Study on Sengon (Falcataria moluccana) Resistance to Boktor Pest (Xystrocera festiva) and Gall Rust (Uromycladium tepperianum) Part III: Optimation of Cloning Protocols and Gene Expression Study
UJ. Siregar, S. Hartati

Source: SEAMEO BIOTROP's Research Grant | 2018


Sengon (Falcataria moluccana Miq.) is a multifunctional and fast growing tree species which has a high economic value. However, the increasing trend of sengon plantation has caused a pest outbreak, especially Xystrocera festiva stem borer. Boktor larvae have trypsin enzyme in their gut to hydrolyse stem of sengon.  Some sengon resistant accessions showed inhibitory activity of trypsin enzyme, which are important aspect for sengon tree improvement program. The objectives of this research are to optimize preparation of cloning method for the construction of genomic library, especially trypsin inhibitor (TI) gene. This research used shotgun cloning and specific gene cloning method to establish genomic library. The genomic library was successfully constructed using pGEM-T Easy vector in E. coli bacteria strain DH5a, generating about 9 colonies by shotgun cloning method and sequences of sengon TI genes were obtained with sizes ranging from 107 to 308 bp. The sequences of the  colonies had high similarity from nuclear, chloroplast and mitochondrial genomes of various tree species and TI sequences were similar to Tamarindus indica, Glycine max, Populus trichocarpa and Populus nigra, however those TI sequences had low E values.

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