Molecules (2019)
Update date:2022-08-25
Topics:
Schramm, Sebastian
Boco, Maria Fe Angela Comia
Manzer, Sarah
K?nig, Oliver
Zhang, Tong
Mony, Fatima Tuz Zohora
Adedeji-Badmus, Adebimpe Nafisat
Poppenberger, Brigitte
Rozhon, Wilfried
Feeding experiments with stable isotopes are helpful tools for investigation of metabolic fluxes and biochemical pathways. For assessing nitrogen metabolism, the heavier nitrogen isotope, [15N], has been frequently used. In plants, it is usually applied in form of [15N]-nitrate, which is assimilated mainly in leaves. Thus, methods for quantification of the [15N]-nitrate/[14N]-nitrate ratio in leaves are useful for the planning and evaluation of feeding and pulse–chase experiments. Here we describe a simple and sensitive method for determining the [15N]-nitrate to [14N]-nitrate ratio in leaves. Leaf discs (8 mm diameter, approximately 10 mg fresh weight) were sufficient for analysis, allowing a single leaf to be sampled multiple times. Nitrate was extracted with hot water and derivatized with mesitylene in the presence of sulfuric acid to nitromesitylene. The derivatization product was analyzed by gas chromatography–mass spectrometry with electron ionization. Separation of the derivatized samples required only 6 min. The method shows excellent repeatability with intraday and interday standard deviations of less than 0.9 mol%. Using the method, we show that [15N]-nitrate declines in leaves of hydroponically grown Crassocephalum crepidioides, an African orphan crop, with a biological half-life of 4.5 days after transfer to medium containing [14N]-nitrate as the sole nitrogen source.
View Morewebsite:http://www.guarson.com
Contact:+86-523-88059600,+86-13805268803
Address:Room B1006,Yafang Building,Jiangyan Avenue,Jiangyan District, Taizhou City,Jiangsu,China
website:http://www.sincerityenterp.com
Contact:+86-571-88111846
Address:Binjiang Area, Hangzhou
ZHONGBEI NORTHLAND BIO-CHEM INDUSTRY CO.,LTD
Contact:+86-452-6055586
Address:5F, C Liandalijing Bulding,No 259 Xinjiang Road, Jianhua District
Linyi Shengxin Pharmaceutical R&D Co., Ltd
Contact:+86-18653953873
Address:West First of Yufeng Road, Yishui County
Yangzhou Zhongbao Pharmaceutical Co.,Ltd
Contact:+86-514-88290838
Address:Jiangsu Baoying county economic develpment zone in baoying avenue91
Doi:10.1002/ejoc.200600293
(2006)Doi:10.1080/15257770802257895
(2008)Doi:10.1248/cpb.38.2015
(1990)Doi:10.1039/b717081e
(2008)Doi:10.1002/anie.200600493
(2006)Doi:10.1039/d0dt03500a
(2021)