Title:
IMPROVED CORN POLLEN MAGNETOFECTION METHOD
Document Type and Number:
WIPO Patent Application WO/2023/093348
Kind Code:
A1
Abstract:
Disclosed in the present invention is an improved corn pollen magnetofection method, comprising the following steps: (1) preparing plasmid DNA for transfection; (2) combining nanomagnetic beads MNP with the plasmid DNA under a room temperature condition to form an MNP-DNA compound; (3) collecting fresh pollen of corn in a blooming stage, and quickly taking the pollen back to a room while preserving in an ice box in a water-proof manner; (4) uniformly mixing a corn pollen transformation liquid with sieved corn pollen, and performing trepanning pretreatment under a low-temperature condition; (5) adding the MNP-DNA compound into a trepanning pretreatment solution, gently and uniformly mixing, and placing on a low-temperature pre-cooled magnetic plate to transfect; and (6) after the completion of transfection, taking a pollen suspension to a field by using an ice box, and directly pollinating corn ears. According to the method of the present invention, the corn planted in the field is directly used to transform, only a conventional refrigeration device is needed, operation is simple and convenient, the cost is low, and the method is suitable for all corn varieties.
Inventors:
ZHANG ZHONGBAO (CN)
WU ZHONGYI (CN)
ZHENG DENGYU (CN)
WANG ZUOPING (CN)
ZHANG CHUN (CN)
LI XIANGLONG (CN)
WU ZHONGYI (CN)
ZHENG DENGYU (CN)
WANG ZUOPING (CN)
ZHANG CHUN (CN)
LI XIANGLONG (CN)
Application Number:
PCT/CN2022/125244
Publication Date:
June 01, 2023
Filing Date:
October 14, 2022
Export Citation:
Assignee:
BEIJING ACAD AGRIC & FORESTRY (CN)
International Classes:
C12N15/82; A01H5/02; A01H6/46
Foreign References:
| CN114085866A | 2022-02-25 | |||
| CN110331167A | 2019-10-15 | |||
| US20200354732A1 | 2020-11-12 | |||
| US20030145346A1 | 2003-07-31 |
Other References:
JIA, YANJING ET AL.: "Preliminary Analysis on Transgenic Technology of Pollen-Mediated by Magnetic Nanoparticles in Tree Peony", MOLECULAR PLANT BREEDING, vol. 18, no. 8, 31 August 2020 (2020-08-31), XP009546695, ISSN: 1672-416X
ZHAO XIANG; MENG ZHIGANG; WANG YAN; CHEN WENJIE; SUN CHANGJIAO; CUI BO; CUI JINHUI; YU MANLI; ZENG ZHANGHUA; GUO SANDUI; LUO DAN; : "Pollen magnetofection for genetic modification with magnetic nanoparticles as gene carriers", NATURE PLANTS, NATURE PUBLISHING GROUP UK, LONDON, vol. 3, no. 12, 27 November 2017 (2017-11-27), London , pages 956 - 964, XP036428622, DOI: 10.1038/s41477-017-0063-z
ZHAO XIANG; MENG ZHIGANG; WANG YAN; CHEN WENJIE; SUN CHANGJIAO; CUI BO; CUI JINHUI; YU MANLI; ZENG ZHANGHUA; GUO SANDUI; LUO DAN; : "Pollen magnetofection for genetic modification with magnetic nanoparticles as gene carriers", NATURE PLANTS, NATURE PUBLISHING GROUP UK, LONDON, vol. 3, no. 12, 27 November 2017 (2017-11-27), London , pages 956 - 964, XP036428622, DOI: 10.1038/s41477-017-0063-z
Attorney, Agent or Firm:
BEIJING HENGLV IP FIRM (CN)
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