An application of a PTRE1 gene in regulation and control of plant heat resistance
A technology of plants and gramineous plants, applied in the fields of botany and genetic engineering, can solve problems such as shortened growth period and reduced water use efficiency
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[0044] As an embodiment of the present invention, the gene encoding the PTRE1 protein is cloned into an appropriate vector by conventional methods, and the recombinant vector with the foreign gene is introduced into the plant cells that can express the PTRE1 protein , making the plant cell express PTRE1 protein. Plants overexpressing PTRE1 protein can be obtained by regenerating said plant cells into plants. Preferably, the gene encoding the PTRE1 protein is transferred into plants using the Agrobacterium transformation method.
[0045] As used herein, the forward connection refers to: the connection between the coding gene of PTRE1 and the expression vector is a positive connection, that is, the coding gene is connected to the vector in the direction of 5'→3'. Usually, the gene encoding PTRE1 is located downstream of the promoter in the expression vector, that is, the 3' end of the promoter is connected downstream to the 5' end of the encoding gene. The coding gene is opera...
Embodiment 1
[0050] Example 1. Isolation and phenotype analysis of PTRE1 gene
[0051] In the research on the regulation of proteasome activity, the inventors found that PTRE1 gene can regulate the heat tolerance of plants.
[0052] Plants with PTRE1 overexpression were obtained by constructing PTRE1 overexpression vector and transfecting Arabidopsis plants.
[0053] 1. Construction of PTRE1 overexpression vector
[0054] The vector construction method is as follows: use the genome of Arabidopsis Col as the amplification template, amplify the PTRE1 cDNA gene by primer P1 and primer P2, and insert it into the polyclonal polyclonal of the vector pCAMBIA1302 (the plasmid has its own GFP coding sequence) by restriction endonuclease ligation. site, a PTRE1 overexpression vector was obtained, called pCAMBIA1302-PTRE1, which could express 35S-driven PTRE1-GFP (p35S::PTRE1-GFP).
[0055] Primer P1: CATG CCATGG CGAATTCTCAGACGGTGA (SEQ ID NO: 4);
[0056] Primer P2: GGACTAGTTATAAAATCTGAACCGCCG ...
Embodiment 2
[0061] Phenotype Analysis of Plants Overexpressed in Embodiment 2, PTRE1 Gene
[0062] In this example, the phenotypic traits of ptre1 mutants and PTRE1 gene overexpressed transgenic plants were detected.
[0063] 1. Establishment of Arabidopsis mutants
[0064] In order to study the physiological function of PTRE1, the inventors searched the Arabidopsis T-DNA insertion mutant library (http: / / signal.salk.edu / cgi-bin / tdnaexpress) and obtained a possible T-DNA insertion mutant Salk_034353. After seed propagation, screening and identification, it was proved that the insertion position was on the first intron. About 300 plants of the progeny population were verified by T-DNA insertion. It was found that in the seedlings of plants heterozygous for T-DNA insertion, there was no significant difference in growth and development status from the wild type except for the abnormal embryonic development in the fruit pod. The development of plants homozygous for the insertion was markedl...
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