Mutated nucleotide molecule, and transformed plant cells and plants comprising the same

Abstract

The present invention relates to a method for producing male sterile plant, a mutated nucleotide molecule comprising a nucleotide sequence of the transcription factor bHLH142 and an inserted T-DNA segment, and a novel transformed plant cell and a male-sterile mutant plant comprising the mutated nucleotide molecule, in which the transcription factor bHLH142 is not expressed. The present invention also relates to a novel reversible male sterile transgenic plant, wherein the transcription factor bHLH142 is overexpressed, and its preparation method. The bHLH gene is tissue specifically expresses in the anther and it plays a pivotal role in pollen development. Both the male sterile and reversible male sterile transgenic plants showed a completely male sterile phenotype, but the fertility of the reversible male sterile transgenic plant can be restored under low temperature.

What is claimed is: 
     
         1 . A mutated nucleotide molecule, comprising a nucleotide sequence of the transcription factor bHLH142 and an inserted T-DNA segment. 
     
     
         2 . The mutated nucleotide molecule according to  claim 1 , wherein the nucleotide sequence of the transcription factor bHLH142 has a DNA sequence of SEQ ID No: 1 or a DNA sequence having at least 60% similarity to SEQ ID No: 1. 
     
     
         3 . The mutated nucleotide molecule according to  claim 1 , wherein the T-DNA segment is inserted in the third intron of the nucleotide sequence of the transcription factor bHLH142. 
     
     
         4 . The mutated nucleotide molecule according to  claim 2 , wherein the T-DNA segment is inserted at +1257 bp of the nucleotide sequence of the transcription factor bHLH142. 
     
     
         5 . A transformed plant cell, which comprises the mutated nucleotide molecule according to  claim 1 . 
     
     
         6 . The transformed plant cell according to  claim 5 , wherein the transcription factor bHLH142 has a DNA sequence of SEQ ID No: 1 or a DNA sequence having at least 60% similarity to SEQ ID No: 1. 
     
     
         7 . The transformed plant cell according to  claim 5 , wherein the T-DNA segment is inserted in the third intron of the nucleotide sequence of the transcription factor bHLH142. 
     
     
         8 . The transformed plant cell according to  claim 6 , wherein the T-DNA segment is inserted at +1257 bp of the nucleotide sequence of the transcription factor bHLH142. 
     
     
         9 . A male sterile mutant plant, which comprises the mutated nucleotide molecule according to  claim 1 , and the transcription factor bHLH142 is not expressed. 
     
     
         10 . The male sterile mutant plant according to  claim 9 , wherein the transcription factor bHLH142 has a DNA sequence of SEQ ID No: 1 or a DNA sequence having at least 60% similarity to SEQ ID No: 1. 
     
     
         11 . The male sterile mutant plant according to  claim 9 , wherein the T-DNA segment is inserted in the third intron of the nucleotide sequence of the transcription factor bHLH142. 
     
     
         12 . The male sterile mutant plant according to  claim 10 , wherein the T-DNA segment is inserted at +1257 bp of the nucleotide sequence of the transcription factor bHLH142. 
     
     
         13 . The male sterile mutant plant according to  claim 9 , which is a homozygous mutant. 
     
     
         14 . The male sterile mutant plant according to  claim 9 , wherein the plant is a monocot plant. 
     
     
         15 . The male sterile mutant plant according to  claim 14 , wherein the monocot plant is rice, maize, wheat, millet, sorghum or  Brachypodium distachyon.    
     
     
         16 . The male sterile mutant plant according to  claim 9 , wherein the plant is a dicot plant. 
     
     
         17 . The male sterile mutant plant according to  claim 16 , wherein the dicot plant is  Arabidopsis  or  Brassica  species. 
     
     
         18 . A transformed plant cell, which comprises a plasmid comprising the sequence of the transcription factor bHLH142 and a strong promoter. 
     
     
         19 . The transformed plant cell according to  claim 18 , wherein the transcription factor bHLH142 has a DNA sequence of SEQ ID No: 1 or a DNA sequence having at least 60% similarity to SEQ ID No: 1. 
     
     
         20 . The transformed plant cell according to  claim 18 , wherein the strong promoter is Ubiquitin promoter, CaMV 35S promoter, Actin promoter, an anther tapetum-specific promoter or a pollen-specific promoter. 
     
     
         21 . A reversible male sterile mutant plant, wherein the transcription factor bHLH142 is overexpressed. 
     
     
         22 . The reversible male sterile transgenic plant according to  claim 21 , wherein the transcription factor bHLH142 has a DNA sequence of SEQ ID No: 1 or a DNA sequence having at least 60% similarity to SEQ ID No: 1. 
     
     
         23 . The reversible male sterile transgenic plant according to  claim 21 , wherein the expression of the transcription factor bHLH142 is controlled by a strong promoter. 
     
     
         24 . The reversible male sterile transgenic plant according to  claim 21 , wherein the fertility of the plant is recovered under low temperature. 
     
     
         25 . The reversible male sterile transgenic plant according to  claim 21 , wherein the plant is a monocot plant. 
     
     
         26 . The reversible male sterile transgenic plant according to  claim 25 , wherein the monocot plant is rice, maize, wheat, millet, sorghum or  Brachypodium distachyon.    
     
     
         27 . The reversible male sterile transgenic plant according to  claim 21 , wherein the plant is a dicot plant. 
     
     
         28 . The reversible male sterile transgenic plant according to  claim 27 , wherein the dicot plant is  Arabidopsis  or  Brassica  species. 
     
     
         29 . A method for preparing the reversible male sterile transgenic plant according to  claim 21 , comprising:
 (a) constructing a plasmid comprising the sequence of the transcription factor bHLH142 and a strong promoter; and   (b) introducing the plasmid into a target plant.   
     
     
         30 . The method according to  claim 29 , wherein the DNA sequence of bHLH142 is SEQ ID No: 1 or a DNA sequence having at least 60% similarity to SEQ ID No: 1. 
     
     
         31 . The method according to  claim 29 , wherein the strong promoter is Ubiquitin promoter, CaMV 35S promoter, Actin promoter, an anther tapetum-specific promoter or a pollen-specific promoter. 
     
     
         32 . The method according to  claim 29 , wherein the plasmid is introduced into calli of the target plant via  Agrobacterium tumefaciens.