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Nat Plants. 2019 Jan;5(1):26-33. doi: 10.1038/s41477-018-0320-9. Epub 2018 Dec 10.

Sensitive detection of pre-integration intermediates of long terminal repeat retrotransposons in crop plants.

Author information

1
The Sainsbury Laboratory, University of Cambridge, Cambridge, UK. jungnamcho@sippe.ac.cn.
2
National Key Laboratory of Plant Molecular Genetics, CAS Center for Excellence in Molecular Plant Sciences, Institute of Plant Physiology and Ecology, Shanghai, China. jungnamcho@sippe.ac.cn.
3
CAS-JIC Centre of Excellence for Plant and Microbial Science, Chinese Academy of Sciences, Shanghai, China. jungnamcho@sippe.ac.cn.
4
The Sainsbury Laboratory, University of Cambridge, Cambridge, UK.
5
School of Biosciences, University of Birmingham, Birmingham, UK.
6
Laboratory of Nematology, Wageningen University, Wageningen, the Netherlands.
7
The Sainsbury Laboratory, University of Cambridge, Cambridge, UK. jerzy.paszkowski@slcu.cam.ac.uk.
8
Radachowka 37, Kolbiel, Poland. jerzy.paszkowski@slcu.cam.ac.uk.

Abstract

Retrotransposons have played an important role in the evolution of host genomes1,2. Their impact is mainly deduced from the composition of DNA sequences that have been fixed over evolutionary time2. Such studies provide important 'snapshots' reflecting the historical activities of transposons but do not predict current transposition potential. We previously reported sequence-independent retrotransposon trapping (SIRT) as a method that, by identification of extrachromosomal linear DNA (eclDNA), revealed the presence of active long terminal repeat (LTR) retrotransposons in Arabidopsis3. However, SIRT cannot be applied to large and transposon-rich genomes, as found in crop plants. We have developed an alternative approach named ALE-seq (amplification of LTR of eclDNAs followed by sequencing) for such situations. ALE-seq reveals sequences of 5' LTRs of eclDNAs after two-step amplification: in vitro transcription and subsequent reverse transcription. Using ALE-seq in rice, we detected eclDNAs for a novel Copia family LTR retrotransposon, Go-on, which is activated by heat stress. Sequencing of rice accessions revealed that Go-on has preferentially accumulated in Oryza sativa ssp. indica rice grown at higher temperatures. Furthermore, ALE-seq applied to tomato fruits identified a developmentally regulated Gypsy family of retrotransposons. A bioinformatic pipeline adapted for ALE-seq data analyses is used for the direct and reference-free annotation of new, active retroelements. This pipeline allows assessment of LTR retrotransposon activities in organisms for which genomic sequences and/or reference genomes are either unavailable or of low quality.

PMID:
30531940
PMCID:
PMC6366555
[Available on 2019-06-10]
DOI:
10.1038/s41477-018-0320-9

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