Efficient transformation of the isolated microspores of Chinese cabbage (Brassica rapa L. ssp. pekinensis) by particle bombardment
Abstract Background The low efficiency of genetic transformation in Chinese cabbage (Brassica rapa L. ssp. pekinensis) is the key problem affecting functional verification. Particle bombardment is a widely used method along with the Agrobacterium-mediated method. As a physical means, it has almost n...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2024-01-01
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| Series: | Plant Methods |
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| Online Access: | https://doi.org/10.1186/s13007-024-01134-1 |
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| author | Yujia Liu Shujiang Zhang Shifan Zhang Hui Zhang Guoliang Li Rifei Sun Fei Li |
| author_facet | Yujia Liu Shujiang Zhang Shifan Zhang Hui Zhang Guoliang Li Rifei Sun Fei Li |
| author_sort | Yujia Liu |
| collection | DOAJ |
| description | Abstract Background The low efficiency of genetic transformation in Chinese cabbage (Brassica rapa L. ssp. pekinensis) is the key problem affecting functional verification. Particle bombardment is a widely used method along with the Agrobacterium-mediated method. As a physical means, it has almost no restrictions on the type of host and a wide range of receptor types, which largely avoids the restriction of explants. The bombardment parameters, which include the number of bombardments, the bombardment pressure, and the bombardment distance, may affect the microspores' genetic transformation efficiency. Results The transformation efficiency was improved using the particle bombardment method under the combination of bombardment shot times (3, 4, 5) × bombardment pressure (900, 1100, 1350 psi) × bombardment distance (3, 6, 9 cm). The average viability of microspores in the treatment group ranged from 74.76 to 88.55%, while the control group was 88.09%. When the number of shot times was 4, the number of embryos incubated in the treatment group ranged from 16 to 236 per dish, and the control group had 117 embryos per dish. When the bombardment parameters of the biolistic method were 4 shot times—1350 psi—3 cm, 4 times—1100 psi—3 cm, and 4 times—900 psi—3 cm, they had high transient expression efficiency, and the average number of transformed microspores was 21.67, 11.67, and 11.67 per dish (3.5 mL), respectively. When the bombardment parameters were 4 times, 900 psi, and 6 cm, the highest genetically transformed embryos were obtained, and the transformation efficiency reached 10.82%. Conclusion A new genetic transformation system with proper parameters for Chinese cabbage microspores was established using particle bombardment. This proper transformation system could provide a useful tool for the improvement of cultivar quality and the investigation of functional genes in Chinese cabbage. |
| first_indexed | 2024-03-07T14:59:30Z |
| format | Article |
| id | doaj.art-7ca2f25558de44f7bc574d56e65c202e |
| institution | Directory Open Access Journal |
| issn | 1746-4811 |
| language | English |
| last_indexed | 2024-03-07T14:59:30Z |
| publishDate | 2024-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Plant Methods |
| spelling | doaj.art-7ca2f25558de44f7bc574d56e65c202e2024-03-05T19:15:17ZengBMCPlant Methods1746-48112024-01-0120111510.1186/s13007-024-01134-1Efficient transformation of the isolated microspores of Chinese cabbage (Brassica rapa L. ssp. pekinensis) by particle bombardmentYujia Liu0Shujiang Zhang1Shifan Zhang2Hui Zhang3Guoliang Li4Rifei Sun5Fei Li6Institute of Vegetables and Flowers, Chinese Academy of Agricultural SciencesInstitute of Vegetables and Flowers, Chinese Academy of Agricultural SciencesInstitute of Vegetables and Flowers, Chinese Academy of Agricultural SciencesInstitute of Vegetables and Flowers, Chinese Academy of Agricultural SciencesInstitute of Vegetables and Flowers, Chinese Academy of Agricultural SciencesInstitute of Vegetables and Flowers, Chinese Academy of Agricultural SciencesInstitute of Vegetables and Flowers, Chinese Academy of Agricultural SciencesAbstract Background The low efficiency of genetic transformation in Chinese cabbage (Brassica rapa L. ssp. pekinensis) is the key problem affecting functional verification. Particle bombardment is a widely used method along with the Agrobacterium-mediated method. As a physical means, it has almost no restrictions on the type of host and a wide range of receptor types, which largely avoids the restriction of explants. The bombardment parameters, which include the number of bombardments, the bombardment pressure, and the bombardment distance, may affect the microspores' genetic transformation efficiency. Results The transformation efficiency was improved using the particle bombardment method under the combination of bombardment shot times (3, 4, 5) × bombardment pressure (900, 1100, 1350 psi) × bombardment distance (3, 6, 9 cm). The average viability of microspores in the treatment group ranged from 74.76 to 88.55%, while the control group was 88.09%. When the number of shot times was 4, the number of embryos incubated in the treatment group ranged from 16 to 236 per dish, and the control group had 117 embryos per dish. When the bombardment parameters of the biolistic method were 4 shot times—1350 psi—3 cm, 4 times—1100 psi—3 cm, and 4 times—900 psi—3 cm, they had high transient expression efficiency, and the average number of transformed microspores was 21.67, 11.67, and 11.67 per dish (3.5 mL), respectively. When the bombardment parameters were 4 times, 900 psi, and 6 cm, the highest genetically transformed embryos were obtained, and the transformation efficiency reached 10.82%. Conclusion A new genetic transformation system with proper parameters for Chinese cabbage microspores was established using particle bombardment. This proper transformation system could provide a useful tool for the improvement of cultivar quality and the investigation of functional genes in Chinese cabbage.https://doi.org/10.1186/s13007-024-01134-1Chinese cabbageMicrosporeParticle bombardmentTransformation |
| spellingShingle | Yujia Liu Shujiang Zhang Shifan Zhang Hui Zhang Guoliang Li Rifei Sun Fei Li Efficient transformation of the isolated microspores of Chinese cabbage (Brassica rapa L. ssp. pekinensis) by particle bombardment Plant Methods Chinese cabbage Microspore Particle bombardment Transformation |
| title | Efficient transformation of the isolated microspores of Chinese cabbage (Brassica rapa L. ssp. pekinensis) by particle bombardment |
| title_full | Efficient transformation of the isolated microspores of Chinese cabbage (Brassica rapa L. ssp. pekinensis) by particle bombardment |
| title_fullStr | Efficient transformation of the isolated microspores of Chinese cabbage (Brassica rapa L. ssp. pekinensis) by particle bombardment |
| title_full_unstemmed | Efficient transformation of the isolated microspores of Chinese cabbage (Brassica rapa L. ssp. pekinensis) by particle bombardment |
| title_short | Efficient transformation of the isolated microspores of Chinese cabbage (Brassica rapa L. ssp. pekinensis) by particle bombardment |
| title_sort | efficient transformation of the isolated microspores of chinese cabbage brassica rapa l ssp pekinensis by particle bombardment |
| topic | Chinese cabbage Microspore Particle bombardment Transformation |
| url | https://doi.org/10.1186/s13007-024-01134-1 |
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