Application of rice protein CHR1 in adjusting content of plant nitrate
A nitrate content, nitrate technology, applied in the direction of application, plant products, botany equipment and methods, etc., can solve the problems of increasing agricultural production costs, environmental pollution, etc.
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Embodiment 1
[0030] Example 1, rice protein CHR1 is a nitrate transporter
[0031] 1. Screening of rice mutants
[0032] Studies have shown that chlorate, as a toxic analog of nitrate, can be used to identify Arabidopsis mutants with nitrate uptake defects (literature: Tsay YF, Schroeder JI, Feldmann KA, & Crawford NM (1993) The Herbicide Sensitivity Gene Chl1of Arabidopsis Encodes a Nitrate-Inducible Nitrate Transporter. Cell72(5):705-713.Wilkinson JQ& Crawford NM(1991) Identification of the Arabidopsis CHL3Gene as the Nitrate Reductase Structural Gene NIA2.Plant Cell3(5):461-47 .). In this experiment, chlorate was used to screen the rice T-DNA mutant library created by the inventor's research team using rice Zhonghua 11 as the background material, and a rice mutant chr1 with significant resistance to chlorate was obtained.
[0033] The method and results of the above-mentioned chlorate screening test are as follows:
[0034]The mutant seeds and wild-type rice Zhonghua 11 were subjecte...
Embodiment 2
[0065] Example 2, overexpression of protein CHR1 increases the nitrate content of rice or improves the utilization rate of rice to nitrate
[0066] 1. Construction of plant recombinant expression vector
[0067] Use primer 5'- CCCGGG CGCCGTCTTACTCTCTCTG-3' (the slash part of the base is the XmaI restriction recognition sequence, and the rest of the sequence is the same as the 44th-62nd position of the sequence listing sequence 2) and 5'- TCTAGA AATTATACCCGGCGAAATAC-3' (the slashed base is the recognition sequence for XbaI digestion, and the rest of the sequence is reverse complementary to the 2068-2087 position of sequence 2 in the sequence listing), the cDNA obtained by reverse transcription of the total RNA of rice Zhonghua 11 is Template, carry out PCR amplification, recover and purify the fragment of 1.8kb, after double digestion with XmaI and XbaI, connect with the carrier backbone of the vector pCAMBIA2300-Actin1 after double digestion with XmaI and XbaI, it is confir...
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