The changes to the law took effect on Saturday in Uzbekistan obliging the citizens to put their right hand over heart to honor the national anthem, RIA Novosti reported.
Until recently, Uzbeks were supposed to be standing while the national anthem is playing.
The changes to the law were approved by the Central Asian country's parliament earlier this month and signed by President Islam Karimov on Friday. The law was published in national media on Saturday.
The amendments also require men in uniform to give the salute at the first note of the anthem.
The Uzbek national anthem, My sunny free land, has a tune of the old Soviet anthem and the new lyrics, calling Uzbekistan the "beacon of independence and guardian of peace."
The law obliging citizens to place hand over heart during the singing of the anthem has been introduced in many countries, including the United States.
Several years ago the similar law was approved by Uzbekistan's neighbor, Kyrgyzstan.