Biotechnology Bulletin ›› 2022, Vol. 38 ›› Issue (10): 124-131.doi: 10.13560/j.cnki.biotech.bull.1985.2022-0069

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Identification and Gene Mapping of a Seedling Lethal Mutant in Rice

TANG Yue-hui(), ZHAO Yu-fan, LIN Jin, WANG Yin, CAO Bo-yuan, CHE Yi-fan, YANG Wen-jie, BAO Xin-xin, YANG Tong-wen   

  1. College of Life Science and Agronomy,Zhoukou Normal University,Zhoukou 466001
  • Received:2022-01-13 Online:2022-10-26 Published:2022-11-11
  • Contact: TANG Yue-hui E-mail:yhtang2005@163.com

Abstract:

Peptide release factor 1(RF1)plays a key role in the regulation of the translation termination by recognizing stop codons. Rice seedling lethal mutant was obtained,the expression profile of OseRF1-1 was clarified,and a basis for further function analysis of OseRF1-1 was provided. Using the Japonica rice cv. Huazhiwu and T-DNA insertion mutant as materials,Tail-PCR,paraffin section technology,Southern blotting and Northern blotting technologies were used to identify a T-DNA single-copy insertion mutant of the peptide release factor eRF1-1,named ls. DNAMAN6.0 software and SMART were applied to analyze the OseRF1-1’s bioinformatics characteristics,and RT-qPCR was used to analyze the tissue-specific expression of OseRF1-1. Transforming Arabidopsis thaliana by a PEG-mediated approach was to clarify the subcellular localization of OseRF1-1. The results suggested that the ls mutant rapidly died at the 3rd to 5th leaf tillering stages. Paraffin section showed that the junction region between leaf sheath and tiller node in ls mutant was browning,and the symptoms were more serious in the lower part of tiller nodes. Tail-PCR analysis showed that the right border of the T-DNA and the NOS terminator sequence in ls mutant were lost,resulting in a fusion transcript of GUS and OseRF1-1 in ls mutant. However,the stop codon remained in GUS,which led to the OseRF1-1 not translated normally. OseRF1-1 was expressed in all tissues tested,especially highly expressed in panicles. This OseRF1-1 gene was localized in nucleus and cytoplasm. Above results indicate that ls mutant causing rice seedlings lethal may be resulted from the abnormal transcription of OseRF1-1 gene by T-DNA insertion.

Key words: Oryza sativa L., peptide release factor 1, ls mutant, seedling lethal