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Tissue Culture and Dihydroflavonol-4-reductase (DFR) Gene Cloning in Saussurea Involucrata
Author: PangHongXia
Tutor: ZuoJianBing
School: Xinjiang Normal University
Course: Botany
Keywords: Snow lotus Callus Plant hormones Total flavonoids DFR gene Separate Clone
CLC: S567.239
Type: Master's thesis
Year: 2010
Downloads: 56
Quote: 0
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Abstract
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Snow lotus (Saussurea involucrata Kar et Kir) are a class of rare Chinese herbal medicine, medicinal active ingredients flavonoids. In this paper, through the study of different plant hormones on the snow lotus callus induction and differentiation, explore different plant hormones and incubation time on the snow lotus callus growth and total flavonoid content, and isolated the snow lotus DFR gene fragment, the use of biological software DFR gene and protein structure and function analysis, forecasts. Transforming based on callus carried genetic provide reference data. The main results are as follows: snow lotus seed germination temperature, medium composition and penetration potential significantly affect the germination of lotus seeds, in which the temperature of snow lotus seed germination affect very significantly, the highest seed germination rate of 20/10 ° C variable temperature conditions; osmotic potential and medium composition on the snow lotus seed germination significantly affected, the most suitable medium for seed germination 1/2MS0. Polyethylene glycol (PEG) when the above-0.9MPa, the seeds can not germinate. Plant hormones on the snow lotus callus induction and differentiation of plant hormones on the snow lotus callus induction and differentiation. Single factor induced NAA induced effect was significantly better than other hormones, induced effect: Root gt; the petiole gt; leaves; NAA and 6-BA induced by hormone combination, there is a very significant difference between the blade and petiole induction rate, and there was a significant difference between the blade root, petiole and root induction rate and induced effect: blades gt; the petiole gt; roots. Leaf explants, the best induction medium MS 6-BA1.00mg / L NAA2.00mg / L; the best budding medium of the MS-BA1.00mg / of L NAA0.20 mg / L; Best rooting medium was 1/2MS0 NAA0.30mg / L KT0.03mg / L. 3 the snow lotus callus flavonoids Determination of total flavonoid content and yield of following the generation of media and incubation time on the amount of snow lotus callus growth, the results showed that: (1) Following the generation of medium to MS NAA 2.00 mg / of L-BA1.00mg / L Callus Growth and yield of total flavonoids highest; highest total flavonoid content of the callus in MS NAA 4.00mg / L 6-BA2.00mg / L; (2) callus the amount of tissue growth followed by generation of prolonged incubation time increased, its total flavonoids unstable, subculture the first 30d achieve the highest. 4. The snow lotus DFR gene fragment separation Cloning and sequence analysis by landing a combination of PCR and gradient PCR method, isolated snow lotus DFR gene fragment length 1464bp, a molecular weight of about 44 kDa. The high degree of similarity of the DFR gene fragment obtained by the separation with the the medusa DFR gene DNA (EF672726), having homology. Sequence Analysis of prediction can be seen, the DFR gene derived protein does not exist a coiled-coil structure, without the signal peptide, is a hydrophobic transmembrane protein, high-level structure containing six alpha-helix, 3? - Slice.
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CLC: > Agricultural Sciences > Crop > Economic crops > Medicinal crops > Herbal > Perennial > Other
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