Transcription factor for improving plant seed aliphatic acid content and application thereof

A technology of plant seeds and fatty acids, applied in the direction of plant genetic improvement, application, plant peptides, etc., to achieve the effect of increasing the content of fatty acids in seeds

Active Publication Date: 2012-11-21
CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

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Method used

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  • Transcription factor for improving plant seed aliphatic acid content and application thereof
  • Transcription factor for improving plant seed aliphatic acid content and application thereof
  • Transcription factor for improving plant seed aliphatic acid content and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0083] Embodiment 1, the acquisition of the coding gene GhWRI1 of cotton transcription factor GhWRI1

[0084] 1. Total RNA extraction: Upland cotton (Gossypium hirsutum L.) variety Coker201 (Wu Xiuming, Liu Chuanliang, Zhang Chaojun, Li Fuguang. Research progress in cotton somatic embryogenesis. Bulletin of Botany, 2008,25(4):469-475 ) as the experimental material, the immature embryo material 20 days after flowering was taken, quick-frozen in liquid nitrogen, and CTAB method was applied (references: Liu Yang, He Xinyao, Ma Hongbo, Wu Yongli, Yang Youming. The total RNA of each tissue of cotton was extracted by CTAB-PVP method Research. Journal of China Agricultural University, 2006, 11(1):53-56) to extract RNA.

[0085] 2. Use Promaga's RT-PCR reaction system to reverse transcribe the RNA extracted in step 1 to synthesize cDNA.

[0086] 3. RT-PCR amplification:

[0087] According to the EST information of cotton in the NCBI database, spliced ​​into contig, the following prime...

Embodiment 2

[0095] Example 2, Functional Analysis of Transcription Activation of GhWRI1 Transcription Factor

[0096] In this example, pGBT9 (Meng Hongyan, Du Xiongming, Zhang Chunyi, etc. Cloning of cotton transcription factor gene GhMS3 and identification of its promoter function. Chinese Agricultural Sciences, 2010, 43 (17)) was used as the original vector to construct the following three expression vectors ( figure 2 Middle A): (1) Recombinant expression vector pGBT-GhWRI1 containing the full-length CDS of the GhWRI1 gene (sequence 1, referred to as GhWRI1-CDS); (2) containing the N-terminal of the GhWRI1 gene (1-654 core (GhWRI1-N for short) recombinant expression vector pGBT-GhWRI1-N; (3) recombinant expression vector pGBT containing the C-terminal of the GhWRI1 gene (nucleotides 661-1317 of sequence 1, GhWRI1-C for short) -GhWRI1-C. In order to analyze the transcription activation function of GhWRI1 transcription factor.

[0097] 1. Construction of recombinant expression vector...

Embodiment 3

[0112] Embodiment 3, expression analysis of GhWRI1 gene

[0113] Upland cotton (Gossypium hirsutum L.) varieties 09F9083, 09F9077, 09F9077, 09E1021 and 09E1022 with different oil contents were extracted (all varieties were purchased from Hebei Jifeng Seed Industry Co., Ltd.; the oil content of seeds was extracted by Soxhlet The results are shown in Table 1) The RNA of immature embryos 20 days after flowering was reverse-transcribed into cDNA by Promaga's RT-PCR reaction system, and the primer 5'-GGCATAGATGGACTGGGAGAT-3' (the 251st part of Sequence 1) was designed for the GhWRI1 gene -271 position) and 5'-GCTGCTGCTTCCTCTTGTGT-3' (reverse complementary sequence of positions 637-656 of Sequence 1), Ghactin was used as an internal reference gene, and primers 5'-ATCCTCCGTCTTGACCTTG-3' and 5'-TGTCCGTCAGGCAACTCAT-3 were designed ', using semi-quantitative RT-PCR to analyze the expression of GhWRI1 gene in oily materials.

[0114] The result is as Figure 4 As shown, the expression ...

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Abstract

The invention discloses a transcription factor related to the improvement of plant seed aliphatic acid content, a coding gene and an application thereof. The transcription factor provided by the invention is (a) or (b) as follows, wherein (a) is any protein fragment which has a carboxyl terminal that at least contains the 221st-438th amino acid sequence of a sequence 2 in a sequence table, has an amino terminal extending from the 221st of the sequence 2 along the amino acid sequence direction of the sequence 2, and has a length of 218-438 amino acids; the protein fragment has transcription activity; (b) is protein which is obtained by substitution and/or deletion and/or addition of one ore more than one amino acid residues in the amino acid sequence defined by (a), has transcription activity, or is derived from the amino acid sequence that is related to the improvement of plant seed aliphatic acid content and is defined by (a). The transcription factor GhWRI 1 provided by the invention can significantly improve the plant seed aliphatic acid content, increase the thousand seed weight, increase the size of seed cells, and has good application prospects.

Description

technical field [0001] The invention relates to a transcription factor related to increasing the fatty acid content of plant seeds, its encoding gene and application, in particular to a transcription factor derived from cotton and related to increasing the fatty acid content of plant seeds, its encoding gene and application. Background technique [0002] Cotton is an important economic crop and oil crop in my country. my country has 6 million hectares of cotton planted all the year round, with an output of about 7.6 million tons of lint and 13 million tons of cottonseed. As the main by-product of cotton production, cottonseed is rich in oil and protein (Xu Hongxia, Yang Weihua, Wang Yanqin, Zhou Dayun, Kuang Meng, Feng Xinai. Analysis of the quality of cottonseed for oil in my country. China Cotton, 2009,36(7):2-3) , the fat content of cottonseed after shelling is 30% to 40%, which can produce oil. In recent years, with the increasing shortage of oil resources and energy re...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/10C12N1/19C12N1/21C12N15/82C12N15/84A01H5/00
Inventor 华金平刘正杰张园赵鹏赵清翠李玉华王玉美
Owner CHINA AGRI UNIV
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