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Acta Agron Sin ›› 2012, Vol. 38 ›› Issue (03): 549-555.doi: 10.3724/SP.J.1006.2012.00549

• RESEARCH NOTES • Previous Articles     Next Articles

Molecular Cloning and Express Analysis of Δ1-Pyrroline-5-Carboxylate Synthetase (P5CS) Gene in Ramie

ZHOU Jing-Hua1,XING Hu-Cheng1,2, JIE Yu-Cheng1,2,ZHONG Yin-Li1,3,ZHU Shou-Jing1,JIANG Jie1,WANG Liang1   

  1. 1 Institute of Ramie, Hunan Agriculture University, Changsha 410128, China; 2 Hunan Provincial Key Laboratory of Corp Germplasm Innovation and Utilization, Changsha 410128, China; 3 College of Bioscience and Biotechnology, Changsha 410128, China
  • Received:2011-06-25 Revised:2011-10-12 Online:2012-03-12 Published:2011-12-01
  • Contact: 揭雨成, E-mail: ibfcjyc@vip.sina.com, Tel: 13874940038

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Abstract: Proline is one of osmotic regulators in plant, which plays an important role in regulating ramie in response to drought stress. The synthesis of proline has been reported to be restricted by the enzyme of Δ1-pyrroline-5-carboxylate synthetase (P5CS). In this study, taking Xiangzhu 3 with drought tolerance as an experimental material, we obtained a ramie unigene highly identified with P5CS gene from the ramie transcription group sequencing, and the gene length was 1 448 bp. According to the known fragment, the 5' and 3' RACE PCR nest primers were designed, and a 590 bp 5′ terminal sequence and a 293 bp 3′ terminal sequence were amplified by RACE and RT-PCR, separately. The results of bioinformatics analysis showed that the full length of the P5CS gene was 2 318 bp, the open reading frame was 2 154 bp, coding 717 amino acids, the isoelectric point and the molecular weight of coded protein were 6.57 and 77.56 kD, respectively. Nucleotide sequence Blast indicated that the unigene of ramie shared an identity of 81%, 81%, 75% and 72% with P5CS genes of cotton, leprosy tree,Arabidopsis thaliana and rice, respectively, and the similarity in protein Blast was 86%, 85%, 78%, and 77%, respectively. The result showed it was a homologous gene of P5CS in ramie, and named BnP5CS1. Semi-quantitative RT-PCR analysis indicated that the expression of BnP5CS1 had a higher level in stem tip than in root, and the gene induced by the drought stress.P5CS is a key enzyme gene in the synthesis of proline in plants, so the cloning of BnP5CS1 gene might lay a foundation for the adversity resistance molecular breeding of ramie and further function analysis.

Key words: Boehmeria nivea, P5CS, Proline, Drought, Expression analysis

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