Orica-GreenEDGE have confirmed Gerrans and Matthews will be two of their main weapons for the 2-24 July Tour, which starts next Sunday at the Normandy coast in northern France.
After Colombian Esteban Chavez finished an historic second overall at the May Giro d'Italia, the team's main aim at the 2016 Tour will be stage wins.
Gerrans and Matthews are two of their main hopes for big results while rising British star Adam Yates will be their other protected rider.
While the two Australians are big threats on flat or undulating stages, the mountains are Yates's terrain.
Matthews finished runner-up to Peter Sagan (Tinkoff) at last September's world road championships and Gerrans was sixth.
Matthews later said Gerrans was sprinting against him at the finish and his compatriot should have been working for him.
The pair later said separately that the issue had been discussed at the Australian team's post-race debrief.
The two Australians have usually competed in different races so far this season, but they were in the same lineup in April at the Amstel Gold one-day classic.
After Matthews finished fifth and Gerrans 11th, there was more talk about whether they were working together properly.
But team manager Matthew White vehemently denied there was any issue, saying the two team leaders simply weren't good enough on the day.
Next Sunday will be the first time since Amstel Gold that Matthews and Gerrans have competed in the same team.
There is rising speculation that Matthews will leave Orica-GreenEdge at the end of this season.
Matthews made his Tour debut last year and had a torrid race, bravely finishing despite fracturing ribs in the horrific pileup during stage three.
"Even with these unfortunate problems, Michael soldiered on bravely which ultimately gave him valuable experience to show how far he could push certain barriers," White said.
"He is super motivated to perform well here for the first time on the biggest stage in the world."
Gerrans is a member of the select Australian cycling club of riders who have worn the yellow jersey as the Tour's leader and has won two stages in the three-week classic.
But he was among three Orica-GreenEdge riders who had to withdraw from the Tour because of their injuries in the stage-three crash.
"Simon has had great preparation throughout June with stints at altitude and racing at the Dauphine," White said.
"He performs very well under pressure and has the ability to come up with the goods when the big goals are set."
Paris-Roubaix winner Mat Hayman will ride in his second Tour as team captain.
Luke Durbridge is the other Australian in the team, along with South African Daryl Impey, Swiss stalwart Michael Albasini, Spaniard Ruben Plaza and Dane Chris Juul Jensen.
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