CN-116355953-B - Plant expression vector for improving artemisinin content in sweet wormwood herb, recombinant transformant, sweet wormwood herb and application thereof

Abstract

The invention provides a plant expression vector for improving the content of artemisinin in sweet wormwood, a recombinant transformant, sweet wormwood and application thereof. The first aspect of the invention provides a plant expression vector, which comprises a gene encoding a transcription factor AabHLH, wherein the gene encoding the transcription factor AabHLH39 has a nucleotide sequence shown as SEQ ID NO. 1. According to the invention, the plant expression vector containing the gene for encoding the transcription factor AabHLH39 is constructed and transformed into the sweet wormwood, so that the sweet wormwood overexpresses the transcription factor AabHLH39, and the content of artemisinin in the sweet wormwood is improved.

Inventors

• ZHAO JINGYA
• FU XUEQING
• SUN XIAOFEN
• TANG KEXUAN

Assignees

• 上海交通大学

Dates

Publication Date

20260512

Application Date

20230302

Claims (10)

1. 1. The plant expression vector is characterized by comprising a gene encoding a transcription factor AabHLH, and the gene encoding the transcription factor AabHLH39 has a nucleotide sequence shown as SEQ ID NO. 1.
2. 2. The plant expression vector of claim 1, wherein the plant expression vector is a pHB vector comprising a gene encoding a transcription factor AabHLH.
3. 3. A recombinant transformant comprising the plant expression vector of claim 1 or 2.
4. 4. The recombinant transformant according to claim 3, wherein the recombinant transformant is Agrobacterium tumefaciens comprising the plant expression vector.
5. 5. The sweet wormwood herb is characterized in that the sweet wormwood herb overexpresses a transcription factor AabHLH39, and a gene encoding the transcription factor AabHLH39 has a nucleotide sequence shown as SEQ ID NO. 1.
6. 6. The application of the transcription factor AabHLH39 in improving the artemisinin content in sweet wormwood herb is characterized in that the sweet wormwood herb is used for overexpressing the transcription factor AabHLH39, and a gene for encoding the transcription factor AabHLH39 has a nucleotide sequence shown as SEQ ID NO. 1.
7. 7. A method for increasing the content of artemisinin in artemisia annua, which is characterized by comprising the following steps: Constructing a plant expression vector containing a gene encoding a transcription factor AabHLH, wherein the gene of the transcription factor AabHLH has a nucleotide sequence shown as SEQ ID NO. 1; Transferring the plant expression vector into agrobacterium tumefaciens to obtain a recombinant transformant; and (3) transforming the recombinant transformant into sweet wormwood, and screening to obtain a transgenic plant.
8. 8. The method of claim 7, wherein constructing a plant expression vector comprising a gene encoding a transcription factor AabHLH, specifically comprises: cloning a gene encoding a transcription factor AabHLH from a cDNA library of sweet wormwood; Operably linking the gene encoding transcription factor AabHLH to a pHB vector results in a plant expression vector comprising the gene encoding transcription factor AabHLH 39.
9. 9. The method of claim 7, wherein the agrobacterium tumefaciens is EH105.
10. 10. The method according to claim 7, wherein the artemisinin content is above 14.6 mg/g based on the mass of the leaves of artemisia annua.

Description

Plant expression vector for improving artemisinin content in sweet wormwood herb, recombinant transformant, sweet wormwood herb and application thereof Technical Field The invention relates to a plant expression vector for improving the content of artemisinin in sweet wormwood, a recombinant transformant, sweet wormwood and application thereof, and relates to the technical field of genetic engineering. Background Artemisia annua (ARTEMISIA ANNUA L.) is an annual herb of Artemisia of Compositae, and sesquiterpene lactone oxide-artemisinin extracted from aerial parts thereof and containing peroxy bridge is currently the most widely used antimalarial drug with the best curative effect, and is particularly more effective on cerebral malaria and chloroquine malaria. Although the microbial synthesis of the arteannuic acid has been successful, the arteannuic acid still belongs to the semi-synthesis of the arteannuin, and has the problems of large investment in the early stage and difficult equipment maintenance, while the chemical synthesis of the arteannuin has large difficulty and cannot meet the market demand. At present, the artemisinin mainly comes from plant extraction, but the content of the artemisinin in the artemisia annua is low and only accounts for (0.1-1%) of the dry weight of the artemisia annua leaves, so that the market supply and the value of the artemisinin are limited. Therefore, how to increase the content of artemisinin in artemisia annua by means of genetic engineering is one of the hot spots continuously focused by the person skilled in the art. Disclosure of Invention The invention provides a plant expression vector, which comprises a gene sequence capable of encoding a transcription factor AabHLH and a transcription factor AabHLH, wherein the transcription factor AabHLH belongs to bHLH transcription factors, can positively regulate and control the synthesis of artemisinin and improve the content of artemisinin in artemisia annua. The invention also provides a recombinant transformant comprising the plant expression vector, sweet wormwood herb, application of the transcription factor AabHLH in improving the content of artemisinin in sweet wormwood herb and a method for improving the content of artemisinin in sweet wormwood herb. The first aspect of the invention provides a plant expression vector, which comprises a gene encoding a transcription factor AabHLH, wherein the gene encoding the transcription factor AabHLH39 has a nucleotide sequence shown as SEQ ID NO. 1. Further, the plant expression vector is a pHB vector including a gene encoding a transcription factor AabHLH 39. In a second aspect, the invention provides a recombinant transformant comprising a plant expression vector as described in any one of the preceding claims. Further, the recombinant transformant is agrobacterium tumefaciens comprising the plant expression vector. In a third aspect, the invention provides a sweet wormwood herb, which can overexpress a transcription factor AabHLH and the gene encoding the transcription factor AabHLH39 has a nucleotide sequence shown as SEQ ID NO. 1. In a fourth aspect, the invention provides the use of transcription factor AabHLH39 in increasing artemisinin content in sweet wormwood, so that sweet wormwood overexpresses transcription factor AabHLH39, and the gene encoding transcription factor AabHLH39 has a nucleotide sequence shown as SEQ ID NO. 1. In a fifth aspect, the present invention provides a method for increasing the content of artemisinin in artemisia annua, comprising the steps of: constructing a plant expression vector comprising a gene encoding a transcription factor AabHLH; Transferring the plant expression vector into agrobacterium tumefaciens to obtain a recombinant transformant; and (3) transforming the recombinant transformant into sweet wormwood, and screening to obtain a transgenic plant. Further, constructing a plant expression vector comprising a gene encoding a transcription factor AabHLH, specifically comprising: cloning a gene encoding a transcription factor AabHLH from a cDNA library of sweet wormwood; Operably linking the gene encoding transcription factor AabHLH to a pHB vector results in a plant expression vector comprising the gene encoding transcription factor AabHLH 39. Further, the agrobacterium tumefaciens is EH105. Further, the content of artemisinin is above 14.6mg/g based on the mass of the leaves of Artemisia annua. According to the invention, the plant expression vector containing the gene for encoding the transcription factor AabHLH39 is constructed and converted into the sweet wormwood, so that the sweet wormwood can overexpress the transcription factor AabHLH39, and the content of artemisinin in the sweet wormwood is improved. Drawings FIG. 1 shows the analysis of the expression pattern of transcription factor AabHLH, wherein a is the relative expression level of transcription factor AabHLH39 in the stems, roots, old leaves, flower buds 1, flowe