转Bt基因棉杀虫蛋白含量时空分布及对棉铃虫产生抗性的影响

采用ＥＬＩＳＡ法（酶链免疫法）、室内初孵棉铃虫生测法和田间棉铃虫为害调查方法，研究和分析了中国构建的 Ｂｔ（Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）基因抗虫棉品系ＧＫ３和美国构建的Ｂｔ基因棉品系新棉３３Ｂ的不同生育期以及花铃期不同器官的杀虫蛋白含量、校正死亡率和田间受害率变化趋势以及它们之间的关系。结果表明，转Ｂｔ基因棉Ｂｔ杀虫蛋白含量在棉花生育过程中呈时空动态变化，在时间分布上，各生育期顶尖平展叶表现为：初花期＞蕾期、花铃期＞苗期＞吐絮期；在花铃期，各器官表现为：功能叶、茎尖＞小蕾、幼铃＞花蕊、花瓣、苞叶、老叶。室内生测幼虫校正死亡率与棉株Ｂｔ杀虫蛋白含量高度一致；田间表现与Ｂｔ杀虫蛋白含量有一定的差异，除主要受Ｂｔ杀虫蛋白影响外，棉铃虫取食选择性，以及残存高龄幼虫为害和棉株各部位营养结构等也影响其抗虫性，造成后期棉铃虫主要为害花、蕾和幼铃，两材料抗虫性表现较为一致。     

棉铃虫对Bt毒蛋白(Cry1 Ac)抗性品系的选育

1 997～2000年,用含Bt毒蛋白(Cry1Ac)的人工饲料对来自本所试验棉田的棉铃虫经过室内21代中16代次的筛选,筛选后F19代LC50值(4.3646g@L1)比筛选前F2代LC50值(0.2972 g@L-1)提高了14.7倍.试验还发现雌性棉铃虫对Bt毒蛋白的敏感性大于雄性.对Bt毒蛋白抗性品系筛选前后分别测定了其不同菌系(商品制剂Dipel和Xentari)的剂量-死亡率回归线,发现Bt毒蛋白抗性品系与其不同菌系间不存在明显的交互抗性.     

First results on synchrony between seasonal pattern of pheromone trap captures of cotton bollworm, <Emphasis Type="Italic">Helicoverpa armigera</Emphasis> and appearance of freshly emerged larvae on developing cobs of corn hybrids

Synchrony between development of five corn hybrid varieties of various seasonal growing rates (FAO numbers), seasonal flight pattern of male cotton bollworm, Helicoverpa armigera Hb. (Lepidoptera: Noctuidae), as monitored by pheromone traps, and the subsequent appearance of newly hatched larvae on developing cobs were studied at Mezőhegyes, South-Hungary, in 2003 and 2004. The phenological stages of corn hybrids were evaluated using the Iowa State University Scale (R₁–R₅), the flight of male moths was monitored by large capacity, funnel type of pheromone traps and the appearance of freshly hatched larvae on developing cobs were counted by visual inspection. The synchrony between the flight peak of male moths and the peak apperance of L₁ larvae on cobs was investigated by cross-correlation. In 2003 (average daily temperatures 20.7°C, average daily relative humidity 59.9% for the period of 4–18 July) “DK 391”, “DK 443” and “Maraton” hybrid varieties already reached silking stage (R₁) by the time when pheromone traps indicated a peak, at 7 July (peak capture at 3-day intervals 755.5 males/traps). The number of L₁ larvae peaked on developing cobs of these varieties also at 7 July (7.0, 4.0 and 3.8 larvae/50 cobs, respectively). The synchrony between the flight peak of male moths and the peak appearance of L₁ larvae on cobs was proven (LAG = 0). A rather similar trend was observed in “Vilma” hybrid variety: it reached R₁ stage at 7 July, and L₁ larvae appeared only a few days later (11 July, 3.8 larvae/50 cobs) (LAG = −1). However, “Maxima” hybrid variety reached R₁ stage 1 week later (14 July) than the time of peak captures. Here L₁ larvae peaked as late as at 18 July (0.8 larvae/50 cobs), i.e., only after the cob had reached R₁ stage (LAG = −3). A reverse order of dates of R₁ stage and peak capture was observed in “Maxima” in 2004 (average daily temperatures; 22.5°C, average daily relative humidities 72.6% for the period of 15 July–6 August): it reached R₁ stage at 19 July, while peak trap captures were recorded at 6 August (peak capture at 3-day intervals 20.5 males/traps). L₁ larvae were found in the highest numbers on 2 August (1.5 larvae/50 cobs), practically in synchrony with peak caputres (LAG = 0). “Káma” reached R₁ stage in 16 July, and L₁ larvae peaked at 2 August (1.3 larvae/50 cobs) (LAG = 0). On the rest of the corn varieties larvae were found only in too numbers for performing statistical analysis. We conclude that in order to predict the appearance of L₁ larvae, the phenological stage of the corn variety and the seasonal flight pattern of moths, as measured by large capacity pheromone traps, should be considered in combination. If the corn variety already reached R₁ stage, L₁ larvae appear on cobs as early as the time of peak flight of moths. However, no young larvae appear on cobs despite of high trap captures, until the corn reaches the R₁ stage. These findings are discussed in the view of specifying optimal timing of a pesticide application.     

Inheritance and cross-resistance of <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> insecticidal crystal protein Cry1Ac resistance in cotton bollworm <Emphasis Type="Italic">Helicoverpa armigera</Emphasis> Hubner population from Tamil Nadu,India

Cotton bollworm, Helicoverpa armigera Hubner selected for five generations with Bacillus thuringiensis insecticidal crystal protein Cry1Ac in the laboratory developed 12.98-fold resistance. Resistance and susceptible populations were mass crossed to study the dominance of resistance. The Cry1Ac—selected (BCR) population showed 5.8-fold resistance to Cry1Aa and 5.04-fold resistance to Cry1Ab. The degree of dominance (D) was 0.34 and 0.40 for the R × S and S × R hybrids, respectively, which indicates incomplete recessive character of Cry1Ac resistance in the population. The estimated realized heritability (h ²) and response quotient (Q) of resistance for Cry1Ac were 0.52 and 0.15, respectively. This indicated the lower phenotypic variation in the selected population. The resistance risk assessment based on h ² indicated that the resistance would increased tenfold after <9 generations for Cry1Ac in the resistant population. The results show the ability of H. armigera to develop resistance against Cry1Ac and cross-resistance to Cry1Aa and Cry1Ab.     

Perennial Wild Relatives of Chickpea as Potential Sources of Resistance to Helicoverpa armigera

The legume pod borer, Helicoverpa armigera (Hübn.), is one of the major constraints to chickpea production, and host plant resistance is an important component for the management of this pest. The levels of resistance in the cultivated chickpea are low to moderate, and therefore, we evaluated 17 accessions of perennial Cicer along with three cultivated chickpea genotypes for resistance to H. armigera. There was a significant reduction in both leaf feeding and larval weights when the larvae were fed on the leaves of Cicer microphyllum Benth. accessions ICC 17146, ICC 17236, ICC 17240, and ICC 17248. Relative resistance index based on leaf feeding, larval survival, and larval weight indicated that C. microphyllum accessions ICC 17146, ICC 17236, ICC 17234, ICC 17240, ICC 17243, and ICC 17248 were highly resistant to H. armigera. Under natural infestation, accessions belonging to C. microphyllum, C. canariense Santos Guerra et Lewis, and C. macracanthum M. Pop suffered a damage rating of <2.0 compared to 4.0 in C. judaicum Boiss. accession ICC 17148 (annual species) and 8.5–9.0 in the cultivated chickpeas (1 = <10% leaf area damaged, and, 9 = >80% leaf area damaged). There was considerable diversity in the accessions belonging to perennial wild species of chickpea, and these can be exploited to increase the levels and diversify the basis of resistance to H. armigera in the cultivated chickpea.     

Congruence of conventional and molecular studies to locate genes that control flavone synthesis in maize silks

The presence of maysin, a flavone glycoside, and its analogues in the silks of corn is an important defence against invasion of the ear by corn earworm in the southeastern USA. Three dent maize inbreds with high silk‐maysin concentrations were evaluated for chromosomal location of major genes controlling synthesis of three antibiotic chemicals by crossing them to a series of waxy‐marked chromosome 9 reciprocal translocations. The data indicated that genes for maysin and its analogues are probably present on the short arms of chromosomes 1 and 10, and long arms of chromosomes 5 and 8 for inbred line GT114, the short arms of chromosome 1 and 6, and the long arms of chromosome 5 and 8 for inbred line GE37, and the short arms of chromosomes 1 and 10, and the long arm of chromosome 4 for inbred line SC102. These results are in general agreement with other translocation studies on corn earworm injury to sweet corn inbreds and gene and restriction fragment length polymorphism marker studies to locate quantitative trait loci (QTL) for maysin synthesis, with the exception that QTL on chromosome 9 have been found to be strongly associated with flavone synthesis. The most significant concordance between conventional and molecular techniques for locating chromosome regions influencing synthesis of antibiotic chemicals from silks of high silk antibiosis sources is found for the short arm of chromosome 1. This region is the most likely one on which to place emphasis during the initial stages of transferring high‐maysin silks to elite inbred lines. A chromosome 1 marker plus at least one more from any chosen high‐maysin inbred source should be sufficient to transfer high maysin silks to other lines. Other markers may be needed for transfer of specific traits when recovering recurrent parent genome types in a backcrossing procedure.     

Cloning and Sequencing of a Gene Encoding Aminopeptidase N in the Midgut of Helicoverpa armigera (Hübner)

A cDNA encoding aminopeptidase N was cloned by degenerated PCR combined with RACEtechnique in this paper. The full-length of APN-Harm is 3 043 bp. Open reading frame is 2 856 bp inlength, encoding 951 amino acid residues. Its predicted molecular weight and isoelectric point are 108.3kDa and 5.29, respectively. This deduced amino acid sequence shares some common structural features withaminopeptidase N from several moth species, including the consensus zinc-binding motif HEXXHX18 E and the GAMEN motif common to gluzincin aminopeptidases. The first 20 amino acid residues at N-termini ishydrophobic transmembrane helix. The sequence of APN-Harm was deposited in GenBank and the accessionnumber is AY181026.     

棉铃虫膨胀线粒体基因组的鉴定与分析

以已公布的棉铃虫线粒体DNA序列对来自4头棉铃虫雄蛹的DNA的三代测序数据进行筛选,获得了11条与线粒体DNA有同源性的三代read序列,并根据其中的read 66003鉴定出了一种膨胀的线粒体基因组。该线粒体基因组大小为27 113 bp,其保守区域包含13个蛋白编码基因、2个rRNA基因、22个tRNA基因以及1个AT富集区,与已公布的棉铃虫线粒体基因组的结构相似。膨胀区域位于cox1基因编码区内部,大小为11 467 bp,经预测含有一个完整的真核基因(依赖ATP的RNA解旋酶)以及多种转座元件的片段,但与线粒体DNA无同源性,也无I类或Ⅱ类内含子存在的证据。对田间和室内棉铃虫DNA样品的PCR扩增未能检测到膨胀线粒体基因组的存在。以上结果表明膨胀片段可能是细胞核DNA序列通过偶然的水平转移事件而整合到线粒体基因组中的,且该种膨胀方式的发生概率极低。本文报道的膨胀线粒体基因组为日后动物线粒体基因组学的研究提示了一种独特的变异方式。     

中红侧沟茧蜂对棉铃虫核型多角体病毒的传播作用

中红侧沟茧蜂Microplitis mediator与核型多角体病毒(nucleopolyhedrovirus, NPV)是棉铃虫Helicoverpa armigera的两种重要生物防治因子。中红侧沟茧蜂传播NPV对于利用二者协同防治棉铃虫具有重要意义。本研究探讨了给中红侧沟茧蜂饲喂含病毒蜂蜜水、体表喷洒病毒液、中红侧沟茧蜂在染毒宿主体内产卵、从染毒宿主体内发育、蜂茧浸泡病毒5种带毒方式的传播病毒效率,以及饲喂带病毒蜂蜜水方式下的传播机制。结果表明,饲喂带病毒蜂蜜水和体表喷洒病毒时中红侧沟茧蜂传毒率较高,在连续传毒的3 d内传毒效率分别为15.1%、9.1%～9.3%、2.4%～4%,其他3种方式传毒效率较低。在田间防治时可以利用中红侧沟茧蜂的传毒作用采用病毒与寄生峰的协同防控策略。     

抗三氟氯氰菊酯棉铃虫种群相对适合度研究

应用经三氟氯氰菊酯汰选的棉铃虫种群,在室内条件下分别研究了抗性、敏感及杂交Ｆ_１代（ＲＳ和ＳＲ）基因型相对适合度。结果表明,抗性基因型在产卵量、卵孵化率、幼虫及蛹历期和成虫存活期等方面与敏感基因型相比较存在明显的不利性;与敏感基因型相比,抗性基因型只有０.４５的适合度,ＲＳ和ＳＲ基因型的相对适合度分别为１.１４和１.０８。