Rice yellow-green leaf mutation gene YGL8, protein coded by rice yellow-green leaf mutation gene YGL8, and application of rice yellow-green leaf mutation gene YGL8
A technology for mutating genes and yellow-green leaves, applied in the field of rice yellow-green leaf mutant gene YGL8, can solve problems such as yield reduction, abnormal plant growth and development, and decreased photosynthetic efficiency
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Embodiment 1
[0018] Example 1, Obtaining and Morphological Observation of Rice Yellow-green Leaf Mutant ygl8
[0019] In the rice EMS mutant bank of the Rice Research Institute of Southwest University, a mutant with yellow-green leaves during the whole growth period was found, and it was named ygl8 according to the phenotypic characteristics ( figure 1 ). This trait has been observed through multiple generations, showing stable inheritance. The ultrastructure of the leaf cells of the wild type Jinhui 10 and the ygl8 mutant was observed by transmission electron microscopy. figure 2 ); major agronomic traits such as panicle length and grain number per panicle were significantly reduced, however, there was no significant difference in thousand-grain weight ( image 3 ).
Embodiment 2
[0020] Embodiment 2, mutation ygl8 gene genetic analysis and localization
[0021] When the phenotypically normal sterile line Xinong1A (Xinong1A) was crossed with the ygl8 mutant, all F 1 The plants were all phenotypically normal. f 2 There were obvious segregation phenomenon of parental traits in the population. Among the 9473 populations, there were 7095 normal plants and 2378 yellow-green mutant plants. The chi square test showed that the segregation ratio was 3:1, indicating that the mutant trait was affected by a pair of recessive genes. control.
[0022] Preliminary mapping: 400 pairs of SSR markers evenly distributed on the 12 rice chromosomes were selected to amplify the parental and gene pool DNA. It was found that the markers RM6141, RM12233 and RM6321 located on the first chromosome may be linked to YGL8. Further analysis of F using linkage markers RM6141, RM12233 and RM6321 2 452 individuals with recessive mutations were found in the generation population, and...
Embodiment 3
[0027] Embodiment 3, clone Os01g73450 gene
[0028] According to the rice Nipponbare gene Os01g73450 sequence registered in GenBank, use VectorNTI software to design and amplify mRNA-specific primers for wild-type Jinhui 10 and ygl8 mutant Os01g73450 sequence: upstream primer YGL8-F: 5'-gtcctcatcgtcatcatctcgt-3' (SEQIDNo. 13); downstream primer YGL8-R: 5'-agcaataggcaatgcagctac-3' (SEQ ID No. 14). Take 2 g of young leaves of wild-type Jinhui 10 and mutant ygl8 cultured in light for two weeks, put them into liquid nitrogen quickly and grind them into powder, and extract total RNA according to the instructions of the Trizol kit. The results of electrophoresis of total RNA of wild-type Jinhui 10 and mutant ygl8 showed that the main band was clear and complete, and the band brightness ratio of 28S and 18S was about 2:1, indicating that the concentration and purity of RNA met the experimental requirements and could be used for synthesis double-stranded cDNA. Then use the obtained ...
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