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Novel regulation protein

A benzene ring-type, polynucleotide technology, applied in the direction of anti-plant immunoglobulin, plant phenotype improvement, introduction of foreign genetic material using vectors, etc., can solve the problem of not identifying regulatory genes and other issues

Inactive Publication Date: 2008-04-23
KEYGENE NV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Only a few structural genes have been cloned and characterized, and no regulatory genes have been identified to date

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Example 1: Identification and Expression of Transcription Factors Involved in the Regulation of Petunia Flower Odor

[0067] To identify elements involved in floral odor regulation in petunia, a directed transcriptomics approach was used. Using specialized, highly specific microarrays, the transcriptomes of odorous flowers were compared to those of soon-to-be-scented flowers and non-scented petunia cultivar flowers. Transcription factors were selected for transcription factors that were elevated immediately before odor production and were very low in odorless petunias. One transcription factor, ODO1 (ODORANT 1), is described in detail herein.

[0068] The transcript level of ODO1 increased when volatile benzene ring compounds started to be emitted between noon and 14.00h, which was consistent with its role in regulating flower odor. Transcript levels of ODO1 rose briefly, returning to their lowest levels early the next morning. ODO1 expression is restricted to flower...

Embodiment 2

[0069] Example 2: Characterization of transcription factors involved in regulation of flower odor

[0070] Sequencing of ODO1 showed that it encodes a 294-amino acid putative protein (SEQID NO.1), which has high homology with R2R3-type MYB family members and has no nuclear localization signal. Although the N-terminal R2R3 domain (amino acids 1-128 of SEQ ID NO.1) contains highly conserved motifs and contains presumably involved in DNA binding to certain variable coremotives (variable coremotives), forming helices- The amino acid of the turn-helix structure, but the C-terminus has no homologous sequence in the Genbank database. Phylogenetic dendrogram analysis showed that ODO1 was most closely related to MYBs from Pimpinella brachicarpa and two MYBs from Arabidopsis thaliana, AtMYB42 and AtMYB85, but the functions of these MYBs were unknown. Seventeen more variable amino acids in the R2R3 domain are conserved among the three proteins.

Embodiment 3

[0071] Example 3: Silencing the ODO1 gene to demonstrate its role in the regulation of flower odor

[0072] To investigate the role of ODO1 in the regulation of flower odor, a transgenic approach was employed. Expression of ODO1 in Mitchell was suppressed by RNAi. Since ODO1 is only expressed in flower tubes and petals and not in any other tissue, we used a constitutive promoter in the RNAi construct. This promoter drives the sequence encoding the C-terminus of ODO1 (which has no homology to other genes in the database), and thus is able to inhibit the accumulation of ODO1 transcripts. We used the ODO1 intron in the RNAi construct and also to transform the Mitchell line as a negative control. At 17.00h, the transcript level of ODO1 in the flowers of each independent transformant was analyzed (the parental Mitchell line had a higher transcript level at 17.00h). To rapidly study the volatiles produced by individual flowers of each transgenic line, we used a directed metabolom...

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Abstract

The present invention relates to a polypeptide which belongs to the R2R3-type MYB family and which regulates the shikimate pathway towards the production of benzenoids. The shikimate pathway is a biosynthesis pathway through which the three essential aromatic amino acids tyrosine, phenylalanine and tryptophane are synthesized in plants, bacteria and fungi. The present inventiont provides for the first time a regulatory protein in the shikimate pathway and a means to regulate the biosynthesis of these three essential amino acids which cannot be produced by mammals. At the same time, it opens up the way for the regulation of the biosynthesis of aromatic and-non- aromatic compounds which are derived from these essential amino acids. A polypeptide or polynucleotide of the invention may be used in a method for manipulating the transcript levels of the genes of the shikimate pathway towards benzenoids for instance the genes of 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase (DAHPS), 5-enol-pyruvylshikimate-3-phosphate synthase (EPSPS), L- phenylalanine ammonia-lyase (PAL) and chorismate mutase (CM). Through these enzymes, biosynthetic processes at lower levels may be influenced. For instance, compounds of the invention may be used in a method to regulate scent in flowers or to regulate resistance to pest insects or pathogenic organisms.

Description

technical field [0001] The present invention relates to myb regulatory proteins in plants. More specifically, the present invention relates to the R2R3 type myb protein, the gene encoding the protein, and the application of the protein and the gene encoding the protein. Background technique [0002] Odor is a very important trait in plants for a number of reasons. For example, flowers produce volatile compounds that play an important role in attracting pollinating insects and successfully producing large numbers of seeds during reproduction. Alternatively, volatile compounds produced by plants in their reproductive or vegetative parts also attract harmful insects or their predators. During this process, the condition of the volatiles determines the susceptibility or resistance to harmful organisms such as harmful insects, nematodes or fungi. Interfering with and altering the synthesis and release of volatiles can be an effective way to intervene in the plant / insect relati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/11C12N15/63C07K14/415C07K16/16C12N1/15C12N1/21C12N5/10A01H3/00A01H5/02C12P21/00C12N15/113
CPCC12N15/1135C07K14/415C12N15/8243C12N2310/14C12N15/8254C12N15/8251C12N2310/111
Inventor M·A·哈林R·C·斯凯瑞克J·C·弗登科A·J·范图楠
Owner KEYGENE NV
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