Spain has long sought to regain control of Gibraltar, an enclave of 32,000 people on the southern tip of the Iberian Peninsula that was ceded to Britain in 1713.
Yesterday, the EU suggested that Spain would get a veto over the territory's future relationship with the trading bloc.
Gibraltar reacted angrily to the EU position, saying Spain was using the UK's departure from the EU to force negotiations that it has wanted on the status of the territory.
"As ever, the UK remains implacable and rock-like in our support for Gibraltar," Johnson tweeted after the conversation.
British Prime Minister Theresa May failed to mention Gibraltar in her letter triggering formal negotiations on Britain's departure from the EU causing dismay in the territory.
The EU fuelled that anxiety yesterday when it released a nine-page draft roadmap for the talks, which included a statement that no future agreement after Brexit between Britain and the bloc would apply to Gibraltar unless both the UK and Spain agreed.
Picardo expressed outrage at being singled out, describing the guidelines as "a disgraceful attempt by Spain to manipulate the European Council for its own narrow political interests.
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