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Callus Induction and Secondary Metabolite Analysis of Hairy Roots in Eruca Sativa Mill

Author: PuFan
Tutor: WangXinYu
School: Lanzhou University
Course: Cell Biology
Keywords: Yunjie Hairy root Secondary metabolites Glucosinolate
CLC: S565.4
Type: Master's thesis
Year: 2009
Downloads: 11
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Abstract


Yunjie (Eruca Sativa Mill) is a cruciferous arugula case, so far has been found the most drought-resistant, resistant barren cruciferous oil crops. Like other cruciferous plants, Yunjie contain secondary metabolites, the substance called glucosinolate (glucosinolate) under the action of the mustard seeds enzymes (myrosinase) will occur degradation and generates anticancer effects different thiocyanate (isothiocyanates, or ITCs), Yunjie is not only important genetic resources of cruciferous oil crops the glucosinolates and genetic breeding research, but also as a mode to study the mechanism of plants to drought, disease and other crops . Hairy roots (hairy roots) plant injured parts by Agrobacterium rhizogenes infection (Agrobateriumrhizogenes) after being induced by a root-like organs. Under in vitro conditions, the hairy roots can be induced to produce callus, and then differentiate to form the complete plant. May get some good traits of the new plant. The purpose of this study is the use of my brothers RADIATION PROCESSING Master has established the Department of Hairy root cultures, to establish a plant regeneration system. At the same time, a variety of factors, including the medium, cadmium chloride, and salicylic acid on hairy root biomass and glucosinolate content. The main findings are as follows: 1) morphology was observed by five different Hairy roots (C1-C5) found in 1/2MS solid or liquid culture conditions C1 Department of biomass accumulation, taproot length, a The number of branches and the number of secondary branches are maximum. 2) cultured in 1/2MS liquid medium for 15 days, C1, biomass (0.3478g.FW / bottle) than the C5 biomass (0.2348g.FW / bottle); in Roser Tolra (2006), a liquid culturing for 15 days, the amount of bio for C1 (0.1852g.FW / bottle) is slightly lower than the C5 biomass (0.1895g.FW / bottle). In the Roser Tolra liquid medium cadmium chloride or salicylic acid are added, the lines C1 and C5 biomass with cadmium chloride or salicylic acid concentration increased the trend is decreasing. 1/2MS liquid medium was added salicylic acid, with the addition of the salicylic acid concentration increases, the lines C1 and C5 biomass was decreased. 3) thiourea UV measured after 15 days of culture in the liquid medium of 1/2 MS, C1, and C5-based glucosinolate content were 3.853 mg / g.FW and 1.902 mg / g.FW were approximately wild root glucosinolate content (4.706 mg / g.FW) of 81.8% and 40.4%, respectively, 84.9% and 41.9% of the wild leaves glucosinolate content (4.536mg/g.FW). Cultured in Roser Tolra liquid medium after the same time, C1, and C5-based glucosinolate content of 3.799 mg / g.FW and 1.773 mg / g.FW, significantly lower than in the 1/2 MS liquid medium glucosinolate content. CdCl 2 (50,100 μm) the Roser Tolra liquid medium is added after 15 days of culture, the the C1 lines glucosinolate content (4.177mg/g.FW and 6.851mg/g.FW) the C5 lines were higher than glucosinolate content (2.812 mg / g.FW, and 4.261mg/g.FW), and when CdCl 2 concentration of 100 μm in C1 glucosinolate content of the highest (6.851mg/g.FW). Added salicylic acid (0.5, 2 and 4 μM) the Roser Tolra liquid medium, cultured for 15 days, C1 lines glucosinolate content of 2.178,1.216, and 0.304 mg / g.FW; glucosinolate content of the C5-based 4.710,2.330 and 0.557 mg / g.FW. Salicylic acid (0.5,2 4μM), added 1/2MS medium cultured for 15 days, the the C1 lines of glucosinolate content 2.182,1.236 and 0.312mg/g.FW; the C5 system glucosinolate content is 4.710,2.331 and a 0.569 mg / g.FW. C1 and C5 lines containing salicylic acid 1/2MS medium is slightly higher than in the Roser Tolra containing salicylic acid medium glucosinolate content and glucosinolate content downward trend with the higher concentration of salicylic acid . When the medium is 1/2MS medium, the concentration of salicylic acid is 0.5 μm, the glucosinolate content of the C1, C5-based highest (4.710 mg / g.FW). 4) Hairy root in containing 1.0-2.0 ml / 1 NAA 0.1-0.5 ml / 1 6-BA on MS medium can form a callus induction rate was 100%. Containing 0.1-1.5ml / l 2,4-D can be formed on MS medium granular embryogenic callus, and increased with the concentration of 2,4-D, callus color gradually deepened from light yellow to yellow, callus gradually increases.

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CLC: > Agricultural Sciences > Crop > Economic crops > Oil crops > Rapeseed ( Brassica )
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