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Triploid eastern oysters are an important component of the Maryland aquaculture industry because of their fast growth and sustained high meat yield. Commercially, triploids are produced by mating tetraploid oysters with normal diploid oysters. Developing tetraploid stock is crucial to meeting the growing demand for Maryland triploid oysters. However, it is challenging to produce and maintain excellent tetraploid lines for the benefit of industry. In short, there is a clear and pressing need for triploid and tetraploid lines that have region-specific beneficial characteristics, especially tolerance to low-salinity environments. In this project, we will establish the first generation of tetraploid stock derived from Maryland local oyster populations. Several intermediate triploid and diploid lines used to produce the tetraploid will be developed through mass selection and deployed at three sites for performance evaluation in different geographic areas. This tetraploid stock will enable the production of new triploid seed with improved low-salinity tolerance and rapid growth, providing a foundation that can be used for the development of additional superior lines. The tetraploid line will be produced and distributed in collaboration with the Maryland Department of Natural Resources Piney Point Aquaculture Center. This collaboration will provide new triploid seed to Maryland growers. All lines will be maintained by MSU PEARL and new lines will be developed based on these stocks to provide continued industry support. Through the development of new triploid and tetraploid lines, this project will enhance the sustainability, profitability and resilience of the Maryland aquaculture industry.