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KURT, DURSUN/0000-0001-6697-3954; WOS: 000493446300016 Tubers of the tuberous orchids have been collected for centuries, and used for medicinal purposes as well as in beverage and ice-cream industry. These orchids (Sahlep orchids) cannot be propagated vegetatively as they generally do not yield more than one tuber in a year. Seeds do not have microscopic endosperms; they have quite small chance of germination and successful establishment. These orchids are not cultivated and are usually collected from natural populations. Therefore, they are under threat of extinction and thus are placed under protection worldwide with national and international treaties. In this study, specimens of the threatened orchid taxa Ophrys sphegodes subsp. mammosa (Desf.) Soo ex E. Nelson orchid, were stimulated to grow tuber twice in a single vegetation period. Plants' ability to develop new tubers was tested and a new propagation method was proposed for the first time worldwide. Plants harvested at the beginning, middle, and end of flowering period were re-planted. It was observed that the plants harvested at early flowering were able to develop new tubers when they were re-planted in their original places. At the end of their annual life cycle, the number of tubers that were produced by the plants harvested at the three subsequent harvest periods were 2.21, 2.10, and 1.04. Quality of the tubers of the 2nd harvest was also tested in this study to use them as propagation material for the subsequent season, and positive outcomes were achieved. According to the present findings, growth of two tubers in a single vegetation period was achieved for the first time. Possible sustainable cultivation of Sahlep orchids was proved by this new propagation method.