![]() ![]() The T-DNA removal was further validated by sequencing in one plant line. Six plant lines with reduced susceptibility to the pathogen were heat-treated and screened by real-time PCR to quantify the exogenous DNA elimination. Moreover, our construct contained a heat-shock inducible FLP/FRT recombination system designed specifically to remove the T-DNA harbouring the expression cassettes for CRISPR/Cas9, the marker gene and the FLP itself. Sequencing of five potential off-target sites revealed no mutation event. Highly significant reduction in susceptibility was observed compared to control plants. Seven edited lines with a loss-of-function mutation were inoculated with the pathogen. An editing efficiency of 75% was obtained. Fifty-seven transgenic lines were screened to identify CRISPR/Cas9-induced mutations. In this work, MdDIPM4 knockout has been produced in two Malus × domestica susceptible cultivars using the CRISPR/Cas9 system delivered via Agrobacterium tumefaciens. ![]() This resource outlines susceptible hosts, and identification, management, life cycle, and preventative measures related to fireblight in Texas.The bacterium Erwinia amylovora, the causal agent of fire blight disease in apple, triggers its infection through the DspA/E effector which interacts with the apple susceptibility protein MdDIPM4. Some sources recommend the use of streptomycin, however, the use of antibiotics leads to resistance and is not a suitable form of control for fireblight. Also, because new shoots and succulent tissue are more susceptible, excessive watering and fertilizing should be avoided. When removing infected tissue, shears should be dipped in a bleach solution between each cut to prevent cross infection of wounds. Infected tissues need to be pruned and destroyed to prevent further infection. Once a tree is infected, the only way to get rid of the disease is by eradication. MANAGEMENT METHODS: The best management methods currently available are simple cultural practices. Fire blight, the disease caused by Erwinia amylovora, causes necrosis in plant tissues and can cause entire orchards to appear scorched by fire. The pathogen is most often spread by pollinators as they pick up bacteria from infected flowers and transfer them to neighboring trees. Infection begins in a localized fashion at nectaries and grafting wounds and becomes systemic once the bacteria enter the xylem where they travel through the vasculature of the plant. It affects plants of the Rosaceae family, particularly apples and pears. It is rod-shaped and flagellated, capable of causing infection through the nectaries in flowers and wounds. ![]() Often times, infected tissues will exude an amber colored bacterial ooze.īIOLOGY: Erwinia amylovora is a Gram-negative member of the Enterobacteria family. The underlying wood will become discolored. When branches are infected the bark will appear water soaked. Infected shoots turn black and curl giving a ‘Shepherd’s Crook’ appearance. Infected leaves will have a blackened midvein and will eventually shrivel and die while remaining on the tree. These infected tissues will eventually turn black. Symptoms include water soaking on the petals, peduncle, and ovary. SYMPTOMS: Symptoms of fireblight can appear on flowers, buds, shoots, leaves, and mature stems and branches. SCIENTIFIC NAME of causal agent: Erwinia amylovora ![]()
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