A protein related to male and female fertility in rice, its coding gene and application thereof
A rice, male and female technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve problems such as unclear molecular basis
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0069] Example 1 Phenotype of XF2 gene mutation
[0070] A sterile mutant was found in the rice material XF2 (preserved in the Resource Center of the Rice Research Institute of Sichuan Agricultural University). After forward and reverse hybridization and cytological observation, it was found that the mutant was sterile in both male and female organs ( Figure 1-Figure 3 ), further using techniques such as paraffin section and DAPI, it was found that the root cause of its sterility was the abnormal meiosis of mutant pollen mother cells ( Figure 4 ) and structural abnormalities of the embryo sac ( Figure 5 ),Depend on image 3 It can be seen that the pollen of the mutant is inactive (active pollen can be dyed purple by iodine solution because of starch accumulation, and inactive pollen cannot); Figure 5 It can be seen that the mature embryo sac of fertile plants forms a normal cyst cavity, and polar nuclei, egg cells, antipodal cells, and synergid cells can be seen in the c...
Embodiment 2
[0071]Cloning of embodiment 2 XF2 gene mutation
[0072] 1. Map-based cloning of XF2 gene mutation
[0073] Use Guichao 2 as the female parent to hybridize with the heterozygote containing the mutant gene, construct a segregation population, locate the gene, and realize fine mapping. The marker encryption and sequence comparison analysis of the fine positioning interval show that in the mutant, the gene in this region A gene has a deletion of 2 bases ( Figure 6-Figure 8 ),Depend on Figure 6 It can be seen that the gene is located between the two markers INDEL17 and R8 by the method of map-based cloning, and the total number of genes in the positioning interval is as follows: Figure 7 genes; by Figure 7 It can be seen that by sequencing the 7 genes in the interval, it is found that there are two base deletions in the exon of the gene LOC_Os02g42910, indicating that this gene is a candidate target gene, Figure 8 The sequence map of the deletion site is indicated.
[00...
Embodiment 3
[0087] Example 3 XF2 Genetic Rice Transgenic Test
[0088] In this example, the expression vector PHB (Mao et al., 2005, PNAS102: 12270-12275, commercially available) was used as the complementation and overexpression vector of the rice transgene. The vector encodes a bacterial origin of replication (ori), kanamycin resistance gene (Kanr), hygromycin resistance gene (Hygr), herbicide resistance gene (Barr), 2×35S promoter, NOS gene termination Signal sequence and multiple cloning site (MCS) for ligation of fragments of interest. The XF2 gene or its fragment can be inserted into the restriction endonuclease site to construct a transgenic plasmid. In this example, VK005-01 (commercially purchased) is used as the rice transgenic knockout vector, which encodes Ubi promoter, mpCas9, Ru6-gDNA, 2×35S promoter, hygromycin resistance gene (Hygr), kana Mycin resistance gene (Kanr).
[0089] 1. Construction of XF2 gene complementary expression vector
[0090] In this example, pBS-PXF...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More