ame-miR-34 Modulates the Larval Body Weight and Immune Response of <i>Apis mellifera</i> Workers to <i>Ascosphara apis</i> Invasion
MiRNAs are critical regulators of numerous physiological and pathological processes. <i>Ascosphaera apis</i> exclusively infects bee larvae and causes chalkbrood disease. However, the function and mechanism of miRNAs in the bee larval response to <i>A. apis</i> infection is p...
| Main Authors: | , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-01-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/2/1214 |
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| author | Ying Wu Yilong Guo Xiaoxue Fan Haodong Zhao Yiqiong Zhang Sijia Guo Xin Jing Zhitan Liu Peilin Feng Xiaoyu Liu Peiyuan Zou Qiming Li Zhihao Na Kuihao Zhang Dafu Chen Rui Guo |
| author_facet | Ying Wu Yilong Guo Xiaoxue Fan Haodong Zhao Yiqiong Zhang Sijia Guo Xin Jing Zhitan Liu Peilin Feng Xiaoyu Liu Peiyuan Zou Qiming Li Zhihao Na Kuihao Zhang Dafu Chen Rui Guo |
| author_sort | Ying Wu |
| collection | DOAJ |
| description | MiRNAs are critical regulators of numerous physiological and pathological processes. <i>Ascosphaera apis</i> exclusively infects bee larvae and causes chalkbrood disease. However, the function and mechanism of miRNAs in the bee larval response to <i>A. apis</i> infection is poorly understood. Here, ame-miR-34, a previously predicted miRNA involved in the response of <i>Apis mellifera</i> larvae to <i>A. apis</i> invasion, was subjected to molecular validation, and overexpression and knockdown were then conducted to explore the regulatory functions of ame-miR-34 in larval body weight and immune response. Stem-loop RT-PCR and Sanger sequencing confirmed the authenticity of ame-miR-34 in the larval gut of <i>A. mellifera</i>. RT-qPCR results demonstrated that compared with that in the uninfected larval guts, the expression level of ame-miR-34 was significantly downregulated (<i>p</i> < 0.001) in the guts of <i>A. apis</i>-infected 4-, 5-, and 6-day-old larvae, indicative of the remarkable suppression of host ame-miR-34 due to <i>A. apis</i> infection. In comparison with the corresponding negative control (NC) groups, the expression level of ame-miR-34 in the larval guts in the mimic-miR-34 group was significantly upregulated (<i>p</i> < 0.001), while that in the inhibitor-miR-34 group was significantly downregulated (<i>p</i> < 0.01). Similarly, effective overexpression and knockdown of ame-miR-34 were achieved. In addition, the body weights of 5- and 6-day-old larvae were significantly increased compared with those in the mimic-NC group; the weights of 5-day-old larvae in the inhibitor-miR-34 group were significantly decreased in comparison with those in the inhibitor-NC group, while the weights of 4- and 6-day-old larvae in the inhibitor-miR-34 group were significantly increased, indicating the involvement of ame-miR-34 in modulating larval body weight. Furthermore, the expression levels of both <i>hsp</i> and <i>abct</i> in the guts of <i>A. apis</i>-infected 4-, 5-, and 6-day-old larvae were significantly upregulated after ame-miR-34 overexpression. In contrast, after ame-miR-34 knockdown, the expression levels of the aforementioned two key genes in the <i>A. apis</i>-infected 4-, 5-, and 6-day-old larval guts were significantly downregulated. Together, the results demonstrated that effective overexpression and knockdown of ame-miR-34 in both noninfected and <i>A. apis</i>-infected <i>A. mellifera</i> larval guts could be achieved by the feeding method, and ame-miR-34 exerted a regulatory function in the host immune response to <i>A. apis</i> invasion through positive regulation of the expression of <i>hsp</i> and <i>abct</i>. Our findings not only provide a valuable reference for the functional investigation of bee larval miRNAs but also reveal the regulatory role of ame-miR-34 in <i>A. mellifera</i> larval weight and immune response. Additionally, the results of this study may provide a promising molecular target for the treatment of chalkbrood disease. |
| first_indexed | 2024-03-09T12:23:46Z |
| format | Article |
| id | doaj.art-72a7725979044405b44d40c3cdf85652 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T12:23:46Z |
| publishDate | 2023-01-01 |
| publisher | MDPI AG |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-72a7725979044405b44d40c3cdf856522023-11-30T22:37:04ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-01-01242121410.3390/ijms24021214ame-miR-34 Modulates the Larval Body Weight and Immune Response of <i>Apis mellifera</i> Workers to <i>Ascosphara apis</i> InvasionYing Wu0Yilong Guo1Xiaoxue Fan2Haodong Zhao3Yiqiong Zhang4Sijia Guo5Xin Jing6Zhitan Liu7Peilin Feng8Xiaoyu Liu9Peiyuan Zou10Qiming Li11Zhihao Na12Kuihao Zhang13Dafu Chen14Rui Guo15College of Animal Sciences (College of Bee Science), Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Animal Sciences (College of Bee Science), Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Animal Sciences (College of Bee Science), Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Animal Sciences (College of Bee Science), Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Animal Sciences (College of Bee Science), Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Animal Sciences (College of Bee Science), Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Animal Sciences (College of Bee Science), Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Animal Sciences (College of Bee Science), Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Animal Sciences (College of Bee Science), Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Animal Sciences (College of Bee Science), Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Animal Sciences (College of Bee Science), Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Animal Sciences (College of Bee Science), Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Animal Sciences (College of Bee Science), Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Animal Sciences (College of Bee Science), Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Animal Sciences (College of Bee Science), Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Animal Sciences (College of Bee Science), Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaMiRNAs are critical regulators of numerous physiological and pathological processes. <i>Ascosphaera apis</i> exclusively infects bee larvae and causes chalkbrood disease. However, the function and mechanism of miRNAs in the bee larval response to <i>A. apis</i> infection is poorly understood. Here, ame-miR-34, a previously predicted miRNA involved in the response of <i>Apis mellifera</i> larvae to <i>A. apis</i> invasion, was subjected to molecular validation, and overexpression and knockdown were then conducted to explore the regulatory functions of ame-miR-34 in larval body weight and immune response. Stem-loop RT-PCR and Sanger sequencing confirmed the authenticity of ame-miR-34 in the larval gut of <i>A. mellifera</i>. RT-qPCR results demonstrated that compared with that in the uninfected larval guts, the expression level of ame-miR-34 was significantly downregulated (<i>p</i> < 0.001) in the guts of <i>A. apis</i>-infected 4-, 5-, and 6-day-old larvae, indicative of the remarkable suppression of host ame-miR-34 due to <i>A. apis</i> infection. In comparison with the corresponding negative control (NC) groups, the expression level of ame-miR-34 in the larval guts in the mimic-miR-34 group was significantly upregulated (<i>p</i> < 0.001), while that in the inhibitor-miR-34 group was significantly downregulated (<i>p</i> < 0.01). Similarly, effective overexpression and knockdown of ame-miR-34 were achieved. In addition, the body weights of 5- and 6-day-old larvae were significantly increased compared with those in the mimic-NC group; the weights of 5-day-old larvae in the inhibitor-miR-34 group were significantly decreased in comparison with those in the inhibitor-NC group, while the weights of 4- and 6-day-old larvae in the inhibitor-miR-34 group were significantly increased, indicating the involvement of ame-miR-34 in modulating larval body weight. Furthermore, the expression levels of both <i>hsp</i> and <i>abct</i> in the guts of <i>A. apis</i>-infected 4-, 5-, and 6-day-old larvae were significantly upregulated after ame-miR-34 overexpression. In contrast, after ame-miR-34 knockdown, the expression levels of the aforementioned two key genes in the <i>A. apis</i>-infected 4-, 5-, and 6-day-old larval guts were significantly downregulated. Together, the results demonstrated that effective overexpression and knockdown of ame-miR-34 in both noninfected and <i>A. apis</i>-infected <i>A. mellifera</i> larval guts could be achieved by the feeding method, and ame-miR-34 exerted a regulatory function in the host immune response to <i>A. apis</i> invasion through positive regulation of the expression of <i>hsp</i> and <i>abct</i>. Our findings not only provide a valuable reference for the functional investigation of bee larval miRNAs but also reveal the regulatory role of ame-miR-34 in <i>A. mellifera</i> larval weight and immune response. Additionally, the results of this study may provide a promising molecular target for the treatment of chalkbrood disease.https://www.mdpi.com/1422-0067/24/2/1214<i>Ascosphaera apis</i>honey bee<i>Apis mellifera</i>miR-34larvabody weight |
| spellingShingle | Ying Wu Yilong Guo Xiaoxue Fan Haodong Zhao Yiqiong Zhang Sijia Guo Xin Jing Zhitan Liu Peilin Feng Xiaoyu Liu Peiyuan Zou Qiming Li Zhihao Na Kuihao Zhang Dafu Chen Rui Guo ame-miR-34 Modulates the Larval Body Weight and Immune Response of <i>Apis mellifera</i> Workers to <i>Ascosphara apis</i> Invasion International Journal of Molecular Sciences <i>Ascosphaera apis</i> honey bee <i>Apis mellifera</i> miR-34 larva body weight |
| title | ame-miR-34 Modulates the Larval Body Weight and Immune Response of <i>Apis mellifera</i> Workers to <i>Ascosphara apis</i> Invasion |
| title_full | ame-miR-34 Modulates the Larval Body Weight and Immune Response of <i>Apis mellifera</i> Workers to <i>Ascosphara apis</i> Invasion |
| title_fullStr | ame-miR-34 Modulates the Larval Body Weight and Immune Response of <i>Apis mellifera</i> Workers to <i>Ascosphara apis</i> Invasion |
| title_full_unstemmed | ame-miR-34 Modulates the Larval Body Weight and Immune Response of <i>Apis mellifera</i> Workers to <i>Ascosphara apis</i> Invasion |
| title_short | ame-miR-34 Modulates the Larval Body Weight and Immune Response of <i>Apis mellifera</i> Workers to <i>Ascosphara apis</i> Invasion |
| title_sort | ame mir 34 modulates the larval body weight and immune response of i apis mellifera i workers to i ascosphara apis i invasion |
| topic | <i>Ascosphaera apis</i> honey bee <i>Apis mellifera</i> miR-34 larva body weight |
| url | https://www.mdpi.com/1422-0067/24/2/1214 |
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