Use of soybean qne1 gene

Abstract

The application of soybean QNE1 gene relates to the field of genetic engineering, and particularly relates to a new application of the soybean QNE1 gene. The purpose of the present application is to provide a new function of the soybean QNE1 gene. The present application determines the regulation of the soybean QNE1 gene on the petiole angle of soybean by overexpressing the soybean QNE1 gene in soybean, and the overexpression of the QNE1 gene makes the petiole angle of soybean smaller. The soybean plant with smaller petiole angle is beneficial to optimizing the close planting of soybean, thereby improving the yield of the soybean plant. Therefore, the soybean QNE1 gene can be applied to optimizing the petiole angle of soybean and changing the plant structure of soybean to improve the yield through molecular or genetic engineering breeding.

Description

Technical Field

[0001] This invention relates to the field of genetic engineering, specifically to soybeans. QNE1 New applications of genes. Background Technology

[0002] Soybean (Glycine max) is one of the most important food and oilseed crops, providing humans with edible oils and proteins, and is a major renewable feedstock for global biodiesel production. With the rapid increase in human consumption and industrial use, the demand for soybean yields has increased significantly and will continue to increase. Therefore, improving soybean yield potential has become a long-term breeding goal. Soybean yield potential can be improved by creating an ideal plant structure, which is mainly determined by the leaf and stem composition. The petiole angle (LPA), the inclination between the petiole and stem, is particularly important in determining the plant's plant architecture; a smaller LPA is crucial for optimizing planting density and increasing yield.

[0003] soybeans QNE1 The gene encodes a transcription factor, which is located in the cell nucleus. QNE1 Genes that delay soybean flowering are present, but so far, few genes have been reported that regulate soybean petiole angle. The molecular mechanism regulating soybean petiole angle is not yet fully understood, and the regulatory network among various genes remains unclear. Further exploration of genes related to soybean petiole angle regulation is of great significance for a deeper understanding of the regulatory mechanism of petiole angle, thereby enabling effective control of soybean plant architecture. Summary of the Invention

[0004] The purpose of this invention is to provide soybeans QNE1 New functions of genes.

[0005] This invention provides soybeans QNE1 Application of genes in regulating soybean petiole angle.

[0006] The soybean QNE1 The nucleotide sequence of the gene is shown in SEQ ID NO: 1 in the sequence listing.

[0007] The soybean QNE1 The amino acid sequence of the gene-encoded protein is shown in the sequence listing SEQ ID NO: 2.

[0008] Furthermore, the soybeans QNE1 Genes are introduced into soybean cells, tissues, or organs, and then the transformed soybean cells, tissues, or organs are cultivated into plants, enabling the... QNE1 When the gene is expressed in soybeans, soybean plants with smaller petiole angles are obtained.

[0009] Furthermore, the soybeans QNE1 Genes are used to optimize high-density planting of soybeans.

[0010] Furthermore, the soybeans QNE1 Genes are used to increase soybean yield.

[0011] The beneficial effects of this invention are: This invention uses soybeans QNE1 The gene was overexpressed in soybean, which determined the soybean QNE1 The regulatory role of genes on soybean petiole angle QNE1 Gene overexpression reduces the petiole angle in soybeans. Obtaining soybean plants with smaller petiole angles is beneficial for optimizing soybean planting density, thereby increasing soybean yield.

[0012] Therefore, soybeans QNE1 Genes can be used to optimize soybean petiole angle and alter soybean plant structure through molecular or genetic engineering breeding to increase yield. Attached Figure Description

[0013] Figure 1 for QNE1 RT-qPCR analysis of genes in overexpressed transgenic soybeans and non-transgenic soybeans; Figure 2 for QNE1 Photographs of petiole angle phenotypes in transgenic and non-transgenic soybeans overexpressing the gene; Figure 3 for QNE1 Statistical analysis of petiole angle phenotype data in transgenic and non-transgenic soybeans overexpressing genetically modified soybeans; Figure 4 for QNE1 CT scan results of the petiole angle microstructure of overexpressing transgenic soybeans and non-transgenic soybeans. Detailed Implementation

[0014] The technical solution of the present invention is not limited to the specific embodiments listed below, but also includes any combination of the specific embodiments.

[0015] Specific Implementation Method 1: Soybeans in this Implementation Method QNE1 Application of genes in regulating soybean petiole angle.

[0016] This invention uses soybeans QNE1 The gene was overexpressed in soybean, which determined that soybean QNE1 The regulatory role of genes on soybean petiole angle QNE1 Gene overexpression reduces the petiole angle in soybeans. Obtaining soybean plants with smaller petiole angles is beneficial for optimizing soybean planting density, thereby increasing soybean yield.

[0017] Specific Implementation Method Two: This implementation method differs from Specific Implementation Method One in that the soybeans... QNE1The nucleotide sequence of the gene is shown in SEQ ID NO: 1 in the sequence listing. Everything else is the same as in Specific Implementation Method 1.

[0018] Specific Implementation Method Three: This implementation method differs from Specific Implementation Method One in that the soybeans... QNE1 The amino acid sequence of the gene-encoded protein is shown in SEQ ID NO: 2. Everything else is the same as in Specific Implementation Method 1.

[0019] Specific Implementation Method Four: This implementation method differs from Specific Implementation Methods One to Three in that: the soybeans... QNE1 Genes are introduced into soybean cells, tissues, or organs, and then the transformed soybean cells, tissues, or organs are cultivated into plants, enabling the... QNE1 The gene is expressed in soybeans, resulting in soybean plants with smaller petiole angles. Other aspects are the same as in any of the specific implementation methods one to three.

[0020] Specific Implementation Method Five: Soybeans described in this implementation method QNE1 Genes are used to optimize high-density planting of soybeans.

[0021] Specific Implementation Method Six: Soybeans described in this implementation method QNE1 Genes are used to increase soybean yield.

[0022] The embodiments of the present invention will be described in detail below. The following embodiments are implemented based on the technical solution of the present invention, and detailed implementation schemes and specific operation processes are given. However, the protection scope of the present invention is not limited to the following embodiments.

[0023] Example 1: Soybeans QNE1 Construction of gene overexpression vectors Using cDNA from Heinong 51 soybean plant leaves as a template, QNE1-F1 and QNE1-R1 Using these as primers, PCR amplification was performed using the high-fidelity enzyme FastPfu Fly DNA Polymeras purchased from Beijing TransGen Biotechnology Co., Ltd. PCR reaction conditions were as follows: 94℃ pre-denaturation for 5 min, followed by 6 cycles of (94℃ denaturation for 30 s, 58℃ annealing for 30 s, 72℃ extension for 60 s), then 94℃ denaturation for 30 s, 68℃ annealing for 30 s, 72℃ extension for 60 s, for 26 cycles, followed by a final extension at 72℃ for 5 min. Gel recovery was performed using the Wizard® SV Gel and PCR Clean Up System purchased from Promega. The desired DNA was obtained through PCR amplification. QNE1 Gene amplification bands ( QNE1 The nucleotide sequence of the gene is shown in SEQ ID NO.1 of the sequence listing, and it was amplified by this PCR. QNE1 upstream introduction of genesBamH The restriction enzyme site I is introduced downstream. Sac The restriction enzyme site of I. The amplified PCR product and the vector pBA:35S:6×cmyc (this vector has been published in the paper Wenfeng Ning, et al. 2017) were simultaneously subjected to... BamH Ⅰ and Sac After double digestion, the DNA was ligated overnight using T4 DNA ligase, transformed into competent E. coli cells, and pBA:35S:6×cmyc: was constructed. QNE1 Vector. pBA:35S:6×cmyc: QNE1 The carrier contains Bar Selection marker gene (glufosinate resistance), driven by the 35S promoter QNE1 Gene.

[0024] The constructed vector pBA:35S:6×cmyc: QNE1 The bacteria were transferred into the genome of soybean variety Williams 82 via Agrobacterium tumefaciens-mediated transformation of cotyledonary nodes. QNE1 Gene overexpression transgenic soybean lines.

[0025] Example 2: QNE1 Molecular identification of transgenic soybeans with overexpressed genes 1. Through genetic transformation of soybean, using a 160 mg / L glufosinate application experiment, two transformation events (35S:QNE1#L1, 35S:QNE1#L2) were obtained in the T1 generation to acquire glufosinate resistance. RNA was extracted from mature soybean leaves of these two transformation events. The RNA extraction method is described in the Invitrogen TRIzol kit manual.

[0026] 2. The total RNA extracted in step 1 was treated with DNase. The RNA content and quality of the treated total RNA were determined using NanoDrop, purchased from Toyobo.

[0027] 3. Following step 2, 2µg of total RNA was collected after reaching the standard using NanoDrop assay. This was then analyzed using TransScript RNA PCR technology purchased from Beijing TransGen Biotech Co., Ltd. ® One-Step gDNA Removal and cDNA Synthesis SuperMix Kit, and the dT included with the kit 18 Primers were used to synthesize cDNA according to the kit's instruction manual.

[0028] 4. Take 1 µl of the cDNA diluted 30 times in step 3 as a qPCR template, and use q QNE1-F1 and qQNE1-R1As primers, soybean internal reference gene TUA5 The primer sequences are qTUA5 The TransStart Top Green qPCR Super Mix Kit was used according to the manufacturer's instructions, employing SYBR Green I as the dye and a three-step PCR reaction. The qPCR reaction conditions were: 95℃ pre-denaturation for 30 s, 95℃ denaturation for 5 s, 60℃ annealing for 15 s, 72℃ extension for 20 s, 40 cycles, using 2... –ΔΔCT Algorithm, computation QNE1 The relative expression levels of genes, the results are as follows: Figure 2 As shown, compared to non-GMO soybeans, QNE1 Gene expression levels were significantly higher in both independent transformation events than in non-GMO soybeans. QNE1 The gene was successfully overexpressed in soybeans.

[0029] Example 3: QNE1 Phenotypic analysis of transgenic soybeans with gene overexpression T2 generation QNE1 Gene-overexpressing transgenic soybeans and the non-transgenic control variety Williams 82 were sown in pots, with 10 plants per line, and planted under natural light (long day). A detailed phenotypic analysis of the plants was conducted. The petiole angles of both the non-transgenic control variety Williams 82 and the overexpressing transgenic soybean were observed using a soil microstructure scanning CT system (an existing system at our facility). QNE1 Photographs of the petiole angle phenotypes of overexpressing transgenic soybeans and non-transgenic soybeans are shown below. Figure 2 As shown, QNE1 Statistical data on petiole angle phenotypes of overexpressed transgenic soybeans and non-transgenic soybeans are as follows: Figure 3 As shown, QNE1 CT scan results of the petiole angle microstructure of overexpressing transgenic soybeans and non-transgenic soybeans are as follows: Figure 4 As shown. The results show that, QNE1 The petiole angle of transgenic soybeans with overexpressed genes was significantly reduced.

[0030] By soybeans QNE1 Genes are introduced into soybean cells, tissues, or organs, and then the transformed soybean cells, tissues, or organs are cultivated into plants, enabling the... QNE1 The gene is expressed in soybeans, resulting in soybean plants with smaller petiole angles. This is beneficial for optimizing dense planting of soybeans, thereby increasing soybean yield.

Claims

1. Soybeans QNE1 The application of genes in regulating soybean petiole angle is characterized by, soybeans QNE1 Gene overexpression reduces the petiole angle of soybean; the soybean QNE1 The amino acid sequence of the gene-encoded protein is shown in the sequence listing SEQ ID NO:

2.

2. The application according to claim 1, characterized in that... The soybean QNE1 The nucleotide sequence of the gene is shown in SEQ ID NO: 1 in the sequence listing.

3. The application according to claim 1, characterized in that... The soybean QNE1 Genes are introduced into soybean cells, tissues, or organs, and then the transformed soybean cells, tissues, or organs are cultivated into plants, enabling the... QNE1 When the gene is expressed in soybeans, soybean plants with smaller petiole angles are obtained.

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