A method of promoting plant growth

A technology for plant expression vectors and seeds, applied in the fields of RNA epigenetics and RNA methylation modification

Active Publication Date: 2020-12-01
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] The use of transgenic technology to introduce exogenous genes to change plant traits has been studied for a long time, but so far, none of the exogenous genes studied, reported or patented involves changing RNA methylation modification m 6 A gene

Method used

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  • A method of promoting plant growth
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  • A method of promoting plant growth

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Introduction of hFTO gene into Arabidopsis thaliana (Ecotype Columbia) by transgenic technology

[0041] The transgenic technology that the present invention adopts is the conventional Agrobacterium-mediated method used in the laboratory, and the brief experimental process is as follows:

[0042] 1. Construction of plant expression vector containing human FTO (hFTO) gene

[0043] Using subcloning technology, the hFTO gene was constructed into a plant expression vector (pCAMBIA1307, containing CaMV35S promoter, kanamycin and hygromycin resistance).

[0044] The plant expression vector pCAMBIA1307 is a CaMV35S promoter, resistant to kanamycin and hygromycin, and the hFTO gene is inserted into BamHI and SalI by subcloning technology.

[0045] a) Duplicate the hFTO gene into an insert with BamHI and SalI restriction endonucleases by PCR

[0046] The PCR primers used are respectively the following sequences (as shown in SEQ ID NO: 1-2):

[0047] hFTO-p1 (with ...

Embodiment 2

[0065] Example 2 Screening of Transgenic Arabidopsis and Identification of Homozygous Lines

[0066] 1. Screening of transgenic Arabidopsis T0 generation

[0067] Disinfect the collected T0 generation seeds with sodium hypochlorite and plant them on hygromycin-resistant MS medium plates. After vernalization, culture them under normal conditions. Yellowing can be seen in most seedlings in about 2 weeks. , only a few seedlings did not yellow, and the seedlings that did not yellow may be positive plants.

[0068] 2. Screening of T1 generation of positive plants

[0069] Transplant the above-mentioned few seedlings into flower pots and grow under normal conditions. After the seedlings grow up and pod, the seeds are collected, and the T2 generation is screened, as in the previous step.

[0070] 3. Identification of the T2 generation of homozygous lines

[0071] Transplant the T2 generation seedlings into flower pots and grow under normal conditions. After growing up, collect t...

Embodiment 3

[0072] Example 3 hFTO promotes the growth of Arabidopsis

[0073] Seeds of the T2 generation homozygous lines identified in Example 2 were planted on normal MS medium. After identification and observation, compared with the pCAMBIA1307 empty vector control group, hFTO transgenic Arabidopsis grew faster, had longer root systems, and grew more Larger, please refer to the attached picture for details: figure 1 It is the test result of the 10th day of root growth (the medium plate is placed obliquely during the experiment), and the root length of the seedlings in the transgenic group is 2-3 times that of the control group. figure 2 and image 3 They are photographed from the top and side of the plant, respectively, and are the test results on the 28th day (seeds were transplanted into soil after growing on MS medium for 10 days). From figure 2 It can be clearly seen that the rosette leaves of the transgenic plants are much larger than those of the control group. image 3 ...

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Abstract

The invention relates to a method for promoting plant growth. The method comprises the following steps: constructing a plant expression vector comprising FTO or other homologous demethylated protease gene; converting the plant expression vector into acceptor bacteria; converting the acceptor bacteria into target plants; culturing the obtained target plants, collecting and culturing the T0 generation seeds, screening positive plants from the seedlings; culturing the screened positive plants, after the growth of the seedlings, collecting the T1 generation seeds; culturing the T1 generation seeds, choosing positive plants from the seedlings; culturing the selected positive plants, after the growth of the seedlings, collecting the T2 generation seeds; culturing the T2 generation seeds, screening homozygous strain lines from the seedlings; and culturing the T1 generation seeds of the plants of the T2 generation homozygous strain lines so as to obtain the target plants. An exogenous gene that can modulate the RNA methylation modification of m6A in a cell is introduced into a plant through a transgene technology so as to modulate the m6A on the RNA of a plant in order to promote the plant growth.

Description

technical field [0001] The present invention relates to the fields of RNA epigenetics and RNA methylation modification, in particular to regulating the growth of plants by regulating the methylation modification on mRNA, other nuclear RNA or non-coding RNA in plants. Background technique [0002] More than 100 kinds of chemically modified bases have been found in various RNAs, including ribosomal RNA (rRNA), transfer RNA (tRNA), messenger RNA (mRNA) and small nuclear RNA (snRNA), to help complete various RNA function. In living organisms, the main type of modification is methylation modification. Among them, the methylation modification on the mRNA of eukaryotes is mainly: N7-methylguanine (m 7 G), the 2'-O-methylated base located at the two bases at the 5' end or the middle part (N m ), 5-methylcytosine (m 5 C) and N6-methyladenine (m 6 A). [0003] The most widely distributed eukaryotic mRNA is m 6 A, derived from methyltransferase complex at the N6 position of aden...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/84C12N15/52C12N9/00A01H5/00A01H6/20
Inventor 贾桂芳何川段洪超
Owner PEKING UNIV
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