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A method for improving the fertility of rice indica and japonica hybrids by using lrk1 gene transformation

A technology of gene transformation and rice, which is applied in the field of genetic modification, can solve the problems of indica and japonica rice that have not yet been seen, and achieve the effects of overcoming the sterility of indica and japonica hybrids, wide compatibility, and high efficiency

Inactive Publication Date: 2016-12-21
FUDAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] So far, there has been no report on the use of this group of LRK genes to improve the fertility of rice indica and japonica hybrids

Method used

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  • A method for improving the fertility of rice indica and japonica hybrids by using lrk1 gene transformation
  • A method for improving the fertility of rice indica and japonica hybrids by using lrk1 gene transformation
  • A method for improving the fertility of rice indica and japonica hybrids by using lrk1 gene transformation

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Experimental program
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Effect test

Embodiment 1

[0034] Example 1 Functional verification of the 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 pCAMBIA1304 vector downstream of the 35S constitutive promoter; the expression vector structure containing the LRK1 gene was 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, is used as the transgenic recipient; the mature seeds of 9311 are 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 particle gun microprojectile bombardment; the resistant cell line was screened by continuous culture on MS medium supplemented with 30ppm hygromycin for 3-4 wee...

Embodiment 2

[0046] Example 2 Method for transforming rice LRK1 gene into indica rice 9311

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

[0048] (2) Culture medium for inducing rice callus

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

[0050] 2) Hyperosmotic 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) Rooting and strong seedling medium: 1 / 2MS+0.1mg / LNAA;

[0055] Note: ①The above mediums all contain 30g / L sucrose + 2.5g / L agar, pH5.8; ②Callus induction, subculture, and screeni...

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Abstract

The invention belongs to the field of transgenic technology, and relates to a method for improving the fertility of indica and japonica rice hybrids by using LRK1 gene transformation; the method uses a particle gun microprojectile bombardment method to transform rice LRK1 gene into indica rice callus, and obtains the expression of LRK1 through recovery culture Genetic rice plants. Experimental results show that the LRK1 gene can be successfully transformed into rice and japonica rice varieties Nipponbare and Akimitsu, respectively, and the F 1 The seed setting rate of the generation was significantly higher than that of the control, reaching the level of the regular variety. The rice LRK1 gene and its homologous gene of the invention are transformed into rice so that they have broad compatibility and can overcome the indica-japonica hybrid sterility. The invention provides a low-cost and high-benefit new method for utilizing the heterosis between indica and japonica rice subspecies.

Description

technical field [0001] The invention belongs to the field of transgenic technology, and relates to a method for improving the fertility of rice indica and japonica hybrids, in particular to a method for improving the fertility of rice indica and japonica hybrids by using LRK1 gene transformation. Background technique [0002] In recent years, there have been many reports on the causes of hybrid sterility among rice subspecies, and the genetic mechanism of the sterility has also been discussed; studies have shown that there are many reasons for intersubspecies hybrid sterility, among which some Hybrid sterility in combinations is caused by pollen abortion, some other combinations show partial abortion of female gametes with a certain allele, and more combinations have both female gametes abortion and Male gametophytes are aborted. A large number of experiments in the prior art have proved that the sterility of indica and japonica hybrids is controlled not only by a single ge...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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