Application of rice fatty acid hydroxylase gene OsFAH2
A fatty acid hydroxylase and unsaturated fatty acid technology, applied in the field of plant genetic engineering, can solve problems such as reduced seed viability, cell membrane damage, and weakened cell function
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0043] Example 1 Development of OsFAH2 Gene Molecular Marker
[0044]According to the gene sequence variation information of RiceVarMap (http: / / ricevarmap.ncpgr.cn / v2 / ), the 91st nucleotide G / T (SNP site) of OsFAH2 is differentiated in indica / japonica rice, and 97% of the nucleosides in indica rice varieties The acid is T, while the nucleotide of 99% japonica rice varieties is G. The 91st nucleotide G / T variation of gene OsFAH2 was used to design KASP (Competitive Allele Specific PCR) markers for precise selection, and the OsFAH2 near-isogenic line of indica rice variety 9311 background was constructed.
[0045] The KASP detection primers for the above SNP sites are as follows:
[0046] OsFAH2F-F:5'-CGGCCTGGTTCCGCTACTCGGCGG-3' (SEQ ID NO: 6)
[0047] OsFAH2H-F:5'-CGGCCTGGTTCCGCTACTCGGCGT-3' (SEQ ID NO:7)
[0048] OsFAH2-R:5'-GTGGCAGTAGAGGCAGTAGTCCGG-3' (SEQ ID NO:8)
Embodiment 2
[0049] Example 2 KASP marker genotyping of OsFAH2 and construction of near-isogenic lines
[0050] The KASP primer can accurately distinguish OsFAH2 japonica rice and indica rice varieties or genotypes, and can be used in the subsequent molecular marker-assisted selection breeding process. KASP marker genotyping effect of OsFAH2 gene see figure 1 . Using the KASP primer of OsFAH2 gene, NIP and 9311 hybrids were continuously backcrossed with 9311 for 4 times for gene selection, and the NIP near-isogenic line of OsFAH2 was constructed with the genetic background of 9311. Finally, a near-isogenic line with OsFAH2 replaced by NIP and a genetic background of 9311 was obtained. The genotype was detected by resequencing, and it was found that the whole genome only contained a 1Mb NIP-introduced fragment at OsFAH2. Near isogenic line NIL-OsFAH2 NIP Chromosomal structure map of genotype see figure 2 .
[0051] 1. The expression level of OsFAH2 in the near isogenic line of 9311 an...
Embodiment 3
[0067] Example 3 OsFAH2 Gene Overexpression Vector Construction and Genetic Transformation
[0068] The 25 bp primers were cut from both ends of the CDS of OsFAH2, and the homologous recombination adapters on both sides of the KpnI restriction site of the pC1301S vector were added to amplify the OsFAH2 coding region of the cDNA isolated clone of 9311.
[0069] The primer sequences are as follows:
[0070] OEK-F: 5'-atgatgatgataaaggtaccATGCTCCCGTACGCGACGGCGGCGG-3'
[0071] OEK-R: 5'-ctagaggatccccgggtaccATCTTGTTTCCCGTTGCCGAATCCA-3'
[0072] To overexpress the vector pC1301S, that is to introduce the CaMV35S promoter in the multiple cloning site region of the pC1301 vector. The OsFAH2 coding region fragment isolated by PCR amplification was connected to the KpnI digested linearized pC1301S vector by homologous recombination. After the sequencing comparison is correct, the OsFAH2 gene overexpression vector is obtained. The OsFAH2 gene overexpression vector was transformed into...
PUM
| Property | Measurement | Unit |
|---|---|---|
| Diameter | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to View More