Rice gene OsNF-YC4 and applications thereof

A gene and rice technology, applied in the rice gene OsNF-YC4 and its application fields, can solve the problems of complex regulation mode, complex physiological and biochemical changes, limited functional genes, etc., and achieve wide application prospects, widened grain width, and grain length lengthening effect

Inactive Publication Date: 2017-07-04
SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the complexity of rice's own genetic basis and physiological and biochemical changes under stress, the identified functional genes are relatively limited, and the related molecular mechanisms of expression regulation still need to be further explored
[0004] In fungi and animals, each subunit of NF-Y is encoded by a single gene, while plant NF-Y has multiple homologous genes, which can form different combinations to participate in transcriptional regulation to exert specific biological functions. The pattern may be more complex; there are few reports on the role of NF-Y in plant stress response at this stage

Method used

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  • Rice gene OsNF-YC4 and applications thereof
  • Rice gene OsNF-YC4 and applications thereof
  • Rice gene OsNF-YC4 and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 The cDNA cloning and identification of rice OsNF-YC4 and its coding gene

[0026] The experimental materials were provided by the Research Office of Plant Hormone Regulation Group, South China Botanical Garden, Chinese Academy of Sciences, and the materials were planted in the greenhouse and managed in the conventional field.

[0027] The method of bioinformatics was used to analyze, design primers, and clone the rice gene OsNF-YC4. The specific method is as follows:

[0028] OsNF-YC4 gene ORF forward primer P1 is shown in SEQ ID NO.3.

[0029] OsNF-YC4 gene ORF reverse primer P2 is shown in SEQ ID NO.4.

[0030] Two-week-old young leaves of rice materials were taken, ground in liquid nitrogen, and RNA extraction was carried out according to TIANGEN total RNA extraction kit RNA Plant Extraction kit DP417. cDNA first-strand synthesis was based on TaKaRa reagent company cDNA first-strand synthesis kit TaKaR a PrimeScript TM 1st strand cDNA Synthesis Kit D61...

Embodiment 2

[0032] Utilize the Nde1 and EcR1 endonucleases of Takara Company and the T4-DNA ligase of Takara Company to positively insert the cloned gene OsNF-YC4 of Example 1 into the expression vector Pubi vector, transform E. Positive clones were screened on LB solid medium with L kanamycin. After sequencing verification, the plasmid was extracted to obtain the plant excess Pubi-OsNF-YC4 expression vector ( figure 1 ). Pubi-OsNF-YC4 was transformed into Agrobacterium tumefaciens strain EHA105 (Biovector Co., LTD) by freeze-thaw method. The wild-type rice ZH11 was respectively transformed by Agrobacterium EHA105. Extract the genomic DNA of transgenic rice as a template, use the wild-type rice genome and water as a negative control, and the Pubi-OsNF-YC4 plasmid as a positive control, and use the internal primers of the target gene to perform PCR to initially identify positive transgenic rice. The method is as follows:

[0033] Design internal primers P3: SEQ ID NO.5 and P4: SEQ ID NO...

Embodiment 3

[0036] Example 3 Functional verification of OsNF-YC4 gene

[0037] Disinfect the transgenic rice seeds of the T1 generation and sow them in tissue culture bottles. The culture temperature is 28°C, and the day and night time is 12 / 12h. After the rice grows for 3 weeks, the wild-type ZH11 and overexpressed rice are treated with 40°C high temperature and 10% PEG to simulate drought. Strains OsNF-YC4-OE-8, OsNF-YC4-OE-17, after 10 days, it can be clearly observed that the leaves of overexpressed rice seedlings remain bright green, and the leaves of wild-type seedlings die, such as Figure 4 .

[0038] Further grain statistics of the overexpressed rice lines grown in the greenhouse showed that compared with wild-type ZH11, the rice grain width of the overexpressed lines OsNF-YC4-OE-8 and OsNF-YC4-OE-17 was significantly wider, In addition, the thousand-grain weight of the two overexpression lines reached a very significant level compared with the wild type ZH11, and the stem lengt...

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Abstract

The invention discloses a rice gene OsNF-YC4 and applications of the rice gene OsNF-YC4. The DNA sequence of an open reading frame of the rice gene OsNF-YC4 is shown as SEQ ID NO:1, and the amino acid sequence of protein encoded by the rice gene OsNF-YC4 is shown as SEQ ID NO:2. According to the invention, for the first time, the rice OsNF-YC4 gene is obtained from a rice genome through cloning, the functions of the rice OsNF-YC4 gene are verified, and meanwhile, a plant transgenic vector is utilized for realizing the overexpression of the rice gene OsNF-YC4 in strains OsNF-YC4-OE-8 and OsNF-YC4-OE-17 and the wild type (ZH11). The result shows that the strains with overexpression of the rice gene OsNF-YC4 provided by the invention become high, the grain width and the thousand seed weight are obviously increased, the stress resistance is obviously enhanced, and therefore, the rice gene OsNF-YC4 plays an important role in plant growth and regulation on the stress resistance, plays an important role in researching rice growth and stress resistance mechanism, and has the wide application prospect in crop breeding.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, more specifically, the invention relates to rice gene OsNF-YC4 and its application. Background technique [0002] Higher plants will be affected by abiotic stresses such as high temperature and drought during their development, which will hinder plant growth and even cause death, restricting the development of agriculture and economy. Rice (Oryza.sativa L.) is an important economic and food crop. Among the many factors that affect the growth and yield of rice, high temperature, drought and saline-alkali land hazards are the most important adversity factors. The improvement of yield and quality is of great significance. [0003] With the development of biotechnology, people's research on the mechanism of plant adversity stress is also deepening, and a large number of stress-related genes have been identified and studied. However, due to the complexity of rice's own genetic basi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/82C12N1/21C07K14/415A01H5/00C12R1/01
Inventor 侯兴亮唐伶俐刘旭
Owner SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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