Method for improving fertility of indica-japonica rice hybrids by virtue of LRK1 gene transformation

A technology for gene transformation and rice, applied in the field of transgenic, can solve the problems such as indica-japonica rice that have not yet been seen, and achieve the effects of overcoming the sterile characteristics of indica-japonica hybridization, low cost, and wide compatibility.

Inactive Publication Date: 2014-02-12
FUDAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0009] So far, there has been no report on the use of this group of L

Method used

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  • Method for improving fertility of indica-japonica rice hybrids by virtue of LRK1 gene transformation
  • Method for improving fertility of indica-japonica rice hybrids by virtue of LRK1 gene transformation
  • Method for improving fertility of indica-japonica rice hybrids by virtue of LRK1 gene transformation

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Experimental program
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Embodiment 1

[0034] Example 1 Functional verification of LRK1 gene

[0035] (1) Construction of LRK1 gene expression vector

[0036] Using pCAMBIA1304 as the expression vector, the Dongxiang wild rice LRK1 gene was inserted into the downstream of the 35S-type promoter of the pCAMBIA1304 vector; the structure of the expression vector containing the LRK1 gene is as follows: figure 1 shown;

[0037] (2) LRK1 gene transformation experiments and results

[0038] 9311, one of the main parents of domestic super hybrid rice, was used as the transgenic receptor; the mature seeds of 9311 were dehulled and sterilized and then inoculated on the dedifferentiated plant tissue culture medium to induce callus; after 2-3 weeks of callus induction, The purified pCAMBIA1304 plasmid DNA containing the LRK1 gene was introduced into 9311 callus cells by the method of microprojectile bombardment; the resistant cell lines were screened by continuous culture on MS medium supplemented with 30 ppm hygromycin for 3...

Embodiment 2

[0046] Example 2 The method of transforming indica rice 9311 with rice LRK1 gene

[0047] (1) Select the full-length ORF segment of LRK1 (see sequence listing), amplify it from Dongxiang wild rice by PCR method, and clone it into the plant expression vector pCAMBIA1304 with a synthetic insert to obtain the LRK1 expression vector pCAMBIA1304 / LRK1;

[0048] (2) Induced rice callus medium

[0049] 1) Induction and subculture medium: MS+2mg / L2,4-D;

[0050] 2) Hypertonic medium: MS+2mg / L2,4-D+46.67g / L sorbitol+46.67g / L mannitol;

[0051] 3) The first round of screening medium: MS+2mg / L2,4-D+30mg / L hygromycin;

[0052] 4) The second round of screening medium: MS+2mg / L2,4-D+50mg / L hygromycin;

[0053] 5) Differentiation medium: MS+3mg / L6-BA+0.5mg / LNAA+50mg / L hygromycin;

[0054] 6) Medium for rooting and strong seedlings: 1 / 2MS+0.1mg / LNAA;

[0055] Note: ①The above medium contains 30g / L sucrose+2.5g / L agar, pH 5.8; ②The culture conditions for callus induction, subculture and sc...

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Abstract

The invention belongs to the field of transgenic technologies, and relates to a method for improving the fertility of indica-japonica rice hybrids by virtue of LRK1 gene transformation. According to the method, rice LRK1 genes are transformed into indica calluses by virtue of a gene gun microprojectile bombardment method, and rice plants for expressing the LRK1 genes are obtained through recovery culture. Experimental results show that the LRK1 genes can be used for successfully transforming rice and hybridizing rice with japonica rice varieties such as Nipponbare and Akihikari respectively, and the F1-generation seed setting rates are greatly higher than the control so as to achieve the seed setting rate level of regular varieties. Rice is provided with wide compatibility and can overcome the infertility characteristic of indica-japonica hybrids after being transformed by the rice LRK1 genes provided by the invention and the homologous genes thereof. The invention provides a low-cost and high-benefit novel method for utilization for the advantages of indica-japonica rice intersubspecific hybrids.

Description

technical field [0001] The invention belongs to the technical field of transgenic, and relates to a method for improving the fertility of rice indica-japonica hybrid, in particular to a method for improving the fertility of rice indica-japonica hybrid by utilizing LRK1 gene transformation. Background technique [0002] In recent years, there have been many reports on the reasons for the sterility of rice inter-subspecies hybrids, and the genetic mechanism of its sterility has also been discussed. Hybrid sterility in the combination is caused by pollen abortion, and in some combinations, the female gamete with a specific allele is partially aborted, and more combinations have both female gamete abortion and Male gametophyte aborted. A large number of experiments in the prior art have proved that the sterility of indica and japonica hybrids is not only controlled by a single gene, but is controlled by multiple sterility genes. [0003] Ikehashi et al. systematically studied ...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N15/29C07K14/415A01H5/00A01H4/00
Inventor 罗小金杨金水孙凡
Owner FUDAN UNIV
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