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Soybean phytochrome chromophore synthetic gene GmHY2 as well as encoded protein and application thereof

A phytochrome and synthetic gene technology, applied in the field of molecular biology, can solve problems such as early flowering, limited soybean planting range, and very sensitive photoperiod response

Active Publication Date: 2021-06-15
NORTHEAST INST OF GEOGRAPHY & AGRIECOLOGY C A S
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Soybean is a short-day crop, that is, flowering is significantly earlier under short-day conditions, and its growth and development are very sensitive to photoperiod responses. This characteristic seriously affects the adaptability of soybean varieties and limits the planting range of soybeans.

Method used

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  • Soybean phytochrome chromophore synthetic gene GmHY2 as well as encoded protein and application thereof
  • Soybean phytochrome chromophore synthetic gene GmHY2 as well as encoded protein and application thereof
  • Soybean phytochrome chromophore synthetic gene GmHY2 as well as encoded protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1: Isolation and structural analysis of GmHY2 gene in soybean

[0046] (1) Isolation of GmHY2 gene

[0047] Total RNA was extracted from soybean variety Williams 82, and the first strand of cDNA was synthesized using total RNA as a template and Oligo(T)17 as a primer. Using the first strand of the cDNA as a template, the forward primer (5'-ATGGGTTTTAGAATTAGCGGT-3') and the reverse primer (5'-CCTAAAATATTCAATAAATTCTCCTC-3') were used for PCR amplification, respectively. A 990bp GmHY2 gene cDNA fragment was obtained, and the fragment was connected with the pEASY-Blunt vector (Quanshijin Company) and named Blunt-GmHY2.

[0048] The gene sequence of the obtained GmHY2 gene is shown in SEQ ID NO: 3, and the total length of the GmHY2 gene CDS is 990 bp; the amino acid sequence of the protein encoded by the gene is shown in SEQ ID NO: 2, with a total of 329 amino acids.

[0049] (2) GmHY2 gene structure analysis

[0050] Extract DNA from the young leaves of soybean v...

Embodiment 2

[0051] Example 2: Isolation, screening and phenotypic identification of soybean GmHY2 gene mutants

[0052] Taking Williams 82 as the background, the present invention constructed a soybean mutant library through EMS mutagenesis. Using this platform, non-synonymous mutations in the GmHY2 protein of 4 strains were screened, and they were named Gmlh1-1, respectively. 2, 3, 4. The nucleotide sequence of the Gmlh1-1 mutant is that the 329th A in the nucleic acid sequence shown in SEQ ID NO:3 is mutated to G, and the mutation causes mis-cutting of the transcript, and its encoded protein sequence is SEQ ID NO Frameshift mutation begins to occur at the 110th position in the amino acid sequence shown in: 2; its nucleotide sequence of the Gmlh1-2 mutant is that the G mutation at the 763rd position in the nucleotide sequence shown in SEQ ID NO: 3 is A, Its encoded protein sequence is that the 255th glycine (G) in the amino acid sequence shown in SEQ ID NO:2 is mutated into glutamic aci...

Embodiment 3

[0057] Example 3. Construction of soybean GmHY2 gene sense expression vector

[0058] Extract the DNA of soybean Williams 82 leaves, amplify the promoter of GmHY2 (shown in SEQ ID NO: 4) and the full length of the gene (shown in SEQ ID NO: 1), and connect it forward to the plant expression vector pCAMBIA3301T (CAMBIA Research Center) In this method, the positive expression vector of GmHY2 was obtained, and the full length of the vector was 18.8Kbp (such as Figure 6 ), the resistance in Escherichia coli is kanamycin resistance, and the resistance in plants is glufosinate-ammonium (Bar) resistance.

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Abstract

The invention relates to the technical field of molecular biology, and provides a soybean phytochrome chromophore synthetic gene GmHY2, an encoded protein thereof and functions of the gene and the encoded protein. According to activity levels of the gene and the encoded protein thereof, leaf chlorophyll content and internode length can be regulated, and the gene and the encoded protein can also be used for regulating photoperiod sensitivity of plants, namely regulating flowering time, can be used for solving the problem of flowering asynchronism in cross breeding, and can provide germplasm resources and theoretical support for breeding eurytopic soybean varieties.

Description

technical field [0001] The invention relates to the technical field of molecular biology, and more specifically relates to soybean phytochrome chromophore synthesis gene GmHY2 and its encoded protein and application. Background technique [0002] Light is an important environmental factor in plant growth and development, and light can regulate the photosynthetic efficiency and photoperiod response of plants. Photoperiod response is an important regulatory link affecting flowering and maturation of plants, which directly affects the yield, quality and adaptability of plants. Soybean is a short-day crop, that is, flowering is significantly earlier under short-day conditions, and its growth and development are very sensitive to photoperiod responses. This characteristic seriously affects the adaptability of soybean varieties and limits the planting range of soybeans. Soybean planted in northern my country has a long growth period and a late maturity period, so changing the pho...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/00C12N15/52C12N15/82A01H5/00A01H5/12A01H6/54
CPCC12N9/00C12N15/825C12N15/8261Y02P60/14
Inventor 杨素欣张芷睿于慧王秋实
Owner NORTHEAST INST OF GEOGRAPHY & AGRIECOLOGY C A S
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