Cucumber CsGL2-LIKE gene and application thereof in regulating partial abortion of male flowers
A cucumber, genetic technology, applied in the direction of application, genetic engineering, plant genetic improvement, etc.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0034] Example 1 Cloning of gene CsGL2-LIKE related to partial abortion of cucumber male flowers
[0035] 1. Materials
[0036] Cucumber 3461 (the variety selected in our laboratory) (Liu et al., 2015; Silencing of the gibberellin receptor homolog, CsGID1a, affects locule formation in cucumber (Cucumis sativus) fruit. NEW PHYTOLOGIST, 210, 551-563.) was used as the test material , the breed belongs to the hermaphroditic line. The cucumber seeds are sown in the solar greenhouse, and the field management is the same as the normal cultivation management. When the male flowers appeared on the plants, the male flower buds were quick-frozen in liquid nitrogen and stored in a -80°C refrigerator for future use.
[0037] 2. Total RNA extraction and cDNA synthesis
[0038] (1) Extraction of total RNA
[0039] The total RNA of cucumber male flower buds was extracted with a plant total RNA extraction kit (purchased from Beijing Huayueyang Biotechnology Co., Ltd.), and the residual DNA...
Embodiment 2
[0067] Embodiment 2, the acquisition of transgenic plants and its detection
[0068] 1. Genetic transformation of cucumber
[0069] (1) Transformation of Agrobacterium with the recombinant expression vector prepared in Example 1: Turn on the Biorad electroporator to preheat, and at the same time place the electroporation cup on ice to precool, and put it into an ultra-clean workbench for ultraviolet sterilization for 30 minutes to sterilize. Add the constructed CsGL2-LIKE-pCAMBIA1008 plasmid (interference expression vector constructed for Example 1 step 5) and pCAMBIA1008 empty plasmid 500ng into 100 μl of C58 Agrobacterium competent cells respectively, flick and mix well (Agrobacterium competent Place the cells on ice in advance to thaw) Add the above-mentioned mixed competent state to the bottom of the electroporation cup, then close the lid and place it on the seat of the electroporation apparatus for electric shock. After the end, add 300ul of liquid YEB medium without an...
Embodiment 3
[0094] Embodiment 3, phenotypic identification of transgenic plants
[0095] Eight transgenic lines were obtained by constructing CsGL2-LIKE RNAi interference vectors to transform cucumbers, among which the lines with more serious interference of CsGL2-LIKE RNAi expression were #5, #7, #21 and #25 (see Figure 4 ). Through careful observation of the transgenic plants, it was found that the fruit thorn density and size of the transgenic plants had no change compared with the wild type (see Figure 5 ). However, in the development of male flowers, there are significant differences between the transgenic lines and the wild-type lines, as follows: (1) The male flower opening time of the interference lines is significantly delayed compared with that of the wild type (see Figure 6A ), the internal structure is not fully developed compared with the wild type (see Figure 6B ); the fresh weight of the male flower was about 50% of the wild type on the day of opening (see Figure 6...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com