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      Effects of midgut bacteria in Hyphantria cunea (Lepidoptera: Erebidae) on nuclear polyhedrosis virus and Bacillus thuringiensis (Bacillales: Bacillaceae)

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          Abstract

          Hyphantria cunea Drury (Lepidoptera: Erebidae) is a quarantine pest in China that can cause damage to hundreds of plants. As biological control agents, Nuclear Polyhedrosis Virus (NPV) and Bacillus thuringiensis Berliner (Bacillales: Bacillaceae) ( Bt) are commonly used to inhibit the prevalence of H. cunea. To investigate the role of midgut bacteria in the infection of NPV and Bt in H. cunea, we performed a series of tests, including isolating the dominant culturable bacteria in the midgut, eliminating intestinal bacteria, and respectively inoculating the dominant strains with NPV and Bt for bioassay. Two dominant bacteria, Klebsiella oxytoca Lautrop (Enterobacterales: Enterobacteriaceae) and Enterococcus mundtii Collins (Lactobacillales: Enterococcaceae), in the midgut of H. cunea were identified, and a strain of H. cunea larvae without intestinal bacteria was successfully established. In the bioassays of entomopathogen infection, K. oxytoca showed significant synergistic effects with both NPV and Bt on the death of H. cunea. In contrast, E. mundtii played antagonistic effects. This phenomenon may be attributed to the differences in the physico-chemical properties of the two gut bacteria and the alkaline environment required for NPV and Bt to infect the host. It is worth noting that the enhanced insecticidal activity of K. oxytoca on NPV and Bt provides a reference for future biological control of H. cunea by intestinal bacteria.

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          16S ribosomal DNA amplification for phylogenetic study.

          A set of oligonucleotide primers capable of initiating enzymatic amplification (polymerase chain reaction) on a phylogenetically and taxonomically wide range of bacteria is described along with methods for their use and examples. One pair of primers is capable of amplifying nearly full-length 16S ribosomal DNA (rDNA) from many bacterial genera; the additional primers are useful for various exceptional sequences. Methods for purification of amplified material, direct sequencing, cloning, sequencing, and transcription are outlined. An obligate intracellular parasite of bovine erythrocytes, Anaplasma marginale, is used as an example; its 16S rDNA was amplified, cloned, sequenced, and phylogenetically placed. Anaplasmas are related to the genera Rickettsia and Ehrlichia. In addition, 16S rDNAs from several species were readily amplified from material found in lyophilized ampoules from the American Type Culture Collection. By use of this method, the phylogenetic study of extremely fastidious or highly pathogenic bacterial species can be carried out without the need to culture them. In theory, any gene segment for which polymerase chain reaction primer design is possible can be derived from a readily obtainable lyophilized bacterial culture.
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            The gut microbiota of insects - diversity in structure and function.

            Insect guts present distinctive environments for microbial colonization, and bacteria in the gut potentially provide many beneficial services to their hosts. Insects display a wide range in degree of dependence on gut bacteria for basic functions. Most insect guts contain relatively few microbial species as compared to mammalian guts, but some insects harbor large gut communities of specialized bacteria. Others are colonized only opportunistically and sparsely by bacteria common in other environments. Insect digestive tracts vary extensively in morphology and physicochemical properties, factors that greatly influence microbial community structure. One obstacle to the evolution of intimate associations with gut microorganisms is the lack of dependable transmission routes between host individuals. Here, social insects, such as termites, ants, and bees, are exceptions: social interactions provide opportunities for transfer of gut bacteria, and some of the most distinctive and consistent gut communities, with specialized beneficial functions in nutrition and protection, have been found in social insect species. Still, gut bacteria of other insects have also been shown to contribute to nutrition, protection from parasites and pathogens, modulation of immune responses, and communication. The extent of these roles is still unclear and awaits further studies. © 2013 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.
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              Multiorganismal insects: diversity and function of resident microorganisms.

              All insects are colonized by microorganisms on the insect exoskeleton, in the gut and hemocoel, and within insect cells. The insect microbiota is generally different from microorganisms in the external environment, including ingested food. Specifically, certain microbial taxa are favored by the conditions and resources in the insect habitat, by their tolerance of insect immunity, and by specific mechanisms for their transmission. The resident microorganisms can promote insect fitness by contributing to nutrition, especially by providing essential amino acids, B vitamins, and, for fungal partners, sterols. Some microorganisms protect their insect hosts against pathogens, parasitoids, and other parasites by synthesizing specific toxins or modifying the insect immune system. Priorities for future research include elucidation of microbial contributions to detoxification, especially of plant allelochemicals in phytophagous insects, and resistance to pathogens; as well as their role in among-insect communication; and the potential value of manipulation of the microbiota to control insect pests.
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                Author and article information

                Contributors
                Role: Subject Editor
                Journal
                J Insect Sci
                J Insect Sci
                jis
                Journal of Insect Science
                Oxford University Press (US )
                1536-2442
                March 2023
                14 March 2023
                14 March 2023
                : 23
                : 2
                : 1
                Affiliations
                Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University , Nanjing 210037, China
                College of Forestry, Nanjing Forestry University , Nanjing 210037, China
                Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University , Nanjing 210037, China
                College of Forestry, Nanjing Forestry University , Nanjing 210037, China
                Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University , Nanjing 210037, China
                College of Forestry, Nanjing Forestry University , Nanjing 210037, China
                Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University , Nanjing 210037, China
                College of Forestry, Nanjing Forestry University , Nanjing 210037, China
                Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University , Nanjing 210037, China
                College of Forestry, Nanjing Forestry University , Nanjing 210037, China
                Author notes
                Corresponding author, mail: djhao@ 123456njfu.edu.cn
                Author information
                https://orcid.org/0000-0001-8241-5581
                https://orcid.org/0000-0001-7743-9968
                https://orcid.org/0000-0001-8541-7857
                https://orcid.org/0000-0002-9676-4069
                https://orcid.org/0000-0002-9525-8033
                Article
                iead009
                10.1093/jisesa/iead009
                10011879
                36916277
                a2da40fb-cd72-471a-8a42-e640288e82cf
                © The Author(s) 2023. Published by Oxford University Press on behalf of Entomological Society of America.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 06 July 2022
                : 05 December 2022
                : 02 February 2023
                Page count
                Pages: 9
                Funding
                Funded by: Forestry Science and Technology Innovation and Promotion Project in Jiangsu Province;
                Award ID: LYKJ[2021]17
                Funded by: Special Fund for Scientific Research of Shanghai Landscaping & City Appearance Administrative Bureau;
                Award ID: G221207
                Categories
                Research
                AcademicSubjects/SCI01382

                Entomology
                hyphantria cunea,intestinal bacteria,synergistic effect,antagonistic effect,nuclear polyhedrosis virus

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