Constitutively active pyr/pyl receptor proteins for improving plant stress tolerance

A technology of stress resistance and plants, applied in the direction of chemicals for biological control, measurement/inspection of microorganisms, angiosperms/flowering plants, etc., can solve unclear problems

Inactive Publication Date: 2017-02-22
RGT UNIV OF CALIFORNIA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Several ABA-binding proteins have been reported, however, it is unclear how they regulate the various effects of ABA because they do not act through known modulators of signal transduction pathways (X. Liu et al., Science 315, 1712 (Mar 23, 2007); F.A. Razem, A. El-Kreamy, S.R. Abrams, R.D. Hill, Nature 439, 290 (2006); Y.Y. Shen et al., Nature 443, 823 (Oct 19, 2006))

Method used

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  • Constitutively active pyr/pyl receptor proteins for improving plant stress tolerance
  • Constitutively active pyr/pyl receptor proteins for improving plant stress tolerance
  • Constitutively active pyr/pyl receptor proteins for improving plant stress tolerance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0277] Example 1: PYR / PYL regulates ABA signal transduction

[0278] Unlike biochemical screens for ABA-binding proteins, genetic analyzes focused on ABA sensing have not yet identified protein-assembling receptors, suggesting that receptors may be functionally redundant, have overlapping functions, or fail to mutate to produce viable gametes or seedlings (P . McCourt, Annual Review of Plant Physiology and Plant Molecular Biology 50, 219 (1999)). As a complementary approach, we adopted a chemogenetic strategy in plants (Y. Zhao et al., Nat ChemBiol 3, 716 (2007)). This method is suitable for organisms with highly redundant genomes, because the variable selectivity of small molecules can produce phenotypes that cannot be manifested by single gene mutations (N. Raikhel, M. Pirrung, PLANT PHYSIOLOGY138, 563 (2005); S. Cutler, P. McCourt, Plant Physiol. 138, 558 (2005)). For example, low-sensitivity antagonists can interfere with the function of entire protein families (as obser...

Embodiment 2

[0304] Example 2: Screening of PYR / PYL agonists

[0305] We next investigated whether other compounds besides ABA and parebactin could act as agonists of PYR / PYL proteins. Yeast two-hybrid strains expressing the ABA receptor and type 2 protein C phosphatase in a suitable vector can be used to monitor the activity of the ABA receptor. Thus, these yeast strains create a convenient screening system for the identification of penetrant compounds that are ABA agonists, ie, compounds that promote the binding of PYR / PYL family members to their target protein phosphatases. When the PYR / PYL protein binds to the target PP2C in yeast two-hybrid, the reporter gene is activated, which, depending on the strain used, can result in the expression of a reporter construct such as a LacZ / β-galactosidase marker or in the Expression of nutritional reporter genes grown on auxotrophic media.

[0306] For these agonist assays, screening compounds are added to microwell plates and the appropriate yea...

Embodiment 3

[0309] Example 3: Phenotypic Analysis of PYR / PYL Overexpression and Loss of Function in Mutant Strains

[0310] Abscisic acid is a multifunctional plant hormone that is involved in a variety of plant protective functions, including bud dormancy, seed dormancy and / or ripening, leaf and fruit abscission, and in response to various biotic stresses (e.g., cold, heat, ). According to independent of CO 2 As a mechanism of concentration, ABA is also responsible for regulating stomatal closure. Since PYR / PYL receptor proteins regulate ABA signaling, these phenotypes can be modulated by modulating the expression of PYR / PYL. However, as discussed above, experiments with individual, triploid, and tetraploid Pyr / Pyl mutants showed that the PYL receptors PYL1, 2, and 4 are functionally redundant with PYR1 under the control of ABA response. In these experiments, we wondered whether other PYR / PYL receptors were functionally redundant with PYR1 in the control of plant protection functions...

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Abstract

The invention relates to constitutively active PYR / PYL receptor proteins for improving plant stress tolerance. The invention provides an expression box, an expression vector comprising the expression box, and a method of using the expression box to enhance the plant resistance. The expression box comprises a heterologous promoter that is operably connected to polynucleotide that encodes PYR / PYL acceptor polypeptides, wherein the PYR / PYL acceptor polypeptides comprise one or more of polyketone cyclase structural domain 2 (PF10604) and polyketone cyclase structural domain 1 (PF03364). By introducing the expression box into plants, the stress resistance of the plants is enhanced, compared with plants without the expression box.

Description

[0001] This application is a Chinese patent with an application date of February 12, 2010, a Chinese patent application number of 201080007672.5, and the title of the invention is "Using Novel ABA Receptor Protein and Synthetic Agonist to Regulate Plant Stress Resistance, Water Use Efficiency and Gene Expression" Divisional application of the application. [0002] Cross References to Related Applications [0003] This application claims priority to US Provisional Application No. 61 / 207,684, filed February 13, 2009, which is hereby incorporated by reference for all purposes. Background technique [0004] Abscisic acid (ABA) has been a hotspot of in-depth research since it was confirmed as an endogenous small molecule growth inhibitor and a physiological regulator of plant stress physiology in the 1960s (K.Ohkuma, J.L.Lyon, F.T. Addicott, O.E.Smith, Science 142, 1592 (1963); C.F. Eagles, P.E. Wareing, Physiologia Plantarum 17,697 (1964); J.W. Cornforth, B.V. Milborrow, G. Rybac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H5/00C12N5/04A01H5/08A01H5/02A01H5/10A01H5/12C12Q1/42A01N43/10A01N43/40A01N43/713A01N43/82A01N37/10A01N37/34A01N43/50A01N47/30A01N37/08A01N37/28A01N37/42A01N41/06A01N43/42A01N37/06A01N43/54A01P21/00
CPCA01N37/06A01N37/08A01N37/10A01N37/28A01N37/34A01N37/42A01N41/06A01N43/10A01N43/40A01N43/42A01N43/50A01N43/54A01N43/713A01N43/82A01N47/30C07K14/415C12N9/16C12N15/8273C12Q1/42C12Y301/03016A01N2300/00A01N45/00A01N43/92A01N25/00
Inventor 肖恩·R·卡特勒相祐·朴安德鲁·德弗里斯
Owner RGT UNIV OF CALIFORNIA
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