CN110184293A - A method of increasing phytomass or yield by improving photosynthetic efficiency - Google Patents
A method of increasing phytomass or yield by improving photosynthetic efficiency Download PDFInfo
- Publication number
- CN110184293A CN110184293A CN201910348836.3A CN201910348836A CN110184293A CN 110184293 A CN110184293 A CN 110184293A CN 201910348836 A CN201910348836 A CN 201910348836A CN 110184293 A CN110184293 A CN 110184293A
- Authority
- CN
- China
- Prior art keywords
- gene
- gdh
- plant
- seq
- overexpressed
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8216—Methods for controlling, regulating or enhancing expression of transgenes in plant cells
- C12N15/8221—Transit peptides
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
- C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
- C12N15/8262—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield involving plant development
- C12N15/8269—Photosynthesis
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0006—Oxidoreductases (1.) acting on CH-OH groups as donors (1.1)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/10—Transferases (2.)
- C12N9/1025—Acyltransferases (2.3)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y101/00—Oxidoreductases acting on the CH-OH group of donors (1.1)
- C12Y101/99—Oxidoreductases acting on the CH-OH group of donors (1.1) with other acceptors (1.1.99)
- C12Y101/99014—Glycolate dehydrogenase (1.1.99.14)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y203/00—Acyltransferases (2.3)
- C12Y203/03—Acyl groups converted into alkyl on transfer (2.3.3)
- C12Y203/03009—Malate synthase (2.3.3.9)
Abstract
Increase the method for phytomass or yield by improving photosynthetic efficiency the invention discloses a kind of, the method is that overexpression ethyl alcohol acidohydrogenase (GDH) gene and malate synthase (MS) gene are realized in plant chloroplast.The present invention improves plant photosynthesis efficiency using GDH gene and MS gene is overexpressed, and then the method for improving biomass or yield, under the premise of being overexpressed MS gene at the same time, the GDH of Escherichia coli is overexpressed in plant chloroplast compared with the GDH for being overexpressed green alga source in plant chloroplast, biomass or yield increase 3%-50%.
Description
(1) technical field
It is the present invention relates to a kind of method for improving plant products, in particular to a kind of to improve plant products using photosynthetic efficiency
Method.
(2) background technique
The increase and improvement of living standard of mankind's quantity, need to consume more grains and feed, and this requires having
More grains are harvested on the soil of limit.Therefore, it is extremely important to cultivate new high yield plant variety.
The photosynthate of plant entirety all derives from enzymatic CO2It is converted into Organic Compounds.Bis- phosphorus of 1,5-
Sour ribulose Carboxylase/oxygenase (RubisCO) is the carboxylase in Calvin cycle (Calvin-Benson (CB) cycle).
Due to RubisCO and CO2Or O2It can react, RubisCO and O2Reaction generates phosphoglycollic acid, recycles into light respiration, and light is exhaled
Inhale the waste for leading to carbon and nitrogen fixed in plant.In the world, this process is every year by the fresh assimilation of about 29GT
Carbon is re-released into atmosphere (Anav A, etal.Spatiotemporal patterns of terrestrial gross
primary production:a review.Rev Geophys 2015,53:785-818.)。
In order to reduce loss caused by light respiration, the photosynthetic efficiency of plant is improved, currently used method is by new
Light respiration branch recycles the CO in glycolic2, to achieve the purpose that reducing light respiration improves photosynthetic efficiency
(Peterhansel C,Blume C,Offermann S.Photorespiratory bypasses:how can they
Work? [J] .Journal of Experimental Botany, 2013,64 (3): 709-715.).
Glycolic can be converted into glyoxalic acid by ethyl alcohol acidohydrogenase (glycolate dehydrogenase, GDH).Mesh
The preceding ethyl alcohol acidohydrogenase studied and applied for plant transgene arises primarily at rudimentary plant green alga
(Chlamydomonas reinhardtii) or Escherichia coli.Ethyl alcohol acidohydrogenase in green alga is encoded by a gene, and
Ethyl alcohol acidohydrogenase in escherichia coli is made of tri- subunits of D, E, F that 3 genes encode respectively.There is registration by potato
After being overexpressed the fusion of the encoding gene of tri- subunits of D, E, F, the DEFp fusion protein expression in plant increases, grape
The sugars such as sugar, fructose and sucrose are multiplied, biomass also dramatically increase (Nolke G, Houdelet M, Kreuzaler F,
et al.The expression of a recombinant glycolate dehydrogenase polyprotein in
potato(Solanum tuberosum)plastids strongly enhances photosynthesis and tuber
yield[J].Plant Biotechnology Journal,2014,12(6):734-742.).But Escherichia coli and
All there were significant differences in terms of function and activity for the ethyl alcohol acidohydrogenase in green alga source, so performance in transgenic plants
There is very big difference.
Malate synthase (malate synthase, MS) can be catalyzed acetyl-CoA and glyoxalic acid be converted into malic acid with
CoA.Malate synthase takes part in glyoxalic acid circulation, is widely present in different plants.Studies have reported that crossing table in tobacco
Its photosynthetic efficiency and biomass (PF South, AP can be improved in MS up to the GDH and pumpkin (C.maxima) of green alga
Cavanagh,HW Liu,etal.Synthetic glycolate metabolism pathways stimulate crop
growth and productivity in the field,Science,2019:363(6422):eaat9077.)。
But the function and active difference of the GDH due to Escherichia coli and the GDH in green alga source, it has been found that
In terms of the effect of biomass or yield for increasing plant, the GDH gene of Escherichia coli is than being overexpressed green alga source
GDH gene is more preferable.Under the premise of being overexpressed MS gene at the same time, the GDH of Escherichia coli is overexpressed in plant chloroplast
Compared with the GDH for being overexpressed green alga source in plant chloroplast, biomass or yield increase 3%-50%.Therefore, it is planting
The GDH and another MS that Escherichia coli is overexpressed in object chloroplaset are to increase plant biological by improving photosynthetic efficiency
The best method of amount or yield.
(3) summary of the invention
It is an object of the present invention to provide a kind of reduction plant light respiration, improve plant photosynthesis efficiency, improve phytomass or
The method of yield, the method are the GDH that Escherichia coli source is overexpressed in plant chloroplast and the MS in any source.The present invention
Solve the problems, such as to be used cooperatively by the GDH gene and MS gene of high activity to optimize Plant Light respiratory pathways, obtains biology
Amount or yield increase more genetically modified plants.
The technical solution adopted by the present invention is that:
The present invention provides a kind of method for increasing phytomass or yield by raising photosynthetic efficiency, and the method is
Overexpression ethyl alcohol acidohydrogenase (GDH) gene and malate synthase (MS) gene are realized in plant chloroplast.It is preferred that described
Ethyl alcohol acidohydrogenase GDH gene source in Escherichia coli, the GDH be made of three independent subunits (nucleotide sequence are as follows:
3126522bp-3128021bp in GenBank:CP029238.1, the amino acid of coding are as follows: GenBank:QBP03082.1；
3125470bp-3126522bp in GenBank:CP029238.1, the amino acid of coding are as follows: GenBank:QBP03081.1；Core
Nucleotide sequence are as follows: 3124236bp -3125459bp in GenBank:CP029238.1, the amino acid of coding are as follows: GenBank:
QBP03080.1), the more preferably GDH gene order is as shown in 125bp-4009bp in SEQ ID NO.1, amino acid sequence
In SEQ ID NO.2 shown in 48-1333.
Malate synthase MS gene source of the present invention is in prokaryotes or eucaryote, such as MS base shown in table 1
Cause, preferably MS gene nucleotide series such as SEQ ID NO.3, SEQ ID NO.4 (11bp-1852bp) or SEQ ID NO.5 institute
Show, amino acid sequence is as shown in SEQ ID NO.6, SEQ ID NO.7 and SEQ ID NO.8.
Table 1: malate synthase (MS) gene in different plant species source
Further, the GDH gene and MS gene be overexpressed in chloroplaset by signal it is peptide-mediated, it is described mediation be by
Chloroplast signal peptide sequence merges the N-terminal in GDH or MS albumen；The signal peptide derives from plant RuBisCO small subunit
(RbcS) or phosphoglucomutase transit peptide sequence, the amino acid sequence of preferred signals peptide such as SEQ ID NO.9 or SEQ
Shown in ID NO.10.
Further, it further includes promoter that the GDH gene and MS gene are overexpressed in chloroplaset, the promoter source
In eucaryote or prokaryotes, artificial synthesized acquisition can also be passed through；Preferred promoter constitutive promoter or specificity open
Mover, the nucleotides sequence of the more preferable promoter are classified as UBI promoter (4879bp- in GenBank:KR297238.1
Shown in 6876bp), Act1 promoter (in GenBank:AY452735.1 shown in 2428bp-3797bp) or 35S promoter
(in GenBank:MG719235.1 shown in 848bp-1628bp).
Further, it further includes terminator that the GDH gene and MS gene are overexpressed in chloroplaset, the terminator source
It can also be SEQ by artificial synthesized acquisition, the preferably described terminator nucleotides sequence in eucaryote or prokaryotes
Shown in ID NO.11 or SEQ ID NO.12.
Overexpression of the present invention can be realized by the method for molecule aggregation or hybridization polymerization；Molecule aggregation refer to by
T-DNA is transferred on the T-DNA of the same carrier by transgene method by the expression frame construction of GDH gene and MS gene
Recipient plant genome in so that being overexpressed GDH gene and MS gene simultaneously in target plant.Hybridization polymerization refers to point
It Huo get not be overexpressed the plant of GDH and MS gene in chloroplaset, then with the method for traditional breeding method, GDH gene will be expressed respectively
Hybridized with the plant of MS gene, obtains while being overexpressed the plant of GDH and MS gene.
Further, the method that the GDH gene and MS gene are overexpressed in chloroplaset are as follows: (1) with commercialized double base
Based on carrier pCambia1300, by XhoI restriction enzyme site hptII (hygromycin resistance) base of carrier
Because being replaced as the bar gene (GenBank:MG719235.1 (287bp-837bp)) of resistance to glufosinate-ammonium, the carrier after displacement is named as
pCambia1300-bar；(2) promoter and GDH gene pCambia1300-bar is connected by the site EcoRI and KpnI to carry
In body, transition vector pCambia1300-bar-GDH is obtained；(3) again by the site KpnI and HindIII promoter and MS base
Because being connected into transition vector pCambia1300-bar-GDH, whole carrier pCambia1300-bar-GDH-MS, i.e. T-DNA are obtained
Plasmid.(4) finally, T-DNA plasmid is transferred in Agrobacterium LB4404 by the method that electricity turns, by containing 15 Fourth Rings μ g/ml
The YEP solid medium of the kanamycins of element and 50 μ g/mL filters out positive colony, and screening obtains light respiration and reduces, photosynthetic effect
Rate improves, the plant of biomass or output increased.
Plant of the present invention includes corn, rice and soybean.
Compared with prior art, beneficial effect of the present invention is mainly reflected in:
Plant photosynthesis efficiency is improved using GDH gene and MS gene is overexpressed the present invention provides a kind of, and then is improved
The method of biomass or yield under the premise of being overexpressed MS gene at the same time, is overexpressed Escherichia coli in plant chloroplast and comes
For the GDH in source compared with the GDH for being overexpressed green alga source in plant chloroplast, biomass or yield increase 3%-50%.
(4) specific embodiment
The present invention is described further combined with specific embodiments below, but protection scope of the present invention is not limited in
This:
The building of embodiment 1, carrier
In order to construct conversion carrier, the GDH gene of artificial synthesized Escherichia coli and corresponding terminator sequence,
Comprising chloroplast signal peptide, GDH encoding gene and terminator, nucleotide sequence (coding histone amino as shown in SEQ ID NO.1
Acid sequence is shown in SEQ ID NO.2), the end 5' and the end 3' are respectively arranged with the site BamHI and KpnI.
The artificial synthesized MS gene of rice includes chloroplast signal peptide, MS encoding gene and terminator, nucleotide sequence
As shown in SEQ ID NO.4, the end 5' and the end 3' are respectively arranged with the site BamHI and HindIII.
Meanwhile the 35S promoter sequence of artificial synthesized cauliflower mosaic virus (CaMV) is (in GenBank:MG719235.1
Shown in 848bp-1628bp), the end 5' and the end 3' are respectively arranged with the site KpnI and BamHI.Artificial synthesized corn Ubi promoter sequence
It arranges (in GenBank:KR297238.1 shown in 4879bp-6876bp), the end 5' and the end 3' are respectively arranged with EcoRI and BamHI
Point.
It can be used for binary vector used in Agrobacterium approach conversion plant to construct, with commercialized binary vector
Based on pCambia1300, hptII (hygromycin resistance) gene before is set by XhoI restriction enzyme site
Changing the bar gene of resistance to glufosinate-ammonium into, (GenBank:MG719235.1 (287bp-837bp), the carrier after displacement are named as
pCambia1300-bar。
Ubi promoter and GDH gene are connected into pCambia1300-bar carrier by the site EcoRI and KpnI, obtained
Transition vector pCambia1300-bar-GDH.35S promoter and MS gene were connected by the site KpnI and HindIII again
It crosses in carrier pCambia1300-bar-GDH, obtains T-DNA plasmid pCambia1300-bar-GDH-MS.
As control, the artificial synthesized CrGDH gene in green alga source includes chloroplast signal peptide, CrGDH encoding gene
And terminator, for nucleotide sequence as shown in SEQ ID NO.13, the end 5' and the end 3' are respectively arranged with the site BamHI and KpnI.
Ubi promoter and CrGDH gene are connected into pCambia1300-bar carrier by the site EcoRI and KpnI, obtained
Obtain transition vector pCambia1300-bar-CrGDH.35S promoter and MS gene are connected by the site KpnI and HindIII again
Enter in transition vector pCambia1300-bar-CrGDH, obtains T-DNA plasmid pCambia1300-bar-CrGDH-MS.
Finally, T-DNA plasmid is transferred in Agrobacterium LB4404 by the method that electricity turns, by containing 15 Fourth Rings μ g/ml
The YEP solid medium of the kanamycins of element and 50 μ g/mL filters out positive colony, and protects bacterium, turns for next plant
Change.
YEP solid medium composition: beef extract 5g/L, yeast extract 1g/L, peptone 5g/L, sucrose 5g/L,
MgSO4·H2O 0.5g/L, solvent are water, pH 7.0.
Embodiment 2, rice conversion
The preparation method of transgenic paddy rice is using the prior art (Lu Xiongbin, Gong ancestral an ancient egg-shaped, holed wind instrument (1998) life science 10:125-
131；Liu Fan etc. (2003) Molecular Plant Breeding 1:108-115).Choose " elegant water -134 " seed decladding of mature and plump, induction
Callus is generated as converting material.Built in Example 1 respectively containing pCambia1300-bar-GDH-MS,
The Agrobacterium of pCambia1300-bar-CrGDH-MS and pCambia1300-bar-GDH plasmid draws plate.Single colonie inoculation is chosen, it is quasi-
Agrobacterium is used in standby conversion.Callus to be transformed is put into the (system of Agrobacterium bacterium solution in the Agrobacterium bacterium solution that OD600 is 0.6
Standby: by Agrobacterium inoculation to culture medium, culture to OD600 is 0.6；Culture medium composition: 3g/L K2HPO4、1g/LNaH2PO4、
1g/LNH4Cl、0.3g/L MgSO4·7H2O、0.15g/L KCl、0.01g/L CaCl2、0.0025g/L FeSO4·7H2O、
5g/L sucrose, 20mg/L acetosyringone, solvent are water, pH=5.8), allow Agrobacterium to be integrated to callus surface, then
Callus is transferred to co-culture medium (MS+2mg/L 2,4-D+30g/L glucose+30g/L sucrose+3g/L agar
(sigma 7921)+20mg/L acetosyringone) in, it co-cultures 2-3 days.Callus after being converted with aseptic water washing, is transferred to
Screening and culturing medium (MS+2mg/L2,4-D+30g/ sucrose+3g/L agar (sigma 7921)+20mg/L acetosyringone+2mM grass
Sweet phosphine (Sigma)) on, screening and culturing two months (intermediate subculture is primary).After screening, the good callus of growth vigor is transferred to
Pre- differential medium (MS+0.1g/L inositol+5mg/L ABA+1mg/L NAA+5mg/L 6-BA+20g/L sorbierite+30g/L sugarcane
Sugar+2.5g/L gelrite) on cultivate 20 days or so, then the callus broken up in advance is moved on on differential medium, often
Its illumination in 14 hours differentiation germination.After 2-3 weeks, resistance regeneration plant is transferred to root media (1/2MS+0.2mg/L NAA
+ 20g/L sucrose+2.5g/L gelrite) on strengthening seedling and rooting, regeneration plant is finally washed away into agar transplanting in greenhouse, selection produces
Amount is high, seed is big or biomass is high etc. can be improved the transgenic line of rice yield, cultivate new varieties.It is obtained respectively containing upper
State the transgenic rice plant of conversion carrier.
3. transformation of soybean of embodiment
The step of acquisition genetically engineered soybean used herein from existing technology (Deng et al., 1998, Plant
Physiology Communications 34:381-387；Ma et al.,2008,Scientia
AgriculturaSinica 41:661-668；Zhou et al.,2001,Journal of Northeast
Agricultural University 32:313-319).Healthy, full, mature " grand No. 1 of day " soybean is chosen, with 80%
Ethanol disinfection 2 minutes, then with sterile water wash, be then placed within and (react life with the dense HCl of 2ml by 50mlNaClO full of chlorine
At) drier in sterilize 4-6 hour.Soybean after sterilizing is sowed in superclean bench into B5 medium, and 25 DEG C
Under the conditions of cultivate 5 days, with optical densities in 90-150 μm of ol photon/m2S is horizontal.When cotyledon greening and top is broken in the seed coat, sterile
Bean sprouts will grow.The bean sprouts for eliminating hypocotyl is cut into fifty and fifty percent in length, so that two panels explant all has son
Leaf and epicotyl.It is cut at the node of cotyledon and epicotyl at explant about 7-8, that is, can be used as the destination organization infected.
It takes respectively and contains carrier pCambia1300-bar-GDH-MS, pCambia1300- by what embodiment 1 constructed
The monoclonal Agrobacterium of bar-CrGDH-MS and pCambia1300-bar-GDH is stand-by by separated culture.Ready explant
It is immersed in agrobacterium suspension and co-cultures 30 minutes or so.Then, the extra cell suspending liquid of the tissue infected is inhaled
Water paper absorbs cleanly, and is then transferred to 25 DEG C dark culture 3-5 days in 1/10B5 co-culture medium.
The plant tissue of co-cultivation is cleaned with B5 fluid nutrient medium, and to remove extra Agrobacterium, it is solid to be then placed into B5
It is cultivated 5 days at 25 DEG C in body culture medium, to its germination.The embryonic tissue that induction occurs is transferred to the sieve of the B5 containing 0.1M glyphosate
Select in culture medium, 25 DEG C illumination cultivation 4 weeks, during which replace a subculture every two weeks.The embryonic tissue screened retransfers
Into B5 solid medium, 25 DEG C of cultures grow up to seedling to it.Then, transgenic plant seedling is transferred in 1/2B5 culture medium
Carry out rooting induction.Finally, being planted in the greenhouse after the cleaned removal agar of the plantlet grown up to.
Embodiment 4: the identification of transgenic paddy rice
Carrier pCambia1300-bar-GDH-MS (GM), pCambia1300-bar- are obtained respectively by embodiment 2
The transgenic rice plant of CrGDH-MS (CGM) and pCambia1300-bar-GDH (G).Above-mentioned transgenic plant turns base with non-
Because control all increased compared to Biomass and yield, also, increasing degree of the GM plant in terms of biomass or yield is maximum.
In order to further identify the performance variation of GM transgenic plant, we to the biomass of above-mentioned transgenic plant and seed production into
Assessment is gone, as a result as shown in table 2 below.Biomass or yield the ratio CGM of GM rice plants and G plant increase 5%-50%.
Table 2
| GM | CGM | G | |
| Biomass amplitude more increased than non-transgenic control | 25% | 15% | 9% |
| Yield amplitude more increased than non-transgenic control | 18% | 12% | 6% |
Embodiment 5: the identification of genetically engineered soybean
Carrier pCambia1300-bar-GDH-MS (GM), pCambia1300-bar- are obtained respectively by embodiment 3
The Transgenic soybean plants of CrGDH-MS (CGM) and pCambia1300-bar-GDH (G).Above-mentioned transgenic plant turns base with non-
Because control all increased compared to Biomass and yield, also, increasing degree of the GM plant in terms of biomass or yield is maximum.
In order to further identify the performance variation of GM transgenic plant, we to the biomass of above-mentioned transgenic plant and seed production into
Assessment is gone, as a result as shown in table 3 below.Biomass or yield the ratio CGM of GM bean plant and G plant increase 5%-50%.
Table 3
| GM | CGM | G | |
| Biomass amplitude more increased than non-transgenic control | 21% | 12% | 7% |
| Yield amplitude more increased than non-transgenic control | 14% | 8% | 5% |
Claims (7)
1. a kind of method for increasing phytomass or yield by improving photosynthetic efficiency, it is characterised in that the method is to plant
What the ethyl alcohol acidohydrogenase GDH gene and malate synthase MS gene that Escherichia coli are overexpressed in object chloroplaset were realized.
2. the method as described in claim 1, it is characterised in that the ethyl alcohol acidohydrogenase GDH gene nucleotide series such as SEQ
In ID NO.1 shown in 125bp-4009bp.
3. method according to claim 2, it is characterised in that the amino acid sequence of the ethyl alcohol acidohydrogenase GDH gene coding
As shown in 48-1333 in SEQ ID NO.2.
4. the method as described in claim 1, it is characterised in that the malate synthase MS gene nucleotide series such as SEQ ID
In NO.3, SEQ ID NO.4 shown in one of 11bp-1852bp or SEQ ID NO.5.
5. the method as described in claim 1, it is characterised in that the GDH gene and MS gene are overexpressed in chloroplaset and pass through
Signal is peptide-mediated；The amino acid sequence of the signal peptide is as shown in SEQ ID NO.9 or SEQ ID NO.10.
6. method as claimed in claim 5, it is characterised in that the GDH gene and MS gene are overexpressed in chloroplaset and also wrap
Terminator is included, the terminator nucleotides sequence is shown in SEQ ID NO.11 or SEQ ID NO.12.
7. the method as described in claim 1, it is characterised in that the plant includes corn, rice and soybean.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910348836.3A CN110184293B (en) | 2019-04-28 | 2019-04-28 | Method for increasing plant biomass or yield by improving photosynthetic efficiency |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910348836.3A CN110184293B (en) | 2019-04-28 | 2019-04-28 | Method for increasing plant biomass or yield by improving photosynthetic efficiency |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110184293A true CN110184293A (en) | 2019-08-30 |
| CN110184293B CN110184293B (en) | 2021-06-22 |
Family
ID=67715208
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910348836.3A Active CN110184293B (en) | 2019-04-28 | 2019-04-28 | Method for increasing plant biomass or yield by improving photosynthetic efficiency |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110184293B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110527689A (en) * | 2019-09-20 | 2019-12-03 | 武汉大学 | A kind of application of rice light controlling gene PSB27 |
| CN115896145A (en) * | 2022-11-30 | 2023-04-04 | 上海市农业科学院 | Method for creating completely degraded 1,2-dichloroethane engineering bacteria by multigene series method and application |
| US11926833B2 (en) | 2022-01-25 | 2024-03-12 | Living Carbon PBC | Compositions and methods for enhancing biomass productivity in plants |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102016012A (en) * | 2008-02-21 | 2011-04-13 | 科隆大学 | Means for improving agrobiological traits in a plant by providing a plant cell comprising in its chloroplasts enzymatic activities for converting glycolate into malate |
| US20180258440A1 (en) * | 2017-03-07 | 2018-09-13 | The United States Of America, As Represented By The Secretary Of Agriculture | Plants With Increased Photorespiration Efficiency |
-
2019
- 2019-04-28 CN CN201910348836.3A patent/CN110184293B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102016012A (en) * | 2008-02-21 | 2011-04-13 | 科隆大学 | Means for improving agrobiological traits in a plant by providing a plant cell comprising in its chloroplasts enzymatic activities for converting glycolate into malate |
| US20180258440A1 (en) * | 2017-03-07 | 2018-09-13 | The United States Of America, As Represented By The Secretary Of Agriculture | Plants With Increased Photorespiration Efficiency |
Non-Patent Citations (4)
| Title |
|---|
| NOLKE G等: "The expression of a recombinant glycolate dehydrogenase polyprotein in potato (Solanum tuberosum) plastids strongly enhances photosynthesis and tuber yield", 《PLANT BIOTECHNOLOGY JOURNAL》 * |
| PAUL F. SOUTH等: "Synthetic glycolate metabolism pathways stimulate crop growth and productivity in the field", 《SCIENCE》 * |
| 姚振等: "马铃薯多基因质体转化载体及其在光呼吸支路构建中的应用", 《分子植物育种》 * |
| 胥华伟等: "集胞藻PCC6803乙醇酸脱氢酶基因的克隆及其叶绿体定位分析", 《核农学报》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110527689A (en) * | 2019-09-20 | 2019-12-03 | 武汉大学 | A kind of application of rice light controlling gene PSB27 |
| US11926833B2 (en) | 2022-01-25 | 2024-03-12 | Living Carbon PBC | Compositions and methods for enhancing biomass productivity in plants |
| CN115896145A (en) * | 2022-11-30 | 2023-04-04 | 上海市农业科学院 | Method for creating completely degraded 1,2-dichloroethane engineering bacteria by multigene series method and application |
| CN115896145B (en) * | 2022-11-30 | 2023-12-01 | 上海市农业科学院 | Method for creating completely-degradable 1,2-dichloroethane engineering bacteria by polygene series method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110184293B (en) | 2021-06-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101948867B (en) | Agrobacterium-mediated jatropha curcas gene transformation method | |
| CN102154364A (en) | Method for agrobacterium tumefaciens-mediated genetic transformation of sugarcane | |
| CN110564760B (en) | Method for improving drought tolerance of plants through light respiration | |
| CN103484436B (en) | Corn (zea mays) derived growth period related protein ZmHUB2 and coding gene and application thereof | |
| WO2022135246A1 (en) | R gene for controlling matching of soybean-rhizobium, protein and use thereof | |
| CN110184293A (en) | A method of increasing phytomass or yield by improving photosynthetic efficiency | |
| CN102719433B (en) | Application of osa-MIR167a gene for regulating and controlling plant type of paddy rice | |
| CN110358772B (en) | OsEBP89 gene for improving abiotic stress resistance of rice, and preparation method and application thereof | |
| CN108893481A (en) | Tomato SlOAS7 gene and its application | |
| CN106544344A (en) | A kind of application of tissue-specific promoter in regulation and control crop yield | |
| CN102250943A (en) | In-vitro tissue culturing method for soybeans under mediation of agrobacterium tumefaciens | |
| CN112626111B (en) | Herbicide resistance gene expression vector and application thereof | |
| ZA200101818B (en) | Methods for producing genetically modified plants, plant materials and plant products produced thereby. | |
| AU777365B2 (en) | Agrobacterium-mediated transformation of cotton with novel explants | |
| CN110628810B (en) | Method for improving photosynthetic efficiency of plant | |
| CA2341781A1 (en) | Methods for producing genetically modified plants, plant materials and plant products produced thereby | |
| Akashi et al. | High-frequency embryogenesis from cotyledons of bird's-foot trefoil (Lotus corniculatus) and its effective utilization in Agrobacterium tumefaciens-mediated transformation | |
| CN109609541B (en) | Method for improving crop traits | |
| CN113444740B (en) | Recombinant vector capable of improving soybean aphid resistance gene and application thereof | |
| CN117187294B (en) | Application of BnaC5.ACBP4 gene in improving flooding resistance of plants | |
| CN112126653B (en) | 0scpy and application of encoding protein thereof in rice seeding | |
| Curtis | Lettuce (Lactuca sativa L.) | |
| KR100770203B1 (en) | Method to improve plants phosphate uptake by increased Nicotiana tabacum phosphate transporter transgene expression and plants with improved phosphate uptake developed by the application of the method | |
| Shinoyama et al. | Establishment of an efficient genetic transformation system for Tanacetum cinerariifolium | |
| CN116479009A (en) | Novel gene for regulating and controlling plant fruit and seed size and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |