CN117721143A - Gene editing method - Google Patents
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- CN117721143A CN117721143A CN202311750953.5A CN202311750953A CN117721143A CN 117721143 A CN117721143 A CN 117721143A CN 202311750953 A CN202311750953 A CN 202311750953A CN 117721143 A CN117721143 A CN 117721143A
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Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to a gene editing method, belonging to the technical field of genetic engineering and transgenosis. Specifically, the invention discovers that the callus induced by the anther culture of rice is infected by agrobacterium carrying the gene editing vector, and has higher gene editing efficiency than the callus induced by rice seed infection.
Description
Technical Field
The invention belongs to the technical field of genetic engineering and transgenosis, and particularly relates to a gene editing method.
Background
The gene editing technology can carry out accurate genetic operation on the genome of a species, and is an emerging technical means for genetic improvement of crops in recent years. Delivery of the editing system is an important factor affecting gene editing efficiency, and common delivery means in plants include particle bombardment, agrobacterium-mediated, polyethylene glycol (PEG) -mediated, liposome-mediated, microinjection, etc., with agrobacterium-mediated still being the dominant means.
Plant material successfully edited at the target site can be obtained by infecting plant callus with agrobacterium containing an editing system and culturing positive callus into a complete plant. In rice, seeds are often used to induce callus, followed by subsequent processes of agrobacterium infection, co-cultivation, callus screening, differentiation and rooting into seedlings. Although the use of Agrobacterium to infect rice seed-induced callus has become the dominant means for breeding transgenic, genetically edited rice and has enabled higher editing efficiency, more efficient techniques remain desirable.
The rice anther culture is to use tissue culture technology, inoculate the rice anther developed to a certain stage on an artificial culture medium by adopting aseptic technique, induce the dedifferentiation of the anther by changing the development program of pollen grains in the anther, continuously carry out mitosis to form cell clusters, further form a cluster of undifferentiated parenchyma-callus, and then make the callus dedifferentiate into embryoid or differentiate into complete rice plants. Rice anthers are commonly used to cultivate conventional rice varieties, but due to the relatively complex operational procedures and low callus induction efficiency, no one has been used for agrobacterium-mediated genetic transformation.
Disclosure of Invention
In order to solve the problems, the invention tests the editing efficiency of two gene editing modes, namely an agrobacterium-mediated seed callus path and an anther callus path, and unexpectedly discovers that the editing efficiency of the anther callus path is higher.
In order to achieve the above purpose, the invention adopts the following technical scheme:
the invention provides a rice gene editing method which is characterized in that agrobacterium carrying a gene editing vector is used for infecting callus induced by rice anther culture, and successfully infecting callus is cultivated into rice plants.
In some embodiments, the T-DNA region of the above gene editing vector consists of a hygromycin resistance gene expression cassette, a Cas9 gene expression cassette, two gRNA expression cassettes, and the structure of the T-DNA region is shown in fig. 1B.
In some embodiments, the two grnas described above may bind to rice chlorophyll synthesis gene Cao1 (see SEQ ID No.1 for genomic sequence), or gibberellin degradation gene Eui (see SEQ ID No.2 for genomic sequence).
In some embodiments, the successfully infected calli described above are obtained by hygromycin antibiotic screening.
In some embodiments, the rice is a middle flower 11 of the japonica variety.
In some embodiments, the agrobacterium strain described above is EHA105.
The invention also provides application of the method in rice gene editing.
The invention has the advantages and beneficial effects as follows: the invention tests the editing efficiency of two gene editing modes, namely the seed callus path and the anther callus path mediated by agrobacterium, and unexpectedly discovers that the editing efficiency of the anther callus path is higher. This helps to provide a new operational procedure for gene editing of rice.
Drawings
Fig. 1 edits the structure of the carrier.
FIG. 2 genetic transformation scheme for anther-induced callus.
FIG. 3 phenotype of different editors of Eui gene.
FIG. 4 phenotype of different editors of Cao1 gene.
Detailed Description
The following definitions and methods are provided to better define the present application and to guide those of ordinary skill in the art in the practice of the present application. Unless otherwise indicated, terms are to be construed according to conventional usage by those of ordinary skill in the relevant art. All patent documents, academic papers, industry standards, and other publications cited herein are incorporated by reference in their entirety.
As used herein, "rice" is any rice plant and includes all plant varieties that can be bred with rice, including whole plants, plant cells, plant organs, plant protoplasts, plant cell tissue cultures from which plants can regenerate, plant callus, whole plant cells in plants or plant parts such as embryos, pollen, ovules, seeds, leaves, flowers, branches, fruits, roots, root tips, anthers, and the like. Unless otherwise indicated, nucleic acids are written in the 5 'to 3' direction from left to right; the amino acid sequence is written in the amino to carboxyl direction from left to right. Amino acids may be represented herein by their commonly known three-letter symbols or by the single-letter symbols recommended by the IUPAC-IUB biochemical nomenclature committee. Likewise, nucleotides may be referred to by commonly accepted single letter codes. The numerical range includes the numbers defining the range.
The following examples are illustrative of the invention and are not intended to limit the scope of the invention. Modifications and substitutions to methods, procedures, or conditions of the present invention without departing from the spirit and nature of the invention are intended to be within the scope of the present application. Examples follow conventional experimental conditions, such as the molecular cloning laboratory manual of Sambrook et al (Sambrook J & Russell D W, molecular cloning: alaboratory manual, 2001), or conditions recommended by the manufacturer's instructions, unless otherwise indicated. Unless otherwise indicated, all chemical reagents used in the examples were conventional commercial reagents, and the technical means used in the examples were conventional means well known to those skilled in the art.
Examples
EXAMPLE 1 vector construction
The inventors selected chlorophyll synthesis gene Cao1 (genomic sequence see SEQ ID NO. 1) and gibberellin degradation gene Eui (genomic sequence see SEQ ID NO. 2) as target editing genes to test editing efficiency. The Cao1 and Eui gene structures and engineered target information are shown in FIG. 1A. 7 targets (T1-T7) are respectively designed for each gene, and T1+T2, T1+T3, T1+T4, T1+T5, T1+T6 and T1+T7 of each gene are used for combination to create gene editing materials with different size fragments knocked out, wherein Cao1 is designed to knock out 98bp, 248bp, 554bp, 1046bp, 1460bp and 2120bp respectively; eui intends to knock out 115bp, 244bp, 565bp, 1010bp, 1554bp, 2013bp, respectively. The dual-target vector was used to express sgrnas that bind to both targets (vector structure as in fig. 1B), yielding a total of 12 editing vectors.
The gene editing vector is obtained using conventional construction methods (e.g., the methods disclosed in CN110093349a patent), and the constructed editing vector is transferred to agrobacterium strain EHA105 for storage.
EXAMPLE 2 genetic transformation
The seeds of flower 11 and anther-induced callus in rice varieties were used, respectively, for genetic transformation.
Among them, procedures for inducing callus and agrobacterium-mediated transformation using seeds are described with reference to procedures conventional in the art (e.g., chen Qiugong, chen Tai, lin Yongjun, chen Hao. (2018.) agrobacterium-mediated genetic transformation of japonica rice. Bio-101e 1010174.).
The flow of anther-induced callus and agrobacterium-mediated transformation is shown in fig. 2, and the specific steps are as follows:
1) Sampling
And (3) observing external characteristics and checking by an iodine-potassium iodide staining method in a microscopic examination way, selecting young spikes of anther development in the middle and later stages of the seventh stage of young spike differentiation (the leaf pillow distance between a sword leaf and the next leaf is about 5 cm-10 cm) in the middle and late stages of single-core (single-core approach period), wherein most glumes are close to the mature size, and the glumes are light green, and the stamens extend to 1/3-1/2 of glumes.
2) Pretreatment of
Wiping and sterilizing the surface of the rice ears She Daosui with buds by using 75% alcohol, wrapping the rice ears with wet gauze, putting the rice ears into a fresh-keeping bag, attaching a label, marking information such as material names and sampling time, and putting the rice ears in a refrigerator for low-temperature pretreatment. The treatment is generally carried out at 8 ℃ for 8 to 12 days.
3) Disinfection
The pretreated rice ears are subjected to bract stripping, the middle rice ears are sheared, soaked in 0.1% mercuric chloride solution for 10min or soaked in newly prepared 3% sodium hypochlorite solution for 25min, then washed for 4-5 times by sterile water, and the water is sucked by sterile filter paper.
4) Inoculating anther
On a sterile operating table, cutting off 1/5 of the glume base, clamping the glume by forceps, knocking down the cut opening at the triangular bottleneck, shaking off anthers onto an induction culture medium, and inoculating 80-100 anthers in each 100mL triangular flask. The anther is not cut or bruised when the anther is inoculated.
5) Inducing callus
And (3) placing the inoculated material into an incubator at 28 ℃ for dark culture until the diameter of the callus is 2-3 mm.
After the callus induction is completed, the steps of callus subculture, agrobacterium infection, co-culture, screening, differentiation, rooting and seedling hardening which are the same as the genetic transformation of the seed-induced callus are adopted.
The preparation method of the related culture medium comprises the following steps:
preparation of induction medium (1L of medium was prepared for example): weighing 3g of plant gel, placing in a pot, adding 800mL of distilled water, and boiling until the gel is completely melted and has no foam; boiling the gel, weighing 50mL of each N6 macroelement I, II, 10mL of each N6 microelement III, 10mL of each ferric salt IV and N6 organic matter V in a beaker, adding 1mg/L KT 1mL, 1mg/L NAA 3mL and 1 mg/L2, 4-D2 mL, mixing uniformly, and adding 600mg of proline in a mother solution mixed solution to dissolve; after the gel is completely melted, 50g of sucrose is added for dissolution, then the mother liquor mixed solution is mixed with the gel, and distilled water is added for volume fixation to 1L. Mixing must be accomplished while stirring to prevent local solidification of the medium. After mixing evenly, the pH value of the culture medium is adjusted to 5.8 by using a precise pH test paper. Subpackaging the culture medium into 100mL triangular bottles while the culture medium is hot, filling 30-35 mL of the culture medium into each bottle, sealing the bottles, sterilizing the bottles by high-pressure steam, and cooling the bottles for later use.
Preparation of differentiation Medium (1L preparation of Medium is taken as an example): weighing 3g of plant gel, placing in a pot, adding 800mL of distilled water, and boiling until the gel is completely melted and has no foam; measuring 50mL of each MS macroelement I, II, 10mL of each MS microelement III, ferric salt IV and MS organic matter V in a beaker while boiling the gel, and then adding 0.5mL of 1 mg/L6-BA, 2mL of 1mg/L KT and 0.5mL of 1mg/L NAA for uniform mixing; after the gel is completely melted, 30g of sucrose is added for dissolution, then the mother liquor mixed solution is mixed with the gel, and distilled water is added for volume fixation to 1L. Mixing must be accomplished while stirring to prevent local solidification of the medium. After mixing evenly, the pH value of the culture medium is adjusted to 5.8 by using a precise pH test paper. Subpackaging the culture medium into 100mL triangular bottles while the culture medium is hot, filling 30-35 mL of the culture medium into each bottle, sealing the bottles, sterilizing the bottles by high-pressure steam, and cooling the bottles for later use.
Preparation of rooting medium (1L of preparation medium is taken as an example): weighing 3g of plant gel, placing in a pot, adding 800mL of distilled water, and boiling until the gel is completely melted and has no foam; measuring 25mL of each MS macroelement I, II, 5mL of each MS microelement III, 5mL of each ferric salt IV and 10mL of MS organic matter V in a beaker while boiling the gel, and then adding 1mg/L IBA and mixing uniformly; after the gel is completely melted, 30g of sucrose is added, the mother liquor mixture is mixed with the gel, and distilled water is added to fix the volume to 1L. Mixing must be accomplished while stirring to prevent local solidification of the medium. After mixing evenly, the pH value of the culture medium is adjusted to 5.8 by using a precise pH test paper. Adding 0.5g of active carbon powder into the culture medium, stirring uniformly, subpackaging the culture medium into 100mL triangular bottles while the culture medium is hot, packaging 30 mL-35 mL of the culture medium into each bottle, sealing, sterilizing by high-pressure steam, and cooling for later use.
The preparation method of the culture medium mother solution and the plant growth substance comprises the following steps:
the mother solution of the culture medium is prepared into a composition table (unit: mg. L-1)
Commonly used plant growth substances
Example 3 comparison of editing effects
Target gene editing conditions of T0 generation transformed seedlings obtained by detecting agrobacterium infection seed induced callus and anther induced callus are shown in table 1. The ratio of homozygous fragment deletion plants obtained by anther-induced callus is higher than that of seed-induced callus by gene editing genetic operation of knocking out two target genes with different fragment sizes. This indicates that the genetic editing operations performed in anther-induced calli are more efficient than the traditional operations performed in seed-induced calli.
TABLE 1 comparison of the efficiency of genetic manipulation for Gene editing in seed and anther-induced calli
The phenotypes of the different editors of the Eui gene and the Cao1 gene are shown in fig. 3 and 4.
While the invention has been described in detail in the foregoing general description and with reference to specific embodiments thereof, it will be apparent to one skilled in the art that modifications and improvements can be made thereto. Accordingly, such modifications or improvements may be made without departing from the spirit of the invention and are intended to be within the scope of the invention as claimed.
Claims (6)
1. A method for editing rice gene is characterized in that agrobacterium carrying a gene editing vector is used for infecting callus induced by rice anther culture, and successfully infecting callus is cultivated into rice plants.
2. The method of claim 1, wherein the T-DNA region of the gene editing vector consists of a hygromycin resistance gene expression cassette, a Cas9 gene expression cassette, and two gRNA expression cassettes.
3. The method of claim 1, wherein the successfully infected callus is obtained by hygromycin antibiotic screening.
4. The method according to claim 1, wherein the rice is japonica rice variety middle flower 11.
5. The method of claim 1, wherein the agrobacterium strain is EHA105.
6. Use of the method according to any one of claims 1-6 in gene editing in rice.
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Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2347675A1 (en) * | 1998-11-03 | 2000-05-11 | Syngenta Participations Ag | Dna comprising rice anther-specific gene and transgenic plant transformed therewith |
| KR20000031106A (en) * | 1998-11-03 | 2000-06-05 | 정명식 | Transfected rice with vector containing anther specific gene and anther specific expression regulator |
| CN1712537A (en) * | 2004-06-24 | 2005-12-28 | 华中农业大学 | Culture of indica rice tran-gene tree from agrobacterium medium |
| KR20110026054A (en) * | 2009-09-07 | 2011-03-15 | 대한민국(농촌진흥청장) | Method for short term production of elite transgenic rice lines by using anther culture and backcross |
| CN103461141A (en) * | 2013-09-27 | 2013-12-25 | 江苏丘陵地区镇江农业科学研究所 | Method for improving culturing efficiency of japonica rice anther |
| CN103740635A (en) * | 2013-12-24 | 2014-04-23 | 北京大北农科技集团股份有限公司 | Method for transforming rice |
| CN104711262A (en) * | 2015-04-13 | 2015-06-17 | 安徽农业大学 | Corn anther specific promoter and application thereof |
| CN105505979A (en) * | 2015-11-28 | 2016-04-20 | 湖北大学 | Method for acquiring aromatic rice strain by targeting Badh2 gene via CRISPR/Cas9 gene editing technology |
| CN105647925A (en) * | 2016-03-25 | 2016-06-08 | 安徽省农业科学院水稻研究所 | Rice anther high-expression promoter OsAnth4 and application thereof |
| CN108396035A (en) * | 2017-12-29 | 2018-08-14 | 青岛袁策生物科技有限公司 | A kind of agriculture bacillus mediated rice transformation method |
| CN112385544A (en) * | 2020-12-04 | 2021-02-23 | 黑龙江省农业科学院绥化分院 | Hybrid rice anther culture method |
-
2023
- 2023-12-19 CN CN202311750953.5A patent/CN117721143A/en active Pending
Patent Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2347675A1 (en) * | 1998-11-03 | 2000-05-11 | Syngenta Participations Ag | Dna comprising rice anther-specific gene and transgenic plant transformed therewith |
| KR20000031106A (en) * | 1998-11-03 | 2000-06-05 | 정명식 | Transfected rice with vector containing anther specific gene and anther specific expression regulator |
| CN1712537A (en) * | 2004-06-24 | 2005-12-28 | 华中农业大学 | Culture of indica rice tran-gene tree from agrobacterium medium |
| KR20110026054A (en) * | 2009-09-07 | 2011-03-15 | 대한민국(농촌진흥청장) | Method for short term production of elite transgenic rice lines by using anther culture and backcross |
| CN103461141A (en) * | 2013-09-27 | 2013-12-25 | 江苏丘陵地区镇江农业科学研究所 | Method for improving culturing efficiency of japonica rice anther |
| CN103740635A (en) * | 2013-12-24 | 2014-04-23 | 北京大北农科技集团股份有限公司 | Method for transforming rice |
| CN104711262A (en) * | 2015-04-13 | 2015-06-17 | 安徽农业大学 | Corn anther specific promoter and application thereof |
| CN105505979A (en) * | 2015-11-28 | 2016-04-20 | 湖北大学 | Method for acquiring aromatic rice strain by targeting Badh2 gene via CRISPR/Cas9 gene editing technology |
| CN105647925A (en) * | 2016-03-25 | 2016-06-08 | 安徽省农业科学院水稻研究所 | Rice anther high-expression promoter OsAnth4 and application thereof |
| CN108396035A (en) * | 2017-12-29 | 2018-08-14 | 青岛袁策生物科技有限公司 | A kind of agriculture bacillus mediated rice transformation method |
| CN112385544A (en) * | 2020-12-04 | 2021-02-23 | 黑龙江省农业科学院绥化分院 | Hybrid rice anther culture method |
Non-Patent Citations (6)
| Title |
|---|
| 刘清;萧浪涛;吴顺;朱允华;刘素纯;史齐;康朵兰;: "农杆菌介导法将反义蜡质基因转入杂交稻亲本", 分子植物育种, no. 04, 28 July 2006 (2006-07-28) * |
| 李三和;韩光明;闸雯俊;刘凯;胡刚;游艾青;: "根癌农杆菌介导水稻成熟胚及抗褐飞虱基因植株的获得", 湖北农业科学, no. 09, 10 May 2016 (2016-05-10) * |
| 王彩芬;付永彩;安永平;韩国敏;张文银;马静;: "宁夏水稻农杆菌转化组培体系的建立", 宁夏农林科技, no. 03, 10 June 2008 (2008-06-10) * |
| 蒋苏, 陈彩艳, 程祝宽, 蔡润, 翟文学, 朱立煌: "用花药愈伤组织作为转化受体的水稻转基因植株的分析", 遗传学报, no. 12, 10 December 2004 (2004-12-10) * |
| 陈彩艳;肖晗;张文利;王爱菊;夏志辉;李晓兵;翟文学;程祝宽;朱立煌;: "以花药愈伤组织为受体的水稻转化和RNA干扰研究", 中国科学(C辑:生命科学), no. 04, 20 August 2006 (2006-08-20) * |
| 韩光明;李三和;陈志军;游艾青;陈温福;: "根癌农杆菌介导的高效水稻遗传转化影响因素研究进展", 沈阳农业大学学报, no. 06, 15 December 2012 (2012-12-15) * |
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