Application of E1-SOC1 gene model in cultivation of soybean variety with wide adaptability

Abstract

The invention belongs to the technical field of molecular breeding, and particularly relates to application of an E1-SOC1 gene model in cultivation of wide-adaptability soybean varieties. Research finds that three genes (E1 and homologous genes E1La and E1Lb) of an E1 family and two genes (SOC1a and SOC1b) of an SOC1 family are knocked out from soybeans at the same time by utilizing a gene editing technology, and the obtained homozygous five mutants show wide adaptability when planted under field conditions of different latitude regions; and the method has certain wide adaptability in the aspects of plant height, section number, flowering period, mature period and yield. It is indicated that the E1-SOC1 gene module (E1, E1La, E1Lb, SOC1a and SOC1b genes) is expected to be applied to molecular design breeding for breeding wide-adaptability soybean models, and has important application prospects in soybean molecular breeding research.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of molecular breeding, and particularly relates to application of an E1-SOC1 gene model in breeding of a widely adaptable soybean variety. BACKGROUND

[0002] Soybean [Glycine max (L.) Merr.] provides plant protein and plant oil for human beings, provides industrial raw materials, and is closely related to human life. Soybean is an important source of edible plant protein and oil in the world. With the rapid economic development and the improvement of people's living standards, the demand for soybean has risen sharply, so breeding a high-yield soybean variety with wide adaptability to climate and cultivation environment is an urgent need of social development.

[0003] Soybean is a short-day crop sensitive to photoperiod, and the strength of its photoperiod response determines the length of the growth period, which in turn determines the yield of soybean by affecting the number of pods per plant. It takes at least 8-10 years to breed a high-yield soybean variety using traditional breeding methods, but because soybean varieties do not have wide adaptability, this seriously limits the process of expanding the planting area of high-yield varieties, thereby limiting the greater positive benefits of these varieties in breeding applications. For example, Heilongjiang Province is a major soybean producing area in China, spanning about 10 latitudes from north to south, but due to differences in light and temperature, the same high-yield variety is difficult to plant across regions, greatly limiting the application potential of excellent varieties. Therefore, it is very important to create a soybean variety with wide adaptability, which can greatly save breeding time and cost and better meet social needs. SUMMARY

[0004] In order to overcome the deficiencies of the prior art, the present application has found that knocking out the E1-SOC1 gene model (E1, E1La, E1Lb, SOC1a, and SOC1b genes) in soybean can create a soybean variety with wide adaptability, which has important application prospects in soybean molecular breeding research.

[0005] In order to achieve the above-mentioned purpose, the technical scheme adopted by the present application is:

[0006] The present application provides, in a first aspect, application of an E1-SOC1 gene model in breeding of a widely adaptable soybean variety, wherein the E1-SOC1 gene model comprises an E1 gene, an E1La gene, an E1Lb gene, a SOC1a gene, and a SOC1b gene.

[0007] Preferably, the breeding method is knocking out the E1-SOC1 gene model in soybean.

[0008] Preferably, the widely adaptable soybean variety has at least one of the following characteristics:

[0009] (1) Plant height has wide adaptability in different latitude regions;

[0010] (2) Node number has wide adaptability in different latitude regions;

[0011] (3) Flowering period has wide adaptability in different latitude regions;

[0012] (4) Maturation period has wide adaptability in different latitude regions;

[0013] (5) Yield has wide adaptability in different latitude regions.

[0014] Preferably, the soybean includes, but is not limited to, soybean variety Williams 82.

[0015] Preferably, the E1-SOC1 gene model further includes a biological material containing the E1-SOC1 gene model, and the biological material includes one or more of a recombinant expression vector, a plasmid, an expression cassette, a recombinant bacteria or a host cell.

[0016] The second aspect of the present application provides a method for cultivating a soybean variety with wide adaptability, specifically: using gene editing technology to knock out an E1-SOC1 gene model in soybean, wherein the E1-SOC1 gene model includes an E1 gene, an E1La gene, an E1Lb gene, an SOC1a gene and an SOC1b gene.

[0017] Compared with the prior art, the present application has the following beneficial effects:

[0018] The present application has found that E1 and its homologous genes E1La and E1Lb are involved in the regulation of soybean wide adaptability, but there are still great differences in node number and plant height in different latitude regions. Therefore, further identification of the downstream genes directly regulated by E1 family genes found that E1 can directly regulate two genes (SOC1a and SOC1b) of the SOC1 family. Then, using gene editing technology to knock out the three genes of the E1 family and the two genes of the SOC1 family at the same time, it is found that the homozygous five mutants obtained all show wide adaptability under field conditions in different latitude regions in China, and have certain wide adaptability in plant height, node number, flowering period, maturation period and yield. It is shown that the E1-SOC1 gene module (E1, E1La, E1Lb, SOC1a, SOC1b genes) is expected to be applied to the molecular design breeding of cultivating a soybean model with wide adaptability, and has important application prospects in soybean molecular breeding research. BRIEF DESCRIPTION OF DRAWINGS

[0019] Figure 1Phenotypes of W82 and e1 e1la e1lb triple mutants in the field in different latitudes of China; A: Phenotypes of W82 in the field in different latitudes of China; B: Phenotypes of e1 e1la e1lb in the field in different latitudes of China; Phenotype data include flowering time, maturation time, node number, plant height and yield per plant (KS represents Keshan, 48 °N; CC represents Changchun, 44 °N; SJZ represents Shijiazhuang, 38 °N; HF represents Hefei, 31 °N; GZ represents Guangzhou, 23 °N; SY represents Sanya, 18 °N); Different lowercase letters represent P < 0.05.

[0020] Figure 2 Identification of downstream genes directly regulated by E1; A: DAP-seq result analysis of the number of downstream genes directly regulated by E1; B: Cis-acting elements directly bound by E1 protein; C: Expression amount of FT2a gene in W82, e1, e1la e1b and e1 e1la e1lb mutants; D: Expression amount of FT5a gene in W82, e1, e1la e1b and e1 e1la e1lb mutants; E: Expression amount of SOC1a gene in W82, e1, e1la e1b and e1 e1la e1lb mutants; F: Expression amount of SOC1b gene in W82, e1, e1la e1b and e1 e1la e1lb mutants.

[0021] Figure 3A: Plant phenotypes of W82, e1e1la e1lb triple mutant, soc1a soc1b double mutant and e1e1la e1lb soc1a soc1b quintuple mutant in different latitudes of China (KS represents Keshan, 48 °N; CC represents Changchun, 44 °N; SJZ represents Shijiazhuang, 38 °N; HF represents Hefei, 31 °N; GZ represents Guangzhou, 23 °N; SY represents Sanya, 18 °N); B: Flowering time phenotypes of W82, e1e1la e1lb triple mutant, soc1a soc1b double mutant and e1e1la e1lb soc1a soc1b quintuple mutant in different latitudes of China, different lowercase letters represent P<0.05; C: Maturity phenotypes of W82, e1e1la e1lb triple mutant, soc1a soc1b double mutant and e1e1la e1lb soc1a soc1b quintuple mutant in different latitudes of China, different lowercase letters represent P<0.05; D: Node number phenotypes of W82, e1e1la e1lb triple mutant, soc1a soc1b double mutant and e1e1la e1lb soc1a soc1b quintuple mutant in different latitudes of China, different lowercase letters represent P<0.05; E: Yield phenotypes of W82, e1e1la e1lb triple mutant, soc1a soc1b double mutant and e1e1la e1lb soc1a soc1b quintuple mutant in different latitudes of China, different lowercase letters represent P<0.05. DETAILED DESCRIPTION

[0022] The specific embodiments of the present application will be further described below. It should be noted that the description of these embodiments is used to help understand the present application, but does not constitute a limitation on the present application. In addition, the technical features involved in each embodiment of the present application described below can be combined with each other as long as they do not conflict with each other.

[0023] The experimental methods in the following examples are all conventional methods unless otherwise specified. The test materials used in the following examples are all commercially available unless otherwise specified.

[0024] Example 1 E1 and its homologous genes E1La and E1Lb are involved in regulating soybean wide adaptability

[0025] Studies have shown that the legume-specific E1 gene and its homologs E1La and E1Lb play a central role in the photoperiod regulation of soybean flowering, but no research has been reported on whether soybean E1 family genes can participate in the regulation of soybean's broad adaptability. To further investigate the role of the E1 family genes in regulating the wide adaptability of soybean, this study used gene editing and stable transgenic technology to simultaneously knock out E1 (gene ID: Glyma.06G207800), E1La (gene ID: Glyma.04G156400), and E1Lb (gene ID: Glyma.04G143300) in the soybean variety Williams 82 (W82) (the specific knockout method is described in the reference "Dong LD, Li SC, Wang LS, Su T, Zhang CB, Bi YD, Lai YC, Kong LP, Wang F, Pei XX, Li HY, Hou ZH, Du HP, Du H, Li T, Cheng Q, Fang C, Kong FJ, Liu BHe et al. (2023) The genetic basis of high-latitude adaptation in wild soybean. Curr. Biol., 33, 252-262."), obtaining e1 and e1la. The e1lb homozygous triple mutant (named e1-3m) was used to determine whether the e1-3m homozygous triple mutant had broad adaptability. In order to identify whether the triple mutant and the control W82 were widely adaptable, the triple mutant and the control W82 were planted in fields at different latitudes in my country (Keshan, 48°N; Changchun, 44°N; Shijiazhuang, 38°N; Hefei, 31°N; Guangzhou, 23°N; Sanya, 18°N) for phenotypic observation.

[0026] The results showed that, as the planting latitude decreased, the flowering and maturity periods of the control variety W82 significantly advanced, the number of nodes decreased, and the plant height and yield per plant decreased. However, it could not mature normally in Keshan at a high latitude, and its yield was extremely low in Guangzhou and Sanya at low latitudes, indicating that the W82 soybean variety does not have wide adaptability. Figure 1 A). The e1-3m triple mutant exhibits the same flowering and maturity periods regardless of latitude, but there are still significant differences in the number of nodes and plant height. Specifically, the e1-3m triple mutant grown in low-latitude regions has approximately 6 more nodes than that grown in high-latitude regions, and its yield is also increased. Figure 1 B). The above results indicate that the e1-3m triple mutant exhibits some broad adaptability in terms of growth period, but not in terms of node number and yield. It is noteworthy that the phenomenon of increased node number and yield when grown in low-latitude regions compared to high-latitude regions has not been reported. Currently, no soybean species in nature exhibits this phenomenon.

[0027] Example 2: Identification of downstream genes of E1

[0028] To identify downstream genes directly regulated by the E1 family genes and subsequently breed widely adaptable soybean varieties based on network models, this study used DAP-seq technology to identify 435 directly bound downstream genes. Figure 2 A), and for the first time reported the cis-acting element A / TCATTTT combined with E1. Figure 2 B) The specific function of this cis-acting element is unclear because it has not been reported in plants. Subsequently, analysis of 435 downstream genes revealed that E1 can directly bind to the promoter regions of FT2a (gene ID: Glyma.16G150700), FT5a (gene ID: Glyma.16G044100), SOC1a (gene ID: Glyma.18G224500), and SOC1b (gene ID: Glyma.09G266200). Therefore, the expression of these four genes was detected under different E1 family gene mutation backgrounds (specific mutation methods refer to the literature "Dong LD, Li SC, Wang LS, Su T, Zhang CB, BiYD, Lai YC, Kong LP, Wang F, Pei XX, Li HY, Hou ZH, Du HP, Du H, Li T, Cheng Q, Fang C, Kong FJ, Liu BH et al. (2023) The genetic basis of high-latitude adaptation in wild soybean. Curr. Biol., 33, 252-262.").

[0029] The results showed that E1 could inhibit the expression of FT2a, FT5a, SOC1a, and SOC1b. Figure 2 (C-2F). Currently, no studies have been reported on the direct regulation of SOC1a and SOC1b by E1. Furthermore, no studies have been reported on whether E1 and its downstream genes SOC1a and SOC1b can participate in the regulation of broad soybean adaptability. Therefore, future research will continue on the role of E1 and its downstream genes SOC1a and SOC1b in regulating broad soybean adaptability.

[0030] Example 3: Creating a new soybean material with broad adaptability by knocking out the E1-SOC1 module using gene editing technology.

[0031] To verify whether the E1-SOC1 module can participate in regulating the broad adaptability of soybean, this study first used gene editing technology to simultaneously knock out three genes from the E1 family and / or two genes from the SOC1 family (SOC1a and SOC1b) in the W82 background. (The specific knockout method of SOC1 refers to the literature "Kou K, Yang H, Li HY, Fang C, Chen LY, Yue L, NanHY, Kong LP, Li XM, Wang F, Wang JH, Du HP, Yang ZY, Bi YD, Lai YC, Dong LD, Cheng Q, SuT, Wang LS, Li SC, Hou ZH, Lu SJ, Zhang YH, Che ZJ, Yu DY, Zhao XH, Liu BH, Kong FJ. (2022) A functionally divergent SOC1 homolog improves soybean yield and latitude.) adaptation.Curr.Biol.,32,1728–1742.e6.”), obtained homozygous five mutants (e1-3msoc1-2m, i.e., e1 e1la e1lb soc1a soc1b five mutants), homozygous three mutants (e1-3m), and homozygous two mutants (soc1-2m), and then planted them in different latitude regions of my country (Keshan, 48°N; Changchun, 44°N; Shijiazhuang, 38°N; Hefei, 31°N; Guangzhou, 23°N; Sanya, 18°N) for field phenotypic observation.

[0032] The results showed that the W82 and SOC1-2m mutants failed to mature in high-latitude regions and experienced reduced yields in low-latitude regions. In contrast, the e1-3m and e1-3m soc1-2m mutants exhibited the same flowering and maturity periods regardless of latitude. However, unlike e1-3m, whose plant height increased with decreasing latitude, the plant height of e1-3m soc1-2m showed less variation across different latitudes. Furthermore, due to its greater branching, its yield remained almost unchanged across different latitudes. Figure 3 (AE). The above studies demonstrate that the E1-SOC1 module plays a crucial role in regulating the broad adaptability of soybean. Furthermore, the e1-3m soc1-2m pentamutator created using gene editing technology in this study exhibits broad adaptability in flowering, maturity, and yield, a first-time report in soybean. Therefore, the E1-SOC1 gene module holds promise for application in molecular design breeding to develop broadly adaptable soybean models.

[0033] In summary, knocking out three genes from the E1 family (E1 and its homologs E1La and E1Lb) and two genes from the SOC1 family (SOC1a and SOC1b) in soybean can breed soybean varieties that exhibit broad adaptability across different latitudes. These new soybean varieties demonstrate broad adaptability in terms of plant height, number of nodes, flowering time, maturity time, and yield, and have significant application prospects in soybean molecular breeding research.

[0034] The embodiments of the present invention have been described in detail above, but the present invention is not limited to the described embodiments. For those skilled in the art, various changes, modifications, substitutions, and variations can be made to these embodiments without departing from the principles and spirit of the present invention, and these variations still fall within the protection scope of the present invention.

Claims

1. The application of the E1-SOC1 gene model in breeding widely adaptable soybean varieties, characterized in that, The E1-SOC1 gene model includes the E1 gene, E1La gene, E1Lb gene, SOC1a gene, and SOC1b gene.

2. The application according to claim 1, characterized in that, The cultivation method involves knocking out the E1-SOC1 gene model in soybeans.

3. The application according to claim 1, characterized in that, The widely adaptable soybean varieties have at least one of the following characteristics: (1) Plant height has wide adaptability in different latitude regions; (2) The number of segments has wide adaptability in different latitude regions; (3) The flowering period has wide adaptability in different latitude regions; (4) It has a wide range of adaptability to different latitudes during its maturity period; (5) The yield has a wide range of adaptability in different latitude regions.

4. The application according to claim 1, characterized in that, The soybeans mentioned include the soybean variety Williams 82.

5. The application according to claim 1, characterized in that, The E1-SOC1 gene model also includes biological materials containing the E1-SOC1 gene model, wherein the biological materials include one or more of recombinant expression vectors, plasmids, expression cassettes, recombinant bacteria, or host cells.

6. A method for breeding widely adaptable soybean varieties, characterized in that, A gene-editing model was developed to knock out the E1-SOC1 gene in soybean. The E1-SOC1 gene model includes the E1 gene, E1La gene, E1Lb gene, SOC1a gene, and SOC1b gene.

Images