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Acta Agron Sin ›› 2012, Vol. 38 ›› Issue (11): 2024-2033.doi: 10.3724/SP.J.1006.2012.02024

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

Cloning and Expression Analysis of ATP-Citrate Lyase Genes from Sugarcane

LI Chang-Ning1,2,NONG Qian1,TAN Qin-Liang1,SRIVASTAVA Manoj Kumar2,YANG Li-Tao1,2,LI Yang-Rui1,2,*   

  1. 1 Agricultural College / State Key Laboratory of Conservation and Utilization of Subtropical Agro-bioresources, Guangxi University, Nanning 530005, China;
    2 Sugarcane Research Center, Chinese Academy of Agricultural Sciences / Key Laboratory of Sugarcane Biotechnology and Genetic Improvement (Guangxi), Ministry of Agriculture / Guangxi Crop Genetic Improvement and Biotechnology Laboratory / Guangxi Key Laboratory of Sugarcane Genetic Improvement, Nanning 530007, China
  • Received:2012-04-27 Revised:2012-07-05 Online:2012-11-12 Published:2012-09-10
  • Contact: 李杨瑞, E-mail: lyr@gxaas.net

Abstract:

Acetyl-CoA plays an important role in the cytosol of plant cells for the synthesis of a diverse set of phytochemicals, and the cytosolic acetyl-CoA is from the reaction of citrate and CoA catalyzed by ATP-citrate lyase (ACL) coupled with the hydrolysis of ATP. In this research, two genes encoding two distinct subunits of ACL were identified from sugarcane (Saccharum officinarum) by RACE and insilico cloning technology, named SoACLA-1 and SoACLB-1, which contained 1 272 and 1 828 bp open reading frames, and encoded 423 and 608 amino acids, respectively. Sequence analysis indicated that both amino acids sequences showed high homology with ACLs from other species and were close clustered with ACLs from gramineous plants in the phylogenetic tree constructed by MEGA5.0 software using a Neighborhood-Joining bootstrap method. Both sequences showed high conservation in the ATP-grasp domain, citrate binding site, active binding site of histidine phosphorylated by ATP, potential ATP-binding, phosphorylated site and CoA-binding site when compared with ACLs from other species. Quantitative real-time PCR results indicated that SoACLA-1 and SoACLB-1 were both induced by the treatment of control+ABA, water stress, water stress +ABA, and their expression levels were higher in leaves than in roots, with the highest express level in water stress treatment, SoACLA-1 and SoACLB-1 showed a synergetic expression pattern. The expression of SoACLA-1 and SoACLB-1 was close correlated with the ABA and ROS contents, indicating that ACL maybe involved in stress responsive metabolic process triggered by ABA in plants.

Key words: 甘蔗, ATP柠檬酸裂解酶, 克隆, 脱落酸, 表达

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