Application of Double Resonator Piezoelectric Cytometry in Salt Tolerance Evaluation of Rice (Oryza sativa) Varieties
YANG Hou-Peng1,2, ZHOU Tie-An1,2,*, PAN Wei-Song1,2, XIAO Ying-Hui2,3
1 College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha 410128, China; 2 Hunan Engineering Research Center for Cell Mechanics and Functional Analysis, Changsha 410128, China; 3 College of Agronomy, Hunan Agricultural University, Changsha 410128, China
Abstract：The selective breeding of rice (Oryza sativa) varieties and the testing of stress resistance (salt tolerance) are usually conducted on the tested plants, which is not only laborious, time-consuming and destructive, but also susceptible to environmental factors, which greatly limits the efficiency and reliability of selective breeding. The purpose of this study was to establish a new method for evaluating salt tolerance of rice varieties based on double resonant piezoelectric cytometry (DRPC) and cell mechanics testing, using live rice cells as the characterization object. Firstly, 'Haidao 86' and 'Nipponbare' rice varieties with known strong salt tolerance and salt-sensitive characteristics respectively were selected to prepare their adherent cell culture systems, and then DRPC was used to measure the responses and variations of cells generated surface stress (ΔS) and cell viscoelasticity index (CVI) to different concentrations of NaCl stress. The results showed that the ΔS of 'Nipponbare' cells decreased first and then increased under the conditions of 20 and 40 mmol/L NaCl, and decreased monotonously under the conditions of 60~120 mmol/L NaCl; at the same time, the cells became softer and then harder under the conditions of 20 and 40 mmol/L NaCl, and became softer monotonously under the conditions of 60~120 mmol/L NaCl, and reached the limit of resisting salt stress under the condition of 40 mmol/L NaCl. The ΔS of 'Haidao 86' cells decreased first and then increased under the condition of 20 mmol/L NaCl, increased first and then decreased under the conditions of 40 and 60 mmol/L NaCl, fluctuated up and down under the conditions of 80 and 100 mmol/L NaCl, and increased sharply under the condition of 120 mmol/L NaCl. At the same time, the cells became harder under the conditions of 20, 80 and 100 mmol/L NaCl, and became harder first and then softer under the conditions of 40, 60 and 120 mmol/L NaCl, and reached the limit of resisting salt stress under the condition of 100 mmol/L NaCl. By a comprehensive comparison of the changes in cells generated stress and CVI produced by the cells of the 2 varieties under different concentrations of NaCl stress, it could be confirmed that the salt tolerance of 'Haidao86' was higher than that of 'Nipponbare', which was consistent with the result that the salt tolerance of 'Haidao86' was higher than that of Nipponbare' reported at the level of field plants. Therefore, this study not only confirms the accuracy and reliability of the DRPC for the evaluation of salt tolerance of rice varieties, but also provides an important reference for the wide application of this new method to efficiently evaluate crop stress resistance, which is of great significance for crop breeding for abiotic stress resistance.
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