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Rice zebra leaf mutant gene zebra15 and its encoded protein and application

A mutated gene, zebra leaf technology, applied in rice zebra leaf mutant gene ZEBRA15, protein and application fields, can solve problems such as poor traits, difficulties, and reduced agronomic traits

Inactive Publication Date: 2016-11-23
SOUTHWEST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since most of the marker traits at the seedling stage are not good in themselves, they often lead to a significant decrease in other important agronomic traits. When using them, a cross-breeding process is required to overcome the genetic burden of bad traits and achieve the aggregation of excellent traits. This process is often It is very difficult and can only be achieved through a large number of long-term transfers

Method used

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  • Rice zebra leaf mutant gene zebra15 and its encoded protein and application
  • Rice zebra leaf mutant gene zebra15 and its encoded protein and application
  • Rice zebra leaf mutant gene zebra15 and its encoded protein and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1, Obtaining and Morphological Observation of Rice Zebra Leaf Mutant zebra15

[0017] A genetically stable rice "zebra leaf" mutant named zebra15 was obtained by using ethyl methanesulfonate (EMS) to mutate self-fertilizing Jinghui No. 10. In the rice "zebra leaf" mutant zebra15, when the air temperature suddenly drops and rises in a short period of time (that is, a V-shaped change) during the seedling stage to the tillering period, the leaves of the plant will appear regular light yellow vertical to the veins (White in severe cases) and green zebra stripes, and the leaf sheath part is normal green. After the temperature rises steadily for a period of time, the phenotype will slowly return to normal green. When this V-shaped change in temperature is repeated, the "zebra stripe" mutant phenotype recurs. It shows that the mutant zebra15 may mainly respond to extreme temperature and light changes, and then affect the photoprotection system to form zebra leaves. ...

Embodiment 2

[0018] Embodiment 2, mutation zebra15 gene genetic analysis and localization

[0019] F 1 The leaves of the generation plants were all normal green, and then 4350 F 2 In the generation population, two phenotypes of mutant leaves and normal leaves were separated according to the traits of zebra leaves, and 1054 mutant plants were isolated, and the rest were normal plants. It indicated that the mutant trait was controlled by a pair of recessive single genes.

[0020] Preliminary positioning: 480 pairs of SSR primers evenly distributed on 12 rice chromosomes were selected to detect polymorphisms between the parents zebra-15 and Xinong 1A, and 98 pairs of SSR primers showed polymorphisms. These 98 pairs of primers were used for gene linkage analysis in the normal and mutant gene pools, and the SSR markers linked to the gene Zebra-15 were screened. It was found that Zebra-15 was linked with markers RM3322 and RM6082 on the short arm of chromosome 5. The analysis of 150 recessive...

Embodiment 3

[0025] Embodiment 3, clone Os05g12680 gene

[0026] According to the rice Nipponbare gene Os05g12680 sequence registered in GenBank, use Vector NTI software to design mRNA-specific primers for amplifying the zebra-15 mutant and wild-type Jinhui 10 Os05g12680 sequence: upstream primer ZEBRA15F: 5'-atgtcgtcgccgaccgccg-3'(SEQ ID No.10); downstream primer ZEBRA15R: 5'-tgaactcttgtcaggcaactcaaccc-3' (SEQ ID No.11).

[0027] Take 2 g of young leaves of wild-type Jinhui 10 and mutant zebra15 cultured in the 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 zebra15 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-strand...

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Abstract

The present invention discloses the rice zebra leaf mutant gene ZEBRA15 and its encoded protein and its application. The nucleotide sequence of the rice zebra leaf mutant gene ZEBRA15 is shown in SEQ ID No.12, and its amino acid sequence is shown in SEQ ID No.13. Compared with the wild-type rice zebra leaf mutant gene ZEBRA15, the 64th base of the third exon was converted from G to A, and the encoded amino acid at the 22nd position was changed from glycine to aspartic acid. During the period from seedling stage to tillering of rice zebra15 mutant, when the air temperature drops and rises sharply in a short period of time, the leaves of the plant show zebra leaf traits, which are found to be recessive traits through hybridization, so this trait can be used for breeding Identification of new varieties and seed purity is of great significance to rice genetics and breeding.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a rice zebra leaf mutation gene ZEBRA15, and also relates to a protein encoded by the gene and its application. Background technique [0002] Rice (Orvza sativa L.) is one of the most important food crops in the world, and nearly half of the world's population uses rice as a staple food. The planting of hybrid rice has significantly increased the yield of rice, from 200-300 jin per mu in the past to 800 jin per mu now. Together with the second green revolution, it has almost solved the global food crisis. To protect the country and the world Food security has made a huge contribution. However, seed purity is an important factor restricting hybrid rice from playing a greater role. In recent years, the losses caused to farmers due to impure hybrid rice seeds have become more and more serious. This issue has attracted the attention of the government and experts. Seed pro...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/54C12N9/12C12N15/82A01H5/00
Inventor 赵芳明何光华施军琼王秋实桑贤春凌英华李云峰王楠杨正林
Owner SOUTHWEST UNIV