Mutant alleles of zmamp1 gene and their applications
An allele and gene technology, applied in the field of alleles, can solve problems such as dwarfing and lack of breeding application value, and achieve the effects of reducing plant height, improving lodging resistance and yield, and promoting early harvesting.
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Embodiment 1
[0083] Example 1 F2:3 Population Phenotyping and QTL Mapping
[0084] Aiming at the characteristics of semi-dwarf maize mutant grmm and early flowering, Si273 was crossed with grmm to construct F2:3 segregating population. In the F2:3 population, plant height-related traits (including plant height, ear height, and ear height), flowering period-related traits (including heading stage, powder-scattering stage, and silking stage), and panicle traits (ear length) were counted. and ear diameter) and yield-related traits (including number of rows per ear, number of grains per row, 100-grain weight, and yield per plant). For each trait tested, there were significant differences between parents (Table 1). The coefficients of variation of these traits ranged from 3.13% to 25.27%, and the generalized heritability ranged from 0.55 to 0.89, indicating that these traits were mainly controlled by genetic factors. Correlation analysis of the tested traits showed that plant height was signi...
Embodiment 2
[0098] Example 2 qPH1 Fine Mapping and Allelicity Test
[0099] Plant height was used as the target trait for fine mapping of qPH1, which lies in the interval of 273-277 cM ( figure 2 A), which roughly corresponds to the physical location of 281.8-289.3 Mb (B73 reference genome version 3). Preliminary screening of crossover individuals was carried out using markers D2 and D25 (see Table 5 for primer sequences), whose positions are 281.2 and 289.9 Mb, respectively. Using the BC2F2 population, the QTL region was narrowed down to the region between G2 and R18 (see Table 5 for primer sequences), which is 227 kb in length according to the maize B73 reference genome version 3. Using the BC2F3 population, the QTL region between R14 and G13 was further narrowed down to 16 kb (see Table 5 for primer sequences). Two candidate genes (GRMZM2G010353 and GRMZM2G011385) were found in this region ( figure 2 B).
[0100] Table 5 Primer sequence information
[0101]
[0102] According...
Embodiment 3
[0110] Example 3 Mining of ZmAMP1 variant sites
[0111] First, qPH1-based near-isogenic lines (NILs) were screened in the BC3F3 generation, and the NILs containing the grmm fragment were called NILs grmm , containing the Si273 fragment called NIL Si273 , RNA was extracted from young stem tissues of vegetative growth V6 stage of NILs and parents, and ZmAMP1 was compared between NILs (NILs) by transcriptome analysis and real-time quantitative PCR (RT-PCR). Si273 and NIL grmm ) and differences between parents (grmm and Si273).
[0112] qRT-PCR showed that NIL Si273 and NIL grmm There were no significant differences between and between parental lines ( Figure 4 A), suggesting that the semi-dwarf phenotype of grmm is independent of ZmAMP1 expression. Therefore, the genomic sequence of ZmAMP1_grmm (after the ATG transcription start codon) was compared with that of ZmAMP1 from Si273, B73, CML247, EP1 and F7 (the sequences of B73, CML247, EP1 and F7 were from www.maizegdb.org)...
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