Welcome to Chinese Journal of Tropical Crops,

Chinese Journal of Tropical Crops ›› 2019, Vol. 40 ›› Issue (12): 2411-2417.DOI: 10.3969/j.issn.1000-2561.2019.12.014

• Biotechnology and Tissue Culture • Previous Articles     Next Articles

Cloning and Expression of a Chitinase Gene SgGH19-1 from Stylosanthes and Its Enzymatic Properties Analysis

LIU Pandao1,WU Xique2,LUO Jiajia1,XU Wenrong2,*(),LIU Guodao1,*()   

  1. 1. Tropical Crops Genetic Resources Institute, Chinese Academy of Tropical Agriculture Sciences / Key Laboratory of Crop Gene Resources and Germplasm Enhancement in Southern China, Ministry of Agriculture and Rural Affairs, Haikou, Hainan 571101, China
    2. Department of Chemistry, School of Science, Hainan University, Haikou, Hainan 570228, China
  • Received:2019-08-19 Revised:2019-10-08 Online:2019-12-25 Published:2019-12-20
  • Contact: XU Wenrong,LIU Guodao


Chitin is a major component of the cell wall of fungi, exoskeleton of insects, and crustacean shells. Chitinase is one of the main hydrolase that catalyzes the degradation of chitin. Induction of PR-3 chitinase protein accumulation is an adaptive mechanism for plants to enhance the resistance to fungal diseases. Stylo (Stylosanthes spp.) is an important tropical forage legume. Although anthracnose caused by Colletotrichum gloeosporioides is one of the most destructive fungal diseases of stylo, little information is available regarding the responses of stylo chitinase during the pathogen infection. In the study, the PR-3 chitinase of stylo was characterized, and its expression pattern and biochemical properties were also analyzed. The results showed that a chitinase gene of PR-3 group in stylo was the cloned by homologous gene sequence method, and its coding region was 984 bp. The gene belongs to the Class I chitinase of the glycoside hydrolase 19 family, and was named SgGH19-1. Quantitative PCR analysis showed that the expression level of SgGH19-1 gene in the leaves of stylo was significantly increased after C. gloeosporioides infection, and with the increase of chitinase activity. Subsequently, the recombinant protein of SgGH19-1 was expressed and purified in Escherichia coli. Biochemical properties of SgGH19-1 showed both exochitinase and endochitinase activities, and the activities of endochitinase was 9.1 fold higher than that of exochitinase. Furthermore, the optimum pH and temperature for SgGH19-1 activity was 5.0 and 40 ℃ respectively. Taken together, SgGH19-1 is a chitinase that involved in the response of stylo to C. gloeosporioides attack. These results herein suggest that SgGH19-1 could potentially be employed as a new marker gene for developing cultivars of stylo with great tolerance to anthracnose.

Key words: chitin, chitinase, Stylosanthes, protein purification, enzymatic property

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