Analyzing the evolution characteristics of territorially spatial pattern and its eco-environmental effects from the perspective of major functional zones can effectively evaluate the impact of major functional zone planning on the territorial development and protection, which is conducive to providing reference for building a rational and orderly territorially spatial pattern and protecting ecological environment. This paper took Zhejiang Province as an example, the multi-period remote sensing monitoring data of land use were used to analyze the evolution characteristics of territorially spatial pattern in Zhejiang Province before and after the promulgation of the major functional zone planning (2000-2010 and 2010-2020). The eco-environmental quality index and ecological contribution rate were used to quantitatively analyze its eco-environmental effects in different periods. The results show that:(1) Compared with 2000-2010, the conversion degree of territorial space in Zhejiang Province decreased significantly during 2010 and 2020, the spatial agglomeration of territorial space was weakened, and the proportion of high-high clustering areas in key development zones increased from 22.3% to 39.3% during the study period. (2) During 2000 and 2010, the relationships between the increase and decrease of different territorial spaces in Zhejiang Province showed significantly negative spatial correlations, and the negative spatial correlations of the relationships were weakened in 2010-2020. These areas with significant local spatial autocorrelation had obvious regional differences in each major functional zone. (3) Compared with 2000-2010, the declining rate of the eco-environmental quality in Zhejiang Province slowed down in 2010-2020, the eco-environmental enhancement areas increased, while the eco-environmental deterioration areas decreased, and the distribution of eco-environmental deterioration areas transferred from the ecological economic zone and optimized development zone to the key development zone. (4) Compared with 2000-2010, the contribution rate of eco-environmental quality improvement or deterioration due to the transformation between agricultural production space and forest ecological space in each major functional zone increased in 2010-2020. The contribution rate of eco-environmental deterioration due to the transformation of agricultural production to urban and rural living space in optimization development zones and key development zones decreased.