[Objective] The paper was to improve the efficiency and accuracy of early forecast of Lepidopteran oak-infesting pests.[Method] DNA barcoding technique was established for quick species identification using mitochondr...[Objective] The paper was to improve the efficiency and accuracy of early forecast of Lepidopteran oak-infesting pests.[Method] DNA barcoding technique was established for quick species identification using mitochondrial cytochrome C oxidase subunit Ⅰ（COⅠ） as the standard gene.This barcoding technique was used to amplify and sequence genomic DNA samples from eggs and pupae of 11 species of Lepidopteran pests collected from oak.[Result] The DNA barcoding standard genes of 594-708 bp were determined from eggs and pupae of Lepidopteran insects.There were differences of 0-2 bases in DNA barcode sequences between conspecific eggs and pupae,with the sequence identity of 99.7%-100%.The average content of A,T,G and C of DNA barcode sequences from Lepidopteran insects were 30.7%,38.5%,14.9% and 15.9%,respectively.The obtained DNA barcode sequences had 91.4%-100% identity and 0-8.6% difference degree with GenBank-deposited DNA barcode sequences from organisms of the genetically-closest relationship.Among them,DNA barcode sequences from egg and pupa samples of 10 Lepidopteran insects（No.1-20） had 99%-100% identity and 0-1.0% difference degree with homologous sequences in GenBank database,while the remaining samples（No.21-22） had high difference degree（8.6%） with homologous sequences.[Conclusion] The established DNA barcoding technique is an effeetive tool for species identification of Lepidopteran pests using genomic DNA from eggs and pupae of Lepidopteran insects.展开更多
昆虫性别决定机制复杂多样,研究昆虫性别决定机制既能加强对物种进化关系的理解,也能为通过遗传调控控制害虫种群提供新方向。鳞翅目昆虫多为雌雄二型,繁殖方式以两性生殖为主,性染色体系统大多为ZW/ZZ系统,雌性为ZW型,雄性为ZZ型。昆...昆虫性别决定机制复杂多样,研究昆虫性别决定机制既能加强对物种进化关系的理解,也能为通过遗传调控控制害虫种群提供新方向。鳞翅目昆虫多为雌雄二型,繁殖方式以两性生殖为主,性染色体系统大多为ZW/ZZ系统,雌性为ZW型,雄性为ZZ型。昆虫性别决定是通过级联反应调控的,在鳞翅目昆虫ZW/ZZ系统中,位于W染色体上的性别决定初始信号沉默雄性化关键基因Masculinizer(Masc),然后Doublesex(dsx)基因被雌性特异性剪接,从而调控雌性偏向基因的表达;而雄虫中Masc正常表达,调控dsx进行雄性特异剪接,实现雄性分化。本文对鳞翅目昆虫性别决定初始信号、Masc和dsx的研究现状,以及基于基因编辑技术和性别决定机制在鳞翅目昆虫遗传调控中的应用进行了综述。相关研究为鳞翅目昆虫的性别筛选、不育昆虫释放技术(sterile insect technique,SIT)和释放携带雌性特异显性致死基因昆虫技术(female specific release of insects carrying dominant lethal,fsRIDL)奠定理论基础。展开更多
几丁质脱乙酰基酶是几丁质修饰关键酶,在昆虫几丁质组装过程中发挥重要作用,是潜在的绿色农药靶标。课题组近期研究发现家蚕中肠3个几丁质脱乙酰基酶中,BmCDA7对围食膜几丁质具有最高活性,在进食期高表达,可能参与该时期围食膜形成。本...几丁质脱乙酰基酶是几丁质修饰关键酶,在昆虫几丁质组装过程中发挥重要作用,是潜在的绿色农药靶标。课题组近期研究发现家蚕中肠3个几丁质脱乙酰基酶中,BmCDA7对围食膜几丁质具有最高活性,在进食期高表达,可能参与该时期围食膜形成。本研究通过进一步优化分离纯化条件,建立了金属螯合层析和阴离子交换层析两步分离纯化方法,获得了高纯度的BmCDA7重组蛋白。采用SDS-PAGE结合Calcofluor White M2R显色定性确定了BmCDA7对底物乙二醇几丁质的催化活性。通过测定催化反应过程中释放的乙酸量,确定了BmCDA7的最适反应条件、稳定性及底物选择性。BmCDA7催化反应最适温度为60℃,最适pH为8.0,在温度低于60℃以及中性、偏碱性条件下较为稳定。此外,BmCDA7对乙二醇几丁质、胶体几丁质的酶活力分别为2.9926和0.4270μmol/(min·μmol),而对高结晶度的α-几丁质和β-几丁质无活力。这些结果丰富了昆虫几丁质脱乙酰基酶的生物化学基础研究。展开更多
To provide pest technicians with a convenient way to recognize insects,a novel method is proposed to classify insect images by integrated region matching (IRM) and dual tree complex wavelet transform (DTCWT).The wing ...To provide pest technicians with a convenient way to recognize insects,a novel method is proposed to classify insect images by integrated region matching (IRM) and dual tree complex wavelet transform (DTCWT).The wing image of the lepidopteran insect is preprocessed to obtain the region of interest (ROI) whose position is then calibrated.The ROI is first segmented with the k-means algorithm into regions according to the color features,properties of all the segmented regions being used as a coarse level feature.The color image is then converted to a grayscale image,where DTCWT features are extracted as a fine level feature.The IRM scheme is undertaken to find K nearest neighbors (KNNs),out of which the nearest neighbor is searched by computing the Canberra distance of DTCWT features.The method was tested with a database including 100 lepidopteran insect species from 18 families and the recognition accuracy was 84.47%.For the forewing subset,a recognition accuracy of 92.38% was achieved.The results showed that the proposed method can effectively solve the problem of automatic species identification of lepidopteran specimens.展开更多
基金Supported by Natural Science Foundation of Liaoning Province(2014027002)National Special Fund for Development of Cocoon Silk(201601)National Modern Agriculture Industry Technology System Construction Project(CARS-18)
文摘[Objective] The paper was to improve the efficiency and accuracy of early forecast of Lepidopteran oak-infesting pests.[Method] DNA barcoding technique was established for quick species identification using mitochondrial cytochrome C oxidase subunit Ⅰ（COⅠ） as the standard gene.This barcoding technique was used to amplify and sequence genomic DNA samples from eggs and pupae of 11 species of Lepidopteran pests collected from oak.[Result] The DNA barcoding standard genes of 594-708 bp were determined from eggs and pupae of Lepidopteran insects.There were differences of 0-2 bases in DNA barcode sequences between conspecific eggs and pupae,with the sequence identity of 99.7%-100%.The average content of A,T,G and C of DNA barcode sequences from Lepidopteran insects were 30.7%,38.5%,14.9% and 15.9%,respectively.The obtained DNA barcode sequences had 91.4%-100% identity and 0-8.6% difference degree with GenBank-deposited DNA barcode sequences from organisms of the genetically-closest relationship.Among them,DNA barcode sequences from egg and pupa samples of 10 Lepidopteran insects（No.1-20） had 99%-100% identity and 0-1.0% difference degree with homologous sequences in GenBank database,while the remaining samples（No.21-22） had high difference degree（8.6%） with homologous sequences.[Conclusion] The established DNA barcoding technique is an effeetive tool for species identification of Lepidopteran pests using genomic DNA from eggs and pupae of Lepidopteran insects.
文摘昆虫性别决定机制复杂多样,研究昆虫性别决定机制既能加强对物种进化关系的理解,也能为通过遗传调控控制害虫种群提供新方向。鳞翅目昆虫多为雌雄二型,繁殖方式以两性生殖为主,性染色体系统大多为ZW/ZZ系统,雌性为ZW型,雄性为ZZ型。昆虫性别决定是通过级联反应调控的,在鳞翅目昆虫ZW/ZZ系统中,位于W染色体上的性别决定初始信号沉默雄性化关键基因Masculinizer(Masc),然后Doublesex(dsx)基因被雌性特异性剪接,从而调控雌性偏向基因的表达;而雄虫中Masc正常表达,调控dsx进行雄性特异剪接,实现雄性分化。本文对鳞翅目昆虫性别决定初始信号、Masc和dsx的研究现状,以及基于基因编辑技术和性别决定机制在鳞翅目昆虫遗传调控中的应用进行了综述。相关研究为鳞翅目昆虫的性别筛选、不育昆虫释放技术(sterile insect technique,SIT)和释放携带雌性特异显性致死基因昆虫技术(female specific release of insects carrying dominant lethal,fsRIDL)奠定理论基础。
文摘几丁质脱乙酰基酶是几丁质修饰关键酶,在昆虫几丁质组装过程中发挥重要作用,是潜在的绿色农药靶标。课题组近期研究发现家蚕中肠3个几丁质脱乙酰基酶中,BmCDA7对围食膜几丁质具有最高活性,在进食期高表达,可能参与该时期围食膜形成。本研究通过进一步优化分离纯化条件,建立了金属螯合层析和阴离子交换层析两步分离纯化方法,获得了高纯度的BmCDA7重组蛋白。采用SDS-PAGE结合Calcofluor White M2R显色定性确定了BmCDA7对底物乙二醇几丁质的催化活性。通过测定催化反应过程中释放的乙酸量,确定了BmCDA7的最适反应条件、稳定性及底物选择性。BmCDA7催化反应最适温度为60℃,最适pH为8.0,在温度低于60℃以及中性、偏碱性条件下较为稳定。此外,BmCDA7对乙二醇几丁质、胶体几丁质的酶活力分别为2.9926和0.4270μmol/(min·μmol),而对高结晶度的α-几丁质和β-几丁质无活力。这些结果丰富了昆虫几丁质脱乙酰基酶的生物化学基础研究。
基金Project (No.2006AA10Z211) supported by the National High-Tech Research and Development Program (863) of China
文摘To provide pest technicians with a convenient way to recognize insects,a novel method is proposed to classify insect images by integrated region matching (IRM) and dual tree complex wavelet transform (DTCWT).The wing image of the lepidopteran insect is preprocessed to obtain the region of interest (ROI) whose position is then calibrated.The ROI is first segmented with the k-means algorithm into regions according to the color features,properties of all the segmented regions being used as a coarse level feature.The color image is then converted to a grayscale image,where DTCWT features are extracted as a fine level feature.The IRM scheme is undertaken to find K nearest neighbors (KNNs),out of which the nearest neighbor is searched by computing the Canberra distance of DTCWT features.The method was tested with a database including 100 lepidopteran insect species from 18 families and the recognition accuracy was 84.47%.For the forewing subset,a recognition accuracy of 92.38% was achieved.The results showed that the proposed method can effectively solve the problem of automatic species identification of lepidopteran specimens.
