Scientia Agricultura Sinica ›› 2017, Vol. 50 ›› Issue (12): 2199-2208.doi: 10.3864/j.issn.0578-1752.2017.12.001

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS •     Next Articles

Expression and Functional Analysis of Acireductone Dioxygenase Gene in Rice

XIONG Wei, YANG Bo, LIU WeiYin, WANG Quan, KONG XiaoCong, JIN YaJun, LIANG ShanShan, LUAN WeiJiang, ZHANG SiJu   

  1. College of Life Science, Tianjin Normal University/Tianjin Key Laboratory of Animal and Plant Resistance, Tianjin 300387
  • Received:2017-01-12 Online:2017-06-16 Published:2017-06-16

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Abstract: 【Objective】 The tolerance to biotic/abiotic stress is highly associated with grain yield, which is a very important goal in crop breeding. The objective of this study was to analyze the expression pattern of OsARD1, to investigate its function to abiotic stress, and to provide a theoretical basis for rice breeding. 【Method】 To analyze the expression of OsARD1 in different tissues, total RNAs were isolated from different organs. To analyze the expression pattern of OsARD1, RNAs were isolated from wild type seedlings at 14-day-old under different abiotic stress, including PEG, high salinity and mechanical damage. RT-PCR method was used to analyze the expression of OsARD1 in different organs and under abiotic stress. An overexpressing vector of OsARD1 was constructed and OsARD1 overexpression transgenic plants were produced following the Agrobacterium-mediated transformation procedure. Transgenic plants with empty vector were also generated as a control. For drought stress, T1 generation transgenic plants and wild type at 12-day-old were subjected to water deficit treatment and then cultured in nutrition solution for 3 days to recover. For PEG osmotic stress experiment, wild type and OsARD1-overexpressing (OsARD1-OV) seeds were planted in agar medium containing 5% PEG6000 whose osmotic potential was -0.05 MPa. For control group, no PEG was added into agar medium. The phenotypes of wild type and OsARD1-OV plants were observed and recorded. 【Result】The tissue-specific expression analysis showed that OsARD1 expressed highly in root and mature organs, especially in senescent organs. OsARD1 gene transcript levels were strongly induced by PEG6000, high salinity and mechanical damage. Six independent OsARD1-OV lines were generated by Agrobacterium-mediated transformation method. RT-PCR expression analysis showed that the expression level of OsARD1 was significantly increased in OsARD1-OV lines of T0 and T1 generation plant compared with that in wild type, suggesting that overexpression vector worked and inherited stably. The experiments of water deficit of OsARD1-OV of T1 generation and wild type plants were further carried out. The results showed that the wild type plants exhibited severe dehydration symptoms with rolled and withered leaves after five hours of water deficit treatment, while OsARD1-OV plants were growing normally. After 8 hours of water deficit treatment and recovery culture for 3 days, there were only 10% wild type plants survived, while 80% transgenic plants survived. In PEG6000 osmotic stress experiment, it was found that the shoot and root length of wild type and OsARD1-OV plants had no significant difference with that in control group. However, the growth of WT roots was strongly inhibited in 5% PEG6000 treatment group compared with OsARD1-OV lines, and the root length of wide type was shorter than that of OsARD1-OV lines, indicating that the overexpression of OsARD1 improved drought tolerance in rice.【Conclusion】OsARD1 displayed higher expression in roots and mature tissues of rice, and was strongly induced by PEG, high salinity and mechanical damage. Overexpression of OsARD1 significantly improved the tolerance to water deficit and osmotic stress in rice, suggesting that OsARD1 plays a key role in drought tolerance.

Key words: rice, OsARD1, overexpression, drought stress, ethylene

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