Functional analysis of the orphan genes Tssor-3 and Tssor-4 in male Plutella xylostella

doi:10.1016/S2095-3119(21)63655-9

Journal of Integrative Agriculture ›› 2021, Vol. 20 ›› Issue (7): 1880-1888.DOI: 10.1016/S2095-3119(21)63655-9

所属专题：昆虫合辑Plant Protection—Entomolgy；昆虫分子生物学合辑Insect Molecular Biology

收稿日期:2020-11-04 出版日期:2021-07-01 发布日期:2021-06-02

Functional analysis of the orphan genes Tssor-3 and Tssor-4 in male Plutella xylostella

LI Tian-pu^{1, 2, 3}, ZHANG Li-wen^{1, 2, 3}, LI Ya-qing^{1, 2, 3}, YOU Min-sheng^{1, 2, 3}, ZHAO Qian^{1, 2, 3}

¹ State Key Laboratory of Ecological Pest Control for Fujian-Taiwan Crops, Institute of Applied Ecology, Fujian Agriculture and Forestry University, Fuzhou 350002, P.R.China
² Joint International Research Laboratory of Ecological Pest Control, Ministry of Education, Fuzhou 350002, P.R.China
³ Key Laboratory of Integrated Pest Management for Fujian-Taiwan Crops, Ministry of Agriculture and Rural Affairs, Fuzhou 350002, P.R.China

Received:2020-11-04 Online:2021-07-01 Published:2021-06-02
Contact: Correspondence ZHAO Qian, Tel: +86-591-83843724, E-mail: zhaoqian977228@126.com
About author:LI Tian-pu, E-mail: zhkqgs@163.com
Supported by:
This work was supported by the Natural Science Foundation of Fujian Province, China (2020J01525), the National Natural Science Foundation of China (31320103922 and 31230061) and the Major Science and Technology Projects in Fujian Province, China (2018NZ010100130).

摘要/Abstract

摘要：

孤儿基因是指在其他物种中没有同源序列的一类基因。在此，我们在小菜蛾中鉴定了两个孤儿基因，命名为Tssor-3和Tssor-4。这两个基因都含有一个信号肽序列，表明它们具有分泌蛋白的功能。基于实时荧光定量PCR（qPCR）的表达模式分析表明，这两个孤儿基因均在除睾丸外的雄性生殖腺中特异表达；其表达量在雄成虫时期达到顶峰。免疫荧光实验表明，这两种蛋白均为精液蛋白，暗示它们在调节雄性生殖方面具有潜在的作用。为了进一步探索它们的功能，我们通过RNA干扰（RNAi）下调了这两个基因的表达量，结果表明，干扰后24 h，Tssor-3和Tssor-4的表达量均显著低于对照组。生物测定实验表明，当Tssor-3和Tssor-4基因表达量降低时，小菜蛾的产卵量和子代卵的孵化率均显著下降，表明这两个孤儿基因在小菜蛾雄性育性中起作用。我们的结果为孤儿基因参与雄性生殖调节提供了证据，这对雄性在进化过程中的适应性很重要。

Abstract:

Orphan genes are genes with no sequence homologues in other species. Here, we identified two orphan genes, namely, Tssor-3 and Tssor-4, in Plutella xylostella. Both genes contained a signal peptide sequence, suggesting their functions as secreted proteins. Expression pattern analysis based on real-time quantitative PCR (qPCR) showed that both orphan genes were specifically expressed in all male gonads except the testes. The expression of both the orphan genes peaked at the male adult stage. Immunofluorescence assays suggested that the two proteins were seminal proteins, indicating their potential roles in male reproductive regulation. To further explain their functions, we knocked down the expression of these two genes by RNA interference (RNAi). The results showed that the expression of Tssor-3 and Tssor-4 was significantly downregulated at 24 h after injection compared to that of the controls. Biological assays showed that the number of laid eggs and the hatching rate of offspring eggs were significantly reduced when the expression of Tssor-3 and Tssor-4 was reduced, suggesting that the two orphan genes played a role in male fertility in P. xylostella. Our results provide evidence that orphan genes are involved in male reproductive regulation, which is important for male fitness during evolution.

Key words: Plutella xylostella , orphan genes , male fertility , RNA interference

. [J]. Journal of Integrative Agriculture, 2021, 20(7): 1880-1888.

LI Tian-pu, ZHANG Li-wen, LI Ya-qing, YOU Min-sheng, ZHAO Qian. Functional analysis of the orphan genes Tssor-3 and Tssor-4 in male Plutella xylostella[J]. Journal of Integrative Agriculture, 2021, 20(7): 1880-1888.

参考文献

Chapman T. 2001. Seminal fluid-mediated fitness traits in Drosophila. Heredity, 87, 511–521.
Chapman T, Bangham J, Vinti G, Seifried B, Lung O, Wolfner M F, Smith H K, Partridge L. 2003. The sex peptide of Drosophila melanogaster: female post-mating responses analyzed by using RNA interference. Proceedings of the National Academy of Sciences of the United States of America, 100, 9923–9928.
Chen K, Tian Z, Chen P, He H, Jiang F, Long C A. 2020. Genome-wide identification, characterization and expression analysis of lineage-specific genes within Hanseniaspora yeasts. FEMS Microbiology Letters, 367, fnaa077.
Christiaens O, Swevers L, Smagghe G. 2014. DsRNA degradation in the pea aphid (Acyrthosiphon pisum) associated with lack of response in RNAi feeding and injection assay. Peptides, 53, 307–314.
Domazet-Loso T, Tautz D. 2003. An evolutionary analysis of orphan genes in Drosophila. Genome Research, 13, 2213–2219.
Fathi S A A, Bozorg-Amirkalaee M, Sarfaraz R M. 2010. Preference and performance of Plutella xylostella (L.) (Lepidoptera: Plutellidae) on canola cultivars. Journal of Pest Science, 84, 41–47.
Furlong M J, Wright D J, Dosdall L M. 2013. Diamondback moth ecology and management: Problems, progress, and prospects. Annual Review of Entomology, 58, 517–541.
Gao X, Wu G X, Ye G Y. 2008. Physiological functions of male accessory gland secretions of insects. Journal of Yunnan University, 30, 152–159. (in Chinese)
Gong L, Chen Y, Hu Z, Hu M. 2013. Testing insecticidal activity of novel chemically synthesized siRNA against Plutella xylostella under laboratory and field conditions. PLoS ONE, 8, e62990.
Guo W J, Li P, Ling J, Ye S P. 2007. Significant comparative characteristics between orphan and nonorphan genes in the rice (Oryza sativa L.) genome. Comparative and Functional Genomics, 2007, 21676.
Kaessmann H. 2010. Origins, evolution, and phenotypic impact of new genes. Genome Research, 20, 1313–1326.
King M, Eubel H, Millar A H, Baer B. 2011. Proteins within the seminal fluid are crucial to keep sperm viable in the honeybee Apis mellifera. Journal of Insect Physiology, 57, 409–414.
Kumar A, Gates P B, Czarkwiani A, Brockes J P. 2015. An orphan gene is necessary for preaxial digit formation during salamander limb development. Nature Communication, 6, 8684.
Li C H, Cai B, Liang Y H, Jia S L. 2013. Expression analysis and functional conjecture of orphan gene in grape. Genomics and Applied Biology, 32, 240–245. (in Chinese)
Li L, Zheng W, Zhu Y, Ye H, Tang B, Arendsee Z W, Jones D, Li R, Ortiz D, Zhao X, Du C, Nettleton D, Scott M P, Salas-Fernandez M G, Yin Y, Wurtele E S. 2015. QQS orphan gene regulates carbon and nitrogen partitioning across species via NF-YC interactions. Proceedings of the National Academy of Sciences of the United States of America, 112, 14734–14739.
Li Z, Feng X, Liu S S, You M, Furlong M J. 2016. Biology, ecology, and management of the diamondback moth in China. Annual Review Entomology, 61, 277–296.
Liu J, Kolliopoulou A, Smagghe G, Swevers L. 2014. Modulation of the transcriptional response of innate immune and RNAi genes upon exposure to dsRNA and LPS in silkmoth-derived Bm5 cells overexpressing BmToll9-1 receptor. Journal of Insect Physiology, 66, 10–19.
Livak K J, Schmittgen T D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2–??CT method. Methods, 25, 402–408.
Miyatake T, Chapman T, Partridge L. 1999. Mating-induced inhibition of remating in female Mediterranean fruit flies Ceratitis capitata. Journal of Insect Physiology, 45, 1021–1028.
Mohr S E. 2014. RNAi screening in Drosophila cells and in vivo. Methods, 68, 82–88.
Ni F, Qi J, Hao Q, Lyu B, Luo M C, Wang Y, Chen F, Wang S, Zhang C, Epstein L, Zhao X, Wang H, Zhang X, Chen C, Sun L, Fu D. 2017. Wheat Ms2 encodes for an orphan protein that confers male sterility in grass species. Nature Communication, 8, 15121.
Thumecke S, Beermann A, Klingler M, Schroder R. 2017. The flipflop orphan genes are required for limb bud eversion in the Tribolium embryo. Frontiers Zoology, 14, 48.
Wynant N, Verlinden H, Breugelmans B, Simonet G, Vanden Broeck J. 2012. Tissue-dependence and sensitivity of the systemic RNA interference response in the desert locust, Schistocerca gregaria. Insect Biochemistry and Molecular Biology, 42, 911–917.
Xu Y, Wu G, Hao B, Chen L, Deng X, Xu Q. 2015. Identification, characterization and expression analysis of lineage-specific genes within sweet orange (Citrus sinensis). BMC Genomics, 16, 995.
Yang L, Zou M, Fu B, He S J B G. 2013. Genome-wide identification, characterization, and expression analysis of lineage-specific genes within zebrafish. BMC Genomics, 14, 65.
Yang Y, Wan P J, Hu X X, Li G Q. 2014. RNAi mediated knockdown of the ryanodine receptor gene decreases chlorantraniliprole susceptibility in Sogatella furcifera. Pesticide Biochemistry and Physiology, 108, 58–65.
Yoon J S, Shukla J N, Gong Z J, Mogilicherla K, Palli S R. 2016. RNA interference in the Colorado potato beetle, Leptinotarsa decemlineata: Identification of key contributors. Insect Biochemistry and Molecular Biology, 78, 78–88.
Yu B, Li D T, Lu J B, Zhang W X, Zhang C X. 2016. Seminal fluid protein genes of the brown planthopper, Nilaparvata lugens. BMC Genomics, 17, 654.
Zalucki M P, Shabbir A, Silva R, Adamson D, Liu S S, Furlong M J. 2012. Estimating the economic cost of one of the world’s major insect pests, Plutella xylostella (Lepidoptera: Plutellidae): Just how long is a piece of string? Journal of Economic Entomology, 105, 1115–1129.

Functional analysis of the orphan genes Tssor-3 and Tssor-4 in male Plutella xylostella

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 0

编辑推荐

Metrics