The 45-minute "Life in the UK" tests, which all applicants must sit from Tuesday, require immigrants to answer 24 multiple questions about British life ranging from knowledge of Queen Elizabeth to regional dialects.
The tests, which cost 34 pounds and can be taken at about 90 centres across the country, come on top of a need for new citizens to demonstrate a working knowledge of English.
"An understanding of the British language and our way of life is vital," said Immigration minister Tony McNulty. "The measures we are introducing today will help new citizens to gain a greater appreciation of the civic and political dimension of British citizenship and, in particular, to understanding the rights and responsibilities that come with ... British citizenship."
Example questions include "Which TWO telephone numbers can be used to dial the emergency services? 112, 123, 555, 999". The correct answers are 112 and 999.
Those who fail the test will not be able to apply to be British citizens and will have to take it again.
The Immigration Advisory Service, a charity that advises immigrants and asylum seekers, gave the tests a cautious welcome but said the questions needed "a light touch".
"The danger is that this will be seen as a way of excluding people from British citizenship," said chief executive Keith Best.
"The perversity of our education system is such that new immigrants who seek naturalisation may end up having more knowledge about life in the UK through these tests and their associated teaching than many who are born British citizens."