物歧化酶SOD、过氧化物酶POD和过氧化氢酶CAT)活力的影响,以期明确该类害虫对大气CO2浓度升高及转Bt水稻的生理响应。研究结果表明:(1)AChE受CO2浓度水平影响显著,与对照CO2处理相比,高CO2条件下该酶活力显著降低;(2)高CO2水平及水稻中Bt毒素蛋白的存在均会导致二化螟幼虫体内POD活力显著降低,SOD初期活力显著升高,但不能维持较长时间,CAT活力表现为先升后降的特点;(3)当受到水稻品种、CO2浓度水平和取食时间的综合影响时,以上代谢酶和保护酶活力会发生显著变化。短时间内二化螟幼虫可通过体内以上生理代谢调节减缓受CO2浓度升高和水稻中Bt毒素蛋白的不利影响,但长时间环境胁迫下这种应对不利环境的响应机制将会失去作用,最终导致种群适合度的降低。     

Diapause development and acclimation regulating enzymes associated with glycerol synthesis in the Shonai ecotype of the rice stem borer larva,Chilo suppressalis walker

Overwintering larvae of the Shonai ecotype of the rice stem borer, Chilo suppressalis, enter diapause in early September and terminate diapause at the end of October. Cold acclimation at 0°C did not influence glycerol, trehalose or glycogen content in larvae collected on 22 September. Acclimation at 0°C increased the glycerol content and reduced the glycogen content significantly in larvae collected on 2 October and 22 November compared with acclimation at 15°C. These results indicate that overwintering larvae at different phases of diapause development respond differently to the low temperature stimulus for glycerol synthesis. Thus, we evaluated the metabolic rearrangements associated with glycerol synthesis during diapause development and after temperature acclimation. Larvae collected on 2 October were acclimated at 15°C for 15 and 60 days. Some of those acclimated at 15°C were then moved to 0°C for 15 days. The larvae acclimated at 15°C for 15 days were in deep diapause and accumulated little glycerol, while larvae acclimated at 15°C for 60 days were nearly ready to emerge from diapause and accumulated glycerol at 155.5 μmol/g. When larvae acclimated to 15°C for 15 days were transferred to 0°C, glycerol accumulation was stimulated to the same extent (ca 140 μmol/g) as it was in larvae that were acclimated to 15°C for 60 days and then transferred to 0°C. These results indicate that low temperature has a cumulative effect on glycerol production in larvae at different phases of diapause development. Glycerol accumulation was accomplished by activation of glycogen phosphorylase and inhibition of fructose-1,6-bisphosphatase, and activation of enzymes associated with glycerol synthesis, mainly glyceraldehyde-3-phosphatase and polyol dehydrogenase with glyceraldehyde activity.     

抗二化螟的水稻品种筛选

【目的】筛选抗二化螟Chilo suppressalis水稻品种资源,明确其抗虫机理,是农业防治的基础。【方法】采用室内人工接虫为害和大田自然受害相结合的方法,鉴定了不同的水稻品种抗性;同时通过目测不同水稻植株外部形态特征、在网罩内供试品种上接入成虫后2 d的产卵量、幼虫孵化后2 d观察稻株内二化螟的存活率,探讨抗虫品种的抗虫机理。【结果】从344个水稻品种资源和当家品种中筛选出"碑田倒"和"银间杂糯"2个抗虫品种,以及"农飞"和"三香糯"等65个中抗品种,进一步对鉴定出的2个抗虫品种和65个中抗品种进行大田验证,发现符合率达88.06%;其中,抗虫品种"碑田倒"和"银间杂糯"稻株叶舌、叶耳紧贴,叶鞘紧包,脉间距较窄,其植株上的产卵量和幼虫存活率显著少于感虫品种"香稻"、"赣南早"、"三粒寸"、"赣优晚"及中抗品种"赛湖早"、"TKM6"。【结论】在籼稻、粳稻和糯稻中均可筛选出抗虫和中抗品种。其中,糯稻中的抗虫品种和中抗品种较多,占糯稻品种的33.3%;粳稻品种次之,中抗品种占20%,籼稻品种较少,为18.83%。其抗虫机理主要是稻株外部形态特征不利于二化螟蚁螟侵入、成虫产卵量少和幼虫生长发育不良、存活率低。     

Identification of tissues showing the lowest tolerance to freezing in larvae of the rice stem borer, Chilo suppressalis

Abstract.  Even though overwintering larvae of the rice stem borer, Chilo suppressalis , are freeze-tolerant, they cannot survive below −30 °C. Furthermore, nondiapausing larvae cannot survive freezing. However, the cause of death due to freezing is unclear. To identify the cause of death by freezing in larvae, those tissues most injured by low temperatures are identified using the vital stain trypan blue. In overwintering larvae, the midgut of dead larvae stains blue, and remarkable colour density differences between dead and surviving larvae are observed in the midgut. In nondiapausing larvae incubated at −10 °C for several hours, the fat body of dead larvae is strongly stained. Furthermore, increases in mortality with treatment time correspond with increases in the area of the fat body stained. Sterile nondiapausing larvae with lower supercooling points, below −20 °C, do not freeze at −10 °C and survive the treatment. However, all the larvae die when subjected to inoculative freezing at −10 °C, and the fat body stains blue. These results suggest that the midgut in overwintering larvae and the fat body in nondiapausing larvae have the lowest tolerance to freezing.     

Integrated management of the striped rice stem borer,Chilo suppressalis Walker on the Hashemi Tarom rice under the farm conditions

Abstract

An experiment was carried out in order to study the methods of managing striped rice stem borer, Chilo suppressalis Walker on rice, with 11 treatments including the early planted, late planted, Trichogramma brassicae Bezdenko release, 10% Diazinon GR, Bt microbial insecticide, sex pheromone, T. brassicae?+?Bt, T. brassicae?+?sex pheromone, sex pheromone?+?Bt, T. brassicae?+?sex pheromone?+?Bt and control. In this study, the studied traits included the percentage of brownish leaf sheath (BS), dead heart (DH), white head (WH), yield per hectare and the percentage of converting paddy rice to white rice. The results showed that the best yield in all traits was related to the integrated treatment of T. brassicae+sex pheromone?+?Bt, which was in the highest group and its amount in BS was 1.99%, in DH was 1.37%, in WH was 0.34%, the yield/ha was 5910?kg and the percentage of converting paddy rice to white rice was 63%.     

二化螟人工饲料关键因子的优化及其优化配方的饲养效果

采用5因子4水平正交试验,将作者在以前筛选出的二化螟Chilosuppressalis(Walker)人工饲料的5个关键因子进行了进一步的优化,并得出了二化螟人工饲料的优化配方,其成分如下:稻茎粉18.75g、大豆粉15.00g、麦芽粉15.00g、稻糠粉6.25g、茭白茎粉7.50g、干酪素10.00g、酵母粉18.75g、纤维素7.50g、蔗糖15.00g、葡萄糖7.50g、维生素C7.50g、复合维生素B3.00g、胆固醇0.375g、氯化胆碱0.25g、Beck氏盐2.50g、山梨酸1.35g、琼脂13.49g和自来水809.25g。通过连续3代继代饲养试验,比较了该优化配方和二化螟天然饲料水稻茎的饲养效果。结果表明,在一些关键指标上(如存活率、化蛹率和羽化率等),所优化的二化螟人工饲料配方的饲养效果明显的好于其天然饲料水稻茎的;而在许多其它的指标上(如蛹期、成虫期、产卵量、卵期和孵化率等),该优化配方对二化螟的饲养效果与水稻茎的相当;但该配方饲养的二化螟在第3代时幼虫历期明显的比水稻茎的有所延长。     

二化螟性信息素应用技术:笼罩诱捕器和筒形诱捕器

除目前在我国普遍使用的水盆诱捕器 ,本文还介绍了两种诱捕器———笼罩诱捕器和筒形诱捕器。 3种诱捕器的诱蛾效果比较研究表明 :( 1 )笼罩诱捕器中使用铁纱所制的诱笼的诱蛾量较多 ,同时其锥体的锥角以 60°左右为宜 ;( 2 )筒形诱捕器的诱蛾量略高出笼罩诱捕器 ;( 3) 3种诱捕器中 ,水盆诱捕器的诱蛾效果较好 ,筒形诱捕器次之 ,而笼罩诱捕器较差。     

杀虫单、氟虫腈及其混剂对二化螟的防治效果

20 0 0～ 2 0 0 1年在安徽省庐江县进行了 2次药剂及剂量对二化螟的防治和保苗效果试验。试验1表明 ,40 %氟虫腈·杀虫单WP 42 0g(每hm2 有效成分用量 )处理的虫口减退率为 94 7% ,与 5 40g,660g及 5 %氟虫腈 (锐劲特 )SC 3 0g处理相差不大。试验 2表明 ,3 0 %杀虫单·氟虫腈WP 2 70g喷雾即可达到96 2 %的防效 ,接近 3 60g和 5 %氟虫腈SC 2 2 5g的效果。 2次试验中各处理的保苗效果与虫口减退率基本一致。比较药剂成本 ,可见这 2种新型复配剂对二化螟具有良好的防治和保苗效果 ,且上述剂量是经济高效的。     

Molecular characterization and functional analysis of the vitellogenin receptor in the rice stem borer,Chilo suppressalis

As a member of the low-density lipoprotein receptor (LDLR) superfamily, vitellogenin (Vg) receptor (VgR) is responsible for the uptake of Vg into developing oocytes and is a potential target for pest control. Here, a full-length VgR complementary DNA (named as CsVgR) was isolated and characterized in the rice stem borer, Chilo suppressalis. The composite CsVgR gene contained an open reading frame of 5,484 bp encoding a protein of 1,827 amino acid residues. Structural analysis revealed that CsVgR contained two ligand-binding domains (LBDs) with four Class A (LDLR_A) repeats in LBD1 and seven in LBD2, which was structurally different from most non-Lepidopteran insect VgRs having five repeats in LBD1 and eight in LBD2. The developmental expression analysis showed that CsVgR messenger RNA expression was first detectable in 3-day-old pupae, sharply increased in newly emerged female adults, and reached a peak in 2-day-old female adults. Consistent with most other insects VgRs, CsVgR was exclusively expressed in the ovary. Notably, injection of dsCsVgR into late pupae resulted in fewer follicles in the ovarioles as well as reduced fecundity, suggesting a critical role of CsVgR in female reproduction. These results may contribute to the development of RNA interference-mediated disruption of reproduction as a control strategy of C. suppressalis.     

二化螟抗杀虫单和甲胺磷品系的生化特性

二化螟抗性和敏感品系分别采自浙江和安徽太湖。毒力测定结果表明,抗性品系对杀虫单和甲胺磷分别产生了37.7和52.7倍的抗性。代谢酶活性的测定结果显示,抗性品系多功能氧化酶氧脱甲基活性、氮脱甲基活性分别是敏感品系的3.3和1.34倍,而羧酸酯酶活性和谷胱甘肽转移酶活性两个品系之间没有显著差异。说明多功能氧化酶活性提高可能是二化螟对甲胺磷、杀虫单抗性的一个重要机制。为了研究二化螟可能存在的对沙蚕毒素杀虫剂靶标不敏感机制,采用RT-PCR 等分子生物学技术,分别对5个敏感个体和4抗性个体中克隆杀虫单作用的靶标烟碱型乙酰胆碱受体α1亚基（nAChR α subunit 1）cDNA序列进行了分子克隆。序列比较发现一共存在33个单核苷酸的多态性,其中14个引起了编码氨基酸的改变。但是没有发现与抗性有关的特有的碱基突变。     

湖北江汉稻区优质香稻上二化螟的为害特性

研究了二化螟Chilo suppressalis(Walker)对优质香稻鄂香1号和武香988及优质常规中稻鄂中5号的田间为害特性。结果显示,1代二化螟危害造成的枯心苗数优质香稻略低于优质常规中稻。2代二化螟危害造成的枯孕穗和白穗优质香稻鄂香1号低于常规中稻,平均每15丛7.91株。田间剥查发现优质香稻中4~5龄2代二化螟所占百分比高于常规中稻,武香988中2代二化螟蛹所占比例高于常规中稻,达46.51%。3代二化螟量优质香稻低于常规中稻。结果说明香稻受二化螟的危害集中在2代,因而香稻二化螟的防治应注意消灭1代老熟幼虫和2代幼虫。     

扬州地区水稻二化螟寄生蜂种类及主要寄生蜂发生动态

本文报道了江苏扬州地区水稻二化螟Chilo suppressalis(Walker)寄生蜂主要有10种,即中华钝唇姬蜂Eriborus sinicus(Holmgren)、二化螟盘绒茧蜂Cotesia chilonis(Munakata)、稻螟小腹茧蜂Microgaster russata Haliday、螟甲腹茧蜂Chelonus munakatae Munakata、中华茧蜂Amyosoma chinensis(Szepligeti)、螟黄足盘绒茧蜂Cotesia flavipes(Cameron)、夹色姬蜂Auberteterus alternecoloratus(Cushman)、桑蟥聚瘤姬蜂Iseropus(Gregopimpla)kuwanae(Viereck)、螟蛉瘤姬蜂Itoplectis naranyae(Ashmead)以及二化螟盘绒茧蜂的重寄生蜂绒茧灿金小蜂Trichomalopsis apanteloctena(Crawford)。田间系统调查表明:扬州地区7-9月水稻二化螟寄生蜂的主要种类为二化螟盘绒茧蜂、中华钝唇姬蜂、螟甲腹茧蜂、稻螟小腹茧蜂、螟蛉瘤姬蜂、桑蟥聚瘤姬蜂,但这些寄生蜂对一代水稻二化螟幼虫的寄生率较低,对二代水稻二化螟幼虫的寄生率高。水稻二化螟越冬期的系统调查表明:扬州地区二化螟越冬幼虫的寄生蜂主要有3种,即二化螟盘绒茧蜂、中华钝唇姬蜂和稻螟小腹茧蜂,其中二化螟盘绒茧蜂和中华钝唇姬蜂是寄生水稻二化螟越冬幼虫的优势种。水稻二化螟越冬幼虫的寄生率调查表明,扬州地区水稻二化螟越冬幼虫的寄生率在不同年份间有一定的波动,但一般都在15%以上,有的甚至高达89.88%,因此寄生蜂对二化螟越冬幼虫具有很好的控制作用,能有效压低来年水稻二化螟的田间越冬基数。此外,3种寄生蜂出茧时间在1月和2月之间达到最长,分别达到19.79 d、17.26 d和27.00 d,该结果表明,在扬州地区这3种寄生蜂可能都存在滞育现象。     

用正交试验法优选二化螟人工饲料配方

采用15因子2水平正交试验,优选出1种二化螟Chilo suppressalis(Walker)人工饲料配方,筛选出影响二化螟发育的5个关键因子,分别是稻茎粉、麦芽粉、稻糠粉、酵母粉和蔗糖。其中稻茎粉、麦芽粉、酵母粉和稻糠粉影响二化螟的幼虫成活率,蔗糖和麦芽粉影响二化螟的幼虫历期,稻茎粉影响二化螟的蛹重。     

二化螟滞育生物钟蛋白TIME-EA4基因的克隆及时空和温度诱导表达分析

【目的】生物测时蛋白TIME-EA4是昆虫特有的一种滞育生物钟蛋白。本文旨在克隆二化螟Chilo suppressalis TIME-EA4基因,研究其在二化螟不同发育时期和不同组织中的时空表达模式及在不同田间种群和不同温度诱导下的滞育和非滞育种群中的表达水平。【方法】通过RACE技术克隆二化螟TIME-EA4基因全长c DNA序列,利用荧光定量PCR的方法检测TIME-EA4基因在二化螟不同发育时期、6龄幼虫不同组织中的表达模式,及在不同季节采集的田间种群和不同温度处理1h后的滞育和非滞育种群6龄幼虫中的表达量变化。【结果】从二化螟中克隆获得TIME-EA4的c DNA序列(Gen Bank登录号:KU356855),全长821 bp,开放阅读框516 bp,编码172个氨基酸;TIME-EA4基因在二化螟不同发育时期和不同组织中均有表达,且在早期蛹、雌雄成虫以及6龄幼虫的头部、脂肪体和中肠中表达水平较高,同时在滞育幼虫组织中的表达高于非滞育幼虫。此外,TIME-EA4基因在滞育种群中的表达量是非滞育种群的3倍左右,同时在非滞育种群中,该基因的表达明显受10℃及以下低温的诱导,且在10℃时表达量最高,而在滞育种群中该基因的表达受温度影响不明显。【结论】结果说明TIME-EA4基因与二化螟的抗低温胁迫以及低温滞育相关。本研究为TIME-EA4在农业害虫滞育的分子作用机制研究奠定了基础。     

Influence of soil moisture on supercooling capacity and associated physiological parameters of overwintering larvae of rice stem borer

Influence of soil moisture on the supercooling capacity and associated physiological parameters of overwintering larvae of the rice stem borer Chilo suppressalis was examined by exposing larvae to soil moistures of 25, 50, 75 and 100% of saturated soil water content (SSWC) at ambient temperature for 30 d from December 2007 to January 2008 in Beijing, China. At the end of the exposure, supercooling points (SCPs) varied significantly among the treatments, the lowest being in the larvae exposed to soil moisture of 25% SSWC. Fresh weight was significantly higher in the larvae exposed to soil moisture of 100% SSWC than in those kept at 25 and 50% SSWC. Dry weight and body water content (% fresh weight) were not different among the treatments. Glucose and trehalose contents were markedly lower, and glycerol content was significantly higher in the larvae confined to soil moisture of 25% SSWC than in those exposed to the other soil moisture treatments. It is suggested that variation in body water content (% fresh weight) contributes to the differences in SCPs of the overwintering C. suppressalis larvae in all treatments, but the influence of soil moisture treatments on supercooling capacity are caused through changes in glycerol content.     

高温对二化螟实验种群生长、存活和繁殖的影响

以2 7℃和30℃恒温为对照,研究了平均温度在2 7～2 9.2℃内的5种变温条件下(每天2 0 h的适温2 7℃和12 :0 0～16 :0 0的高温处理,分别为日最高气温2 7℃、30℃、33℃、36℃、4 0℃)二化螟实验种群的发育历期、存活率和繁殖力的影响,组建了特定年龄生命表和繁殖力生命表。结果表明高温可导致二化螟种群的发育速率延缓、存活率和生殖力下降,主要表现在幼虫历期随日最高温度的升高而延长,36℃时延至12 d以上;在日最高气温高于33℃时,其存活率从2 7℃时的30 .3%降至36℃时的13.1%和4 0℃时的5 .5 % ,繁殖力也从30℃时每雌产卵量14 8.7粒减至36℃时的70 .5粒。因此,夏季异常高温可能是二化螟田间种群数量增长的一个重要抑制因子     

二化螟与稻纵卷叶螟幼虫取食诱导的水稻挥发物比较

前期研究结果表明,二化螟和稻纵卷叶螟幼虫为害所诱导的水稻挥发物能对二化螟绒茧蜂Apanteles chilonis Munakata、二化螟Chilo suppressalis(Walker)和稻纵卷叶螟Cnaphalocrocis medinalis(Guene)雌成虫的行为产生影响,并且两者的影响效应存在明显差异,但至今仍不清楚2种害虫危害后水稻挥发物的变化情况。利用动态吸附法对健康水稻苗、二化螟或稻纵卷叶螟幼虫为害苗的挥发物进行捕集与鉴定。在3种处理水稻苗中,共捕集到28种挥发物组分,包括萜类化合物、烷烃类化合物、酮类、醇类、水杨酸甲酯和一些未知化合物。二化螟和稻纵卷叶螟为害均能导致水稻挥发物总量和大多挥发物组分释放量的上升;同时,二化螟幼虫危害能诱导水稻新释放2-heptanonol,(E)-linalooloxide和2种未知挥发物组分,稻纵卷叶螟为害则可诱导水稻产生2-heptanonol,(E)-linalooloxide,MeSA和β-caryophyllene4种新化合物。2种虫害稻株间的比较表明,β-caryophyllene和MeSA是稻纵卷叶螟危害水稻后诱导产生的特有挥发物组分,并且linalool,α-copaene,(+)-cedrol,n-heptadecane和n-nonadecane在二化螟为害苗与稻纵卷叶螟为害苗之间差异显著。这一结果也许可以解释2种害虫诱导水稻挥发物对天敌和害虫行为影响不同的原因。