水稻吸收和转运N15-硝酸盐的活力实验

Yongqiang Liu; Bin  Hu; Chengcai  Chu

doi:10.21769/BioProtoc.1897

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Peer-reviewed

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

YL Yongqiang Liu

BH Bin Hu

Chengcai Chu email

发布: 2016年08月20日第6卷第16期 DOI: 10.21769/BioProtoc.1897 浏览次数: 12999

评审: Tie LiuPooja SaxenaAnonymous reviewer(s)

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Cover of Nature Genetics, featuring study using the protocol.

Jun 2015

Special Issue on Protocols for Organoid Research

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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

English

中文翻译

文章信息

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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.