CN-122012584-A - Application of SiMC gene in regulation and control of anabolism of millet carotenoid

CN122012584ACN 122012584 ACN122012584 ACN 122012584ACN-122012584-A

Abstract

The invention discloses an application of SiMC gene in regulating anabolism of millet carotenoid, belonging to the technical field of genetic engineering breeding. The SiMC gene for regulating and controlling the metabolism of the millet carotenoid is screened, and the nucleotide sequence of the SiMC gene is shown as SEQ ID NO. 1. Experimental results show that compared with a wild plant (WT), the SiMC gene knockout strain (KO) has obviously increased millet yellow color and carotenoid content, and the SiMC gene negatively regulates the synthesis of the millet carotenoid, is a potential efficient millet breeding gene, and can be used for improving the appearance characters and the edible quality of millet. The invention provides a gene modification target and a theoretical basis for screening and cultivating high-quality millet varieties, and is hopeful to obtain new millet germplasm which has appearance and taste and is rich in natural antioxidant nutrition.

Inventors

WANG JUNJIE
ZHANG BIN
MA QI
Zhen Xiaoxi
LI YINGHUI

Assignees

山西农业大学

Dates

Publication Date: 20260512
Application Date: 20260210

Claims (10)

1. Use of SiMC protein in any one of the following: (1) Application in regulating and controlling synthesis of millet carotenoid; (2) Use in increasing the carotenoid content of millet; (3) Use in breeding transgenic millet with increased carotenoid content; (4) Application in regulating millet color; The amino acid sequence of SiMC protein is shown as SEQ ID NO. 2.
2. The use according to claim 1, wherein the expression level of SiMC protein is down-regulated in millet, promoting synthesis of millet carotenoid, increasing the content of millet carotenoid, and yellowing the millet color.
3. Use of a gene encoding the SiMC protein as defined in claim 1 in any one of the following: (1) Application in regulating and controlling synthesis of millet carotenoid; (2) Use in increasing the carotenoid content of millet; (3) Use in breeding transgenic millet with increased carotenoid content; (4) Application in regulating millet color; The nucleotide sequence of the coding gene is shown as SEQ ID NO. 1.
4. The use according to claim 3, wherein the expression level of the coding gene is down-regulated in millet, the synthesis of millet carotenoids is promoted, the content of millet carotenoids is increased, and the millet color is turned yellow.
5. The application of the knockout vector is characterized in that the knockout vector targets the coding gene of SiMC proteins; the nucleotide sequence of the coding gene is shown as SEQ ID NO. 1; The application is any one of the following: (1) Application in regulating and controlling synthesis of millet carotenoid; (2) Use in increasing the carotenoid content of millet; (3) Use in breeding transgenic millet with increased carotenoid content; (4) The application in regulating and controlling the millet color.
6. Use of an engineered bacterium comprising the knockout vector of claim 5 in any one of the following: (1) Application in regulating and controlling synthesis of millet carotenoid; (2) Use in increasing the carotenoid content of millet; (3) Use in breeding transgenic millet with increased carotenoid content; (4) The application in regulating and controlling the millet color.
7. A method for promoting synthesis of a carotenoid from millet, comprising the step of down-regulating the expression level of a gene encoding SiMC protein in millet to promote synthesis of the carotenoid from millet; The nucleotide sequence of the coding gene is shown as SEQ ID NO. 1.
8. A method for increasing the carotenoid content of millet, comprising the step of reducing the expression level of a SiMC protein-encoding gene in millet to increase the carotenoid content of millet; The nucleotide sequence of the coding gene is shown as SEQ ID NO. 1.
9. A method for breeding transgenic millet with increased carotenoid content in millet, comprising the steps of: Knocking out coding genes of SiMC protein in millet cells, then culturing the millet cells, and regenerating millet plants by using the millet cells to obtain transgenic millet with increased millet carotenoid content; The nucleotide sequence of the coding gene is shown as SEQ ID NO. 1.
10. A method for yellowing millet colors, comprising the step of down-regulating the expression level of a gene encoding SiMC protein in millet to cause the millet colors to yellow; The nucleotide sequence of the coding gene is shown as SEQ ID NO. 1.

Description

Application of SiMC gene in regulation and control of anabolism of millet carotenoid Technical Field The invention relates to the technical field of genetic engineering breeding, in particular to application of SiMC gene in regulating and controlling anabolism of millet carotenoid. Background Millet (SETARIA ITALICA) is an important coarse cereal crop in China, and the shelling product of the millet (SETARIA ITALICA) is small Mi Fu and contains carotenoid to present bright yellow color. Millet quality is a comprehensive trait including appearance quality, steamed taste quality, and nutritional quality. The appearance quality of millet refers to the apparent character of the millet, is the most direct and first-line selection index for evaluating the quality of millet by consumers, and is also an important direction for breeding the quality of millet. The rice color affects the preference of consumers to a great extent, and is a main index for evaluating the quality of millet and a main direction for breeding millet quality. Carotenoids (carotenoids) directly affect millet color, are widely used in animals, plants and microorganisms in nature, are terpenes composed of isoprene, and have variable number of conjugated double bonds in the polyene main chain and antioxidant activity. Carotenoids are a generic name for two major pigments of 40 carbon hydrocarbons (carotenes) and their oxidized derivatives (lutein), which mainly include alpha-carotene, beta-cryptoxanthin, lutein, zeaxanthin, lycopene, and the like. The overall pathway of higher plant carotenoid biosynthesis has been clarified, and the whole process includes condensation, dehydrogenation, cyclization, hydroxylation and 5 epoxidation reactions involving PSY, PDS, ZDS, LCYE, LCYB, CHYb, CYP, ZEP, CCD, NCED and other related enzyme genes. Meanwhile, the regulation and control of a plurality of transcription factors are complex processes, and the research on the transcriptional regulation and control mechanism of carotenoid biosynthesis is limited at present. In recent years, many transcription factors involved in regulating structural genes of carotenoid metabolism in plants are identified sequentially, including bHLH, bZIP, MADS-box, NAC, MYB, ERF family members are identified sequentially, however, many of these transcription factors exhibit species-specific regulation features and lack universal regulation rules in the plant kingdom, so that key genes for regulating and clearly regulating carotenoid metabolism in millet are mined and still need to be further studied. Disclosure of Invention The invention aims to provide an application of SiMC gene in regulating and controlling anabolism of millet carotenoid to solve the problems in the prior art, and provides an application of SiMC gene in regulating and controlling millet color to improve the carotenoid content of millet seeds and further improve the quality of millet, so as to provide a molecular mechanism and a theoretical basis for screening and cultivating high-quality millet varieties. In order to achieve the above object, the present invention provides the following solutions: the invention provides an application of SiMC protein in any one of the following: (1) Application in regulating and controlling synthesis of millet carotenoid; (2) Use in increasing the carotenoid content of millet; (3) Use in breeding transgenic millet with increased carotenoid content; (4) Application in regulating millet color; The amino acid sequence of SiMC protein is shown as SEQ ID NO. 2. Further, the expression level of SiMC protein is regulated down in millet, so that the synthesis of millet carotenoid is promoted, the content of millet carotenoid is improved, and the millet color is changed into yellow. The invention also provides application of the SiMC protein coding gene in any one of the following: (1) Application in regulating and controlling synthesis of millet carotenoid; (2) Use in increasing the carotenoid content of millet; (3) Use in breeding transgenic millet with increased carotenoid content; (4) Application in regulating millet color; The nucleotide sequence of the coding gene is shown as SEQ ID NO. 1. Further, the expression level of the coding gene is regulated down in millet, so that the synthesis of millet carotenoid is promoted, the content of the millet carotenoid is improved, and the millet color is changed into yellow. The invention also provides application of the knockout vector, wherein the knockout vector targets the coding gene of SiMC protein; the nucleotide sequence of the coding gene is shown as SEQ ID NO. 1; The application is any one of the following: (1) Application in regulating and controlling synthesis of millet carotenoid; (2) Use in increasing the carotenoid content of millet; (3) Use in breeding transgenic millet with increased carotenoid content; (4) The application in regulating and controlling the millet color. The invention also provides application of the engineeri