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Application of gmcop1a and/or gmcop1b in regulating plant height
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A kind of plant and aspect technology, applied in the application field of regulating plant height, can solve the problems of soybean plant type, yield and quality impact, narrow photoperiod adaptability, etc.
Active Publication Date: 2020-08-07
LONGPING BIOTECHNOLOGY (HAINAN) CO LTD
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As a typical short-day plant, the growth period of most soybean varieties has narrow adaptability to the photoperiod in different latitudes; in addition, the light environment in different ecological regions and cultivation modes also has a significant impact on soybean plant type, yield and quality
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[0099] Single colonies of Agrobacterium K599 and EHA105 were picked and placed in 5 mL of LB liquid medium containing corresponding antibiotics. The resistance of K599 was 100 μg / mL streptomycin; the resistance of EHA105 was 100 μg / mL rifampicin (Rif). Cultivate overnight at 28°C; take 500 μL of the overnight culture and inoculate it into 50 mL LB liquid medium containing the corresponding antibiotics, cultivate at 28°C until the OD600 is about 0.5; place on ice for 30 min; centrifuge at 5,000 rpm for 10 min at 4°C, and pre-cool with 15 mL 10mM CaCl 2 Resuspend the Agrobacterium cells, centrifuge at 5,000 rpm for 10 min at 4°C; use 2 mL of pre-cooled 10 ...
Embodiment 3
[0122] Identification of embodiment 3 transgenic positive strains
[0123] In order to determine the transgenic positive plants, take a piece of fresh and tender leaves, extract DNA, and carry out PCR detection. Among them, for the CRISPR knockout mutant, we detected its Basta resistance gene, Cas9 protein and U6 promoter. The location of the gRNA was amplified by PCR and sequenced.
[0124] According to PCR detection, Gmcop1a-4, Gmcop1a-8, Gmcop1b-8 and Gmcop1a-8 / 1b-9 plants all contained Basta, Cas9 and U6, but Gmcop1b-9 was absent. For specific sequencing results, see figure 1 , 2 and 3.
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Abstract
The invention relates to the technical field of biology and specifically discloses application of GmCOP1a and / or GmCOP1b in an aspect of regulating a plant height. The GmCOP1a and the GmCOP1b come from soybeans; amino acid sequences of the GmCOP1a and the GmCOP1b are correspondingly as shown in SEQ ID NO.2 and SEQ ID NO.4. According to the application of the GmCOP1a and / or the GmCOP1b in the aspect of regulating the plant height, coding genes of the GmCOP1a and / or the GmCOP1b are coded by using a CRISPR-Cas9 system; and it is found that the plant height has no obvious change after the GmCOP1amutates, the plant height is reduced after the GmCOP1b mutates, the plant height is reduced after the GmCOP1a and the GmCOP1b both mutate, and the reduction of the plant height of a plant in which theGmCOP1a and the GmCOP1b both mutate is more obvious than the reduction of the plant height of a plant in which only the GmCOP1b mutates. Therefore, the application of the GmCOP1a and / or the GmCOP1b in the aspect of regulating the plant height, disclosed by the invention, provides a new option for plant height regulation means.
Description
technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the application of GmCOP1a and / or GmCOP1b in regulating plant height. Background technique [0002] COP1 is an E3 ubiquitin ligase. Whether in animals or plants, COP1 is relatively conserved during evolution, and plays a very important role in various growth and development processes. Previous studies have shown that in Arabidopsis, COP1 is the main factor responsible for the rapid degradation of photolabile photoreceptors under light. At the same time, COP1 is also a core inhibitor of photomorphogenesis, which can promote the selective degradation of target proteins. The Arabidopsis COP1 gene is at the core of the light signal transduction pathway. The gene responds to and integrates various light signals to regulate plant photomorphogenesis, flowering induction, and biological rhythms (Lau and Deng, 2012). [0003] Soybean is one of the important food, oil and feed cr...
Claims
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