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The Research on the Optimized Transformation System of the Green Fluorescent Protein Gene to Fusarium Oxysporum from Solanaceae Crop and Its’ Colonization Characteristics

Author: ZhuLei
Tutor: LiuBo
School: Fujian Agriculture and Forestry University
Course: Microbiology
Keywords: Solanaceae Fusarium oxysporum Green Fluorescent Protein Infection Colonization
CLC: S436.41
Type: Master's thesis
Year: 2011
Downloads: 100
Quote: 0
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Abstract


The most devastating diseases the Solanaceae wilt disease caused by Fusarium oxysporum (Fusarium oxysporum), severely restricts the production of Solanaceae crops. Tomato, eggplant and pepper wilt plants collected from various regions of Fujian Province, the separation and identification of the pathogen; Fusarium wilt and a tomato green fluorescent protein (GFP) gene transformation screened fluorescence intensity and stability of the genetic traits transformants, while taking advantage of the green fluorescent protein marker transformants studied Fusarium pathogen colonization within the plant body and rhizosphere soil characteristics, the main findings are as follows: 1. from Zhangzhou, Fuzhou, Xiamen, Fujian Province, Solanaceae The main crop producing areas of acquisition tomato, pepper, eggplant plants show symptoms of wilt Isolation and identification of 40 Fusarium Fusarium oxysporum 35 further pathogenic identified the Fusarium oxysporum this 3 species of Solanaceae crops wilt pathogenic strains. Take the CaCl2-PEG-mediated method, the optimization of the tomato Fusarium oxysporum green fluorescent protein (GFP) transformation system. Optimal conditions for transformation: fresh mycelium cultured for 12-14 h Driselase in 4 mL of lysis buffer (20 mg / mL, 10 mg / mL dissolved wall enzymes), 28 ° C, under the conditions of 70 r / min, enzymatic 2.5-3 h. Studies have shown that the biological characteristics of the resulting transformants, labeled the growth characteristics of the strains in the mycelial morphology, growth rate, PH value is basically the same as with the wild-type strain. The obtained fluorescence intensity, and genetic traits stable transformants screened by stability. Using laser confocal scanning microscope to track strains of the green fluorescent protein was observed in tomato plants infected, the results show that treatment 6 d, visible from the roots of the plant molecular spore germination invasive plant aboveground no significant change; 10 d, the mycelium vascular tissue to reach the roots and stem base of the plants, plant root and stem base browning phenomenon; 17 d plant stem base hyphae increase the amount of the individual branches of the tomato plants wilting, and the stem base there is a clear browning. Tracking observation GFP gene marker strains 32 days of colonization in the different nature of the soil, the results indicate that the marked strain in the different nature of the soil colonization at least one month, with the time of its colonization density gradually decreased. Rice Land the best ability to survive the worst, the ability to survive in the sand.

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CLC: > Agricultural Sciences > Plant Protection > Pest and Disease Control > Horticultural Crops Pest and Disease Control > Vegetable pests > Solanaceous pests and diseases
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