Method for improving rice yield by jointly knocking out aba receptor pyl family genes and use thereof
a technology of pyl and aba receptor, which is applied in the field of agriculture and biotechnology, can solve the problems of little research on pyl in rice, increased stress resistance is often accompanied by growth inhibition, and unclear specific gene functions of its family members
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example 1
Gene Knockout of Members of the Rice PYL Gene Family
[0075]To knock out the PYL gene family, we constructed a CRISPR / Cas9 multiple gene knockout system. In this system, Cas9 was mediated and expressed by the maize Ubiquitin promoter; four sgRNAs were mediated and expressed by the OsU3-1, OsU6-1, OsU3-2, and OsU6-2 promoters respectively (for the promoter sequence, see sequence 1 to sequence 4). Four sgRNA expression cassettes are arranged in tandem on the vector. To specifically target the rice PYL gene, we synthesized a primer that specifically recognized the target gene, which contains a 20 bp target recognition sequence and a 4-5 bp tag sequence. Double-stranded DNA with sticky ends was formed after primer annealing (20 bp double-stranded region). The double-stranded DNA was seamlessly linked to the promoter and downstream sequences under the action of T4 ligase, thereby constructing an sgRNA expression cassette. The sgRNA expression cassettes arranged in tandem were firstly const...
example 2
Subfamily Gene Mutation Promotes Rice Growth
[0078]The homozygous mutant materials obtained in this study were sown at the rice transgenic bases of the Institute of Plant Physiology and Ecology, Chinese Academy of Sciences in Shanghai and Lingshui Li Autonomous County, Hainan Province in mid-June and late-December every year. Field phenotypic observations revealed that there was no significant morphological difference between single-mutant lines or class II gene mutant lines and wild type of all genes (FIG. 2e), while many class I gene mutants showed better growth status than wild type (FIGS. 2b and 2d). At the seedling stage, from pyl1 / 6 to pyl1 / 2 / 3 / 4 / 5 / 6 / 12, most mutants grew more vigorous than the wild type and had stronger growth potential, especially the pyl1 / 4 / 6 triple mutants which had the best growth status (FIGS. 2b and 3a). In order to compare the growth differences between different mutation types and wild types, we calculated the plant height and fresh weight at the seedl...
example 3
Rice PYL Gene on Heading Stage
[0083]Heading stage affects the geographical distribution of rice and its adaptability to the season. In this study, we found that in the class I gene mutants, starting from the four mutants, the heading stage of the mutant lines was significantly delayed (FIG. 2h). Compared to the wild type, the heading stage of pyl1 / 2 / 3 / 4 / 5 / 6 and pyl1 / 2 / 3 / 4 / 5 / 6 / 12 was delayed for about 9 days, and that of pyl1 / 2 / 3 / 4 / 6 was delayed for about 7 days, that of pyl1 / 2 / 3 / 4 was delayed for about 5 days, and that of pyl1 / 4 / 6 was delayed for about 1 day. The heading stage of single mutant, double mutant and other triple mutants of class I gene was not significantly different from that of the wild type. There was no difference between the heading stage of the class II gene mutants and that of the wild type.
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