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Studies on the Chilling Tolerance in Tobaccos Transformed with Ubiquitingene Ta-Ub2 from Wheat

Author: FengYaZuo
Tutor: WangZuo
School: Shandong Agricultural University
Course: Botany
Keywords: Ubiquitin Transgenic Low temperature stress Oxidative damage Photosynthesis Protein carbonyl
CLC: S572
Type: Master's thesis
Year: 2011
Downloads: 50
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Abstract


Ubiquitin / 26S proteasome pathway (ubiquitin/26S proteasome pathway, UPP) is known to be the most important, and highly selective protein degradation pathway. Stress biological cells under non-functional protein accumulation increased, timely removal of these non-functional protein important biological adapt to adversity. Studies have shown that ubiquitin gene expression was under stress conditions increase the short-lived regulatory proteins and abnormal protein degradation rate accelerated indirect role of the ubiquitin system in adaptation to biotic stresses. Our laboratory has cloned wheat monomeric ubiquitin gene, build a sense expression vector, and transformed tobacco multiple transgenic lines overexpressed ubiquitin gene; cloned tobacco ubiquitin gene construct antisense expression vector antisense transforming tobacco multiple transgenic lines inhibit tobacco ubiquitin gene expression. The study found that overexpression of ubiquitin tobacco drought resistance, salt tolerance. In this study, the ubiquitin gene system of justice Ta-Ub2 (T 2 -2, T 2 -11, T 2 -13), turn the ubiquitin gene antisense Nt-Ub1 Department of (AT 2 -1 AT 2 -3 AT 2 -10) and transfected with empty vector control plants (T-GUS) for materials, low-temperature 1-5d, at room temperature 4 ℃ or 10 ℃ stress recovery 3d, study overexpressed of ubiquitin genes on chilling tolerance of transgenic tobacco. External morphological changes under low temperature stress transfer ubiquitin gene tobacco, photosynthetic capacity, penetration ability to regulate various aspects of the degree of membrane injury, the accumulation of reactive oxygen species and clearing activities of antioxidant enzymes, ubiquitin protein expression as well as protein oxidative damage temperature of physiological and biochemical mechanisms of the ubiquitin system affecting plant. The main results are as follows: (1) low temperature stress caused by tobacco photosynthetic apparatus damage, resulting in photosynthetic rate (Pn), PSII maximum photochemical efficiency (Fv / Fm), PSII actual photochemical efficiency (ФPSII) decline. Phase for the control plants (wild-type and turn the empty vector tobacco), excessive expression Ta-Ub2 Pan-hormone gene of the transgenic plants Pn, Fv / Fm ФPSII decline is relatively slow, and that under low temperature stress, excessive expression Ta-Ub2 Pan-hormone gene of The stability of the transgenic plants photosynthetic apparatus to maintain good, especially for the reduction of the low-temperature photoinhibition of PSII. (2) low temperature stress caused by cellular dehydration. Overexpression of the ubiquitin gene in the transgenic lines proline, soluble sugar accumulation, osmotic adjustment ability to effectively prevent the low-temperature dehydration of the cell. Inhibition of the ubiquitin gene expression in transgenic plants, proline, soluble sugar accumulation less osmotic adjustment, water status remained poor. (3) low temperature stress caused by biofilm damage, resulting in membrane lipid peroxidation product malondialdehyde (MDA) content and membrane permeability increased. Overexpression the ubiquitin gene transgenic lines, cell membranes by low temperature injury to a lesser extent, lower MDA content and the amount of electrolyte leakage, effectively protect the stability and integrity of the biofilm; contrast, inhibition of the ubiquitin gene expression low temperature stress under biofilm injury is more serious. (4) low temperature stress leads to superoxide anion radical (O 2 · - ) the rate of formation of hydrogen peroxide (H 2 O 2 ) accumulation increased. Overexpression of the ubiquitin gene in transgenic plants, the main antioxidant enzymes (SOD, CAT, POD, APX) activity is relatively high, and superoxide anion production rates and hydrogen peroxide accumulation less. Inhibition of ubiquitin gene expression in transgenic plants of superoxide anion and hydrogen peroxide accumulation exacerbated. (5) Western blot hybridization of tobacco ubiquitin protein found both under normal growth conditions of low temperature processing conditions, overexpression of the ubiquitin gene transgenic lines and wild-type plants ubiquitin monomeric protein levels were not significantly different, but excessive transgenic plants expressing ubiquitin bound ubiquitin accumulation was significantly higher than that of wild-type plants. (6) low temperature stress leads to oxidative damage of tobacco leaf cell protein, protein carbonylation is an irreversible process, is the main product of protein oxidation. Western blot hybridization analysis of the protein content of the carbonylation of ubiquitin gene overexpression transgenic plants. The results showed that the low temperature treatment and overexpression the ubiquitin gene transgenic plants protein carbonyl degree than the control plants light showed that overexpression of ubiquitin gene to some extent alleviate the low temperature protein oxidative damage. In summary, overexpression of ubiquitin monomers to improve the adaptability of transgenic tobacco plants to low temperature stress, inhibition of the ubiquitin gene expression reduces low temperature tolerance of transgenic plants. Description of the ubiquitin system and plants closely related to the chilling tolerance, the low temperature adversity adaptability of plants can be improved by improving the ubiquitin system.

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CLC: > Agricultural Sciences > Crop > Economic crops > Tobacco ( tobacco )
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