Construction and Analysis of SSH-cDNA Library from Leaves of Susceptible Rubber Clone Resistant to Powdery Mildew Induced by BTH

Chanjuan Luo, Zhiwei Fan, Yide Shen, Xiaoxia Li, Hanting Chang, Qiaoqiao Huang, Lizhen Liu

Environment and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences/Key Laboratory of Integrated Pest Management of Tropical Crops, Ministry of Agriculture/Key Laboratory of Pests Detection and Control for Tropical Agriculture, Haikou, China..

DOI: 10.4236/ajps.2013.43068                

Abstract

To understand the mechanism of benzothiadiazole (BTH)-induced susceptible rubber clone resistance to powdery mildew on gene level, a differentially expressed cDNA library was constructed by suppression subtractive hybridization (SSH) with rubber Reyan 7-33-97 clone. The constructed cDNA library was high integrity through detection of the critical processes of SSH, such as efficiency of adaptor connection, subtraction and conversion, as well as the type of recombinant genes. The positive rate was 99% after identification with random 400 white spots. The size of the cDNA clone inserted fragments was various but most in 400 bp - 1000 bp. There were 23 cDNA sequences matching the function of energy and basic metabolism, signal transduction, membrane and transport, secondary metabolism and so on after detection of the 42 positive clone sequences selected randomly from the cDNA library and comparison on nucleic acid sequences in Genbank. 7 ESTs were logged in Genbank and accession numbers were GW873071 and GW874604- GW874610. The results implicated that BTH could effectively induced rubber tree resistance to powdery mildew through increasing expresses of defense-related genes in leaves of rubber tree susceptible clone. It should provide a new approach for rubber disease management.

Keywords

Benzothiadiazole; ESTs; Hevea brasiliensis; Induced Resistance; Oidium heveae; Suppression Subtractive Hybridization

Conflicts of Interest

The authors declare no conflicts of interest.

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