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Physiological Changes in Response to the Postharvest Water Deficiency and Cloning of Aquaporin Protein Encoding Gene Cmaqp of Chrysanthemum

Author: WangHongBin
Tutor: ChenSuMei
School: Nanjing Agricultural College
Course: Ornamental Plants and Horticulture
Keywords: Cut chrysanthemum Postharvest Water deficit Vase life Aquaporins Salt tolerance
CLC: S682.11
Type: Master's thesis
Year: 2012
Downloads: 38
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Abstract


Chrysanthemum is one of ten traditional famous flowers in China and four cut flowers in the world. The cut chrysanthemum has a great ornamental value and commercial value that accounts for30percent of the total world cut flowers. During the harvest, cut flower had to go through the set of goods, wholesale, transportation, retail and other sectors will inevitably be subjected to water stress. Water deficit stress is the main factors causing postharvest loss. In our country the cut chrysanthemum sales in two main ways, domestic or export to neighboring Japan and Korea. Postharvest storage time away from the water is divided into short term or long term two types.Water transport in plants of short distance between intercellular membrane through the membrane water channel proteins aqurpoins. Water balance participate in a variety of stress response and affect the quality of cut Flowers. Cut chrysanthemum postharvest physiological mechanism of the short term and long term water deficit stress has important theoretical guiding significance for cut chrysanthemum postharvest measures. The research of cut chrysanthemum on postharvest water deficit stress and aquaporins has positive effect on postharvest, resistance molecular breeding and the health development of cut chrysanthemum industry. The research of cut chrysanthemum aquaporins has positive effect on postharvest, resistance molecular breeding and the health development of cut chrysanthemum industry.The study through simulation of postharvest short term and long term storage away from water, then the cut chrysanthemum were treated with short term and long term water deficit stress. We analyzes the changes of leaf water and chlorophyll content, MDA, REC, ROS and defense enzyme activity, hoped to explain the mechanism of the relative component changes in postharvest cut chrysanthemum under various water deficit treated times. Research of CmAQP with other reported PIPs (Plasma membrane intrinsic proteins) aquaporins, in order to obtain new germplasm resistance in chrysanthemum. The main results are follows: 1. In SWD treatment (48h), leaf relative water content decreased slightly. Leaf chlorophyll content increased slightly late in the period of water deficit treatment. REC, MDA, O2-and H2O2content maintained to rise during water deficit treatment, while SOD, POD and APX activity showed a downward trend; After water recovery leaf chlorophyll content first increased then decreased, while REC, MDA, O2-and H2O2content first decreased then increased. POD and APX activity decreased in the later vase period. In SWD the vase life of was3.2days shorter and the maximum diameter was1.1cm smaller than control. In LWD treatment (120h), leaf relative water content rapid decreased while REC, MDA, O2-and H2O2content rapid increased, SOD, POD and APX activity showed a downward trend; After water recovery leaf relative water content rapid increased, REC, MDA, O2-and H2O2content decline slowly, SOD, POD and APX activity increased with water recovery, but declined in the late vase period. In LWD the vase life of was6.9days shorter and the maximum diameter was4.4cm smaller than control.2. We designed the degenerate primers by comsulting the other plant AQP gene’s conserved sequence, and cloned cDNA sequence of AQP gene of cut chrysanthemum using PT-PCR and RACE techniques. The gene was named CmAQP. Sequence analysis showed that, CmAQP gene has1117bp length,906bp open reading frame (ORF), and302amino acids were encoded. CmAQP had a similar sequence with SrAQP (Stevia rebaudiana), PmAQP (Prunus mume), MpPIP2;3(Malus prunifolia) and VvPIP2;4(Vitis vinifera). Phylogenetic tree analysis showed that CmAQP and other PIP2proteins were grouped into the same branch.3. A plant expression vector for CmAQP gene pCAMBIA-1301-CmAQP was constructed in this study, which was introduced into Arabidopsis thaliana via an Agrobacterium tumefaciens EHA105-mediated floral dip method. The chrysanthemum CmAQP has been successfully transferred to Arabidopsis confirmed by RT-PCR detection. The seed germination experiment showed that the germination rate of chrysanthemum CmAQP transgenic arabidopsis was significantly higher than the control. Speculate the chrysanthemum CmAQP can improve plant salt tolerance.

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CLC: > Agricultural Sciences > Gardening > Ornamental Horticulture ( flowers and ornamental trees) > Perennial Flowers > Perennial Flowers Class > Chrysanthemum
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