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。     

二化螟人工饲料研究初报

在温度为(29±1)℃,相对湿度为80%~90%的条件下,分别以3种人工饲料和天然饲料(新鲜水稻茎杆)对二化螟Chilo suppressalisWalker进行连续继代饲养。结果表明,3种人工饲料饲养的二化螟幼虫的生长历期、化蛹率、蛹重、蛹期及羽化率等均与天然饲料的基本接近,而且这3种人工饲料配制简便、成本较低,也不易霉变,是3种较为理想的人工饲料。通过第2代和第3代的继代繁殖表明,3种人工饲料和天然饲料饲养的二化螟的发育情况都稍有下降,但3种人工饲料与天然饲料间不存在显著差异。     

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。     

水稻二化螟抗药性监测方法

本文首先介绍了二化螟Chilo suppressalis(Walker)抗药性监测的方法:点滴法和稻苗浸渍法。其中点滴法已有标准化的生测规程,其适用于以触杀作用为主的杀虫剂抗性监测,但无法准确反映以胃毒作用为主的杀虫剂的毒力效果,因此,本文新建了以胃毒作用为主的双酰胺类或微生物杀虫剂Bt等的抗性监测方法———稻苗浸渍法。     

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。     

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。     

杀虫剂不同处理对二化螟为害、防治效果及水稻产量的影响

近年来,二化螟发生出现各代均偏重或大发生的新特点。为探索杀虫剂防治二化螟的新策略,采用田间小区试验在早晚稻田中分别研究5%氟虫腈加25%灭.水胺的不同施药时期和次数对二化螟幼虫的为害和防治效果及对水稻产量的影响。在试验地,二化螟第1和第2代发生在早稻田,第3和第4代发生在晚稻田。试验结果表明,在第1和第3代二化螟幼虫孵化高峰施药1次、隔10d再施药1次对早稻田和晚稻田的幼虫防效显著,分别达到了92.4%和92.6%。在早、晚稻田分别发生的第1和第3代2龄幼虫高峰期施药1次、同时在次代孵化高峰期再施药1次,均可显著降低虫伤株率和白穗株率,提高保穗效果,并显著提高单位面积上的水稻产量(3.98%~4.20%)。因此,对第1至第4代二化螟幼虫均进行1次药剂防治是二化螟各代均大发生的新形势下的防治策略。     

用甜玉米粒饲养二化螟实验种群

介绍一种能够继代饲养二化螟Chilo suppressalis(Walker)实验种群的天然饲料---"甜玉米粒"及相应饲养方法。结果显示,甜玉米粒和马蹄的饲养成绩皆不同程度地高于稻苗、甘蔗和田间种群。取食甜玉米粒的幼虫存活率达92.0%,显著高于其余3种天然饲料。平均蛹重为60.7mg,单雌产卵量达197.4,除性比为0.9∶1之外,其重要生物学指标和此前报道的饲养成绩最好的马蹄没有显著差异,但成本更低,且更利于清洗消毒等加工处理,因而更适于在实际生产中大量饲养二化螟实验种群。     

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。     

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.     

Nutritional studies of the rice stem borer,Chilo suppressalis Walker,and its mass rearing

The nutritional requirements of the rice stem borer,Chilo suppressalis Walker were studied by rearing on synthetic food media in relation to the constituents of the rice plant. The larvae required amino acids, vitamins, carbohydrates, cholesterol, minerals and water extract of the rice plant for their growth. Among amino acids, arginine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophane and valine were indispensable for the growth, and these amino acids could not be synthesized in the larval body. Among vitamines, thiamine, riboflavin, nicotinic acid, pyridoxine, Ca-pantothenate, folic acid, biotin, cholin chloride and ascorbic acid were essential for the growth. Cholesterol is the only essential nutrient among fat-soluble factors. Fructose, glucose and sucrose showed high nutritive values among carbohydrates. Addition of the water extract of the rice plant to the media improved the development of the insects, but the excess amounts of it resulted inhibition of the growth. Two aromatic acids, benzoic and salicylic acids, were isolated as the growth inhibitory substance. p-methyl acetophenone isolated from the rice plant seemed to play an important role for the attraction of the insect. Mass rearing of the rice stem borer in a laboratory was carried out on two kinds of foods: living rice plant seedlings and synthetic food media. The rearing the insects on rice seedlings is easy if rice seeds are available. The rearing on synthetic food media is modified to simplify from these for the nutritional study. The rearing practices, merit and demerit of both methods are discussed.     

A comparison of the larval overwintering biology of the striped stem borer, Chilo suppressalis (Lepidoptera: Crambidae), in rice and water-oat fields

It has been suggested that the striped stem borer, Chilo suppressalis (Walker), has developed distinct populations inhabiting rice Oryza sativa (Linnaeus) and water-oats Zizania latifolia (Turcz). In this study, we compared several biological traits between overwintering larvae from rice and water-oat fields during the winter of 2010. The parasitism rate was significantly higher in the larvae that overwintered in the rice field than in those that overwintered in the water-oat field. The average body weight of overwintering larvae and the weight of 3-day-old pupae, as well as the number of eggs carried by adult females, were all significantly greater in the samples from the water-oat field. The diapause intensity, defined as the number of days required until pupation by overwintering larvae transferred to 25 °C and L:D 16:8 h, decreased progressively from January to March but decreased more rapidly in the water-oat field. Our results demonstrate differences in the overwintering biology of C. suppressalis larvae from rice and water-oat fields. These differences may contribute to the differentiation and reproductive isolation between host-plant populations in this species.     

In vitro insect-feeding bioassay to determine the resistance of transgenic rice plants transformed with insect resistance genes against striped stem borer (Chilo suppressalis)

Summary To determine the degree of insect resistance in transgenic plants, different bioassays are used which typically use either whole plant or small pieces of leaves or stems of transgenic plants, following culture under greenhouse conditions. An in vitro insect-feeding bioassay is presented which permits the infestation of transgenic plantlets with newly hatched larvae from the striped stem borer. The bioassay consists of the germination of rice seeds in vitro using Murashige and Skoog medium in test tubes, and then infestation of each 3–4 cm long seedling with one neonate larva obtained from surfacesterilized eggs of Chilo suppressalis. The infested in vitro plantlets are kept in culture rooms at 25°C for several days and then the seedling damage and the growth of the larvae are analyzed. Senia (japonica variety) homozygous transgenic rice plants were used for these experiments. The plants were transformed with either the cry1B or the maize proteinase inhibitor (mpi) genes. Both genes confer resistance to Chilo suppressalis. With non-transformed plants the larvae grew and developed normally, feeding on the small rice plantlets. In contrast, with cry1B plants, the neonate larvae died during the first days of the infestation. These plantlets recovered completely and developed similarly to the non-infested control plants. With transgenic plants transformed with the mpi gene, the neonate larvae did not die but grew more slowly compared with the controls. Thus, this in vitro insect-feeding bioassay is a rapid and easy method to detect the resistance of cry and mpi transgenic plants to stem borers such as Chilo suppressalis.     

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.     

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.