A maize arsenic stress resistance gene zmasr1 and its primers, encoded products, linked SNPs and applications
A resistance gene, corn technology, applied in the field of corn genetics and breeding
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Embodiment 1
[0023] Discovery of arsenic stress resistance gene ZmAsR1 in maize
[0024] The material used in this study is a related group consisting of 350 representative maize inbred lines (gifted by Professor Yan Jianbing of Huazhong Agricultural University), of which 151 are from temperate regions and 79 are from tropical and subtropical regions. In Yongcheng (YC) and Yuanyang Henan Agricultural University Base (YY) in Henan, China, at each site, the associated populations were designed using a completely randomized block design with three replicates. Each plot is 3 meters long, with a plant spacing of 0.22 meters and a row spacing of 0.67 meters. The final planting density is 67,500 plants per hectare.
[0025] Harvest of mature ears of associated populations was performed to determine the accumulation and distribution of arsenic in bracts and cobs. The cob and bracts of each inbred line in each environment were collected together and dried naturally. The dried cob and bracts were f...
Embodiment 2
[0038] Development of an intramolecular SNP marker for the arsenic stress resistance gene ZmAsR1 in maize
[0039] dCAPS markers were designed using dCAPS Finder 2.0 (http: / / helix.wustl.edu / dcaps / dcaps.html). Enter the two types of sequences that are the same except for the SNP, and the other sequences are the same,
[0040] The sequence is:
[0041] 5'-CATATTCACTGCTGATCTTTTTCTG(C / T)GGGATGTTTATCACTGTTGATGGC-3' as shown in SEQ ID NO. 5. The SNP is located in the middle of the sequence, with about 25nt bases on both sides. Enter the allowed number of mismatched bases "1" in the third input box, submit the run, and the result is as follows Figure 4 As shown, primers for reference appear.
[0042] like Figure 5 As shown in the figure, if the first base G on the right side of the downstream primer containing the SNP site is replaced with T, the enzyme cleavage recognition site CTGCAG of PstI appears in the material resistant to arsenic stress, and this primer is se...
Embodiment 3
[0045] Application of maize arsenic stress resistance gene ZmAsR1 in maize breeding for heavy metal stress resistance and quality improvement
[0046] The maize natural population DNA was labeled with dCAPS for amplification. The reaction system was: 0.5 μl of upstream and downstream primers, the sequence is shown in SEQ ID NO. 6-7, 5 μl of 2×Taq Master Mix, 1 μl of DNA template, supplemented with ddH2O to 10 μl, reaction program Table 1.
[0047] Table 1 Reaction procedure
[0048]
[0049] Then the PCR product was digested with PstI, and the enzyme digestion system is shown in Table 2. Detected by 4% agarose electrophoresis Figure 4 . Using the developed dCAPS marker, maize inbred lines with high / low arsenic content can be distinguished.
[0050] Table 2 Reaction system
[0051]
[0052] The distribution of PstⅠ digestion in natural population agarose gel electrophoresis is as follows Figure 5 As shown, 1-24 represent different inbred lines, among which, lanes ...
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