Application of zmrph1 gene in regulation of plant height and lodging resistance
A lodging resistance and genetic technology, applied in the fields of biotechnology and plant genetics and breeding, to achieve the effects of improving lodging resistance, improving corn harvest index, and high-yield and stable-yield corn harvest index
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Embodiment 1
[0052] Example 1 Construction and genetic transformation of ZmRPH1 expression vector
[0053] Total RNA was extracted from corn inbred line B73 V3 stage seedlings, and cDNA was synthesized by reverse transcription using cDNASynthesis Kit from Takala Company, and amplified with specific primers to obtain the CDS sequence of ZmRPH1, with a full length of 1650bp (SEQ ID NO: 1). The nucleotide sequence of the primer pair is:
[0054] Upstream primer: ZmRPH1-F:5'-ATGGTGGCTGCCGCGGCG-3'
[0055] Downstream primer: ZmRPH1-R:5'-CTACAATTGTGTCGGGCTCTGCTTT-3'
[0056] The PCR product was recovered, the target fragment was connected to the pBCXUN vector, and Escherichia coli competent cells were transformed to obtain the recombinant vector pBCXUN-ZmRPH1 with correct sequencing.
[0057] The expression cassette is composed of the ZmUbiquitin1 promoter and the 3' transcription termination region from the nopaline synthase (nos) gene, and the selectable marker gene is phosphinothricin. Its...
Embodiment 2
[0060] Embodiment 2 Preparation of hybrids
[0061] Using the material of the inbred line B73-329 background (the transgenic line that overexpresses the ZmRPH1 gene of the homozygous B73-329 background) as the female parent, and the maize inbred line T13 as the male parent, cross breeding to obtain a hybrid that overexpresses ZmRPH1 kind of material.
Embodiment 3
[0062] Example 3 Analysis of tissue expression pattern of ZmRPH1
[0063] The total RNA of multiple tissues and organs (root, node, internode, leaf, ear, filament, tassel) in four different growth stages (V9, V11, V13, V15) of maize inbred line B73-329 were extracted, Reverse transcribed into cDNA. Using ZmRPH1 gene-specific primers (upstream primer: ZmRPH1qPCR-F:5'-CAAAGTTGCAACGCAAGAAC-3'
[0064] Downstream primer: ZmRPH1qPCR-R:5'-GGCTCTGCTTTTGGTTACGTT-3') for real-time fluorescent quantitative PCR amplification, with ZmActin1 gene as an internal reference gene, and its specific primers are: upstream primer: Zmactin1qPCR-F:5'-TTGTCCTCAGTGGGGGATCT-3' Downstream primer: ZmActin1qPCR-R: 5'-CTTTCAGGTGGCGCAATCAC-3'. The result is as figure 1 As shown, it was shown that ZmRPH1 was highly expressed in the internode tissue in the stage of rapid elongation.
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