Use of acyl-CoA ligase or its encoding gene related to pollen development and method for breeding plant sterile lines
A technology of pollen development and ligase, which is applied in the fields of agriculture and biology, can solve the problems of reduction, low purity of seed production, few genetic sterility photothermosensitivity loci, etc.
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Embodiment 1
[0156] Example 1 The sterility phenotypes of Arabidopsis acos5-2 and acos5-3 mutants are restored under low temperature conditions
[0157] The inventors isolated the acos5-2 mutant from Arabidopsis thaliana Ler ecotype by means of EMS chemical mutagenesis. At the same time, an exon-inserted mutant acos5-3 was screened in the Arabidopsis Col ecotype T-DNA insertion mutant library. At normal ambient temperature (24°C), the homozygous acos5-2 mutant had normal vegetative growth, but lost fertility and had only short, seedless pods, such as figure 1 As shown in A. Genetic analysis showed that the acos5-2 mutant was sporophytic male sterility controlled by a single recessive locus. The applicant cultivated the acos5-2 mutant at 24°C until bolting, then moved it to 18°C for continuous culture, and the subsequent fruit pods all recovered their fertility, as shown in figure 1 As shown in A, the fertility recovery of the acos5-3 mutant at low temperature is more difficult than th...
Embodiment 2
[0158] Example 2 Gene mapping of acos5-2 and ACOS5 gene structure
[0159] Using the Arabidopsis thaliana In / Del molecular marker in our laboratory, the ACOS5 gene was located on the first chromosome of Arabidopsis thaliana through the preliminary location of the sterile phenotype plants in the F2 generation of the acos5-2 mutant, and the molecular marker F7A10 Tightly linked. Since then, six pairs of fine-mapping primers have been designed: F24O1, F23N19, F16P17(8), F16P17(Snp-1), F16M19, and F13O11, which further narrowed the mapping interval of the ACOS5 gene to F16P17(23283) and F16P17(23327). Within the 44KB range between molecular markers, this range contains 6 candidate genes, such as figure 2 Shown in A. The At1g62940 gene encodes an acyl-CoA ligase protein. In the acos5-2 mutant, a CCA-to-CTA mutation (proline to leucine) single-nucleotide mutation was detected in the third exon of the gene, as figure 2 Shown in B. For the T-DNA insertion mutant acos5-3, the ap...
Embodiment 3
[0160] Example 3 Low temperature can compensate for microspore development defects in acos5 mutants
[0161] To determine the defect in pollen development of the acos5 mutant, Applicants performed anther semi-thin sections. In wild type, at stages 6 and 7, microspore mother cells undergo meiosis to form tetrads (Sanders et al., 1999), as image 3 Shown in A-B. Subsequently, microspores were released from the tetrad and gradually formed trinucleate pollen grains with normal pollen walls ( image 3 C-D, M-O). In the acos5 mutant at normal temperature (24°C), no visible difference was observed between the mutant and the wild type until the seventh stage of anther development, which indicated that meiosis of the mutant male gametophyte was not affected ( image 3 F). By the eighth stage of anther development, acos5 microspores were released from the tetrad, and some microspores began to vacuolate ( image 3 G). At the 10th stage, most of the acos5 microspores began to degrad...
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