The closure of all but one Japanese port.
Explanation:
Japan was not like the other nations of East Asia when it came to contact with the Europeans, or their own internal affairs. Seeing that the European imperial powers are closing in and managing to occupy the nations in their surroundings, the Japanese were not willing to allow the same to happen to them. The country was unified, had very strong nationalistic core, and quickly started to cut down and move away the Europeans. The Christians were persecuted, while the European ships and merchants were expelled and not allowed to come to Japan.
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