Paddy bacterial leaf streak resistance-related gene OsDRxoc6
A osdrxoc6, bacterial technology, applied in genetic engineering, plant genetic improvement, plant peptides, etc., can solve problems such as yield and quality decline
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Embodiment 1
[0038] Example 1 OsDRxoc6 Gene Structure Prediction and Analysis
[0039] Using the DNA coding sequence SEQ ID No.2 of the OsDRxoc6 gene as a template, the nucleotide database was searched by BLAST method, and it was found that the sequence of SEQ ID No.2 was located on the short arm of chromosome 5 of rice variety Nipponbare, which was consistent with the LOC_05g01380 gene predicted by Genebank At the same locus, it is located in the rice bacterial streak resistance QTL qBlsr5a. According to the coding sequence of SEQ ID No.2, the amino acid sequence of SEQ ID No.3 can be obtained, which consists of 309 amino acids.
Embodiment 2
[0040] Example 2 Analysis of the expression pattern of OsDRxoc6 gene in different rice varieties
[0041] 1. Background expression of OsDRxoc6 gene in resistant variety Acc8558 and susceptible variety Zhonghua 11
[0042] In order to detect whether there is a difference in the background expression level of the OsDRxoc6 gene in the disease-resistant and susceptible varieties, and to verify whether it is related to disease resistance, the present invention uses real-time quantitative RT-PCR technology to analyze the OsDRxoc6 gene in the rice disease-resistant variety Acc8558 and the susceptible variety ZH11. Differences in background expression in . Total RNA was extracted from leaves of rice plants, and 1-5 μg of total RNA was treated with DNaseI (Invitrogen, USA) for 30 minutes to remove genomic DNA contamination, and then oligo(dT)15 was used to oligomerize Primers and SuperScriptII reverse transcriptase (Invitrogen) were used for reverse transcription. Quantitative PCR an...
Embodiment 3
[0045] Example 3 Isolation of OsDRxoc6 Gene and Construction of Plant Expression Vector
[0046] 1. Isolation of OsDRxoc6 gene
[0047] Using the genomic DNA of the susceptible rice variety Zhonghua 11 as a template, by PCR technology, using primers OsDRxoc6F1 / OsDRxoc6R1 to amplify and obtain a partially suppressed expression DNA fragment of the OsDRxoc6 gene, its sequence is shown in the sequence table SEQ ID NO.4 (SEQ ID NO.2 sequence 11th to 580bp), 1% agarose gel electrophoresis detected its size 570bp ( Figure 5 ); Using the genomic DNA of the disease-resistant rice variety Acc8558 as a template, the OsDRxoc6F2 / OsDRxoc6R2 combination primers amplified the complete overexpression fragment, and the target band of 1151bp was detected by 1% agarose gel electrophoresis ( Figure 6 ), whose sequence is shown in the sequence listing SEQ ID NO.1. Primer sequences are shown in Table 1
[0048] Table 1. Related primer sequences
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[0051] 2. Construction ...
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