One Nation's candidate for WA's upper house at the 2025 election, Parminder Singh, was revealed to have made several anti-gay comments in a video on social media.
Mr Singh, who was third on One Nation's upper house ticket at the state election, has since been removed from the party's website, after the video became public.
Mr Joyce said the comments from the candidate did not represent the party.
"People are allowed to have views, of course, but they've got to be within the Australian guardrails," he told ABC TV on Wednesday.
"We're one of the gayest parties in Australia.
"He obviously doesn't know One Nation because we got a surprise for him. We have quite a large proportion of gay people, and they're all the same. They don't want to be glorified. They don't want to be vilified."
Mr Joyce said the comment was highly offensive and Mr Singh must not have been around farms, due to the number of gay animals.
The One Nation recruit said the party would still have teething issues with its vetting processes as the party grows.
"When something's growing like that, you're going to have growing pains, and you're going to have your internal resources won't be able to keep up with the membership as it goes and the applicants as they go," he said.
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