A functional gene for enhancing plant iron deficiency tolerance and iron accumulation and its application
A functional gene, plant technology, applied in the direction of plant gene improvement, application, plant peptides, etc., can solve the problem of unclear regulation and other problems, and achieve the effect of improving absorption
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Embodiment 1
[0024] Example 1 Obtaining of ATMYC1 mutants
[0025] We used Arabidopsis mutant seeds obtained from the American Arabidopsis Germplasm Bank to screen for mutants responding to iron deficiency stress in plants. The seed mutant library of Arabidopsis thaliana was screened using MS medium without iron, and a mutant strain resistant to iron deficiency was obtained. After screening and identification, it was found that the mutant was a mutant of ATMYC1 gene function loss, named atmyc1- 1, whose seed number is SALK-056899. In order to further study the role of ATMYC1 gene in plant response to iron deficiency stress, we obtained another ATMYC1 gene loss-of-function mutant named atmyc1-2 from the American Arabidopsis Germplasm Bank, and its seed number is SALK-057388. Because the seed is a T-DNA insertion mutation, it is amplified by PCR with specific primers and sequenced. The sequencing results are compared in the NCBI database Blast, and the T-DNA insertion site information is ob...
Embodiment 2
[0027] Embodiment 2, cultivating iron-deficiency phenotype sensitive Arabidopsis
[0028] 1, ATMYC1 Gene Overexpression Transgenic Lines ATMYC1 -Acquisition of OE1 and OE5
[0029] In order to further verify the function of this gene in the regulation of plant iron deficiency stress, we constructed an overexpression vector of ATMYC1 gene (35S: ATMYC1). Firstly, the target fragment is amplified. The wild-type Arabidopsis was normally cultured on MS medium for two weeks, the total RNA of the plant was extracted, reverse-transcribed to synthesize cDNA, and the synthesized cDNA was used as a template for PCR to amplify a sufficient amount of the target product, see Figure 5Middle A. Using the PCR product as a template, the second amplification was carried out for the purpose of introducing enzyme cutting sites. The PCR product and vector pCAMBIA1301 were digested and recovered. Then, the recovered and purified target DNA fragment and the vector were ligated overnight with...
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