Application of osgsl5 protein in controlling plant fertility
A plant and fertility technology, applied in the field of OsGSL5 protein in the control of plant fertility, can solve the problems of microspore development process interference, few reports on the molecular regulation mechanism of callose degradation, and abnormal deposition of the outer wall
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Embodiment 1
[0040] Example 1, OsGSL5 gene mutant gsl5-1 acquisition and phenotype
[0041] 1. Obtaining of gsl5-1 mutant
[0042] 1. Obtaining the gsl5-1 mutant
[0043] The rice T-DNA insertion mutant library used was constructed by the Army Biotechnology Research Institute of Chinese Agricultural Sciences using the vector pFX-E24.2-15R (the method for creating the mutant library has been published in a paper, see Wan et al., Activation tagging, an efficient tool for functional analysis of the rice genome. Plant Mol Biol. 2009 Jan; 69(1-2):69-80).
[0044] 10,000 seeds of the transgenic rice variety "Nihonbare (a sequenced japonica variety maintained by the Biotechnology of the Chinese Academy of Agricultural Sciences)" were selected, soaked and accelerated germination, sowed in the seedling field, and transplanted to the field 35 days later. Two rows of each material, 10 plants per row, constitute a family. The planting location is the experimental field in the high-tech industrial pa...
Embodiment 2
[0056] Embodiment 2, the application of the substance that suppresses the expression of OsGSL5 in cultivating rice sterile material
[0057] 1. Preparation of DNA molecules
[0058] Primers were designed according to the CDS sequence (sequence 1) of the OsGSL5 gene as follows: 5'-GCT TAT CTG AAA TCA TCTTGT CTC TC-3' and 5'-CCA AGA TGC GGG AGA TCT G-3'.
[0059] Total RNA was extracted from rice Nipponbare seedlings, and cDNA was synthesized from RNA using reverse transcriptase. Using the cDNA obtained by reverse transcription as a template, PCR amplification was performed with the above primers, and the PCR product was detected by 0.8% agarose gel electrophoresis, and a band with a molecular weight of about 540 bp was obtained. This fragment was recovered using an agarose gel recovery kit (TIANGEN). The recovery fragment was connected with pMD19-TSimple (Takara), the connection product was transformed into Escherichia coli DH5α competent cells, positive clones were screened ...
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