Development and application of molecule marker for corn with considerable number of kernels and excellent allele function in low-nitrogen adverse environment
A corn and seed technology, applied in the fields of application, plant gene improvement, DNA/RNA fragments, etc., can solve the problems of environmental effects of seed number, inability to select corn seed number traits, etc., achieve high development value, speed up breeding, and reduce costs cheap effect
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Embodiment 1
[0020] The 180 maize inbred lines (see Table 1) used for the identification of superior alleles in Example 1 are all parental inbred lines frequently used in maize breeding in my country; the public can obtain them from the Institute of Crop Science, Chinese Academy of Agricultural Sciences. For the genetic background analysis results of each inbred line, see (Chuanxiao Xie, Shihuang Zhang, Minshun Li, Xinhai Li, Zhuanfang Hao, Li Bai, Degui Zhang, Yehong Liang. (2007) Inferring genome ancestry and estimating molecular relatedness among 187 Chinese maize inbredlines .J.Genet.& Genomics 34(8):738-748), and as the basis for the main genetic groups.
[0021] Table 1 Association analysis of natural population inbred lines and their pedigree sources
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[0027] The genetic analysis of embodiment 1, corn kernel number
[0028] 1. Field experiment design and plant testing
[0029] 180 maize inbred lines were designed with α-l...
Embodiment 2
[0042] Example 2, Discovery of Molecular Markers and Design of Specific Primer Pairs
[0043] 1. Discovery of molecular markers
[0044] Analysis of the sequencing results of the glutamine synthetase gene Gln1-3 of 180 inbred lines revealed a 12 bp insertion / deletion (Indels) site located at positions -274 to -285 in the upstream regulatory region of the gene, with the sequence as follows:
[0045] 5'-CCGCGTCGAGGA-3'.
[0046] The gene with the above sequence is the wild type, and the gene lacking the above sequence is the mutant type.
[0047] 2. Design of specific primer pairs
[0048] Design specific primer pairs according to the sequence as follows:
[0049] Forward primer: 5'-ATGTAACTGACAGGTGGGCCT-3' (sequence 2 of the sequence listing);
[0050] Reverse primer: 5'-TTGAGATTGAGGACGAAGGGA-3' (sequence 3 of the sequence listing).
[0051] Using the wild-type glutamine synthetase gene Gln1-3 as a template, use the above primer pair to PCR amplify to obtain the DNA fragme...
Embodiment 5
[0100] Embodiment 5, application of specific primer pair in breeding
[0101] With deletion (D) excellent genotype CA156 as donor and insertion (I) genotype Wu314 as recipient, F 1 generation; the F 1 32 generations of F 2 generation groups. to F 2 Genomic DNA was extracted from 32 individual strains in the generation population, PCR was performed with the specific primer pair designed in Example 2, and the PCR amplification product was detected by electrophoresis, the method was the same as step 2 in Example 3.
[0102] see results figure 2 . 6 homozygous deletion (Deletion) genotype individuals (D), 12 insertion / deletion heterozygous co-dominant (Co-dominant) genotype individuals (C), and 14 homozygous insertion (Insertion) genotype individuals were obtained (I). Among them, the above-mentioned 6 strains of homozygous deletion individuals can be used as excellent homozygous individuals of this genotype directly for breeding applications, and the offspring of 12 inser...
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