Application of abc transport protein related to pollen development or its encoding gene, method for cultivating plant sterile line and plant breeding method
A technology for pollen development and transporter, which can be used in agriculture and biology, and can solve the problems of lowering, fewer temperature-sensitive genetic sites of nuclear sterility, and low purity of seed production.
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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 inventor has isolated the abcg26-1 and abcg26-2 mutants in 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 half-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 up to stage 7 of anther development, suggesting that meiosis of the mutant male gametophyte is not affected as 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) Microspor...
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 transcript 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 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 inflorescence of the wild type and the mutant, and carried out the detection of 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, as Figure 5 . At the same time, in order to identify whether there is a compe...
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