水稻螟虫神经肽PBAN及其受体序列的生物信息学分析

【目的】性信息素合成激活肽(PBAN)是控制昆虫产生性信息素的激素,本文旨在分析水稻螟虫神经肽PBAN及其受体的序列。【方法】通过t Blastn同源检索从水稻螟虫基因组和转录组数据库中鉴定水稻螟虫PBAN神经肽及其受体序列,在此基础上进行序列比对及系统发生分析。【结果】发现二化螟Chilo suppressalis、三化螟Tryporyza incertulas和大螟Sesamia inferens的PBAN成熟肽序列均含有33个氨基酸残基,其C端五肽序列完全相同,3种水稻螟虫PBAN多肽相似度为54.55%~63.64%;发现二化螟PBAN受体3个异构体全长氨基酸序列(PBANR-A、PBANR-B和PBANR-C),均含有7个跨膜区域。【结论】进化树分析发现不同昆虫PBAN神经肽及其受体存在一定的保守性和多样性,并且在进化树上的位置几乎与昆虫系统发育分类一致,推测PBAN神经肽和PBAN受体在昆虫系统进化过程中可能存在协同进化现象。本研究为水稻螟虫PBAN神经肽及其受体的结构和功能分析提供基础。     

水稻钻蛀性螟虫田间调查及测报技术

长期以来,水稻钻蛀性螟虫都是我国水稻上的重要害虫。近年来,随着水稻栽培制度的变更及全球性气候的变化,种群数量逐渐回升,为害日趋严重。因此,科学的调查方法和准确的预测预报,对该类害虫的综合治理具有重要意义。本文根据大螟Sesamia inferens(Walker)、二化螟Chilo suppressalis(Walker)和三化螟Scirpophaga incertulas(Walker)的生物学特性,总结了这3种水稻钻蛀性螟虫的田间调查方法;阐述了它们的发生期预测方法;探讨了在调查取样过程中的注意事项,以期为我国水稻钻蛀性螟虫的综合防治提供可靠的数据支撑。     

Enhanced resistance to two stem borers in an aromatic rice containing a synthetic cryIA(b) gene

Rice cultivars of isozyme group V include high-quality, aromatic rices that are difficult to improve by traditional methods because of the loss of quality characters upon sexual hybridization. Their low-tillering plant type predisposes them to economic loss from attack by stem borers, a group of insects to which they are susceptible. We report here the enhancement of stem borer resistance in cv. Tarom Molaii through transformation by microprojectile bombardment. Embryogenic calli derived from mature seeds were bombarded with gold particles coated with plasmid pCIB4421, carrying a synthetic truncated toxin gene based on the cryIA(b) gene from Bacillus thuringiensis, and plasmid pHygII, carrying the hygromycin phosphotransferase (hpt) selectable marker gene. Inclusion of 50 mg/l hygromycin B in culture media from bombardment through to rooting of plantlets eliminated escapes. The procedure generated three independent hpt transformants of which two also contained the cryIA(b) gene. One such line (No. 827) produced truncated (67 kDa) CryIA(b) protein equivalent to about 0.1% of total soluble protein. The cryIA(b) gene was controlled by the promoter of the maize C₄ PEP carboxylase gene and was expressed in leaf blades but was not expressed to a detectable level in dehulled mature grain. Line 827 contained about 3 copies of the cryIA(b) gene which segregated as a single dominant Mendelian locus in the second (T₁) and third (T₂) generations and co-segregated with enhanced resistance to first-instar larvae of striped stem borer (Chilo suppressalis) and yellow stem borer (Scirpophaga incertulas). T₂ line 827-6 homozygous for the cryIA(b) gene showed no dead hearts or whiteheads after infestation with stem borers, whereas T₂ line 827-25 lacking the gene averaged 7 dead hearts per plant and 2.25 whiteheads per plant. These results establish that transformation of high-quality rices of group V is a feasible alternative to sexual hybridization.     

越桔两种蛀干害虫的发生与防治

咖啡豹蠹蛾Zeuzera coffeae Nietner和樟泥色天牛Uraecha angusta(Pascoe)是江苏地区的美国兔眼越桔上2种主要蛀干害虫。前者在枝干中向顶端方向蛀食,排出圆粒状虫粪;后者向植株基部方向蛀食,排出粉末状虫粪,均造成枝干枯死。咖啡豹蠹蛾发生量较大,是主要虫种。防治这2种蛀干害虫,应采用保护利用天敌、改善果园生态环境、促使树体健壮生长等综合管理措施。     

Seasonal infestations of two stem borers (Lepidoptera: Crambidae) in noncrop grasses of Gulf Coast rice agroecosystems

Infestations of two stem borers, Eoreuma loftini (Dyar) and Diatraea saccharalis (F.) (Lepidoptera: Crambidae), were compared in noncrop grasses adjacent to rice (Oryza sativa L.) fields. Three farms in the Texas rice Gulf Coast production area were surveyed every 6-8 wk between 2007 and 2009 using quadrat sampling along transects. Although D. saccharalis densities were relatively low, E. loftini average densities ranged from 0.3 to 5.7 immatures per m(2) throughout the 2-yr period. Early annual grasses including ryegrass, Lolium spp., and brome, Bromus spp., were infested during the spring, whereas the perennial johnsongrass, Sorghum halepense (L.) Pers., and Vasey's grass, Paspalum urvillei Steud., were infested throughout the year. Johnsongrass was the most prevalent host (41-78% relative abundance), but Vasey's grass (13-40% relative abundance) harbored as much as 62% of the recovered E. loftini immatures (during the winter). Young rice in newly planted fields did not host stem borers before June. April sampling in fallow rice fields showed that any available live grass material, volunteer rice or weed, can serve as a host during the spring. Our study suggests that noncrop grasses are year-round sources of E. loftini in Texas rice agroecosystems and may increase pest populations.     

Field evaluation of resistance of transgenic rice containing a synthetic cry1Ab gene from Bacillus thuringiensis Berliner to two stem borers

Two transgenic rice (Oryza sativa L.) lines, KMD1 and KMD2 at the R4 generation, transformed with a synthetic cry1Ab gene from Bacillus thuringiensis Berliner, were first evaluated for stem borer resistance in the field during the rice growing season of 1998 in two areas of Zhejiang Province, China. Both KMD1 and KMD2 were highly resistant to the stem borers Chilo suppressalis (Walker) and Scirpophaga incertulas (Walker), and were completely undamaged during the whole rice growing season. In contrast, damage to the plants of the untransformed parental control (Xiushui 11) was in the form of deadhearts or whiteheads. Under natural infestation by the C. suppressalis, the damage to control plants reached a peak of 88.7% of plants and 20.1% of tillers encountered with deadhearts. Under artificial and natural infestation of neonate striped stem borers at the vegetative stage and booting stage, 100% of plants and 25.6% of tillers, 78.9% of plants and 15.6% of productive tillers among artificially infested control plants were observed with the symptom of deadhearts and whiteheads, respectively. Damage to the control plants from artificial infestation by the S. incertulas reached a peak of 97.0% of plants and 22.9% of tillers damaged. The field research indicated that both KMD1 and KMD2 show great potential for protecting rice from attack by these two stem borers.     

亚致死剂量茚虫威和氯虫苯甲酰胺对水稻螟虫营养指标的影响

水稻钻蛀性害虫主要包括二化螟Chilo suppressalis和大螟Sesamia inferens。目前,对于二化螟和大螟的防治仍主要采用化学药剂。但是,在杀虫药剂的胁迫下,部分螟虫会接触到低剂量的杀虫药剂,导致部分个体产生亚致死效应。为明确亚致死剂量茚虫威和氯虫苯甲酰胺是否对水稻螟虫取食有刺激作用,本实验通过稻苗浸渍法测定这两种农药对水稻螟虫的毒力,确定亚致死剂量LC_(10)和LC_(25),在此基础上分析了两种药剂亚致死剂量对螟虫取食水稻3 d后营养指标的影响。结果表明,茚虫威对二化螟和大螟的LC_(10)分别为5.420、4.707 mg/L;LC_(25)分别为10.246、8.462 mg/L。氯虫苯甲酰胺对二化螟和大螟的LC_(10)分别为7.601、5.517 mg/L;LC_(25)分别为14.361、10.981 mg/L。生长发育和营养指标测定发现,亚致死剂量茚虫威处理二化螟幼虫后,除YLC_(10)处理组的近似消耗率(AD)显著高于对照组外,其余指标与对照组比均无显著差异。亚致死剂量氯虫苯甲酰胺处理二化螟幼虫后,除LLC_(25)处理组的体重增加量与对照组无显著差异外,其他处理组食物消耗量和体重增加量均显著上升;除LLC_(25)组的粗生长率(ECI)与对照组无显著差异外,其他处理组近似消耗率和ECI均显著下降。LLC_(10)处理组二化螟幼虫的相对消耗速率(RCR)显著高于对照组。亚致死剂量茚虫威和氯虫苯甲酰胺处理大螟幼虫后,体重增加量显著高于对照组;茚虫威YLC_(10)和氯虫苯甲酰胺LLC_(25)处理组的相对生长速率(RGR)、相对消耗速率(RCR)显著高于对照组。对比各组干物质积累曲线,我们发现除氯虫苯甲酰胺LLC_(10)处理组外,其余处理组的二化螟幼虫干/鲜比大于对照组,说明这些处理组幼虫取食水稻后干物质积累比对照组快;除茚虫威LC_(25)处理组外,其余处理组的大螟幼虫取食水稻后干物质积累比对照组快。研究结果初步证明了亚致死剂量(LC_(10)、LC_(25))茚虫威和氯虫苯甲酰胺对水稻螟虫取食、营养、消耗等指标存在一定影响,某种程度上能够刺激水稻螟虫的取食和生长发育。     

Resistance to stem borers (Lepidoptera: Crambidae) among Texas rice cultivars

A 4-yr field study was conducted to assess the resistance of rice, Oryza sativa L., cultivars to injury from the sugarcane borer, Diatraea saccharalis (F.), and the Mexican rice borer, Eoreuma loftini (Dyar) (both Lepidoptera: Crambidae). Several cultivars showed reduced levels of injury ('CLXL8', 'XL8', 'Wells', 'Cheniere', and 'XP710' in 2003; CLXL8, XP723, Cheniere, and 'CL161' in 2004) and lower stem borer yield loss (CLXL8 in 2004) than the more susceptible 'Priscilla'. The resistant CLXL8 also had less injury and yield loss in 2004 and higher yield in 2003 than 'Cocodrie', currently the most popular rice cultivar in Texas and Louisiana. Linear regression analyses between whiteheads and main crop rice yield showed steeper negative slopes for the more resistant cultivars than the more susceptible cultivars, indicating a greater yield loss per whitehead on the resistant cultivars. Compensation from insect injury likely explains the positive associations established in our study between main crop yield and whiteheads for some cultivars. Cultivar resistance is anticipated to become a major control tactic in reducing infestations of E. loftini and D. saccharalis in the Texas and Louisiana rice industries.     

Transgenic rice plants expressing a trypsin inhibitor are resistant against rice stem borers, Chilo suppressalis

A synthetic gene, mwti1b, coding for a winged bean trypsin inhibitor WTI-1B, has been introduced and expressed in rice plants, Oryza sativa. Protein extracts from transgenic rice plants expressing the trypsin inhibitor inhibited the gut proteases of larvae of the serious insect pest, the rice stem borer, Chilo suppressalis (Lepidoptera: Pyralidae) in vitro. The growth of larvae reared on transgenic rice plants expressing WTI-1B at more than 1 ng/10 g total protein was significantly retarded compared to that on non-transgenic control plants.     

Bacillus thuringiensis delta-endotoxin binding to brush border membrane vesicles of rice stem borers

The receptor binding step in the molecular mode of action of five delta-endotoxins (Cry1Ab, Cry1Ac, Cry1C, Cry2A, and Cry9C) from Bacillus thuringiensis was examined to find toxins with different receptor sites in the midgut of the striped stem borer (SSB) Chilo suppressalis (Walker) and yellow stem borer (YSB) Scirpophaga incertulas (Walker) (Lepidoptera: Pyralidae). Homologous competition assays were used to estimate binding affinities (K(com)) of (125)I-labelled toxins to brush border membrane vesicles (BBMV). The SSB BBMV affinities in decreasing order was: Cry1Ab = Cry1Ac > Cry9C > Cry2A > Cry1C. In YSB, the order of decreasing affinities was: Cry1Ac > Cry1Ab > Cry9C = Cry2A > Cry1C. The number of binding sites (B(max)) estimated by homologous competition binding among the Cry toxins did not affect toxin binding affinity (K(com)) to both insect midgut BBMVs. Results of the heterologous competition binding assays suggest that Cry1Ab and Cry1Ac compete for the same binding sites in SSB and YSB. Other toxins bind with weak (Cry1C, Cry2A) or no affinity (Cry9C) to Cry1Ab and Cry1Ac binding sites in both species. Cry2A had the lowest toxicity to 10-day-old SSB and Cry1Ab and Cry1Ac were the most toxic. Taken together, the results of this study show that Cry1Ab or Cry1Ac could be combined with either Cry1C, Cry2A, or Cry9C for more durable resistance in transgenic rice. Cry1Ab should not be used together with Cry1Ac because a mutation in one receptor site could diminish binding of both toxins.     

Effect of transgenic Bacillus thuringiensis rice lines on mortality and feeding behavior of rice stem borers (Lepidoptera: Crambidae)

Ten transgenic Bacillus thuringiensis Bt rice, Oryza sativa L., lines with different Bt genes (two Cry1Ac lines, three Cry2A lines, and five Cry9C lines) derived from the same variety Minghui 63 were evaluated in both the laboratory and the field. Bioassays were conducted by using the first instars of two main rice lepidopteran insect species: yellow stem borer, Scirpophaga incertulas (Walker) and Asiatic rice borer, Chilo suppressalis (Walker). All transgenic lines exhibited high toxicity to these two rice borers. Field evaluation results also showed that all transgenic lines were highly insect resistant with both natural infestation and manual infestation of the neonate larvae of S. incertulas compared with the nontransformed Minghui63. Bt protein concentrations in leaves of 10 transgenic rice lines were estimated by the sandwich enzyme-linked immunosorbent assay. The cry9C gene had the highest expression level, next was cry2A gene, and the cry1Ac gene expressed at the lowest level. The feeding behavior of 7-d-old Asiatic rice borer to three classes of Bt transgenic rice lines also was detected by using rice culm cuttings. The results showed that 7-d-old larvae of Asiatic rice borer have the capacity to distinguish Bt and non-Bt culm cuttings and preferentially fed on non-Bt cuttings. When only Bt culm cuttings with three classes of different Bt proteins (CrylAc, Cry2A, and Cry9C) were fed, significant distribution difference of 7-d-old Asiatic rice borer in culm cuttings of different Bt proteins also was found. In the current study, we evaluate different Bt genes in the same rice variety in both the laboratory and the field, and also tested feeding behavior of rice insect to these Bt rice. These data are valuable for the further development of two-toxin Bt rice and establishment of appropriate insect resistance management in the future.     

Differential metal selectivity and gene expression of two zinc transporters from rice

Zinc is an essential mineral for a wide variety of physiological and biochemical processes. To understand zinc transport in cereals, we identified putative zinc transporters in gene databases. Three full-length cDNAs were identified and characterized from rice (Oryza sativa). Two of the cDNAs partially complemented a yeast (Saccharomyces cerevisiae) mutant deficient in zinc uptake at low concentrations. The two transporters showed many similarities in function but differed in ionic selectivity and pH optimum of activity. Expression patterns also differed between the two genes. One gene was broadly expressed under all conditions, and the other gene was mainly induced by zinc deficiency to higher levels in roots than in leaves. Although the timing of expression differed between the two genes, localization of expression overlapped in roots. Comparisons of the protein sequences, ionic selectivity, and gene expression patterns of the two transporters suggest that they may play different roles in the physiology of the whole plant.     

Differential sensitivity of two insect GABA-gated chloride channels to dieldrin,fipronil and picrotoxinin

In the central nervous system of both vertebrates and invertebrates inhibitory neurotransmission is mainly achieved through activation of γ-aminobutyric acid (GABA) receptors. Extensive studies have established the structural and pharmacological properties of vertebrate GABA receptors. Although the vast majority of insect GABA-sensitive responses share some properties with vertebrate GABAA receptors, peculiar pharmacological properties of these receptors led us to think that several GABA-gated chloride channels are present in insects. We describe here the pharmacological properties of two GABA receptor subtypes coupled to a chloride channel on dorsal unpaired median (DUM) neurones of the adult male cockroach. Long applications of GABA induce a large biphasic hyperpolarization, consisting of an initial transient hyperpolarization followed by a slow phase of hyperpolarization that is not quickly desensitized. With GABA, the transient hyperpolarization is sensitive to picrotoxinin, fipronil and dieldrin whereas the slow response is insensitive to these insecticides.When GABA is replaced by muscimol and cis-4-aminocrotonic acid (CACA) a biphasic hyperpolarization consisting of an initial transient hyperpolarization followed by a sustained phase is evoked which is blocked by picrotoxinin and fipronil. Exposure to dieldrin decreases only the early phase of the muscimol and CACA-induced biphasic response, suggesting that two GABA-gated chloride channel receptor subtypes are present in DUM neurones. This study describes, for the first time, a dieldrin resistant component different to the dieldrin- and picrotoxinin-resistant receptor found in several insect species.     

Diversity and abundance of wild host plants of lepidopteran stem borers in two different agroecological zones of Kenya

A survey was carried out between 2004 and 2005 in two ecologically different locations, Kakamega and Muhaka to assess diversity and abundance of wild host plants of lepidopteran stem borers as compared to maize plots during the cropping and non-cropping seasons. Kakamega in Western Kenya is characterized by a Guineo-Congolian rain forest mosaic and Muhaka at the Kenyan coast by a Zanzibar Inhambane mosaic with secondary grassy and woody vegetation. In Kakamega, wild host plants and maize covered 2 and 43% of the surveyed area. No variation in diversity and relative abundance of wild host plants was observed between both the cropping and non-cropping seasons. In Muhaka, the diversity and relative abundance of wild host plant species differed between seasons, with the Shannon Weaver Index (H) of 1.67 and 0.95 for cropping and non-cropping seasons, respectively. Similarly in this location, wild host plant cover varied between cropping (23%) and noncropping (17.9%). During both seasons, this was higher than the maize cover, with 10.7% and 0% for the cropping and non-cropping seasons, respectively. For both localities, the implication of the differences found in the abundance and diversity between the cropping and non-cropping seasons is discussed.     

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。     

水稻螟虫诱杀植物香根草的发现与应用

香根草Vetiveria zizanioides L.是一种多年丛生的草本植物。近年来研究发现香根草能够有效诱集水稻螟虫(二化螟Chilo suppressalis和大螟Sesamia inferens)雌成虫在其上产卵,但孵化出的幼虫在香根草上不能完成生活史。因此,香根草对于水稻螟虫来讲是一种诱杀植物。开发以香根草为基础的水稻螟虫绿色防控技术,不仅可以有效控制水稻螟虫,减少农药使用,还可以保育寄生蜂等天敌、增强生物多样性。香根草同时具有适应性强、种植管理轻简、无性繁殖不会扩散成为杂草、可供观赏等应用特点。因此,利用诱杀植物香根草为水稻螟虫防治提供了一种生态调控新途径,还可实现农民增收、改善生态环境,具有重要的经济、生态和社会效益。     

Characterization of the complete mitochondrial genome of Chilo auricilius and comparison with three other rice stem borers

The mitogenome of Chilo auricilius (Lepidoptera: Pyraloidea: Crambidae) was a circular molecule made up of 15,367 bp. Sesamia inferens, Chilo suppressalis, Tryporyza incertulas, and C. auricilius, are closely related, well known rice stem borers that are widely distributed in the main rice-growing regions of China. The gene order and orientation of all four stem borers were similar to that of other insect mitogenomes. Among the four stem borers, all AT contents were below 83%, while all AT contents of tRNA genes were above 80%. The genomes were compact, with only 121–257 bp of non-coding intergenic spacer. There are 56 or 62-bp overlapping nucleotides in Crambidae moths, but were only 25-bp overlapping nucleotides in the noctuid moth S. inferens. There was a conserved motif ‘ATACTAAA’ between trnS2 (UCN) and nad1 in Crambidae moths, but this same region was ‘ATCATA’ in the noctuid S. inferens. And there was a 6-bp motif ‘ATGATAA’ of overlapping nucleotides, which was conserved in Lepidoptera, and a 14-bp motif ‘TAAGCTATTTAAAT’ conserved in the three Crambidae moths (C. suppressalis, C. auricilius and T. incertulas), but not in the noctuid. Finally, there were no stem-and-loop structures in the two Chilo moths.     

Differential responses of two rice varieties to salt stress

Two rice varieties, viz. Nonabokra and Pokkali, have been evaluated for their responses to salinity in terms of some physiological and biochemical attributes. During the exposure to salinity (200 mM concentration of sodium chloride for 24, 48, and 72 h), a significant increase in sodium was recorded which was also concomitant with the changes of other metabolic profiles like proline, phenol, polyamine, etc. The protein oxidation was significantly increased and also varied between the two cultivars. The changes in activities of anti-oxidative enzymes under stress were significantly different to the control. The detrimental effects of salinity were also evident in terms of lipid peroxidation, chlorophyll content, protein profiles, and generation of free radicals; and these were more pronounced in Pokkali than in Nonabokra. The assessment and analysis of these physiological characters under salinity could unravel the mechanism of salt responses revealed in this present study and thus might be useful for selection of tolerant plant types under the above conditions of salinity.     

Diversity and abundance of lepidopteran stem borers and their host plants in Ethiopia

Lepidopteran stem borers are among the most important insect pests of maize, sorghum and sugarcane in sub‐Saharan Africa. Except for Chilo partellus, the other stem borer pests in Ethiopia are indigenous to Africa and are assumed to have coevolved with some native grasses and sedges in the natural habitat. In addition to pest species, natural habitats harbour diverse non‐economic stem borer species, some of which are new to science. However, with the growing threats to natural habitats, some non‐economic stem borer species may switch or expand their host ranges to include cultivated crops and evolve as “new” pests. Besides host switch, some of the unknown species currently limited to natural habitats may disappear. We examined the diversity, abundance and interactions of lepidopteran stem borers and their wild host plants in five different vegetation mosaics in Ethiopia. The stem borer species diversity varied among vegetation mosaics and host plants. Forty‐four stem borer species belonging to 14 different genera in the families of Noctuidae, Crambidae, Pyralidae and Tortricidae were recorded from 34 wild host plants and through light trap. Among these families, Noctuidae was the highest in species richness in which 31 species were identified, out of which 15 species and two genera were new to science. This paper discusses the ecological interpretation of host plant–stem borer species interactions, particularly in relation to habitat disturbances.     

Species diversity and distribution of lepidopteran stem borers in South Africa and Mozambique

Country‐wide surveys of lepidopteran stem borers in wild host plants were undertaken between 2006 and 2009 in South Africa and 2005 and 2010 in Mozambique. A total of 4438 larvae were collected from 65 wild host plants in South Africa and 1920 larvae from 30 wild host plants in Mozambique. In South Africa and Mozambique, 50 and 39 stem borer species were recovered, respectively, with four new species and two new genera among noctuids. Less than 5% of the total number of species collected are considered to be economically important in Africa. These species were Busseola fusca (Fuller) (Noctuidae), Chilo partellus (Swinhoe) (Crambidae) and Sesamia calamistis Hampson (Noctuidae). Data from this study and others in East Africa on the very low abundance of stem borers in wild host plants question the putative role of wild host plants as reservoir for stem borer pests. One new host plant family (Prioniaceae), as well as 24 and 13 wild hosts from South Africa and Mozambique respectively, was added to the list of known hosts in Africa.