A maize seedling yellow and white leaf gene and its encoded protein and application

A gene and protein technology, applied to the use of the gene and protein, in the field of yellow and white leaf genes of maize seedlings, can solve the problems of reducing the non-lethal ratio of transgenic pollen, and achieve the effect of obvious characteristics, easy identification and stable inheritance

Active Publication Date: 2022-03-15
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In order to solve this problem, Chinese researchers developed a multi-control sterile system MCS, improved the SPT expression cassette, and increased one pollen lethal gene in SPT to two. The results show that MCS can indeed effectively reduce transgenic pollen. Non-lethal ratio (0.033%-0.263%), but there is still the phenomenon of transgenic drift

Method used

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  • A maize seedling yellow and white leaf gene and its encoded protein and application
  • A maize seedling yellow and white leaf gene and its encoded protein and application
  • A maize seedling yellow and white leaf gene and its encoded protein and application

Examples

Experimental program
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Effect test

Embodiment 1

[0053] Example 1 Obtainment and Genetic Analysis of the Yellow and White Leaf Color Mutant eal1 of Maize Seedlings of the Present Invention

[0054] (1) Obtaining and phenotypic identification of the corn seedling yellow and white leaf mutant eal1 of the present invention

[0055] In 1996, the applicant used a returnable satellite to carry Chuandan No. 9 corn seeds to carry out space mutagenesis. After the space mutagenesis, the Chuandan No. 9 seeds were self-crossed for 8 generations, and a seedling leaf color mutation material was accidentally obtained. , the color of the mutant material is yellow or even white after emergence (hereinafter referred to as: yellow and white leaves), and it quickly changes to normal green in the later stage. The inventor named the mutant material: eal1. The sister plants from the same panicle line with normal leaf color and no separation of leaf color in the selfed offspring were used for conservation and served as the control WT.

[0056] The...

Embodiment 2

[0062] Example 2 Map-based cloning of the zmeal1 mutant gene in maize

[0063] (1) Fine mapping of maize zmeal1 gene

[0064] The F2 population obtained in Example 1 was used as a genetic mapping population, and the genomic DNA of leaves of mutant individual plants in the population was extracted by the CTAB method for genotype identification. By searching the MaizeGDB public database (http: / / www.maizegdb.org / ), 366 pairs of SSR markers covering the entire maize genome were screened. Polyacrylamide gel electrophoresis and agarose gel electrophoresis were used to identify the polymorphisms of these SSR markers between the mutant eal1 and the inbred line B73.

[0065] Firstly, 96 individual plants with leaf color mutation in the F2 population were used for initial mapping, and the mutant gene was located on the short arm of chromosome 4. The mutated gene was positioned between the polymorphic molecular markers InDel2 and InDel19 through encrypted polymorphic markers on chromos...

Embodiment 3

[0072] Example 3 Genotype Identification Test of Maize zmeal1 Mutant Gene Locus Using Specific Primers and Enzyme Cutting Methods

[0073] Example 2 verified the above-mentioned 7 SNP variations and a 3bp deletion at the genomic DNA level. Based on the above variation information, an enzyme-cut amplified polymorphic sequence marker BeCaps1 in the ZmSig2A gene was developed (see Table 1), and the genomic DNA was amplified using the primer BeCaps1, and the reagent used was KOD FX (TOYOBO, Code No.: KFX-101); the amplification program was: pre-denaturation at 94°C for 5 minutes; denaturation at 98°C for 10 seconds, annealing at 59°C for 30 seconds, extension at 68°C for 1 minute, and 34 cycles; extension at 68°C for 5 minutes; end program at 12°C. The resulting amplified PCR product was subjected to agarose gel electrophoresis after being digested with NruI.

[0074] Results (see Figure 7 ) the genotype is dominant homozygous ZmEAL1ZmEAL1, the band size after enzyme digestion is...

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Abstract

The invention discloses a corn seedling yellow and white leaf protein and its coding gene. The protein is composed of the amino acid sequence shown in SEQ ID No.1, and the coding gene is composed of the nucleotide sequence shown in SEQ ID No.2. It also discloses their use in cultivating seedling yellow and white-leaf maize varieties, identifying hybrids or removing impurities, and inducing haploid breeding in hybrids, as marker traits in SPT or MCS seed production technology and other applications. The yellow and white leaves of corn seedlings of the present invention have obvious traits, are easy to identify, and can be stably inherited; secondly, the traits are controlled by a single recessive gene, which can be transferred to inbred lines by backcrossing or transgenic methods, and the breeding method Simple; in addition, the yellow and white leaf traits of corn seedlings of the present invention only appear in a short period of time after emergence, and turn green quickly in the later stage, without affecting the normal growth and development of corn, and have extremely wide application prospects in the fields of corn seed production and hybrid induced haploid breeding .

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a corn seedling yellow and white leaf gene, a protein encoded by the gene, and applications of the gene and protein; the invention also relates to a molecular marker of the gene and its application. Background technique [0002] The occurrence of leaf color mutation is mainly because the mutant gene directly or indirectly interferes with the synthesis and degradation of chlorophyll, and then changes the content and proportion of various pigments in leaf color through various channels, eventually causing leaf color variation. In maize, there are mainly leaf color mutation types such as albino, yellow, light green and striped leaves. Most of the known leaf color mutation materials show obvious leaf color mutation phenotypes throughout the growth period, which not only affect leaf photosynthesis The effect may even be fatal, so it is difficult to apply in production. Curren...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82C12N5/10C12N15/11C12Q1/6895A01H1/02A01H5/12A01H6/46
CPCC07K14/415C12N15/8261C12Q1/6895A01H1/02C12Q2600/13
Inventor 李川曹墨菊余涛胡朝勇王靖文易洪杨夏远燕袁鹏冯开洁
Owner SICHUAN AGRI UNIV
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