Construction Of Poplar SSH CDNA Library Induced By Tetra Lobulifera (Keifer) And Gene Expression Profiling Analysis

Tetra lobulifera (Keifer) was an important pest mite of poplar wide-distributed. it was a new record in our country. There is no detailed report on the mite. Tetra lobulifera (Keifer) is methods of the temporary name in Chinese.We preliminary studied the biological characteristics of the Tetra lobulifera (Keifer). Under the major site in the same conditions, comparisons has been made under different levels of poplar. With the increasing level of Tetra lobulifera (Keifer), the growth potential of the host is weakened, and the more suffered poplar , the more developed its collateral, and slow-growing trunk.A suppression subtractive hybridization (SSH) cDNA library was constructed with populus deltoides induced by Tetra lobulifera (Keifer). The leaf was induced with 48 and 72 hour as tester, the uninduced leaf as driver. Seven hundred and ten cDNA clones were identified, and acquired 496 EST sequences in the end. The BLAST homology search revealed that these genes played varies functions in response to stress, signal transduction, energy metabolism and transportation etc. Real-time PCR was performed to confirm results, ten differentially expressed genes appeared up-regulated in four times.Two full-long cDNA sequences were cloned by the RACE (Rapid Amplification of cDNA Ends), named as PtWRKY and PtERF. The cDNA of PtWRKY was 2053bp long, encoding 588 amino acids, contain two typical WRKY-domain. And the PtERF cDNA was1074bp in length, encoding 210 amino acids, amino acid sequence according to typical characteristics of AP2/ERF transcription factors. Overexpression vetors were constructed and transferred into Populus tremula×P. tremuloides Michx-clone T89. PCR and RT-PCR analysis indicated that the target genes had been integrated into poplar genome.