Glucoside hydrolase gene and promoter for improving cotton yield traits and application thereof
A glycoside hydrolase and gene technology, applied in the field of biotechnology applications, can solve problems such as differences and inability to accumulate flavonoids, and achieve the effects of high sensitivity, small amount of template, and simple and easy test steps
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0034] Example 1 Mining of the glycoside hydrolase gene GHLP1 associated with cotton clothing traits:
[0035] For 486 modern upland cotton varieties or lines, from 2016 to 2017, three replicates of each variety were planted in the fields of Korla, Xinjiang and Shihezi, Xinjiang, and a detailed investigation of the cotton traits was carried out. At the same time, the whole genome resequencing of these 486 cotton varieties (lines) was carried out, and 7.55Tb sequencing data were obtained, with an average sequencing depth of 10.51×. These sequences were compared to the genome sequence of cotton Upland cotton TM-1, and the genome-wide SNPs were identified using bioinformatics software. A total of 4 489 601 high-quality SNPs (minimum gene frequency >0.05) were mined for subsequent analysis . Firstly, genome-wide association analysis was performed, and then according to P-6 Screen for SNP-associated signal sites. Through the analysis of these association loci, we found a SNP sign...
Embodiment 2
[0036] Example 2 Obtaining of glycoside hydrolase gene GHLP1:
[0037] The cDNA sequence, genome sequence and promoter sequence of GHLP1 were obtained from the upland cotton genome sequence, see SEQ ID NO.1, SEQ ID NO.2 and SEQ ID NO.3. Design full-length primers according to the two ends of the cDNA for PCR amplification. The primer sequences are F1: SEQ ID NO.4 and R1: SEQ ID NO.5. The full-length primers of the gene were designed according to the two ends of the promoter, and PCR amplification was carried out. The primer sequences were F2: SEQ ID NO.6 and R2: SEQ ID NO.7. The PCR reaction program was as follows: pre-denaturation at 94°C for 5 min; denaturation at 94°C for 30 sec, annealing at 60°C for 1 min, extension at 72°C for 1 min, 30 cycles; and finally extension at 72°C for 10 min. The PCR amplification product was sequenced, and further compared with the cDNA and promoter sequences to determine the accuracy of the sequence.
Embodiment 3
[0038] Example 3 Analysis of the expression level of GHLP1 in different tissues and developmental stages of cotton:
[0039] In this experiment, RNA samples from different tissues and different developmental stages of cotton TM-1 variety were collected for transcriptome sequencing. Sample material included roots, stems, leaves, ovules and fibers. The ovule tissue includes 3 and 1 days before flowering, the day of flowering, and 1 to 25 days after flowering. Fibrous tissue is included 5 to 25 days after flowering. Illumina HiSeq 2500 platform was used for transcriptome sequencing, and the average sequencing depth of each sample reached 6Gb. The calculation of gene expression level is to compare the reads obtained by sequencing with the upland cotton genome, and the calculated expression level is the number of sequenced fragments contained in every thousand transcript sequenced bases per million sequenced bases (FPKM) express. Experimental results such as Figure 4 As shown...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More