Vascular plants expressing Na+ pumping ATPases

A plant and vascular technology, applied in plant products, plant genetic improvement, botanical equipment and methods, etc., can solve problems such as non-persistent production

Inactive Publication Date: 2007-12-12
植物功能基因组PTY澳大利亚中心有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, despite the improved Na + Tolerant cells can be selected in vitro, but robust Na + Tolerant plants

Method used

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  • Vascular plants expressing Na+ pumping ATPases
  • Vascular plants expressing Na+ pumping ATPases
  • Vascular plants expressing Na+ pumping ATPases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0147] Cloning of Physcomitrella patens ENA1, ENA2 and Saccharomyces cerevisiae ENA1 cDNAs

[0148] The full-length ENA1 and ScENA1 cDNAs in the cloning vectors pCR2.1-TOPO (Invitrogen) and pJQ10, respectively, can be used such as Benito, B., and Rodriguez-Navarro, A. (2003), The Plant Journal 36 : 382-389 and Benito et al. (1997) Biochimica et Biophysica Acta 1328(2) Cloned by the method described in :214-26. The cDNAs were obtained from Alonso Rodriguez-Navarro.

[0149] The nucleotide sequence of Physcomitrella patens ENA1 cDNA is provided in GenBank Accession No. AJ564254, labeled as SEQ ID NO.1. The cDNA encodes a 967 amino acid Na + Pump ATPase, labeled as SEQ ID NO.2.

[0150] The nucleotide sequence of Saccharomyces cerevisiae ENA1 cDNAs is provided in GenBank Accession No. AJ564254, labeled as SEQ ID NO.12. The cDNA encodes a 1091 amino acid Na + Pump ATPase, labeled as SEQ ID NO.13.

[0151]Briefly, cDNAs representing the entire open reading frames of the PpEN...

Embodiment 2

[0161] Generation of Mutant Physcomitrella patens Deleting PpENA1 and / or PpENA2

[0162] Restriction fragments of PpENA1 or PpENA2 can be transferred from the cloned DNA into an appropriate vector such as pGEM-T Easy (Promega). A knockout cassette is then generated by inserting a selectable marker such as a gene that confers resistance to kanamycin, hygromycin, or the herbicide basta, into the full-length gene encoding PpENA1 or PpENA2. This cassette consists of sequences homologous to the PpENA1 or PpENA2 upstream or downstream selectable markers. Anti-G-418 resistance was obtained with the nptll gene behind the 35S-promoter from the pJIT145-Kan plasmid (Figure 6). Hygromycin resistance was obtained using the Hyg gene behind the 35S-promoter from the T-Easy 35S-Hyg plasmid (Figure 7).

[0163] Mutant mosses can be produced by transformation. Protoplasts are generated by treating protonema tissue with an enzyme that removes the cell wall. The protoplasts were treated with ...

Embodiment 3

[0166] Generation of mutant Physcomitrella patens overexpressing PpENA1 and / PpENA2

[0167] A full-length clone of PpENA2 can be obtained by designing primers specific to the 5' and 3' ends of the genomic sequence and performing PCR using cDNA as a template. A suitable overexpression vector is the pTOOL2 vector, as shown in FIG. 1 . This construct can then be used to transform mosses (as described above) and selected mutants (as described above) that overexpress PpENA1 or PpENA2 (or both).

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Abstract

The present invention relates to a vascular plant including cells expressing a Na<+> pumping ATPase.

Description

technical field [0001] The present invention relates to the expression of Na + A vascular plant that pumps ATPase and cells derived from the vascular plant. [0002] The present invention also relates to the expression of Na in vascular plants or cells + Pumps ATPase to increase Na in vascular plants and cells from vascular plants + Secretion and Na + method of tolerance. Background technique [0003] High concentrations of salt in the soil are responsible for greatly reduced yields of many crops around the world. Almost 1 billion hectares of land are affected by soil salinity, accounting for 7% of the total land area. Of the 1.5 billion hectares currently planted, approximately 5% (77 million hectares) are affected by salt. With current farming practices that lead to salinization of water supplies, the problem of soil salinity is only likely to get worse. [0004] Salt solutions have both ionic and osmotic stress effects on plants. These stresses can be discerned at...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H5/00C12N15/29C12N15/55
CPCC12N15/8273
Inventor 安德鲁·雅各布克里斯蒂娜·伦德马克·泰斯特
Owner 植物功能基因组PTY澳大利亚中心有限公司
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