Abstract: The uniformity of growth and mechanical properties of grafted seedlings affect the quality of mechanical grafting operations. The growth uniformity of grafted seedlings in a greenhouse will be poor due to the uneven and unstable light and temperature conditions. Plant factories can cultivate grafted seedlings in the most suitable environment by regulating environmental parameters such as light and temperature. The aim of this study was to investigate the impact of the light conditions on tomato seedlings in plant factory and to develop an optimal cultivation light formula. The effects of light intensity (50, 100, 150, 200, and 250 μmol m−2 s−2) and photoperiod (10, 12, 14, 16, and 18 h a day(h/d)) on the morphological and mechanical properties of tomato seedlings were experimentally investigated. Orthogonal experiments were conducted involving light quality (R:B = 75:25, R:B = 50:50, and R:B = 25:75), light intensity (150 μmol m−2 s−2, 200 μmol m−2 s−2, and 250 μmol m−2 s−2), and photoperiod (14, 16, and 18 h/d) as independent variables to determine the optimal combination. Finally, a comparative grafting experiment was conducted between the seedlings cultivated using the optimal light formula and commercially available seedlings. The result showed that increasing light intensity inhibited hypocotyl length and promoted seedling stem growth, and excessive light intensity decreased seedling mechanical properties. The optimal light intensity for rootstocks is 200 μmol m−2 s−2, and the optimal light intensity for scions is 250 μmol m−2 s−2. Shortening the photoperiod would promote hypocotyl growth and inhibit seedling stem elongation. Different photoperiods had a significant impact on the mechanical properties of tomato seedlings. The most suitable photoperiod for rootstocks was 18 h/d and for scions was 16 h/d. The most suitable light formula was R:B = 50:50, 250 μmol m−2 s−2, 18 h/d. By analyzing the experimental results, the mechanical properties of seedlings grown by the regulated light environment were better than those of commercially available seedlings, and the success rate of mechanical grafting was 7% higher. Overall, in plant factories compared to commercially available tomato seedlings, tomato seedlings cultivated by the regulated light environment were more suitable for mechanical grafting. This research result provides theoretical support for subsequent research on grafting machinery.
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