Response of AtR8 lncRNA to Salt Stress and Its Regulation on Seed Germination in Arabidopsis

doi:10.11983/CBB19244

Abstract

Abstract: Long non-coding RNA (lncRNA) is a type of non-coding RNA that is longer than 200 nucleotides and does not encode proteins. lncRNAs are mainly produced by the transcription of RNA polymerase II and are abundant in the organism and have various biological functions. AtR8 lncRNA is transcribed by RNA polymerase III in Arabidopsis thaliana. Previous studies revealed that Salicylic acid (SA) induces AtR8 lncRNA expression in germinated seeds and that the deletion of AtR8 lncRNA decreases seed germination under SA stress. In this study, we found a conserved salt-stress-responsive element (TCTTCTTCTTTA) in the transcriptional region of AtR8 lncRNA. NaCl treatment inhibited AtR8 lncRNA expression in the germinated seeds. High concentration of NaCl significantly inhibited seed germination of atr8, which had partial deletion of AtR8 lncRNA, compared to that of the wild type, indicating that AtR8 lncRNA plays an important role in regulating seed germination in response to salt stress.

Key words: long non-coding RNA, Arabidopsis thaliana, seed germination, salt stress

Nan Zhang,Ziguang Liu,Shichen Sun,Shengyi Liu,Jianhui Lin,Yifang Peng,Xiaoxu Zhang,He Yang,Xi Cen,Juan Wu. Response of AtR8 lncRNA to Salt Stress and Its Regulation on Seed Germination in Arabidopsis[J]. Chinese Bulletin of Botany, 2020, 55(4): 421-429.

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URL: https://www.chinbullbotany.com/EN/10.11983/CBB19244

https://www.chinbullbotany.com/EN/Y2020/V55/I4/421

Figures/Tables 3

Figure 1 Analysis of AtR8 lncRNA expression during seed germination in Arabidopsis thaliana (A) Northern blotting analysis of AtR8 lncRNA expression during seed germination (expression of 5.8S rRNA serves as loading controls); (B) In situ hybridization of AtR8 lncRNA expression during seed germination (arrow indicates AtR8 lncRNA signal) (Bars=200 μm).

Figure 2 AtR8 lncRNA expression during seed germination of Arabidopsis thaliana after NaCl treatment (A) Sequence comparison between AtR8 lncRNA and UCC salt stress-responsive element (the USE and TATA promoter sequences are capitalized, bloded and framed; the AtR8 lncRNA transcriptional region is capitalized and bolded; and the conserved salt stress-responsive element is marked with asterisk). (B) The location of the salt stress-responsive element in the secondary structure of AtR8 lncRNA predicted by RNAlogo (the salt stress-responsive motif is capitalized, bloded and indicated with an arrow). (C) Northern blotting analysis of AtR8 lncRNA expression in germinating seeds under different NaCl treatments, 5.8S rRNA was used as a loading control. The lower panel shows semi-quantitative analysis of the Northern blotting signals. Two independent experiments gave similar results, and a representative example is shown. Values are means ± SE (t-test, *P<0.05, **P<0.01). (D) Northern blotting analysis of AtR8 lncRNA in germinating seeds under different periods of 150 mmol·L-1 NaCl treatment, 5.8S rRNA was used as a loading control. The lower panel shows semi-quantitative analysis of the Northern blotting signals. Two independent experiments gave similar results, and a representative example is shown. Values are means ± SE (t-test, **P<0.01).

Figure 3 Loss of AtR8 lncRNA inhibits Arabidopsis thaliana seed germination under salt stress (A) Northern blotting analysis of the AtR8 lncRNA loss-of-function mutant atr8 (5.8S rRNA was used as the loading control); (B) Statistical analysis of growth and germination rate of the wild-type and atr8 seeds under different concentrations of NaCl (data are average of three independent experiments, and bars indicate standard error); (C) Fresh and dry weight of the wild-type and atr8 seeds after 7 d of germination under NaCl treatment (the values are average of three independent experiments, and bars indicate standard error (the asterisk indicate significant differences between atr8 and wild type (t-test, * P < 0.05, ** P < 0.01)). WT: Wild type

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