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      2. 蘿卜蚜端粒逆轉錄酶TERT基因的鑒定與表達分析
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        國家自然科學基金項目(31572329)


        Identification and expression of TERT in Lipaphis erysimi (Kaltenbach)
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          摘要:

          為探究端粒酶逆轉錄酶基因(telomerase reverse transcriptase, TERT)在昆蟲生長發育過程中的作用機制,以蘿卜蚜Lipaphis erysimi (Kaltenbach)為研究對象,克隆其TERT基因并分析其時空表達情況。結果發現,蘿卜蚜TERT基因的CDS區長2 658 bp,編碼885個氨基酸,其二級結構含41個α-螺旋、60個卷曲螺旋。系統進化分析結果顯示,蘿卜蚜TERT基因與麥雙尾蚜Diuraphis noxia的TERT 基因關系最近,與桃蚜Myzus persicae、豌豆蚜Acyrthosiphon pisum的TERT基因同源性較高。實時熒光定量PCR顯示,TERT基因在蘿卜蚜生長發育過程中的各個齡期及組織中均有表達,且成蟲TERT基因的相對表達量依次高于1齡、2齡、3齡、4齡若蟲;TERT基因在蘿卜蚜腹部表達量最高,胸部次之,頭部表達量最低。以上研究結果表明,TERT基因在蘿卜蚜的生長發育過程中起重要作用,可以選擇其同源性低的區域作為基因干擾或沉默的靶標區域,用來防治蘿卜蚜在農業上造成的重大危害。

          Abstract:

          The Lipaphis erysimi (Kaltenbach) is one of the agricultural insects endangering cruciferous vegetables,and it has a rapid propagation speed and is distributed all over the world. Telomerase reverse transcriptase (TERT) is a catalytic subunit of telomerase,which not only can induce telomerase activity to affect cell aging and apoptosis,but also has other biological functions besides telomerase dependent. In order to clarify the mechanism of the telomerase reverse transcriptase (TERT) in insect growth and development,the TERT in L. erysimi (Kaltenbach) was cloned and its spatial and temporal expression was analyzed. According to the local transcriptome library,the related sequences of TERT gene of L. erysimi were screened,and the CDs sequences of TERT gene were cloned by gene amplification. The full-length sequence of the CDS region of the gene is 2 658 bp,which can encode 885 amino acid residues. Its secondary structure contains 41 alpha-helixes and 60 coiled coils. After uploading it to the NCBI,it was found that the gene has the highest homology with Diuraphis noxia,and has low homology with other species. The relative expression of TERT in different ages and tissues of L. erysimi was analyzed by real-time PCR. The results showed that the relative expression of TERT gene in the adult stage of L. erysimi was the highest,and there were significant differences with each age. The expression level of 1st instar nymph was significantly lower than that of adults,but significantly higher than that of 2nd,3rd and 4th instar nymphs. The expression level of 4th instar nymph was the lowest,but there was no significant difference with that of 2nd and 3rd instar nymphs. Subsequently,the 4th instar nymphs,which is basically mature and has the lowest expression level of TERT gene,was selected for dissection. We were able to simultaneously detect the expression of TERT gene in the head,chest and abdomen of 4th instar nymphs with the highest expression in the abdomen. These results suggest that the TERT gene plays an important role in the growth and development of L. erysimi,and the region with low similarity can be selected as the target region for gene interference or silence so as to control the major harm caused by L. erysimi in agriculture.

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        張鑫澤,張慧杰,周義成,李昭,王赫遠,朱智慧.蘿卜蚜端粒逆轉錄酶TERT基因的鑒定與表達分析[J].華中農業大學學報,2020,39(6):8-14

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        • 收稿日期:2019-12-05
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        • 在線發布日期: 2020-12-01
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