Objective This study aimed to investigate the current status and influencing factors of healthy diet knowledge, attitude, and practice (KAP) among middle-aged and older adults aged 45–75 in Southwest China. Methods A questionnaire survey was conducted among 1822 middle-aged and older adults in Southwest China (including Guizhou, Sichuan, Yunnan, and Chongqing) from February to May 2021. Results The average score of healthy diet knowledge was (4.82 ± 2.98), with a passing rate of 7.6%. The mean score of healthy diet attitude was (21.26 ± 4.18), with a passing rate of 69.5%. The average score of healthy diet practice was (13.76 ± 2.84), with a passing rate of 55.5%. The score for healthy diet KAP was (39.85 ± 7.21), with a passing rate of 41.2%. Univariate analysis showed that the scores of healthy diet KAP were significantly different among participants of different ages, genders, ethnicities, residences, education levels, monthly household incomes, and regions, as well as varying according to whether several generations have lived in the same area (p < 0.05). The results of multiple linear regression showed that the healthy diet KAP of participants was influenced by age, gender, residence, education level, monthly household income, and region (p < 0.05). Conclusion The healthy diet KAP of middle-aged and older adults aged 45–75 in Southwest China shows room for improvement. The knowledge regarding healthy diet was relatively low, and certain specific healthy diet practices were not up to the standard. However, there was a positive trend in the attitude towards a healthy diet. Healthy diet education should be promoted for middle-aged and older adults.