Application of vitamin B6 content increase in improving resistance of rice to bacterial leaf streak
A vitamin and bacterial technology, applied in the field of plant genetic engineering, can solve problems such as yield and quality decline
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Embodiment 1
[0065] Example 1 Isolation of OsPDX1 Family Genes and Construction of Plant Expression Vectors
[0066] 1. Isolation of OsPDX1 family genes
[0067] Using the genomic DNA of the susceptible rice variety Zhonghua 11 as a template, OsPDX1.1 was amplified by PCR using primers PDX1.1F / PDX1.1R, PDX1.2F / PDX1.2R, and PDX1.3F / PDX1.3R, respectively. , OsPDX1.2, OsPDX1.3 gene DNA fragments, wherein:
[0068] OsPDX1.1, its nucleotide sequence is shown in SEQIDNo.1, its encoded nucleotide sequence is shown in SEQIDNo.4, and its encoded amino acid sequence is shown in SEQIDNo.7;
[0069] OsPDX1.2, its nucleotide sequence is shown in SEQIDNo.2, its encoded nucleotide sequence is shown in SEQIDNo.5, and its encoded amino acid sequence is shown in SEQIDNo.8;
[0070] OsPDX1.3, its nucleotide sequence is shown in SEQIDNo.3, its coded nucleotide sequence is shown in SEQIDNo.6, its coded amino acid sequence is shown in SEQIDNo.9, 1% agarose gel electrophoresis detection its size (such as fig...
Embodiment 2
[0080] Example 2 Functional verification of OsPDX1 family genes
[0081] The invention uses RNA interference (RNAi) technology and exogenous promoter enhanced expression technology to verify the function of the genes by inhibiting and overexpressing OsPDX1.1, OsPDX1.2 and OsPDX1.3 genes in rice varieties. The main mechanism of the RNAi technology approach: Partial fragments of the target gene are connected in the form of inverted repeats to a vector capable of expressing double-stranded RNA (doublestrandRNA, dsRNA), and the vector is introduced into plants through genetic transformation. The obtained transformed plants express a large number of dsRNAs homologous to partial fragments of the target gene, and these dsRNAs rapidly form short interfering RNA (short interfering RNA, siRNA). These siRNAs are complementary to the transcripts (mRNA) of the target gene, and under the action of special enzymes in the cell, the transcripts of the target gene are degraded, thereby inhibiti...
Embodiment 3
[0127] Example 3 Vitamin B6 improves the resistance of rice to bacterial stripe disease
[0128] The experimental results of the above examples show that the changes in the level of vitamin B6 in the transformed plants are closely related to the changes in the resistance of the plants to bacterial stripe disease. The present invention provides representative and reliable results for illustration. The level of vitamin B6 in CD13-22, a typical positive transformation plant with weakened disease resistance, was significantly reduced compared with Hua 11 in the control material ( Figure 15 ). While the expression of OsPDX1 gene was correspondingly increased in CD13-23 and CD13-25 strains ( Figure 14 ). The results indicated that vitamin B6 positively regulates the resistance of rice to bacterial stripe disease.
[0129] In order to further verify the role of vitamin B6 in the anti-bacterial streak bacterium of rice, the present invention adopts vitamin B6 solution (1mM vitam...
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