In order to find out the geographical distribution characteristics of the collected agricultural biological germplasm resources in Guizhou, the external geographical environment and the internal spatial performance of the collected resources were analyzed with the methods of spatial statistical analysis and diversity analysis. Firstly, the characteristics of elevation, temperature, precipitation and other basic geographic environment of germplasm resources are determined by spatial superposition and statistics. Secondly, the spatial distribution, the orientation and difference as well as heterosexuality, density and richness of germplasm resources are analyzed using the optimizing hotspot analysis, the geographic distribution center analysis and spatial diversity analysis.. The results show that the interval of elevation and meteorological factors in the area where the resources are located is large, but most of them are distributed in relatively concentrated geographical environment. 68.7% of the germplasm resources were found to be distributed between 600 and 1400 m of altitude, 65.1% were distributed between 15 and 17 degrees centigrade of average annual temperature, and 61.7% were distributed from 1250 to 1350 mm of annual precipitation. In addition, the 21 selected counties can basically reflect the spatial distribution characteristics of agricultural biological resources in Guizhou from the perspective of geographic space. Germplasm resources of different topics are cross-distributed, with the exception of edible fungi concentrated in the southwest. The resources of food crops, economical crops, vegetables, fruit trees and medicinal plants, which were provided by 14 ethnic group, are relatively evenly distributed in the province. Among them, the Miao ethnic group provided the highest amount (37%) of resources, followed by the Han (16%) and Buyi (15%) ethnic groups. The proportion of resources provided is less than 1% of the total. Thus, this study described the distribution status and characteristics of agricultural biological resources in Guizhou Province, which will provide reference for the formulation of relevant policies for effective protection and efficient utilization of biological resources in Guizhou Province.