Abiotic stress tolerance conferred by J-domain-containing proteins

A technology of structural domains and proteins, applied in the field of plant genetic engineering, can solve problems such as the undocumented functional role of DnaJ protein in abiotic stress tolerance

Inactive Publication Date: 2009-10-07
THE CHINESE UNIVERSITY OF HONG KONG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] To date, there are no reports documenting the functional role of DnaJ proteins in conferring tolerance to abiotic stress (except heat shock) in plants

Method used

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  • Abiotic stress tolerance conferred by J-domain-containing proteins
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  • Abiotic stress tolerance conferred by J-domain-containing proteins

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Experimental program
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Embodiment approach

[0048] In the present invention, it was found that proteins comprising the conserved J-domain motif of DnaJ can successfully confer tolerance to stress factors such as salinity, low osmotic potential and dehydration in plant cells and plants. Plants and plant cells exhibit this tolerance by means of transgenic modification to contain an expression system that produces this protein. Rice and Arabidopsis plants are exemplified below, but are by no means limited to these examples. Any higher plant or cell of a higher plant is a suitable subject for the methods and materials of the invention.

[0049] The method of the present invention is applicable to any plant, preferably higher plants belonging to the class Angiosperms and Gymnosperms. Plants of the subclass Dicotyledonous and Monocotyledonous are particularly suitable. Bileaved plants include the following orders: Magnoliales, Illiciales, Laurales, Piperales, Aristochiales, Nymphaeales, Ranunculales ), Papeverales, Sarrace...

Embodiment 1

[0065] Expression of GmDNJ1 in response to salinity and dehydration stress

[0066] In this example, expression of GmDNJ1 in leaves and roots of two soybean cultivars in response to salinity stress was investigated. With modified Hoagland's solution (4.5mM KNO 3 , 3.6mM Ca(NO 3 ) 2 , 1.2mM NH 4 NO 3 , 3.0 mM MgSO 4 , 1.2mM (NH 4 ) 2 SO 4 , 0.25mM KH 2 PO 4 , 4.5 μM nSO 4 , 4.5 μM ZnSO 4 , 1.5 μM CuSO 4 , 0.4μM (NH 4 ) 6 Mo 7 o 24 , 0.09 mM Fe-EDTA and 1.5 μM H 3 BO 3 ) were irrigated with two soybean germplasms, Wenfeng7 (salt tolerant) and Union (salt sensitive), treated with 125 mM NaCl. Leaf and root samples were collected 0-144 hours after treatment.

[0067] As previously described (Sambrook, J. et al., Molecular Cloning: A Laboratory Manuals (Molecular Cloning: A Laboratory Manual), 3rd Edition, Cold Spring Harbor Laboratory Press, New York, NY, (2001)) Root and leaf extracts were subjected to Northern blot analysis. Antisense single-stranded DNA...

Embodiment 2

[0072] Transgenic Arabidopsis

[0073] Using the vacuum infiltration protocol (Bechtold, N. et al., Arabidopsis Protocols, Humana Press Inc., Totowa, NJ, (1993) 259-266), The recombinant nucleic acid containing GmDNJ1 under the control of the constitutive cauliflower mosaic virus 35S promoter was cloned into a binary vector (Brears, T. et al., Plant Physiol. (1993) 103:1285-1290), introduced into Agrobacterium and transformed into Arabidopsis. After selection of transformants with antibiotic-containing media, individuals with successful integration of the transgene into the genome were verified by PCR using gene-specific primers; Northern blot analysis was performed to confirm transgene expression in the transgenic plant lines. A T containing a single insert was obtained 3 Seeds of homozygous lines were used for subsequent physiological studies. Four homozygous transgenic lines of Arabidopsis thaliana GmDNJ1 were constructed. A-3-4 and M-3-1 with higher expression levels...

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Abstract

The invention provides methods and constructs for enhancing the expression of nucleotide sequences encoding proteins that comprise at least a DnaJ-type J-domain to provide tolerance to plants or plantcells against abiotic stress, especially dehydration and salinity stress.

Description

technical field [0001] The invention relates to the field of plant genetic engineering. More specifically, the present invention relates to conferring tolerance to abiotic stress conditions such as salinity, osmotic and dehydration stress by enhancing the expression of J-domain containing proteins, especially DnaJ-type proteins sexual method. Background technique [0002] Seed maturation proteins, or late embryogenesis abundant (LEA) proteins, are abundantly produced during the late desiccation stage of seed development. Most LEA proteins accumulate in seeds or vegetative tissues exposed to exogenous abscisic acid or subjected to abiotic stress from salinity or dehydration. Some LEA proteins may confer salt and dehydration tolerance to transgenic plants by hydrating macromolecules, chelating ions, and refolding proteins. [0003] Using suppression subtractive technique (suppression subtractive technique), it was found that the expression of 9 genes under abiotic stress co...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415A01H1/00A01H5/00A01P21/00
CPCC07K14/415C12N15/8273C12N15/821
Inventor 林汉明辛世文邵桂花
Owner THE CHINESE UNIVERSITY OF HONG KONG
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