15N-nitrate Uptake Activity and Root-to-shoot Transport Assay in Rice
水稻吸收和转运N15-硝酸盐的活力实验
发布: 2016年08月20日第6卷第16期 DOI: 10.21769/BioProtoc.1897 浏览次数: 11991
评审: Tie LiuPooja SaxenaAnonymous reviewer(s)
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参见作者原研究论文
The authors used this protocol in:
Jun 2015
Abstract
15N is a nonradioactive heavy isotope of nitrogen, widely used for biochemical and physiological research in plants. For instance, 15N-KNO3 was used as the nitrogen source in plants in order to investigate nitrate uptake activity and transport from roots to shoots (Lin et al., 2008). Here, we describe a detailed pipeline used for labeling living rice (Oryza sativa) plants with 15N-KNO3 and determination of net nitrate uptake and transport activity, and this protocol was proved to be valid in Arabidopsis and rice (Lin et al., 2008; Hu et al., 2015).
Keywords: Nitrate uptake (硝酸盐的吸收)
Nitrate transport (硝酸盐转运)
Rice (水稻)
Isotope (同位素)
Materials and Reagents
- 96-well plate
- Rice seeds (Zhonghua11, ZH11)
- NaClO [Sinopharm Chemical Reagent Co.,Ltd (SCRC), catalog number: 7681-52-9 ]
- KNO3 (SCRC, catalog number: 7757-79-1 )
- K15NO3 (Sigma-Aldrich, catalog number: 57654-83-8 )
- CaCl2 (SCRC, catalog number: 10043-52-4 )
- MgSO4·7H2O (SCRC, catalog number: 10034-99-8 )
- KH2PO4 (SCRC, catalog number: 7778-77-0 )
- FeSO4·7H2O (SCRC, catalog number: 7782-63-0 )
- EDTA-Na2 (SCRC, catalog number: 6381-92-6 )
- NaSiO3·9H2O (SCRC, catalog number: 13517-24-3 )
- H3BO3 (SCRC, catalog number: 10043-35-3 )
- CuSO4·5H2O (SCRC, catalog number: 7758-99-8 )
- ZnSO4·7H2O (SCRC, catalog number: 7446-20-0 )
- MnCl2·4H2O (SCRC, catalog number: 13446-34-9 )
- Na2MoO4·2H2O (SCRC, catalog number: 10102-40-6 )
- CaSO4·2H2O (SCRC, catalog number: 10101-41-4 )
- Modified Kimura B solution (see Recipes)
- 5 mM 15N-KNO3 (see Recipes)
- 0.1 mM CaSO4 solution (see Recipes)
Equipment
- Growth chamber (SANYO, model: MLR-351H )
- Isotope ratio mass spectrometer(Thermo Fisher Scientific, model: Finnigan Delta Plus XP) with elemental analyzer (Thermo Fisher Scientific, model: Flash EA 1112 )
Procedure
文章信息
版权信息
© 2016 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Liu, Y., Hu, B. and Chu, C. (2016). 15N-nitrate Uptake Activity and Root-to-shoot Transport Assay in Rice. Bio-protocol 6(16): e1897. DOI: 10.21769/BioProtoc.1897.
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分类
植物科学 > 植物生理学 > 营养
