Applications of pollen development-related ABC transport protein or coding gene of pollen development-related ABC transport protein, method used for culturing plant sterile lines, and plant breeding method
A technology for transporter and pollen development, applied in agriculture and biological fields, can solve the problems of few temperature-sensitive genetic loci of nuclear sterility, low purity of seed production and reduction
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Embodiment 1
[0135] Example 1 The sterility phenotype of the Arabidopsis abcg26 mutant is restored under low temperature conditions
[0136] The inventors isolated the abcg26-1 and abcg26-2 mutants from the T-DNA insertion mutant library of the Arabidopsis Col ecotype purchased from the ABRC center, such as figure 1 Shown in a. At normal ambient temperature (24°C), the homozygous abcg26 mutant had normal vegetative growth, but lost fertility, and had only short, seedless pods, such as figure 1 Shown in c and d. Genetic analysis showed that the abcg26 mutant belonged to sporophytic male sterility and was controlled by a single recessive locus. The applicant cultured the abcg26-1 and abcg26-2 mutants at 24°C until bolting, then moved them to 18°C for continuous culture, and the subsequent fruit pods all recovered their fertility, and then put the mutants back to normal ambient temperature Under the condition of figure 1 Shown in e and f. Under the same low temperature conditions, wild...
Embodiment 2
[0137] Example 2 Low temperature can compensate for the microspore development defect after release from the tetrad in the abcg26 mutant
[0138] To determine the defect in pollen development of the abcg26 mutant, the inventors performed semi-thin sections of anthers. In wild-type stages 6 and 7, microspore mother cells undergo meiosis to form tetrads (Sanders et al., 1999). Subsequently, microspores are released from the tetrad and gradually form normally fertile pollen as image 3 Shown in a-g. In the abcg26 (24°C) mutant at normal temperature (24°C), no visible difference was observed between the mutant and the wild type until stage 7 of anther development, suggesting that meiosis of the mutant male gametophytes is not affected as in image 3 Shown in middle h-i; by the 8th stage of anther development, abcg26 (24°C) microspores were released from the tetrad, showing an irregular swelling phenotype compared with the wild type; at the 9th stage, most of abcg26 (24°C) Micro...
Embodiment 3
[0140] Example 3 ABCG26 gene and other expressions are not induced by low temperature
[0141] In order to elucidate the mechanism of the temperature sensitivity of the abcg26 mutant, the applicant detected the ABCG26 transcription level in wild-type and mutant flower buds under different temperature conditions. Quantitative PCR detection showed that there was no significant difference in the transcription level between the mutant and wild-type ABCG26 under normal temperature (24°C) and low temperature (18°C) conditions, such as Figure 5 . In order to detect whether there is a compensation mechanism for the co-expressed genes, the applicant extracted the total RNA of the inflorescences of the wild type and mutants, and detected the transcription level of the co-expressed genes. The results of semi-quantitative RT-PCR showed that there was no change in expression at low temperature, such as Figure 5 . At the same time, in order to identify whether there is a compensation me...
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