Polypeptide and use thereof for improving stress tolerance in plants

Patent metadata

Field	Value
Publication number	US-2017306348-A1
Application number	US-201715644779-A
Country	US
Kind code	A1
Filing date	Jul 8, 2017
Priority date	Jan 9, 2015
Publication date	Oct 26, 2017
Grant date	—

Abstract

This disclosure provides a stress-responsive polypeptide sequence for fusion with a polypeptide to specifically induce stability of the fusion polypeptide under stress conditions, such as drought, high salt and high temperature, in plants. Also disclosed includes an expression vector for expressing a fusion polypeptide comprising the stress-responsive peptide in plants transformed therewith, and a method for generating a transgenic plant with enhanced tolerance to environmental stresses, comprising introducing into the transgenic plant a polynucleotide encoding a fusion polypeptide which comprises the stress-responsive peptide as disclosed and a plant anti-stress gene, such as the plant senescence-associated gene SSPP. A plant expressing the expression vector that have an enhanced stress tolerance, including Arabidopsis and soybean, is also provided.

First claim

1 . A fusion polypeptide, comprising:
an N-terminal stress-responsive peptide, comprising an amino acid sequence at least 70% identical to the sequence as set forth in SEQ ID NO: 1; a C-terminal polypeptide; and optionally a linker sequence between the N-terminal stress-responsive peptide and the C-terminal polypeptide, said linker sequence having a length of 1-20 amino acid residues; wherein:
the N-terminal stress-responsive peptide is foreign to the C-terminal polypeptide;
and the N-terminal stress-responsive peptide retains an ability to regulate stability of the fusion polypeptide such that the fusion polypeptide is capable of remaining at a low level in absence of, but accumulating at a high level in presence of, at least one environmental stress in the plant.

2 . The fusion polypeptide according to claim 1 , wherein the N-terminal stress-responsive peptide comprises an amino acid sequence at least 90% identical to the sequence as set forth in SEQ ID NO: 1.

3 . The fusion polypeptide according to claim 2 , wherein the N-terminal stress-responsive peptide comprises an amino acid sequence 100% identical to the sequence as set forth in SEQ ID NO: 1.

4 . The fusion polypeptide according to claim 1 , comprising an amino acid sequence as set forth in SEQ ID NO: 3.

5 . An expression vector, comprising a promoter operably linked to a polynucleotide, wherein the polynucleotide encodes the fusion polypeptide according to claim 1 .

6 . The expression vector according to claim 5 , wherein the promoter is a plant-compatible promoter, selected from a group consisting of a constitutive plant promoter, a tissue specific plant promoter, and a promoter of a plant gene.

7 . The expression vector according to claim 6 , wherein the promoter is a constitutive plant promoter, selected from the group consisting of a CaMV35S promoter, an opine promoter, a promoter of ubiquitin (Ubi) gene of a plant species, a promoter of actin 1 (Act-1) gene of a plant species, and a promoter of alcohol dehydrogenase 1 (Adh-1) gene of a plant species.

8 . The expression vector according to claim 7 , wherein the promoter is a CaMV35S promoter.

9 . A plant transformed with a polynucleotide, wherein the polynucleotide encodes, and expresses in the plant, a fusion polypeptide, the fusion polypeptide comprising:
a N-terminal stress-responsive peptide, comprising an amino acid sequence at least 70% identical to the sequence as set forth in SEQ ID NO: 1; a C-terminal polypeptide; and optionally a linker sequence between the N-terminal stress-responsive peptide and the C-terminal polypeptide, said linker sequence having a length of 1-20 amino acid residues; wherein:
the N-terminal stress-responsive peptide is foreign to the C-terminal polypeptide; and
the N-terminal stress-responsive peptide retains an ability to regulate stability of the fusion polypeptide such that the fusion polypeptide is capable of remaining at a low level in absence of, but accumulating at a high level in presence of, at least one environmental stress in the plant.

10 . The plant according to claim 9 , wherein the C-terminal polypeptide is a polypeptide having a function of a plant anti-stress gene, selected from a group consisting ofAREB1, AREB2, rd19, rd22, MYC, MYB, bZIP, AtGolS2, CBF4, DREBla, Adc, HVA1, ZmNF-YB2, IPT, PPC and SSPP.

11 . The plant according to claim 10 , wherein the C-terminal polypeptide is a polypeptide having a function of SSPP.

12 . The plant according to claim 11 , wherein the C-terminal polypeptide comprises an amino acid sequence at least 70% identical to the sequence as set forth in SEQ ID NO: 2.

13 . The plant according to claim 12 , wherein the C-terminal polypeptide comprises an amino acid sequence 100% identical to the sequence as set forth in SEQ ID NO: 2.

14 . The plant according to claim 9 , wherein the polynucleotide comprises a nucleotide sequence at least 70% identical to the sequence as set forth in SEQ ID NO: 4.

15 . The plant according to claim 14 , wherein the polynucleotide comprises a nucleotide sequence 100% identical to the sequence as set forth in SEQ ID NO: 4.

16 . The plant according to claim 9 , exhibiting enhanced tolerance to the at least one environmental stress compared to a plant of a same species not containing the polynucleotide, wherein:
the at least one environmental stress comprises at least one of drought stress, high salt stress, high temperature stress, low temperature stress, water stress, or pathogen stress.

17 . The plant according to claim 16 , wherein the at least one environmental stress comprises at least one of drought stress and high salt stress.

18 . The plant according to claim 9 , wherein the plant is a model plant, a food crop, a cash crop, a vegetable, a fruit, a grass or a flower.

19 . The plant according to claim 17 , wherein the plant is selected from the group consisting of Arabidopsis, corn (maize), sorghum, wheat, sunflower, crucifer, pepper, potato, cotton, rice, soybean, sugarbeet, sugarcane, tobacco, barley, oilseed rape, Brassica sp., alfalfa, rye, millet, safflower, peanut, chestnut, sweet potato, cassava, coffee, coconut, pineapple, apple, orange, pear, watermelon, grape, peach, lemon, cherry, Chinese date, strawberry, blue berry, raspberry, blackberry, papaya, apricot, persimmon, pomegranate, jackfruit, areca nut, kiwi fruit, plum, loquat, cocoa, tea, banana, avocado, fig, guava, mango, olive, papaya, cashew, macadamia, almond, oat, onions, tomato, lettuce, green bean, lima bean, pea, eggplant, zucchini, luffa, mushroom, carrot, spinach, kale, broccoli, pumpkin, white gourd, lotus root, garlic, ginger, chive, yam, cucumber, cantaloupe, muskmelon, azalea, hydrangea, hibiscus, rose, tulip, daffodil, petunia, carnation, poinsettia, chrysanthemum, pulp tree, oil palm, and conifer.

20 . The plant according to claim 18 , wherein the plant is Arabidopsis or soybean.

Assignees

• Univ Nankai

Inventors

• Wang Ningning
• Xiong Li
• Bi Juanjuan
• Liu Sheng
• Xu Wei
• Sun Lifang
• Guo Zhaoxia
• Nasrullah
• Wang Dan
• Mei Yuanyuan

Classifications

• A01H1/00

  Processes for modifying genotypes {; Plants characterised by associated natural traits} (A01H4/00 takes precedence) · CPC title

• C12N15/00

  Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor (mutants or genetically engineered microorganisms, per se C12N1/00, C12N5/00, C12N7/00; new plants per se A01H; plant reproduction by tissue culture techniques A01H4/00; new animals per se A01K67/00; use of medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases, gene therapy A61K48/00) · CPC title

• C12N5/04

  Plant cells or tissues {(culture media C12N5/0025)} · CPC title

• C07K14/415

  from plants · CPC title

• A01H5/00

  Angiosperms, i.e. flowering plants, characterised by their plant parts; Angiosperms characterised otherwise than by their botanic taxonomy · CPC title