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Method for identifying plant gene functions in total genome ranges

A whole genome, gene function technology, applied in the field of plant gene function identification, can solve the problems of high price, low screening throughput, high labor intensity, etc.

Active Publication Date: 2016-07-06
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, TILLING technology is relatively labor-intensive, the screening throughput is small, and the price is expensive
As an effective "forward genetics" method to study gene function, map-based cloning technology needs to establish a large number of mapping groups, which is labor-intensive and screening throughput is small

Method used

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  • Method for identifying plant gene functions in total genome ranges
  • Method for identifying plant gene functions in total genome ranges
  • Method for identifying plant gene functions in total genome ranges

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Example 1 Genome-wide functional identification of maize genes

[0072] 1. Construction of a library of mutants induced by EMS in corn pollen

[0073] On the afternoon before the treatment, corn (Zeamays, corn inbred line—B73,) tassels were bagged after removing the old anthers. On the morning of the treatment, add 1 mL of LEMS (ethyl methanesulfonate) to 99 mL of liquid paraffin oil (Sigma, mineral oil M8410) in the fume hood to make 1% (volume percentage) of EMS (ethyl methanesulfonate) ) mother solution, mix well, then dilute to 0.07% EMS working concentration, and fill in vials. At around 10:00 in the morning, collect fresh corn pollen, pass through a 100-mesh sieve to remove the anthers, add 10 g of collected pollen to a small bottle of treatment solution (0.07% EMS), shake and mix gently, and after 45 minutes, use a brush to remove the pollen. The pollen is applied to the corresponding filaments and tagged. The seeds of the present generation are the M1 generat...

Embodiment 2

[0201] Example 2 Genome-wide functional identification of maize genes

[0202] With reference to the method described in Example 1, it is also possible to directly search for the mutant of the gene of interest in the mutant library to study its function, first to see if there is phenotype statistics in the database, whether these mutants have a common phenotype, Then study the function of the gene according to this commonality. Specifically: first find the M1 generation plant mutants corresponding to all SNP sites in the gene to be tested in the plant mutant genotype library; then find each M1 generation plant mutation in the plant mutant phenotype library All phenotypes of an individual, if the phenotypes are the same, the gene to be tested is a gene related to the phenotype.

[0203] For example, in this example, the gene GRMZM2G093603 encodes a gene related to gibberellin synthesis, and three allelic mutations of the gene were found from the mutants, the mutant numbers are...

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Abstract

The invention relates to a method for identifying plant gene functions in total genome ranges.The method includes 1), mutating single nucleotide of starting plants and establishing plant mutant libraries; 2), detecting phenotypes of mutants in the plant mutant libraries and establishing plant mutant phenotype databases; 3), detecting genotypes of the mutants in the plant mutant libraries and establishing plant mutant genotype databases; 4), determining functions of genes where SNP (single nucleotide polymorphism) sites caused by mutation of the single nucleotide in the starting plants by the aid of the plant mutant phenotype databases and the plant mutant genotype databases.The method has the advantages that the genes related to plant development procedures and biological procedures of stress-tolerant reaction or metabolic pathways or the like can be accurately and quickly found by the aid of the plant mutant phenotype databases and the plant mutant genotype databases which are established by the method, and novel ways can be created for plant functional genome research.

Description

technical field [0001] The invention relates to gene functional omics, in particular to a method for identifying plant gene functions within the whole genome. Background technique [0002] As a "reverse genetics" method, TILLING technology is combined with traditional chemical mutagenesis technology (EMS mutagenesis). It can screen mutant libraries on a large scale without complicated techniques and equipment. In plant functional genomes and The genetic improvement of crops has played a huge role. However, TILLING technology is relatively labor-intensive, the screening throughput is small, and the price is expensive. Map-based cloning technology, as an effective "forward genetics" method to study gene function, needs to establish a large number of mapping populations, which is labor-intensive and screening throughput is small. The rise of second-generation genome sequencing technology provides an effective method for high-throughput detection of single nucleotide polymorph...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C40B50/06
Inventor 张春义路小铎范云六
Owner THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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