Application and method of 5-formyltetrahydrofolate in improving nitrogen utilization rate of plants
A technology of leucovorin and 5-F-THF, which is applied in the agricultural field, can solve the problems of excessive nitrogen fertilizer application and low utilization efficiency, and achieves the improvement of growth condition, the increase of amino acid content, the increase of fresh weight and chlorophyll content. Effect
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Embodiment 1
[0017] 5-Methyltetrahydrofolate (5-M-THF) and 5-formyltetrahydrofolate (5-F-THF) are the two main forms of folic acid in plants. The invention isolates and obtains a folic acid mutant, the main form of folic acid in the folic acid mutant is 5-M-THF, and its content is obviously reduced, and the folic acid mutant shows a phenotype of shortened main root.
[0018] The present invention applies 500 μM 5-M-THF exogenously to the folic acid mutant, such as Figure 8 As shown, the folic acid mutant still maintains the phenotype of tap root development defects; when 500 μM 5-F-THF is applied exogenously to the folic acid mutant, the development of the mutant's tap root is improved, and the tap root length returns to the wild-type level. Therefore, it was shown that 5-F-THF could enter and be utilized in Arabidopsis seedlings.
Embodiment 2
[0020] Using Arabidopsis thaliana and rice as materials, the effects of exogenous application of 5-formyltetrahydrofolate (5-F-THF, 5-formyl-tetrahydrofolate) on alleviating low nitrogen stress in seedlings were analyzed.
[0021] experiment procedure:
[0022] The 5-formyltetrahydrofolate powder was prepared into a 10mM stock solution with sterilized water, and stored at -80°C for future use.
[0023] Arabidopsis thaliana, Columbia-0 wild-type seeds (Arabidopsis thaliana, Columbia-0) were sown on the medium supplemented with 5-formyltetrahydrofolate (working concentration 500 μM) after surface disinfection, and the medium only used NO 3- As the only nitrogen source, other components are the same as 1 / 2MS medium. NO in 9.4N medium 3- The concentration is 9.4mM; NO in 0.3N medium 3- The concentration is 0.3mM, while adding 9.1mM KCl to make K + The concentration was maintained at 9.4 mM. The Arabidopsis plate was placed in a light incubator, the culture conditions were set...
Embodiment 3
[0032] experiment procedure:
[0033] The 5-formyltetrahydrofolate powder was prepared into a 10mM stock solution with sterilized water, and stored at -80°C for future use.
[0034] Arabidopsis thaliana, Columbia-0 wild-type seeds (Arabidopsis thaliana, Columbia-0) were sown on a medium supplemented with 5-formyltetrahydrofolate after surface disinfection, and the medium was only used with NO 3- As the only nitrogen source, other components are the same as 1 / 2MS medium. NO in 9.4N medium 3- The concentration is 9.4mM; NO in 0.3N medium3- The concentration is 0.3mM, while adding 9.1mM KCl to make K + The concentration was maintained at 9.4 mM. The Arabidopsis plate was placed in a light incubator, the culture conditions were set at 22°C, the photoperiod was 16 h light / 8 h dark, and the light intensity was 60 μE m -2 the s -1 . On the 10th day of light culture, the materials were collected, washed with sterilized ultrapure water, and the folic acid extraction and content d...
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