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Acta Agron Sin ›› 2017, Vol. 43 ›› Issue (01): 31-41.doi: 10.3724/SP.J.1006.2017.00031

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Cloning and Expression Analysis of Key Gene ScD27 in Strigolactones Biosynthesis Pathway

WU Zhuan-Di**,LIU Xin-Long**,LIU Jia-Yong,ZAN Feng-Gang,LI Xu-Juan,LIU Hong-Bo,LIN Xiu-Qin, CHEN Xue-Kuan, SU Huo-Sheng,ZHAO Pei-fang,WU Cai-Wen*   

  1. Sugarcane Research Institute, Yunnan Academy of Agricultural Sciences / Yunnan Key Laboratory of Sugarcane Genetic Improvement, Kaiyuan 661699, China
  • Received:2016-01-11 Revised:2016-09-18 Online:2017-01-12 Published:2016-09-28
  • Contact: 吴才文, E-mail: gksky_wcw@163.com
  • Supported by:

    This study was supported by the National Natural Science Foundation of China (31360359), the National Modern Agricultural Industry Technology System Construction Project (CARS-20-1-1), the Young and Middle-aged Academic Technology Leaders Reserve Talented Person in Yunnan Province (2014HB038), the Applied Basic Research Projects in Yunnan Province (2016FB071), the Major Science and Technology Projects – Biology (2015ZA001), and the Science and Technology Innovation Talents Project (2014HC015).

Abstract:

Strigolactones (SLs) is a novel class of plant hormones. D27 regulating reversible metabolic process is located in up-stream of strigolactones biosynthesis pathway. In this study, primers were designed based on the conserved domains from four species inluding Oryza sativa, Zea mays, Sorghum bicolor, and Brachypodium distachyon. Using cDNA from sugarcane cultivar ROC22 as the template, the full-length cDNA sequence of D27 gene from sugarcane was cloned by RT-PCR and RACE method. This gene isnamed as ScD27, with the GenBank accession number of KP987221.1. Its length is 1379bp, and it contains an 867bp open reading frame (ORF), encoding 288 amino acid residues. ScD27 is not a secretory protein and has a molecular weight of 71.58kD, with a theoretical isoelectric point of 5.04. ScD27 is mainly located in chloroplast and the conserved domains of this protein involve two zinc finger protein structures (ZnF_TAZ and ZnF_A20). Amino acid sequences encoded by ScD27 shared more than 70% similarity with the reported amino acid sequences encoded by D27 of Sorghum bicolor, Setaria italica Beauv., Hordeum vulgare subsp. vulgareand Brachypodium distachyon. ScD27 gene was differentially expressed in different parts of sugarcane plant, with higher level of transcript in stem tip and axillary bud but much lower level in leaf, stem and root. Furthermore, the expression of ScD27 could be induced by the stresses of PEG, salt and the deficiencies of phosphorus and nutrition. These results demonstrated that ScD27 might be a key gene participating in the response to abiotic stresses during sugarcane SLs biosynthesis pathway.

Key words: Sugarcane, ScD27, Homology cloning, Bioinformatics, q-PCR

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