Agro-biodiversity plays significant role in food production,ecosystem security and sustainable agricultural development. It is based on agricultural species diversity. Therefore, obtaining precise biodiversity information is a foundation of study and scientific decision of biodiversity conservation. In this paper, taking the survey area (county, township), nationalities and types of biological resource as the basic analysis unit, using biodiversity indexes that were composed of species richness, dominance, diversity index, Pielou index and similarity coefficient to evaluate diversity of agro-biological resource utilized by different nationalities in six townships of Zhenfeng and Songtao County. Results showed that the species diversity of the two counties are high and similar, but the level of species diversity of three towns in Songtao County was higher than the three towns of Zhenfeng County, and species diversity of Miao nationality was higher than those of Buyi, Tujia and Han nationality. In analysis unit of agro-biological resource types, the species diversity of food crop of Buyi and Tujia, food crop of Zhaiying Town in Songtao County, vegetables and annual economic crop of Miao, vegetables and annually economic crop of Panshi Town and Zhengda township in Songtao County, fruit trees and perennially economic crops of Miao and Buyi, fruit trees and perennially economic crops of Lugong Town in Zhenfeng County and Zhaiying Town in Songtao County, medicinal plants of Miao and Panshi Town in Songtao County,was higher than other towns and nationalities.