In this study we will have three tomato variety, 'LA2710', 'micro tom' and 'money maker'. We have already obtained the LePDS silenced plant, which almost have 100% efficiency; in the same time we are working on the GFP overexpression plants. We will choose some gene of interest, especially transcriptional factors from tomato cDNA library, which obtained from Kazusa DNA Research Institute, Tokyo, Japan. For example, bZip, which is related with abiotic stress, like drought and salt (Hsieh TH, 2010); MADS-Box Transcription Factor, which related with fruit ripening (Catherine Martel, 2011). We will use the VIGS Tool in Sol Genomics network to design the target conserved gene sequence with Tomato gene models (ITAG 2.40), which about 300bp. Then we design the primer, amplify the target sequence and insert into the TRV2 vector. The vector will be propagated in E. coli DH5α before transferred into Agrobacterium tumefaciens GV3101. Afterward we will cut the root tips and inoculate the root in the combination of TRV1 and TRV2. After three to four weeks we will observe the phenotype and collect the fresh root tip and extract the RNA from the root tips. The silencing effect will be determined by quantitative real time PCR. Meanwhile, Laser Capture Microdissection (LCM) will be adopted in this experiment for root proteomics study.


Understanding Aluminum Stress in Tomato



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