A deubiquitinase gene ubp5 regulating rice grain shape and leaf color and its application

Abstract

The invention provides a deubiquitinating enzyme gene UPB5 for regulating and controlling a grain shape and leaf color of rice. The deubiquitinating enzyme gene UPB5 has a nucleotide sequence as shown in SEQ ID No.1 in the description. The invention further provides a protein with a UBP5 gene code of the rice, and the protein has an amino acid sequence as shown in SEQ ID No.3 in the description. The experiment proves that the deubiquitinating enzyme gene UPB5 of the rice can cause the leaf blade color and seed shape of the rice to change; the invention further finds that after the ubiquitination enzyme gene UPB5 of the rice mutates, the leaf blades of the rice show the shape of a stripe; the seeds are elongated; the ripening rate becomes lower; the total content of chlorophyll is obviously reduced. The invention provides the application of the UBP5 gene and the mutant of the UBP5 gene to genetic improvement breeding of rice germplasm resources, thereby having a relatively good application prospect.

Description

technical field

[0001] The invention belongs to the field of genetic engineering, and in particular relates to a deubiquitinase gene UBP5 for regulating grain shape and leaf color of rice, its encoded protein and its application in improving rice germplasm resources. Background technique

[0002] Protein ubiquitination and deubiquitination are post-transcriptional modifications that exist widely in eukaryotic cells. The precise regulation of ubiquitination and deubiquitination of substrates has very important biological significance, and is involved in important processes such as protein degradation, autophagy, DNA damage repair, cell cycle, signal transduction, gene expression, and inflammatory immunity. life process.

[0003] The ubiquitination of protein is mainly involved in the degradation of substrates and the removal of abnormal proteins. It is completed by a series of enzymatic reactions, including ubiquitin activating enzyme E1, ubiquitin coupling protein E2 and ub...

Examples

Embodiment 1

[0031] The acquisition and phenotype analysis of embodiment 1 mutant

[0032] The rice ubp5 mutant was derived from the mutation produced by EMS mutagenesis of the japonica rice variety Nipponbare, which was obtained in Beijing, China.

[0033] Under normal sowing conditions, the germination potential of mutant ubp5 seeds was significantly weaker than that of the wild type, and the leaves of the mutant ubp5 showed white stripes after germination; at the tillering and maturity stages, the ubp5 stripe leaf phenotype appeared on some leaves of the plants It is always present and will not disappear in the later stages of childbearing. Compared with the wild type, the chlorophyll content of the mutant decreased in different periods. In addition, the grain length of the mutant increased significantly, while the grain width did not change significantly ( figure 1 ).

Embodiment 2

[0034] Example 2 Isolation and cloning of UBP5 gene

[0035] 1. Genetic analysis of ubp5 mutant traits:

[0036] The ubp5 mutant was crossed with its parent Nipponbare to construct the F 2 Populations were isolated for genetic analysis. The analysis results showed that the F 1 The phenotype of the offspring plants was not significantly different from that of the wild type, and in F 2 218 individual plants were randomly selected in the generation population, and the number of individual plants of the wild-type and mutant phenotypes were 168 and 50, respectively. The chi-square test results (χ 2 3:1 =0.391) in line with the segregation ratio of 3:1, indicating that the traits of the mutant are controlled by a single recessive nuclear gene.

[0037] 2. Gene mapping group construction:

[0038] The ubp5 mutant was crossed with the indica variety Dular, F1 F 2 group. from F 2 1550 ubp5 mutant phenotype plants were selected from the population as the gene mapping population...

Embodiment 3

[0045] Example 3 UBP5 and its homologous gene phylogenetic tree analysis

[0046] Deubiquitinases belong to a large gene family, and each species includes a very large number of deubiquitinase genes. For example, Arabidopsis, a model plant for dicotyledon research, contains 27 deubiquitinase genes, and rice contains 23 deubiquitinase genes. In order to study the evolutionary relationship between UBP5 and other rice deubiquitinase proteins, the phylogenetic tree of UBP5 protein and 22 other rice proteins was constructed using MEGA 4.0 software. The results showed that UBP5 and Os10g07270 (UBP9) had the closest relationship and were on the same branch, while Os07g06610 and Os12g42600 at this site were on a large branch ( image 3 ), indicating that the protein family is relatively conserved in structure and closely related in evolution.

technical field

[0001] The invention belongs to the field of genetic engineering, and in particular relates to a deubiquitinase gene UBP5 for regulating grain shape and leaf color of rice, its encoded protein and its application in improving rice germplasm resources. Background technique

[0002] Protein ubiquitination and deubiquitination are post-transcriptional modifications that exist widely in eukaryotic cells. The precise regulation of ubiquitination and deubiquitination of substrates has very important biological significance, and is involved in important processes such as protein degradation, autophagy, DNA damage repair, cell cycle, signal transduction, gene expression, and inflammatory immunity. life process.

[0003] The ubiquitination of protein is mainly involved in the degradation of substrates and the removal of abnormal proteins. It is completed by a series of enzymatic reactions, including ubiquitin activating enzyme E1, ubiquitin coupling protein E2 and ub...

Examples

Embodiment 1

[0031] The acquisition and phenotype analysis of embodiment 1 mutant

[0032] The rice ubp5 mutant was derived from the mutation produced by EMS mutagenesis of the japonica rice variety Nipponbare, which was obtained in Beijing, China.

[0033] Under normal sowing conditions, the germination potential of mutant ubp5 seeds was significantly weaker than that of the wild type, and the leaves of the mutant ubp5 showed white stripes after germination; at the tillering and maturity stages, the ubp5 stripe leaf phenotype appeared on some leaves of the plants It is always present and will not disappear in the later stages of childbearing. Compared with the wild type, the chlorophyll content of the mutant decreased in different periods. In addition, the grain length of the mutant increased significantly, while the grain width did not change significantly ( figure 1 ).

Embodiment 2

[0034] Example 2 Isolation and cloning of UBP5 gene

[0035] 1. Genetic analysis of ubp5 mutant traits:

[0036] The ubp5 mutant was crossed with its parent Nipponbare to construct the F 2 Populations were isolated for genetic analysis. The analysis results showed that the F 1 The phenotype of the offspring plants was not significantly different from that of the wild type, and in F 2 218 individual plants were randomly selected in the generation population, and the number of individual plants of the wild-type and mutant phenotypes were 168 and 50, respectively. The chi-square test results (χ 2 3:1 =0.391) in line with the segregation ratio of 3:1, indicating that the traits of the mutant are controlled by a single recessive nuclear gene.

[0037] 2. Gene mapping group construction:

[0038] The ubp5 mutant was crossed with the indica variety Dular, F1 F 2 group. from F 2 1550 ubp5 mutant phenotype plants were selected from the population as the gene mapping population...

Embodiment 3

[0045] Example 3 UBP5 and its homologous gene phylogenetic tree analysis

[0046] Deubiquitinases belong to a large gene family, and each species includes a very large number of deubiquitinase genes. For example, Arabidopsis, a model plant for dicotyledon research, contains 27 deubiquitinase genes, and rice contains 23 deubiquitinase genes. In order to study the evolutionary relationship between UBP5 and other rice deubiquitinase proteins, the phylogenetic tree of UBP5 protein and 22 other rice proteins was constructed using MEGA 4.0 software. The results showed that UBP5 and Os10g07270 (UBP9) had the closest relationship and were on the same branch, while Os07g06610 and Os12g42600 at this site were on a large branch ( image 3 ), indicating that the protein family is relatively conserved in structure and closely related in evolution.

Images