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Cotton is an important economic crop; therefore, enhancing cotton yield and cultivating superior varieties are key research priorities. The seedling stage, a critical phase in cotton production, significantly influences the subsequent growth and yield of the crop. Therefore, breeding experts often choose to measure phenotypic parameters during this period to make breeding decisions. Traditional methods of phenotypic parameter measurement require manual processes, which are not only tedious and inefficient but can also damage the plants. To effectively, rapidly, and accurately extract three-dimensional phenotypic parameters of cotton seedlings, precise segmentation of phenotypic organs must first be achieved. This paper proposes a neural network-based segmentation algorithm for cotton seedling organs, which, compared to the average precision of 75.4% in traditional unsupervised learning, achieves an average precision of 96.67%, demonstrating excellent segmentation performance. The segmented leaf and stem point clouds are used for the calculation of phenotypic parameters such as stem length, leaf length, leaf width, and leaf area. Comparisons with actual measurements yield coefficients of determination R 2 of 91.97%, 90.97%, 92.72%, and 95.44%, respectively. The results indicate that the algorithm proposed in this paper can achieve precise segmentation of stem and leaf organs, and can efficiently and accurately extract three-dimensional phenotypic structural information of cotton seedlings. In summary, this study not only made significant progress in the precise segmentation of cotton seedling organs and the extraction of three-dimensional phenotypic structural information, but the algorithm also demonstrates strong applicability to different varieties of cotton seedlings. This provides new perspectives and methods for plant researchers and breeding experts, contributing to the advancement of the plant phenotypic computation field and bringing new breakthroughs and opportunities to the field of plant science research. [ABSTRACT FROM AUTHOR]