Spanish explorers seem to have traveled north seeking the Northwest Passage as far as what we now call Virginia and tried to establish a Mission in the area that they called Ajacan. Accounts are unclear and the location of the Mission is uncertain.
In the early 1500s, Spanish explorers discovered a bay naming it Bahia de Santa Maria and looked for sites on which to establish settlements. This region is now called Chesapeake Bay. Spain tried to establish settlements around Georgia and South Carolina about 1526 but it was several decades before further attempts were made in Virginia.
In 1561 (or maybe 1560), Pedro de Menendez de Aviles sailed into the Bahia de Santa Maria and either convinced or forced Paquiquino, a 17-year old, possibly the son of a weroance (chief), to travel back to Mexico and then Spain. The Spanish named the boy Don Luis. Don Luis was baptized and educated and met many prominent people in Spain. Eventually he convinced the Spanish to let him return to his native home and help convert others of his people to Christianity.
In August 1570, Father Juan Bautista de Segura, Father Luis de Quiros, and six Jesuit brothers left Havana to found a Mission in Ajacan. Don Luis was part of the party acting as guide and a young Spanish boy, Alonso Olmos, called Aloncito, joined the group to serve Mass. They arrived in Ajacan on September 10, 1570 and held a Mass near present-day Newport News. They then continued up the James River to a spot believed to be a few miles from present-day Jamestown Island where they met with members of Don Luis' tribe. Continuing inland, the missionaries established a small Mission between the James and York Rivers, the exact location is unknown. Possible sites for Mission Santa Maria are Queen's Creek on the north side of the Lower Peninsula near the York River, it may have been in the village of Axacam on the New Kent side of Diascund Creek near its confluence with the Chickahominy River, or possibly Deleware Bay. Mission Santa Maria consisted of a chapel and small residence, made of wood and other materials that have not survived.
Don Luis wasn't as committed to converting his people as the Spanish had believed and he left them to rejoin his people. The Spanish were dependent on Don Luis to translate and provide supplies and were upset by this turn. Don's people were uninterested in helping the Spanish who, out of desperation, chastised Don in February 1571. In retaliation, Don lead a party of warriors who killed all the priests taking only an alter boy, Alonso de los Olmos, as prisoner.
That spring a Spanish supply ship arrived and found nothing of the mission except a few robes worn by native people. The Spanish took several captive but learned little from them. The Spanish Governor in Cuba sent another expedition to punish Don Luis and recover Alonso which arrived in August 1572. The Spanish military force killed and captured several Algonquian and recovered Alonso but were unable to learn the location of Don Luis. They hung the Algonquian captives and left Ajacan. This was the end of the Spanish Mission Santa Maria in Virginia, but 35 years later the English established Jamestown in the same area.
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