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Studies on Abiotic Stress Resistance of Tomato Ascorbic Acid Metabolism Related Genes-DHAR1 and MDHAR1

Author: DengChunZuo
Tutor: OuYangBo
School: Huazhong Agricultural University
Course: Olericulture
Keywords: Tomato (Solanum lycopersicum) L-ascorbic acid RD29A DHAR MDHAR ROS abiotic stress genetic transformation
CLC: S641.2
Type: Master's thesis
Year: 2009
Downloads: 43
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Abstract


Abiotic stress including drought, chilling and salinity etc. is one of the major threats to crop production and quality. Reactive oxygen stress generally accompanies with various abotic stresses. Plants have developed their own antioxidant defense systems to alleviate ROS stress and maintain normal physiological process. Ascorbate acid （AsA） is a major antioxidant in plants. Dehydroascorbate reductase （DHAR） and Monodehydroascorbate reductase （MDHAR） are key enzymes in AsA regeneration and essential for maintaining the redox state of AsA. The stress-inducible promoter, such as RD29A promoter which is induced upon drought, salt or cold stress, may be useful to minimize side effects caused by constitutively expressing of stress tolerance-related genes in plant. This study aims to decipher the functions of DHAR1 and MDHAR1 by analyzing of their roles in abiotic stress using strategies of over expression, inducible expression and RNA inference, which may also lead to creation of tomato breeding materials with multiple stress tolerance. The main results were as follows:1. The 823 bp upstream regulatory region（ pRD29A） of RD29A gene（AT5G52310） from Arabidopsis thaliana genome was amplified using PCR technique. pK2GW7I, a binary vector with pRD29A was constructed.2. The tomato inducible expression vectors- pKDI、pKMDI and over expression vectors- pKOED, pKOEMD were constructed for tomato DHAR1 and MDHAR1, respectively.3. A total of 44 putative transgenic tomato plants were obtained. PCR analysis results indicated that T-DNA was integrated into the tomato genome.4. RT-PCR analysis showed that DHAR1, MDHAR1 were constitutively expressed in different tissue of tomato, whereas the expression levels were different.5. Tolerance to oxidative stress for transgenic plants was investigated in T0 generation. Five lines for each construct from T1 transgenic plants were further chosen for function analysis of the DHAR1 and MDHAR1. It was found that that overexpression of DHAR1, MDHAR1 or inducible expression of DHAR1 could improve salt, drought and chilling stress tolerance on tomato. The preliminary results indicated pRD29A functions in tomato. Several transgenic tomato lines with tolerance to salt, drought and chilling stress were obtained and could serve as materials for future research on abiotic stress tolerance.

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CLC: > Agricultural Sciences > Gardening > Vegetable gardening > Solanaceous > Tomatoes ( tomatoes )
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