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Nicotiana benthamiana

ABSTRACT: Transient expression of intron-containing transgenes generates non-spliced aberrant pre-mRNAs that are processed into siRNAs

PROVIDER: PRJNA481240 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR7535030.fastq.gz	Fastqsanger.gz
	SRR7535031.fastq.gz	Fastqsanger.gz
	SRR7535032.fastq.gz	Fastqsanger.gz
	SRR7535033.fastq.gz	Fastqsanger.gz
	SRR7535034.fastq.gz	Fastqsanger.gz

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Project description:Nonsense mediated decay (NMD) is a translation termination coupled quality control system. NMD identifies and degrades aberrant mRNAs containing premature termination codons, thereby preventing the accumulation of detrimental truncated proteins. NMD also controls the expression of several normal mRNAs and non-coding transcripts. RNA-seq assays were conducted to identify the NMD regulated genes in Arabidopsis. However, NMD targets have not been studied in other plants. To identify the conserved NMD targets, we wanted to identify the NMD regulated genes in Nicotiana benthamiana model species. Virus induced gene silencing was used to inactivate UPF1 NMD key factor in N. benthamiana, and then comparative RNA-seq experiment was carried out using UPF1-silenced and control–silenced plants. We found that in N. benthamiana significantly more genes are controlled by NMD than in Arabidopsis and that, surprisingly few conserved NMD targets can be recognized. These results suggest that while the mechanism of NMD is highly conserved, the set of NMD controlled genes can be quickly changed in plants.

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Profile of small interfering RNAs derived from Beet black scorch virus and its associated satellite RNA in Nicotiana benthamiana

Project description:Co-infection of none-coding satellite RNAs (sat-RNAs) usually inhibits replication and attenuates disease symptoms of helper viruses. Here we find that the sat-RNA of Beet black scorch virus (BBSV) enhances both the accumulation and the systemic infection of its helper virus in the Nicotiana benthamiana (N. benthamiana). Besides, the infection of BBSV is higher efficient in low temperature (18ºC) than room temperature (25ºC), suggesting that BBSV is temperature sensitive. To explore the effect of temperature and sat-RNA on the silencing induced by BBSV, we cloned and sequenced the total small RNAs produced during infection of BBSV alone or with its sat-RNA under different temperature. Our results shows that the size profile of the siRNAs derived from either BBSV or coinfection with sat-RNA are 22 nucleotide (nt), followed by 21-nt reads and other length reads. Notably, more than 80% of the total small RNAs sequenced from plants infected with BBSV alone were BBSV-siRNAs, whereas the abundance of sat-RNA-derived siRNAs were higher than BBSV-siRNAs in the co-infected plants, indicating sat-RNA involved in gene silencing as a surrogacy of BBSV. Interestingly, the 5′ termini of siRNAs derived from BBSV and sat-RNA were dominated by Uridines (U) and Adenines (A), respectively. Consistently, the infection of BBSV alone or with sat-RNA induces high accumulation of Argonaute 1 (AGO1) and AGO2, respectively. Our work reveals the profiles of siRNAs of BBSV and sat-RNA and provides a new clue to investigate the complicated interaction between the helper virus and sat-RNA.

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