阅读说明：本技术 大豆糖转运体基因GmSWEET39的应用 (Application of soybean sugar transporter gene GmSWEET39 ) 是由 李艳 苗龙 盖钧镒 张凯 杨松楠 于 2019-10-10 设计创作，主要内容包括：本发明公开了大豆糖转运体基因GmSWEET39的应用。大豆糖转运体基因GmSWEET39在培育高含油量的植物新品种或转基因植物中的应用。该基因在栽培豆中具有3种启动子序列Pro1,Pro2,和Pro3；两种编码序列CDS1和CDS2。Pro3和CDS2为控制大豆籽粒油脂含量的优异变异。过量表达GmSWEET39优异变异序列的拟南芥和大豆发根,转化植株的含油量显著提高。对大豆重组自交系群体中具有不同启动子或CDS类型的后代家系进行含油量比较,证实Pro3和CDS2为GmSWEET39的优异变异,该基因的优异变异类型将作为大豆育种计划中油脂含量遗传改良的宝贵基因资源。(The invention discloses application of a soybean sugar transporter gene GmSWEET 39. Application of soybean sugar transporter gene GmSWEET39 in breeding new plant varieties or transgenic plants with high oil content. The gene has 3 promoter sequences Pro1, Pro2 and Pro3 in cultivated beans; two coding sequences CDS1 and CDS 2. Pro3 and CDS2 are excellent variations for controlling the oil content of soybean seeds. Arabidopsis thaliana and soybean with over-expression GmSWEET39 excellent variant sequence are rooted, and the oil content of the transformed plant is obviously improved. The oil content comparison of the progeny families with different promoters or CDS types in the soybean recombinant inbred line population proves that Pro3 and CDS2 are excellent variation of GmSWEET39, and the excellent variation type of the gene can be used as a precious gene resource for genetic improvement of oil content in a soybean breeding plan.)

1. Application of soybean sugar transporter gene GmSWEET39 in breeding new plant varieties or transgenic plants with high oil content.

2. The use of claim 1, wherein the soybean sugar transporter gene GmSWEET39 promoter sequence is selected from one of SEQ ID No. 1-SEQ ID No. 3.

3. The use of claim 2, wherein the soybean sugar transporter gene GmSWEET39 promoter has the sequence shown in SEQ ID NO. 3.

4. The use according to claim 1, wherein the CDS sequence of the soybean sugar transporter gene GmSWEET39 is selected from one of SEQ ID No.4 or SEQ ID No. 5.

5. The use of claim 4, wherein the CDS sequence of the soybean sugar transporter gene GmSWEET39 is shown in SEQ ID NO. 5.

6. The recombinant expression vector containing the soybean sugar transporter gene GmSWEET39 is applied to culturing new plant varieties or transgenic plants with high oil content.

7. The use according to claim 2, characterized in that the promoter sequence of the soybean sugar transporter gene GmSWEET39 is selected from one of SEQ ID No. 1-SEQ ID No.3, preferably as shown in SEQ ID No. 3.

8. The use according to claim 4, characterized in that the CDS sequence of the soybean sugar transporter gene GmSWEET39 is selected from one of SEQ ID No.4 or SEQ ID No.5, preferably as shown in SEQ ID No. 5.

9. The application of the promoter and/or CDS sequence of soybean sugar transporter gene GmSWEET39 in screening plants with high grain oil content.

10. The use of claim 9, wherein the promoter sequence of the soybean sugar transporter gene GmSWEET39 is shown as SEQ ID NO.3, and the CDS sequence of the soybean sugar transporter gene GmSWEET39 is shown as SEQ ID NO. 5.

Technical Field

The invention belongs to the field of genetic engineering, and discloses application of a soybean sugar transporter gene GmSWEET 39.

Background

Soybean (Glycine max) is a crop with important strategic significance in China and also is the most important leguminous crop in feed and food in the world, because high-quality soybean seeds contain more than 40% of protein and 20% of oil content. Although the protein content of seeds is usually about twice that of oil, soybeans are the oil crops with the highest yield in the world, and contribute 29% of the share of edible oil in the world. Soy oil and its derivatives are ubiquitous ingredients in packaged foods, especially cookies and some snack foods. With the increasing demand of people for vegetable oil and the increasing awareness of consumers about dietary quality fatty acids, the oil content and quality are attracting attention of soybean breeding workers. Therefore, the discovery and utilization of the soybean seed oil favorable genes have important significance for improving the soybean quality and meeting the production requirements of human beings.

Sugars are the main carbon and energy source of organisms and play an important role in plant growth and development. Sweet (sugar wing eventually be expressed transporters) is a class of sugar transporters identified in recent years, a seven-helix structure consisting of tandem repeats of three transmembrane domains, linked by inverted linker transmembrane domains. SWEET can promote sugar to pass through cell membranes along concentration gradient, provides for long-distance transportation of sugar and play a role in non-photosynthetic tissues, and has important significance for molecular improvement of soybean varieties with excellent grain oil content by analyzing the relation between SWEET protein-mediated sugar transportation and oil accumulation and cloning excellent SWEET genes.

Disclosure of Invention

The invention aims to provide a function and application of a sugar transporter gene GmSWEET39(Glyma.15g049200) in the aspect of oil accumulation aiming at the improvement and breeding of oil and fat properties of soybean grains.

The purpose of the invention can be realized by the following technical scheme:

application of soybean sugar transporter gene GmSWEET39 in breeding new plant varieties or transgenic plants with high oil content.

The GmSWEET39 gene has 3 promoter sequences (Pro1, Pro2 and Pro3) in cultivated beans, and the nucleotide sequences are SEQ ID NO.1, SEQ ID NO.2 and SEQ ID NO. 3; two coding sequences (CDS1 and CDS2) having the nucleotide sequences SEQ ID NO.4 and SEQ ID NO. 5. Pro3 and CDS2 are excellent variations for controlling the oil content of soybean seeds.

The recombinant expression vector containing the soybean sugar transporter gene GmSWEET39 is applied to culturing new plant varieties or transgenic plants with high oil content.

The soybean sugar transporter gene GmSWEET39 promoter sequence is preferably selected from one of SEQ ID NO. 1-SEQ ID NO.3, and is further preferably shown as SEQ ID NO. 3.

The CDS sequence of the soybean sugar transporter gene GmSWEET39 is preferably selected from one of SEQ ID NO.4 or SEQ ID NO.5, and is further preferably shown as SEQ ID NO. 5.

The application of the promoter and/or CDS sequence of soybean sugar transporter gene GmSWEET39 in screening plants with high grain oil content.

The promoter sequence of the soybean sugar transporter gene GmSWEET39 is preferably shown as SEQ ID NO.3, and the CDS sequence of the soybean sugar transporter gene GmSWEET39 is preferably shown as SEQ ID NO. 5.

Has the advantages that:

the differential expression analysis of the sugar transporter gene GmSWEET39 in high-oil and low-oil soybean varieties shows that the gene plays an important role in soybean seed oil accumulation. The expression abundance of the gene is found to be in positive correlation with the oil content in 80 soybean varieties with different oil contents. By performing promoter and CDS sequencing on GmSWEET39 genes of different varieties, the genes have 3 promoter sequences and two Coding (CDS) sequences. Comparing oil contents of varieties with different promoters or CDS types, and finding that Pro3 and CDS2 are excellent variation for controlling the oil content of soybean grains. The excellent GmSWEET39 variant sequence is inserted into an over-expression vector and is further subjected to genetic transformation to generate over-expressed Arabidopsis and soybean hairy roots, so that the oil content of a transformed plant can be remarkably improved. Meanwhile, the oil content of the progeny families with different promoters or CDS types in the soybean recombinant inbred line population is compared, so that Pro3 and CDS2 are proved to be excellent variation of GmSWEET39, and the excellent variation type of the gene can be used as a precious gene resource for genetic improvement of the oil content in the soybean breeding plan.

Drawings

FIG. 1 is a relative expression analysis of GmSWEET39 gene in grains of different periods of high oil varieties and low oil varieties; wherein, DAF: days after flowering, days after flowering.

The oil content of 280 soybean varieties and the expression level of GmSWEET39 gene in soybean seeds of different varieties are shown in the figure; and fitting and drawing the trend line by using a local weighted scatter point smoothing method.

FIG. 3 shows the variation of the promoter and CDS sequence of GmSWEET39 gene in 80 soybean varieties.

FIG. 4 compares the oil content of soybean varieties containing different promoter types and CDS types of the GmSWEET39 gene.

FIG. 5 comparison of translated amino acid sequences of CDS1 type, CDS2 type and the well-known and commonly used Williams 82 reference genome GmSWEET39 gene.

FIG. 6 transgenic Arabidopsis thaliana examined the allelic effect of GmSWEET39 gene: influence of overexpression of different CDS type GmSWEET39 genes on the fatty acid content of Arabidopsis seeds.

FIG. 7 transgenic soybean hairy roots test the allelic effect of GmSWEET39 gene.

FIG. 8 inspection of GmSWEET39 gene allelic effect by soybean recombinant inbred line population

A, picture A: the oil content frequency distribution of 155 offspring families; wherein ZYD2612 and NN92-128 indicate amphipathic oil content, respectively; and B, drawing: and comparing the oil content of the offspring pedigree grains containing different promoter types and different CDS types of the GmSWEET39 gene.

Detailed Description