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Expression of transcription regulators that provide heat tolerance

A tolerant, transgenic plant technology, applied in genetic engineering, plant genetic improvement, angiosperms/flowering plants, etc., can solve problems such as iron deficiency tolerance

Active Publication Date: 2012-05-30
PERFORMANCE PLANTS INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has been found that overexpression of Arabidopsis basic helix-loop-helix (bHLH) 38 or Arabidopsis basic helix-loop-helix (bHLH) 39 by FIT can alter the expression pattern of FRO2 and IRT1, Produces constitutive activation and leads to resistance to iron deficiency (see Yuan et al. 2008)

Method used

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  • Expression of transcription regulators that provide heat tolerance
  • Expression of transcription regulators that provide heat tolerance
  • Expression of transcription regulators that provide heat tolerance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0107] Example 1: Identification of homologous genes

[0108] Basic helix-loop-helix (bHLH) 39 and Basic local alignment search for its Arabidopsis homologues basic helix-loop-helix (bHLH)38, basic helix-loop-helix (bHLH)100, and basic helix-loop-helix (bHLH)101 Tool (Blast) search. Genome sequence databases are for varieties for which the complete sequence is known, such as rice and sorghum. To confirm that the sequence does indeed belong to the ortholog of the target sequence, a basic local alignment search tool (blast) search of all putative homologues in the complete Arabidopsis protein database needs to be performed thereafter , and any sequence that hits Arabidopsis first and is not one of the four target sequences will be filtered out without any further analysis. To minimize the redundancy that exists between multiple hits, all EST sequence hits were assembled using the cap3 assembly program to ensure that they had a minimum of 40 nucleotides in length. 90% ident...

Embodiment 2

[0113] Embodiment 2: the construction of carrier

[0114] The binary vector pBI121 is optimized for the transformation of Arabidopsis and different crops. The GUS gene present in pBI121 was deleted by digestion and religation of SmalI and EcolCR1, thereby obtaining the vector pBI121ΔGUS. This vector is used to clone genes for overexpression. The 1.1 Kb portion of the C-terminus of the GUS gene was isolated from pBI121 as an EcoR V-Sac I fragment (position 6613-7715 on pBI121) and the Sma I on pBI121ΔGUS and clone it at the Sac I site to obtain pBI121tGUS (the N-terminal portion of the GUS gene has been deleted). The vector is used to make a hairpin construct for gene deregulation, wherein the construct is constructed using a part of the GUS sequence as the loop or spacer.

[0115] The NPTII gene and its derivatives present in the vector pBI121 contain a site-directed mutagenesis (a G to T mutagenesis at position 3383 of pBI121, amino acid substitution E182D). The above-m...

Embodiment 3

[0118] Example 3: Overexpression constructs of basic helix-loop-helix (bHLH) subgroup 1b body building

[0119] Construct 35S - Arabidopsis basic helix-loop-helix (bHLH) 39:

[0120] The Arabidopsis basic helix-loop-helix (bHLH) 39 (At3g56980) coding sequence was amplified from Arabidopsis by reverse transcription-polymerase chain reaction (RT-PCR), wherein Forward primer BHLH039FW-XbaI and reverse primer BHLH039RV-BamH I were used in the addition. The polymerase chain reaction (PCR) product (0.8Kb) was cloned at the Xba I and BamH I sites of the binary vectors pBI300ΔGUS and pBI800ΔGUS to establish the construct 35S-Arabic Mustard basic helix-loop-helix (bHLH)39-pBI300 and 35S-Arabidopsis basic helix-loop-helix (bHLH)39-pBI800.

[0121] Construct 35S - Arabidopsis basic helix-loop-helix (bHLH) 101:

[0122] The Arabidopsis basic helix-loop-helix (bHLH) 101 (At5g04150) coding sequence was amplified from Arabidopsis by reverse transcription-polymerase chain reaction (RT-...

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Abstract

The invention relates to methods of producing a desired phenotype in a plant by manipulation of gene expression within the plant. The method relates to means to increase expression level of a bHLH subgroup Ib gene expression or activity, wherein a desired phenotype such as increased heat tolerance relative to a wild type control plant following heat stress results in reduced flower abortion and increased yield. Included are plants produced by said methods. The invention also relates to nucleic acid sequences and constructs useful such methods and methods of generating and isolating plants having increased expression of a bHLH subgroup Ib expression or activity.

Description

[0001] References to related applications [0002] This application claims priority to US Provisional Application No. 61 / 221813, filed June 30, 2009. The contents of the above-mentioned documents are incorporated by reference in the present invention as a whole. field of invention [0003] The present invention belongs to said field of plant molecular biology and relates to transgenic plants with novel phenotypes, methods for producing such plants and polynucleotides and polypeptides which can be effectively used in such methods . More specifically, the present invention relates to the expression of a transcriptional regulator and transgenic plants, wherein the transgenic plant has enhanced activity of the transcriptional regulator, thereby enabling the production of plants with beneficial phenotypes. Background of the invention [0004] Environmental stress is responsible for a significant increase in the yield of crops. A study of the relationship between corn and soybe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H5/00A01H5/10C12N15/00C12N15/29C07K14/415
CPCC12N15/8273C12N15/8271
Inventor 万江欣黄雅凡
Owner PERFORMANCE PLANTS INC
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