Feiyan Huang1, Farhat Abbas2, Dylan O'Neill Rothenberg3, Muhammad Imran4, Sajid Fiaz5, Naveed Ur Rehman6, Sikandar Amanullah7, Afifa Younas8, Yan Ding9, Xianjie Cai10, Xiaolong Chen11, Lei Yu1, Xianwen Ye12, Lin Jiang13, Yanguo Ke14,15, Yuansheng He16. 1. College of Agriculture and Life Sciences, Yunnan Urban Agricultural Engineering & Technological Research Center, Kunming University, Kunming, China. 2. The Research Center for Ornamental Plants, College of Forestry and Landscape Architecture, South China Agricultural University, Guangzhou, 510642, China. 3. College of Horticulture Science, South China Agricultural University, Guangzhou, 510642, China. 4. Department of Crop Science and Technology, College of Agriculture, South China Agricultural University, Guangzhou, 510642, China. 5. Department of Plant Breeding and Genetics, University of Haripur, Khyber Pakhtunkhwa, Pakistan. 6. State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangdong Provincial Key Laboratory of Plant Molecular Breeding, South China Agricultural University, Guangzhou, 510642, China. 7. College of Horticulture and Landscape Architecture, Northeast Agricultural University, Harbin, 150030, Heilongjiang, China. 8. Department of Botany, Lahore College for Women University, Lahore, Pakistan. 9. Material Procurement Center, Shanghai Tobacco Group Co., Ltd, Shanghai, 200082, Yunnan, China. 10. Material Procurement Center, Shanghai Tobacco Group Co., Ltd, Shanghai, 200082, Yunnan, China. caixj@sh.tobacco.com.cn. 11. Tobacco Leaf Purchase Center, China Tobacco Henan Industrial Co., Ltd., Zhengzhou, 450000, China. 12. Kunming Tobacco Corporation of Yunnan Province, Kunming, 650021, China. 13. Honghe Tobacco Corporation of Yunnan Province, Honghe, 661400, China. 14. College of Agriculture and Life Sciences, Yunnan Urban Agricultural Engineering & Technological Research Center, Kunming University, Kunming, China. keyanguo@126.com. 15. College of Economics and Management, Kunming University, Kunming, China. keyanguo@126.com. 16. Lincang Company of Yunnan Tobacco Company, Lincang, 677000, China.
Abstract
BACKGROUND: 12-oxophytodienoic acid (OPDA) is a signaling molecule involved in defense and stress responses in plants. 12-oxophytodienoate reductase (OPR) is involved in the biosynthesis of jasmonic acid and trigger the conversion of OPDA into 3-oxo-2(2'[Z]-pentenyl)-cyclopentane-1-octanoic acid (OPC-8:0). METHODS AND RESULTS: Sequence analysis revealed that Nicotiana tabacum 12-oxophytodienoate reductase 1 (OPR1) and OPR2 encoded polypeptides of 375 and 349 amino acids with molecular masses of 41.67 and 39.04 kilodaltons (kDa), respectively, while the deduced protein sequences of NtOPR1 and NtOPR2 showed high homology with other 12-oxophytodienoate reductases. BLAST (Basic local alignment search tool) analysis revealed that both NtOPRs belong to the family of Old Yellow Enzymes (OYE), and analysis of genomic DNA structure indicated that both genes include 5 exons and 4 introns. Phylogenetic analysis using MEGA X showed that NtOPR1 and NtOPR2 shared a close evolutionary relationship with Nicotiana attenuata 12-oxophytodienoate reductases. In silico analysis of subcellular localization indicated the probable locations of NtOPR1 and NtOPR2 to be the cytoplasm and the peroxisome, respectively. Tissue-specific expression assays via qRT-PCR revealed that NtOPR1 and NtOPR2 genes were highly expressed in Nicotiana tabacum roots, temperately expressed in leaves and flowers, while low expression was observed in stem tissue. CONCLUSIONS: Presently, two 12-oxophytodienoate reductase genes (NtOPR1 and NtOPR2) were cloned and comprehensively characterized. Our findings provide comprehensive analyses that may guide future deep molecular studies of 12-oxophytodienoate reductases in Nicotiana tabacum.
BACKGROUND: 12-oxophytodienoic acid (OPDA) is a signaling molecule involved in defense and stress responses in plants. 12-oxophytodienoate reductase (OPR) is involved in the biosynthesis of jasmonic acid and trigger the conversion of OPDA into 3-oxo-2(2'[Z]-pentenyl)-cyclopentane-1-octanoic acid (OPC-8:0). METHODS AND RESULTS: Sequence analysis revealed that Nicotiana tabacum 12-oxophytodienoate reductase 1 (OPR1) and OPR2 encoded polypeptides of 375 and 349 amino acids with molecular masses of 41.67 and 39.04 kilodaltons (kDa), respectively, while the deduced protein sequences of NtOPR1 and NtOPR2 showed high homology with other 12-oxophytodienoate reductases. BLAST (Basic local alignment search tool) analysis revealed that both NtOPRs belong to the family of Old Yellow Enzymes (OYE), and analysis of genomic DNA structure indicated that both genes include 5 exons and 4 introns. Phylogenetic analysis using MEGA X showed that NtOPR1 and NtOPR2 shared a close evolutionary relationship with Nicotiana attenuata 12-oxophytodienoate reductases. In silico analysis of subcellular localization indicated the probable locations of NtOPR1 and NtOPR2 to be the cytoplasm and the peroxisome, respectively. Tissue-specific expression assays via qRT-PCR revealed that NtOPR1 and NtOPR2 genes were highly expressed in Nicotiana tabacum roots, temperately expressed in leaves and flowers, while low expression was observed in stem tissue. CONCLUSIONS: Presently, two 12-oxophytodienoate reductase genes (NtOPR1 and NtOPR2) were cloned and comprehensively characterized. Our findings provide comprehensive analyses that may guide future deep molecular studies of 12-oxophytodienoate reductases in Nicotiana tabacum.
Authors: H. Nojiri; M. Sugimori; H. Yamane; Y. Nishimura; A. Yamada; N. Shibuya; O. Kodama; N. Murofushi; T. Omori Journal: Plant Physiol Date: 1996-02 Impact factor: 8.340
Authors: Jürgen Engelberth; Irmgard Seidl-Adams; Jack C Schultz; James H Tumlinson Journal: Mol Plant Microbe Interact Date: 2007-06 Impact factor: 4.171