中国农业科学2025，Vol.58Issue(14)：2720-2738,19.DOI:10.3864/j.issn.0578-1752.2025.14.002
黑麦活性着丝粒特异反转录转座子的筛选及其对逆境胁迫的响应
Screening of Active Centromeric Retrotransposons of Rye and Their Response to Stress
摘要
Abstract
[Objective]Centromeric retrotransposons(CR)play an important role in maintaining chromosomal stability of Poaceae species.Rye(Secale cereale L.cv.Imperial),a valuable genetic donor for wheat improvement,showed enhanced stress tolerance and disease resistance,the centromeric regions of which are enriched with CR.Screen active rye specific CR(CR of Rye,CRR)and study their transposition patterns under stress conditions,would help elucidate the mechanisms of the possible influence of transposable elements(TEs)transposition on genome stability of rye under stress conditions.[Method]To identify intact CRR,the rye genome was subjected to de novo annotation of TEs using bioinformatics tools.Semi-quantitative analysis was applied to screen highly expressed CRR in both leaves and roots of rye.Quantitative real-time PCR(qRT-PCR),methylation-specific PCR(MSP),and transposon display(TD)techniques were used to analyze the expression and methylation level,and transposition activity of the screened CRR in leaves and roots of abiotic stressed rye seedlings(at the one-tip-two-leaf stage),including salt,ABA,H2O2,PEG,low temperature,and high temperature.[Result]Seventeen CRR were identified,and seven have intact structure(CRR1,CRR2,CRR3,CRR4,CRR5,CRR7,and CRR11).Semi-quantitative analysis revealed CRR2,CRR4,and CRR7 were highly expressed in both leaves and roots.Structural analysis of the three CRR indicated that they could encode all the enzymes necessary for TE transposition(reverse transcriptase,ribonuclease H,and integrase),with CRR7 also encoded a Gag protein.Under normal conditions,CRR2,CRR4,and CRR7 were basically expressed,which were upregulated by stress treatments,the methylation level of CRR7 changed most under stressed conditions,followed by CRR2 and CRR4.Additionally,the copy number of the three CRR was dynamically changed under stress conditions.Under different stress conditions,the insertion and excision frequency of CRR was different under different stress conditions,but the overall excision frequency was higher than the insertion frequency.[Conclusion]The higher of the sequence homology between the 5'and 3'LTRs of CRR,the higher of the transcriptional activity of CRR;active CRR have basical transcriptional level under normal conditions,the transcription and transposition activity of which were upregulated by stress stimuli,which were primarily regulated by post-transcriptional regulatory mechanisms.Genomic rearrangement might be the main factor affecting the copy number of CRR in stress conditions.关键词
黑麦/着丝粒/转座子元件/逆境胁迫/甲基化/转座活性Key words
rye/centromere/transposon element/adversity stress/methylation/transposition activity引用本文复制引用
吴翠翠,赵丽丽,孙吉,方正武,程玲,朱展望,刘易科,张迎新,王书平,王若禹,马驰,何美月,尹晓康,冯佳宜,周宾寒,姜宇凡,金汉炳..黑麦活性着丝粒特异反转录转座子的筛选及其对逆境胁迫的响应[J].中国农业科学,2025,58(14):2720-2738,19.基金项目
湖北省重点研发计划(2024BBB004)、植物细胞与染色体工程国家重点实验室开放课题(PCCE-KF-2023-10)、粮食作物种质创新与遗传改良湖北省重点实验室开放课题(2024lzjj15) （2024BBB004）
