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Application of ga3ox1 protein in regulation of alfalfa plant type
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A technology of alfalfa and plant type, applied in the biological field, can solve the problems of limiting the effectiveness and convenience of agricultural activities, plant height, etc.
Active Publication Date: 2022-04-19
CHINA AGRI UNIV
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However, most alfalfa cultivars are erect and have a high plant height, which limits the effectiveness and convenience of the above agricultural activities to a large extent; while the prostrate trait is more conducive to interplanting while maintaining other production advantages , intercropping application
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Embodiment 1
[0122] Example 1. Obtaining and Phenotypic Detection of Medicago truncatula MtGA3ox1 Gene Knockout Mutant
[0125] The coding sequence (CDS) of Medicago truncatula R108 is the MtGA3ox1 coding gene shown in sequence 4 in the sequence listing, which encodes the protein MtGA3ox1 whose amino acid sequence is shown in sequence 2 in the sequence listing.
[0126] Through the online target prediction website ( http: / / crispor.tefor.net / ) to generate a target list, select target 1 as 5'-ACACCATCTGGGGAACGAA-3', and target 2 as 5'-AAGCCATTCGTTCCCCAGA-3', such as figure 1 shown.
[0127] 1.b, sgRNA module construction
[0128] As shown in Table 1, the primers MtGA3ox1-BsF, MtGA3ox1-F0, MtGA3ox1-R0, MtGA3ox1-BsF and MtGA3ox1-BsR were synthesized by BGI.
[0129] The p5CBC-Target amplification reaction was carried out by bridging PCR, and the sgRNA module p5CBC-MtGA3ox1 with the target sequence...
Embodiment 2
[0161] Example 2. Obtaining and phenotypic detection of alfalfa MsGA3ox1 gene knockout mutants
[0162] 1. Construction of alfalfa MsGA3ox1 gene CRISPR / Cas9 system vector
[0164] The coding sequence (CDS) contained in Medicago sativa is the MsGA3ox1 coding gene shown in sequence 3 in the sequence listing, and the protein MsGA3ox1 whose amino acid sequence is shown in sequence 1 in the sequence listing is encoded.
[0165] Through the online target prediction website ( http: / / crispor.tefor.net / ) to generate a target list, select target 1 as 5′-ACACCATCCGGGGAACGAA-3, and target 2 as 5′-AAGCCATTCGTTCCCCGGA-3′.
[0166] 1.b, sgRNA module construction
[0167] Primers MsGA3ox1-BsF, MsGA3ox1-F0, MsGA3ox1-R0 and MsGA3ox1-BsR were synthesized by BGI Corporation.
[0168] The p5CBC-Target amplification reaction was carried out by bridging PCR, and the sgRNA module p5CBC-MsGA3ox1 with the target sequence was amplified. The reaction system ...
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Abstract
The invention discloses a method for cultivating dwarf plant type and / or compact plant type and / or creeping plant type alfalfa by gene editing technology. The invention provides the application of GA3ox1 protein or its encoding gene in regulating the plant type of alfalfa. Knocking out the gene encoding GA3ox1 protein in alfalfa will reduce or inactivate the protein expression and / or activity, and the gibberellin content in alfalfa will decrease. Experiments have shown that the plant height and internode length of alfalfa GA3ox1 knockout plants are significantly reduced compared with the wild type, showing a dwarf and / or compact and / or creeping plant type. Using gene editing technology to target GA3ox1 to quickly and efficiently obtain new plant types of alfalfa is of great significance for the application of alfalfa interplanting and intercropping.
Description
technical field [0001] The invention relates to the field of biotechnology, in particular to the application of GA3ox1 protein in regulating alfalfa plant type. Background technique [0002] Alfalfa is a perennial leguminous plant, which has the characteristics of high yield, high quality, rich nutritional value and strong adaptability, and is known as the "king of forage grass". Alfalfa has important agricultural production value. In addition to being a high-quality foragecrop, it can also be interplanted with silage corn to enhance the activity of soil nitrogen-fixing microorganisms, improve the utilization efficiency of rhizospheresoil nutrients, improve soil fertility, and promote plant growth and development; interplanting in orchards Alfalfa is conducive to the formation of a virtuous cycle ecosystem in orchards, improves the economic effects of fruit tree planting, promotes feed production, and then promotes the development of animal husbandry; intercropping alfalfa...
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