Rice grain length and graining shattering associated proteins and their coding genes and use
A coding and shattering technology, applied in the field of protein and its coding genes and applications, can solve the problems of little knowledge of population genetic structure and evolutionary history, lack of research on African cultivated rice, etc. The effect of long grain length
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Embodiment 1
[0071] Example 1, Regulating the location of African rice grain size and shattering genes
[0072] African rice variety IRGC102305 was crossed with African wild rice W1411, and then IRGC102305 was used as the recurrent parent, and a line GIL25 with obvious difference in grain size from IRGC102305 was obtained through multiple backcrosses, and its grain was easy to shed ( figure 1 ), crossed it with IRGC102305 to construct a F2 segregation population containing 186 individuals. In this F2 population, both kernel length and kernel shattering were closely linked to markers RM3335 and RM5608, and the GL4 gene was located between these two genes. Then the segregation population was expanded to 6500 individual plants, and it was found that shattering and kernel size co-segregated. And according to the sequence design primers between the markers RM3335 and RM5608, the GL4 gene is finally located on the 5.9kb between the markers M3 and M4 ( figure 2 ). And all the lines with longe...
Embodiment 2
[0073] Example 2, Obtaining and functional verification of GL4 transgenic African rice
[0074] 1. Construction of expression vector
[0075] Primers were designed according to the gene ORGLA04G0254300 and the promoter sequence of African wild rice W1411, and restriction endonuclease XbaI and HindIII recognition sites and protective bases were introduced at both ends of the primers respectively. The primer sequences are as follows:
[0076] pGL4-1F: 5'- TGCTCTAGA GCTGGCCGTAGAAGTCAAG-3' (the underlined base is the restriction endonuclease XbaI recognition site and the protection base, and the sequence thereafter is the 1-19th position of sequence 4);
[0077] pGL4-1R: 5'- ACAAAGCTT GTATAGCTCAGCACGAGC-3' (the underlined base is the restriction endonuclease HindIII recognition site and the protection base, and the following sequence is the reverse complementary sequence of the 4876-4893 position of sequence 4).
[0078] The whole genome DNA of African wild rice W1411 was ext...
Embodiment 3
[0092] Embodiment 3, the acquisition and verification of key variation
[0093] 1. Acquisition of key variants
[0094] Using qPCR to compare the expression levels of GL4 between African wild rice W1411 and African cultivated rice variety IRGC102305, it was found that there was no significant difference in the expression of GL4 between the two materials. Sequence comparison of the coding region of the GL4 gene of African wild rice W1411 and African cultivated rice variety IRGC102305 found that there were five mutations: G294A, C766T, C1466T, A1515G and GCCGCC681-686 deletions (the variation is relative to the African cultivated rice variety IRGC102305 African wild rice W1411). By comparing the GL4 gene sequences of 16 wild rice and 67 African cultivated rice, it was found that C766T had the highest correlation with grain length and grain shattering.
[0095] 2. Construction of expression vector
[0096] Using the genome of the African cultivated rice variety IRGC102305 as a...
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