The 2016 AON Open Challenger was a professional tennis tournament played on clay courts. It was the fourteenth edition of the tournament which was part of the 2016 ATP Challenger Tour. It took place in Genoa, Italy between 5 and 11 September 2016.
| Country | Player | Rank1 | Seed | |
|---|---|---|---|---|
 Spain | Nicolás Almagro | 48 | 1 | |
 Argentina | Horacio Zeballos | 71 | 2 | |
| Argentina | Carlos Berlocq | 76 | 3 | |
| Spain | Roberto Carballés Baena | 103 | 4 | |
 Russian Federation | Teymuraz Gabashvili | 105 | 5 | |
 Uzbekistan | Denis Istomin | 107 | 6 | |
 Czech Republic | Adam Pavlásek | 112 | 7 | |
 Slovakia | Jozef Kovalík | 126 | 8 |
The following players received wildcards into the singles main draw:
Nicolás Almagro
Gianluigi Quinzi Denis Istomin Edoardo EreminThe following player received entry into the singles main draw as an alternate:
The following players received entry from the qualifying draw:
See main article: 2016 AON Open Challenger – Singles.
Jerzy Janowicz def. Nicolás Almagro 7–6(7–5), 6–4See main article: 2016 AON Open Challenger – Doubles.
Horacio Zeballos def. 
Aliaksandr Bury / Andrei Vasilevski, 6–4, 6–3.