Application of SPL18 gene in enhancing plant yield
A gene and plant technology, applied in the application field of SPL18 gene, can solve the problem of unknown function of OsSPL gene
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0039] Example 1 Construction of Rice SPL18 Gene OsSPL18 Overexpression Vector
[0040] OsSPL18 gene expression box genome sequence (nucleotide sequence shown in SEQIDNO.5) acquisition: design PCR primers:
[0041] OsSPL18-geno-F (5'TGGTACCGATCTCACGTAAATACAATTCTCC) and OsSPL18-geno-R (5'CGGGTACCAATGATATATTTCTGCATAAGTTT) were amplified by PCR using the commercial rice variety Xiushui-134 genome as a template, including the putative OsSPL18 gene promoter, expression sequence and termination The size of the subsequence is a genome fragment of 7.4kb, and the sequence is shown in SEQ ID NO:5. The PCR reaction conditions were: 95°C for 3 minutes; 95°C for 15 seconds, 58°C for 15 seconds, 72°C for 7.5 minutes, repeating 33 cycles; and then 72°C for 10 minutes.
[0042] Construction of the Agrobacterium T-DNA vector: the binary vector pCambia1300-p35S-G10 vector (InternationalPat.No.PCT / CN2012 / 087069:SEQIDNO.47) contains a glyphosate-resistant gene (EPSPS) (as a marker gene for trans...
Embodiment 2
[0043] The construction of embodiment 2 rice SPL18 gene cDNA overexpression vector
[0044] 1. Cloning of cDNA
[0045] Cloning of rice OsSPL18 gene cDNA: PCR primers OsSPL18-CF (5'GGATCCAACAATGGATTGGGATCTCAAGATG) and OsSPL18-CR (5'GAGCTCCTACTGCCACGAGAATGGGAGCG) were designed, using the cDNA reverse-transcribed from the total RNA of inbred line 9311 as a template, and obtained by PCR amplification cDNA sequence of OsSPL18. The PCR reaction conditions were: 95°C for 3 minutes; 95°C for 15 seconds, 58°C for 15 seconds, 72°C for 90 seconds, repeating 33 cycles; and then 72°C for 10 minutes. The obtained PCR product of about 1.5 kb was cloned into the T-vector pMD19. Then, the corresponding cDNA (SEQ ID NO: 6) was obtained by double digestion with BamHI and SacI, and DNA sequence determination showed that the nucleotide sequence of the cDNA was correct. Then use pCambia1300 as a transition vector, double digest with BamHI and SacI to obtain the sequenced correct OsSPL18cDNA fra...
Embodiment 3
[0063] Embodiment 3, transformation of rice
[0064] The method of obtaining transgenic rice is to adopt the existing technology (Lu Xiongbin, Gong Zuxun (1998) Life Science 10: 125-131; Liu Fan et al. (2003) Molecular Plant Breeding 1: 108-115). The mature and plump seeds of "Xiushui-134" were dehulled, and callus was induced as transformation materials. Take the vectors constructed in Example 1 and Example 2 (pCambia1300-pOsSPL18-OsSPL18-p35S-pZmUbi-1174(geno);
[0065] pCambia1300-p35S-OsSPL18-pZmUbi-1174;
[0066] pCambia1300-pOsSPL18-OsSPL18-ter-p35S-pZmUbi-1174,
[0067] pCambia1300-pOsGA20ox1-OsSPL18-ter-p35S-pZmUbi-1174;
[0068] pCambia1300-pOsTEL-OsSPL18-ter-p35S-pZmUbi-1174;
[0069] Agrobacterium plating of pCambia1300-pOsARGOS-OsSPL18-ter-p35S-pZmUbi-1174). Pick a single colony to inoculate and prepare Agrobacterium for transformation. The callus to be transformed is put into the Agrobacterium bacterium liquid that OD is about 0.6 (preparation of Agrobacteri...
PUM

Abstract
Description
Claims
Application Information

- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com