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Nucleotide sequence for mediating plant male fertility and application method thereof

A nucleotide sequence and nucleotide technology, applied in the field of producing male sterile plants, can solve the problem of not finding MS45 male sterile lines, etc.

Active Publication Date: 2014-03-26
BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the MS45 gene was cloned, this gene was mainly used to restore the fertility of MS45 mutants, and so far there has been no report on the construction of MS45 male sterile lines using RNAi technology

Method used

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  • Nucleotide sequence for mediating plant male fertility and application method thereof
  • Nucleotide sequence for mediating plant male fertility and application method thereof
  • Nucleotide sequence for mediating plant male fertility and application method thereof

Examples

Experimental program
Comparison scheme
Effect test

no. 1 example

[0056] The first embodiment, the synthesis of rZmMS45 nucleotide sequence

[0057] According to the known ZmMS45 nucleotide sequence (as shown in SEQ ID NO: 1 in the sequence listing), the RZmMS45 nucleotide sequence of the present invention is shown in SEQ ID NO: 2 in the sequence listing; the rZmMS45 nucleotide sequence is shown in As shown in SEQ ID NO:3 in the sequence listing, the rZmMS45 nucleotide sequence was synthesized by Nanjing KingScript Biotechnology Co., Ltd.; the 5' end of the synthesized rZmMS45 nucleotide sequence (SEQ ID NO:3) was also A RsrII restriction site is connected, and the 3' end of the rZmMS45 nucleotide sequence (SEQ ID NO: 3) is also connected with a BsaI restriction site.

no. 2 example

[0058] The second embodiment, construction of recombinant expression vector and transformation of recombinant expression vector into Agrobacterium

[0059] 1. Construction of recombinant cloning vector DBN01-T containing rZmMS45 nucleotide sequence

[0060] The synthetic rZmMS45 nucleotide sequence was ligated into the cloning vector pGEM-T (Promega, Madison, USA, CAT: A3600), and the operation steps were carried out according to the instructions of the pGEM-T vector produced by Promega Company to obtain the recombinant cloning vector DBN01-T, which Build process such as figure 1 Shown (wherein, Amp represents the ampicillin resistance gene; f1 represents the replication origin of phage f1; LacZ is the LacZ start codon; SP6 is the promoter of SP6 RNA polymerase; T7 is the promoter of T7 RNA polymerase; rZmMS45 is the nucleoside of rZmMS45 acid sequence (SEQ ID NO: 3); MCS is a multiple cloning site).

[0061] Then, the recombinant cloning vector DBN01-T was transformed into ...

no. 3 example

[0068] The third embodiment, the acquisition and verification of maize plants transferred to the rZmMS45 nucleotide sequence

[0069] 1. Obtaining maize plants transferred to the rZmMS45 nucleotide sequence

[0070] According to the commonly used Agrobacterium infection method, the immature embryos of the aseptically cultured maize variety Zong 31 (Z31) were co-cultured with the Agrobacterium described in 3 in the second example, so that the The T-DNA in the recombinant expression vector DBN100348 (including the rZmMS45 nucleotide sequence, the promoter sequence of the maize Ubiquitin gene, the PMI gene and the Nos terminator sequence) was transferred into the maize genome, and the rZmMS45 nucleotide sequence was obtained maize plants; meanwhile, wild-type maize plants were used as controls.

[0071] For Agrobacterium-mediated transformation of maize, briefly, immature immature embryos are isolated from maize and the immature embryos are contacted with a suspension of Agrobac...

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Abstract

The invention relates to a nucleotide sequence for mediating plant male fertility and an application method thereof. The nucleotide sequence is encoded with an RNA (Ribonucleic Acid) molecule; the RNA molecule is used for inhibiting the expression of plant MS45 genes and comprises at least 19 continuous nucleotides represented by SEQ ID NO:1 and / or complementary sequences thereof. According to the nucleotide sequence for mediating the plant male fertility and the application method thereof, an MS45 male sterility line is constructed by using an RNAi (Ribonucleic Acid interference) technology for the first time; the MS45 male sterility line is used for inhibiting the expression of corn MS45 genes and plants which are completely male sterile can be obtained; the crossbreeding period can be shortened and a current situation that the resource of a current male sterility line is lacked and the crossbreeding purity is not high can be solved pointedly; a castration step in the crossbreeding process is saved and the mechanical loss and the output influences on corn plants by mechanical castration are reduced; the purity and the output of crossbreeding germplasms are guaranteed so that the economic benefits can be realized to the greatest extent.

Description

technical field [0001] The present invention relates to a nucleotide sequence that mediates the male fertility of plants and a method for using it, in particular to a genetic method for producing male sterile plants by using RNAi technology to specifically delete or close the expression of fertility genes. Background technique [0002] Heterosis refers to the performance of F1 hybrids that are superior to those of the F1-producing parents. Heterosis in crop plants is often manifested in production as larger plants, stress tolerance, disease resistance, uniformity and increased yield, collectively referred to as hybrid vigor. [0003] Heterosis has been observed and documented in many important crop species, and the development of hybrids has become routine by plant breeders. However, hybrids can only be used in such plants when an effective and economical method of pollination control exists in the plant to ensure cross-pollination and prevent self-pollination. Mechanisms ...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12N15/82A01H5/00
Inventor 于彩虹李建勇李梅杨进孝陈淑阳
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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