This study investigated physical activity and fitness of midlife and older rural women. Random-digit dialing was used to recruit 225 women (57.9 ± 5.6 years old). Self-reported activity (moderate activity, flexibility, and strength) and fitness (body composition, flexibility, strength, and estimated VO2max) were assessed. The women demonstrated low daily energy expenditure (30.74 ± 10.63 kcal · kg−1 · day−1) and estimated VO2max (20.12 ± 7.81 ml · kg−1 · min−1), with 51.5% reporting fair or poor health. Few women reported meeting Healthy People 2010 targets for moderate activity (43.1%), flexibility (28.9%), or strength (14.2%). When classified by estimated VO2max into three categories, differences were observed for body-mass index, percent body fat, sit and reach, and timed chair stands, with the poorest performance by those with low cardiorespiratory fitness. Adherence to Healthy People 2010 targets for moderate activity and strengthening was associated with higher cardiorespiratory fitness. These rural women are targets for physical activity interventions because of their sedentary behaviors and low cardiorespiratory fitness.