CN105198967A - Application of protein VdAL to improvement of plant product quality - Google Patents
Application of protein VdAL to improvement of plant product quality Download PDFInfo
- Publication number
- CN105198967A CN105198967A CN201510691295.6A CN201510691295A CN105198967A CN 105198967 A CN105198967 A CN 105198967A CN 201510691295 A CN201510691295 A CN 201510691295A CN 105198967 A CN105198967 A CN 105198967A
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- vdal
- plant
- protein
- dilution
- sequence
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Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
-
- 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
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- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Wood Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biophysics (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Zoology (AREA)
- Biomedical Technology (AREA)
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Abstract
The invention discloses application of protein VdAL to improvement of plant product quality. In the application provided by the invention, the protein VdAL is A1, A2 or A3 as follows: A1) protein with an amino acid sequence of 1; A2) protein having the same function and derived from A1 by substituting and/or deleting and/or adding one or a plurality of amino acid residues in the amino acid sequence 1; A3) fused protein obtained by connecting labels at the N-terminal or/and C-terminal of A1 or A2. As proved by experiments, the protein VdAL provided by the invention and biological preparations thereof can improve the plant product quality.
Description
Technical field
The present invention relates to protein VdAL in biological technical field and improve the application in plant prod quality.
Background technology
The Sustainable high yield of current China big area grain and cash crop mainly relies on and uses chemical fertilizer, prevention and elimination of disease and pests mainly to rely in a large number to spray chemical pesticide in a large number.Chinese Academy of Agricultural Sciences's rural economy and Institute of Development Studies researcher Hu Dinghuan, when referring to agricultural pollution problem, he points out, our country is present still quite serious, the excessive use of chemical fertilizer and agricultural chemicals, and in aquaculture, a large amount of microbiotic of using and heavy metal additive all will cause serious pollution to agriculture production.
This province usage quantity of current chemical fertilizer year is added up more than 30,000 tons according to Liaoning Province, unit surface chemical fertilizer application intensity about 340 kilograms of per hectares, country's ecotope construction requirements per hectare limit value is then per hectare 250 kilograms, the fertile light general problem of fertilization lack of standardization of phosphate fertilizer potash fertilizer phenomenon of diazonium is also comparatively outstanding, and Liaoning Province's agricultural chemicals annual consumption is more than 1.4 ten thousand tons and in ascendant trend year by year.Remains of pesticide is by very easily causing Environment Pollution Event in the form entered environment of atmospheric precipitation and rain drop erosion and agricultural-food.The environmental protection Room, Liaoning Province rural area department director Wang Jun represents in recent years along with rural environmental pollution problem outstanding day by day environment letters and calls case is situation occurred frequently.According to statistics the every annual of counties and districts concerning farmers of 74, Jin Liangnian Liaoning Province environment letters and calls case 4310 wherein nearly 2795 of case concerning farmers account for about 65% of letters and calls case.
Strengthen antagonistic action between ion when excessive fertilization can cause farm crop to absorb nutritive element, if the existence of certain nutrient ion high density can suppress another kind of or the activity of multiple nutrient ion, thus affect the absorption of farm crop to another kind of nutrient ions.As in acid soil, the sowing amount of nitrogenous fertilizer is too much, Crop calcium ion is just very difficult.If excessively use the trace element deficiency such as zinc, boron, iron, magnesium, manganese that calcareous fertilisers can bring out farm crop.Potassium Fertilizer too much also can affect the absorption of farm crop to calcium ion, magnesium ion; And excessive fertilization easily causes farm crop poisoning, because applying a large amount of fertilizer, add the concentration of the soil solution, make farm crop Root Absorption moisture difficulty, cause overground part to wilt, plant is withered.In addition, excessive fertilization also affects the quality of agricultural-food, the crop growth intermediary and later stages, if apply chemical fertilizer in a large number, agricultural-food organ sugar degree can be made to reduce, not storage tolerance, thus affect commodity value, reduces the economic benefit of farm crop.
Summary of the invention
Technical problem to be solved by this invention how to improve plant prod quality.Described plant prod is the product that the necessary for human of mankind cultivated plant production is wanted.The judgement criteria of described quality is different because of the purposes difference of product.Quality as the product of food can be withdrawn deposit in the nutritional quality and eating quality of product; The quality being used as the product of clothing raw material can be withdrawn deposit on the fibrous quality of product.Evaluate product quality, general adopt two kinds of indexs, one is the content of biochemical component and objectionable impurities, as nutrition content and micronutrient levels; Two is physical indexs, as the shape, size, flavour, fragrance, color and luster, seed coat thickness, reguarity, staple length, fibre strength etc. of product, as the ginning outturn of cotton.
For solving the problems of the technologies described above, the present invention provide firstly protein and is improving the application in plant prod quality.
Protein provided by the present invention is improving in the application in plant prod quality, and the name of described protein is called VdAL, is following A1) or A2) or A3):
A1) aminoacid sequence is the protein of sequence 1;
A2) in the aminoacid sequence of sequence 1 through replacement and/or disappearance and/or add that one or several amino-acid residue obtains have identical function by A1) derivative protein;
A3) at A1) or N end A2) or/and C end connects the fused protein that obtains of label.
Wherein, sequence 1 is made up of 297 amino acid.
In order to make A1) in protein be convenient to purifying, in sequence table, the N-terminal of the protein shown in sequence 1 or C-terminal can connect label as shown in table 1.
The sequence of table 1, label
| Label | Residue | Sequence |
| Poly-Arg | 5-6 (being generally 5) | RRRRR |
| Poly-His | 2-10 (being generally 6) | HHHHHH |
| FLAG | 8 | DYKDDDDK |
| Strep-tag II | 8 | WSHPQFEK |
| c-myc | 10 | EQKLISEEDL |
Above-mentioned A2) in VdAL can synthetic, also can first synthesize its encoding gene, then carry out biological expression and obtain.Above-mentioned A2) in the encoding gene of VdAL by the codon by lacking one or several amino-acid residue in the DNA sequence dna shown in sequence in sequence table 2, and/or carry out the missense mutation of one or several base pair, and/or the encoding sequence connecting the label shown in table 1 is held to obtain at its 5 ' end and/or 3 '.
Above-mentioned VdAL is improving in the application in plant prod quality, and described raising plant prod quality can be the content that improves described plant prod nutritive ingredient and/or improves the content of described plant prod medium trace element and/or improve the palatability of described plant prod.Described plant prod nutritive ingredient specifically can be vitamins C, soluble sugar and/or protein.Described trace element specifically can be Cu, Fe, K, Mn and/or Zn.Described palatability may be embodied in dry rate and/or the decline of coarse-fibred content.
Above-mentioned VdAL is improving in the application in plant prod quality, and described raising plant prod quality can be the quality improving described plant prod by promoting the maturation of described plant prod.In one embodiment of the invention, VdAL uses the maturation promoting cotton production, and before the frost of cotton, flower output increases, and then improves cotton quality.
Above-mentioned VdAL is improving in the application in plant prod quality, and described plant can be dicotyledons or monocotyledons.
Above-mentioned VdAL is improving in the application in plant prod quality, described dicotyledons can be following a), b), c) or d):
A) cucurbitaceous plant;
B) watermelon;
C) Malvaceae plant;
D) cotton.
For solving the problems of the technologies described above, present invention also offers the biomaterial relevant to VdAL and improving the application in plant prod quality.
The biomaterial relevant to VdAL provided by the present invention is improving the application in plant prod quality;
Described biomaterial is following B1) to B20) in any one:
B1) nucleic acid molecule of coding VdAL;
B2) containing B1) expression cassette of described nucleic acid molecule;
B3) containing B1) recombinant vectors of described nucleic acid molecule;
B4) containing B2) recombinant vectors of described expression cassette;
B5) containing B1) recombinant microorganism of described nucleic acid molecule;
B6) containing B2) recombinant microorganism of described expression cassette;
B7) containing B3) recombinant microorganism of described recombinant vectors;
B8) containing B4) recombinant microorganism of described recombinant vectors;
B9) containing B1) the transgenic plant cells system of described nucleic acid molecule;
B10) containing B2) the transgenic plant cells system of described expression cassette;
B11) containing B3) the transgenic plant cells system of described recombinant vectors;
B12) containing B4) the transgenic plant cells system of described recombinant vectors;
B13) containing B1) Transgenic plant tissue of described nucleic acid molecule;
B14) containing B2) Transgenic plant tissue of described expression cassette;
B15) containing B3) Transgenic plant tissue of described recombinant vectors;
B16) containing B4) Transgenic plant tissue of described recombinant vectors;
B17) containing B1) the transgenic plant organ of described nucleic acid molecule;
B18) containing B2) the transgenic plant organ of described expression cassette;
B19) containing B3) the transgenic plant organ of described recombinant vectors;
B20) containing B4) the transgenic plant organ of described recombinant vectors.
The above-mentioned biomaterial relevant to VdAL is improving in the application in plant prod quality, B1) described nucleic acid molecule can be following b1) b2) or b3) gene:
B1) nucleotide sequence is cDNA molecule or the DNA molecular of sequence 2 in sequence table;
B2) nucleotide sequence and b1) limited has more than 75% or 75% identity, and the cDNA molecule of coding VdAL or genomic DNA molecule;
B3) under strict conditions with b1) nucleotide sequence hybridization that limits, and the cDNA molecule of coding VdAL or genomic DNA molecule.
Wherein, described nucleic acid molecule can be DNA, as cDNA, genomic dna or recombinant DNA; Described nucleic acid molecule can be also RNA, as mRNA or hnRNA etc.
Wherein, sequence 2 is made up of 894 Nucleotide, the protein shown in encoding sequence 1.
Those of ordinary skill in the art can adopt known method easily, the method for such as orthogenesis and point mutation, suddenly change to the nucleotide sequence of coding VdAL of the present invention.Those are through manually modified, have and be separated the nucleotide sequence 75% of the VdAL obtained or the Nucleotide of higher identity with the present invention, as long as coding VdAL and have VdAL function is all be derived from nucleotide sequence of the present invention and be equal to sequence of the present invention.
Term used herein " identity " refers to the sequence similarity with native sequence nucleic acid.The nucleotide sequence that " identity " comprises the protein formed with the aminoacid sequence shown in the VdAL of coding of the present invention has 75% or higher, or 85% or higher, or 90% or higher, or the nucleotide sequence of 95% or higher identity.Identity can with the naked eye or computer software evaluate.Use computer software, the identity between two or more sequence can represent with per-cent (%), and it can be used for evaluating the identity between correlated series.
The above-mentioned biomaterial relevant to VdAL is improving in the application in plant prod quality, described stringent condition is at 2 × SSC, in the solution of 0.1%SDS, hybridize at 68 DEG C and wash film 2 times, each 5min, again in the solution of 0.5 × SSC, 0.1%SDS, hybridize at 68 DEG C and wash film 2 times, each 15min; Or, in the solution of 0.1 × SSPE (or 0.1 × SSC), 0.1%SDS, hybridize under 65 DEG C of conditions and wash film.
More than above-mentioned 75% or 75% identity, can be the identity of more than 80%, 85%, 90% or 95%.
The above-mentioned biomaterial relevant to VdAL is improving in the application in plant prod quality, the expression cassette (VdAL expression casette) of the nucleic acid molecule containing coding VdAL B2), refer to the DNA that can express VdAL in host cell, this DNA not only can comprise the promotor starting VdAL genetic transcription, also can comprise the terminator stopping VdAL genetic transcription.Further, described expression cassette also can comprise enhancer sequence.Promotor used in the present invention includes but not limited to: constitutive promoter, the promotor that tissue, organ and growth are special, and inducible promoter.The example of promotor includes but not limited to: the constitutive promoter 35S of cauliflower mosaic virus: from the wound-inducible promoter of tomato, leucine aminopeptidase (" LAP ", the people such as Chao (1999) PlantPhysiol120:979-992); From tobacco chemical inducible promoter, pathogeny be correlated with 1 (PR1) (by Whitfield's ointment and BTH (diazosulfide-7-carbothioic acid S-methyl ester) induction); Tomato proteinase inhibitor II promotor (PIN2) or LAP promotor (all available methyl jasmonate induction); Heat-shock promoters (United States Patent (USP) 5,187,267); Tetracycline inducible promoter (United States Patent (USP) 5,057,422); Seed specific promoters, as Millet Seed specificity promoter pF128 (CN101063139B (Chinese patent 200710099169.7)), the special promotor of seed storage protein matter (such as, the promotor (people (1985) EMBOJ.4:3047-3053 such as Beachy) of phaseollin, napin, oleosin and soybean betaconglycin).They can be used alone or are combined with other plant promoter.All reference cited herein all quote in full.Suitable transcription terminator includes but not limited to: Agrobacterium nopaline syntase terminator (NOS terminator), cauliflower mosaic virus CaMV35S terminator, tml terminator, pea rbcSE9 terminator and nopaline and octopine synthase terminator (see, such as: the people (I such as Odell
985) Nature313:810; The people such as Rosenberg (1987) Gene, 56:125; The people such as Guerineau (1991) Mol.Gen.Genet, 262:141; Proudfoot (1991) Cell, 64:671; The people GenesDev. such as Sanfacon, 5:141; The people such as Mogen (1990) PlantCell, 2:1261; The people such as Munroe (1990) Gene, 91:151; The people such as Ballad (1989) NucleicAcidsRes.17:7891; The people such as Joshi (1987) NucleicAcidRes., 15:9627).
Available existing expression vector establishment contains the recombinant vectors of described VdAL expression casette.Described plant expression vector comprises double base agrobacterium vector and can be used for the carrier etc. of plant micropellet bombardment.As pAHC25, pBin438, pCAMBIA1302, pCAMBIA2301, pCAMBIA1301, pCAMBIA1300, pBI121, pCAMBIA1391-Xa or pCAMBIA1391-Xb (CAMBIA company) etc.Described plant expression vector also can comprise 3 ' end untranslated region of foreign gene, namely comprises the DNA fragmentation of polyadenylation signals and any other participation mRNA processing or genetic expression.The bootable polyadenylic acid of described polyadenylation signals joins 3 ' end of mRNA precursor, as Agrobacterium crown-gall nodule induction (Ti) plasmid gene (as rouge alkali synthetase gene Nos), plant gene (as soybean storage protein genes) 3 ' hold the non-translational region of transcribing all to have similar functions.When using gene constructed plant expression vector of the present invention, also enhanser can be used, comprise translational enhancer or transcriptional enhancer, these enhanser regions can be ATG initiator codon or neighboring region initiator codon etc., but must be identical with the reading frame of encoding sequence, to ensure the correct translation of whole sequence.The source of described translation control signal and initiator codon is widely, can be natural, also can be synthesis.Translation initiation region can from transcription initiation region or structure gene.For the ease of identifying transgenic plant cells or plant and screening, can process plant expression vector used, the coding can expressed in plant as added can produce the enzyme of colour-change or the gene (gus gene of luminophor, luciferase genes etc.), antibiotic marker gene is (as given the nptII gene to kantlex and associated antibiotic resistance, give the bar gene to herbicide phosphinothricin resistance, give the hph gene to microbiotic hygromycin resistance, with the dhfr gene given methotrexate resistance, give EPSPS gene to glyphosate) or chemical resistance reagent marker gene etc. (as anti-weedkiller gene), the mannose-6-phosphate isomerase gene of metabolism seminose ability is provided.From the security consideration of transgenic plant, any selected marker can not be added, directly with adverse circumstance screening transformed plant.
The above-mentioned biomaterial relevant to VdAL is improving in the application in plant prod quality, and described carrier can be plasmid, glutinous grain, phage or virus vector.Described plasmid can be carrier pET42a (+).
The above-mentioned biomaterial relevant to VdAL is improving in the application in plant prod quality, and described microorganism can be yeast, bacterium, algae or fungi, as intestinal bacteria.Described intestinal bacteria can be e. coli jm109.
The above-mentioned biomaterial relevant to VdAL is improving in the application in plant prod quality, and described transgenic plant cells system, Transgenic plant tissue and transgenic plant organ all do not comprise reproductive material.
In an embodiment of the invention, the encoding gene of VdAL is imported in e. coli jm109 by the recombinant vectors of the expression cassette of the encoding gene containing VdAL and obtains recombinant microorganism.Described recombinant vectors for the sequence between NdeI and the KpnI recognition site of carrier pET42a (+) is replaced with sequence 2 in sequence table 1-894 position Nucleotide shown in DNA molecular (i.e. VdAL gene), the recombinant vectors obtained.VdAL shown in described recombinant vectors expressed sequence 1.VdAL shown in described recombinant microorganisms express sequence 1.
The above-mentioned biomaterial relevant to VdAL is improving in the application in plant prod quality, and described raising plant prod quality can be the content that improves described plant prod nutritive ingredient and/or improves the content of described plant prod medium trace element and/or improve the palatability of described plant prod.Described plant prod nutritive ingredient specifically can be vitamins C, soluble sugar and/or protein.Described trace element specifically can be Cu, Fe, K, Mn and/or Zn.Described palatability may be embodied in dry rate and/or the decline of coarse-fibred content.
The above-mentioned biomaterial relevant to VdAL is improving in the application in plant prod quality, and described raising plant prod quality can be the quality improving described plant prod by promoting the maturation of described plant prod.In one embodiment of the invention, VdAL uses the maturation promoting cotton production, and before the frost of cotton, flower output increases, and then improves cotton quality.
The above-mentioned biomaterial relevant to VdAL is improving in the application in plant prod quality, and described plant can be dicotyledons or monocotyledons.
The above-mentioned biomaterial relevant to VdAL is improving in the application in plant prod quality, described dicotyledons can be following a), b), c) or d):
A) cucurbitaceous plant;
B) watermelon;
C) Malvaceae plant;
D) cotton.
For solving the problems of the technologies described above, present invention also offers a kind of method improving plant prod quality.
The method of raising plant prod quality provided by the present invention, comprises and uses VdAL, described biomaterial or biotechnological formulation to object plant, improves described object plant prod quality; The activeconstituents of described biotechnological formulation is VdAL or described biomaterial.
In the method for above-mentioned raising plant prod quality, described biotechnological formulation can according to method 1) preparation, described method 1) be: cultivate described recombinant microorganism, the encoding gene of VdAL is expressed, obtains described biotechnological formulation.
In the method for above-mentioned raising plant prod quality, described method 1) specifically can be following 11) and 12):
11) cultivate described recombinant microorganism, described encoding gene is expressed, obtain the recombinant microorganism culture of expressing described protein;
12) thalline in broken described microorganisms cultures, obtains described biotechnological formulation.
In the method for above-mentioned raising plant prod quality, described method 1) also can comprise and drying is carried out to described biotechnological formulation.
In the method for above-mentioned raising plant prod quality, described encoding gene can be B1) described nucleic acid molecule.
In an embodiment of the invention, described recombinant microorganism is that the recombinant vectors of the expression cassette of the encoding gene containing VdAL is imported the recombinant microorganism obtained in e. coli jm109.Described recombinant vectors for the sequence between NdeI and the KpnI recognition site of carrier pET42a (+) is replaced with sequence 2 in sequence table 1-894 position Nucleotide shown in DNA molecular (i.e. VdAL gene), the recombinant vectors obtained.VdAL shown in described recombinant vectors expressed sequence 1.VdAL shown in described recombinant microorganisms express sequence 1.
In the method for above-mentioned raising plant prod quality, describedly VdAL, described biomaterial or biotechnological formulation are used to object plant specifically can be VdAL, described biomaterial or described biotechnological formulation are used to described object plant leaf, as to object plant-spraying VdAL, as described in biomaterial or as described in biotechnological formulation, also can use the seed of object plant described in VdAL, described biomaterial or described biotechnological formulation process, as with VdAL, as described in biomaterial or as described in biotechnological formulation liquid preparation to as described in the seed of object plant soak seed.The liquid preparation of VdAL, described biomaterial or described biotechnological formulation can be directly with the liquid that clear water dissolves or suspension VdAL, described biomaterial or described biotechnological formulation obtain.Amount object plant being used to VdAL, described biomaterial or biotechnological formulation can be determined according to the kind of described object plant and/or growth period.
In the method for above-mentioned raising plant prod quality, described raising plant prod quality can be the content that improves described plant prod nutritive ingredient and/or improves the content of described plant prod medium trace element and/or improve the palatability of described plant prod.Described plant prod nutritive ingredient specifically can be vitamins C, soluble sugar and/or protein.Described trace element specifically can be Cu, Fe, K, Mn and/or Zn.Described palatability may be embodied in dry rate and/or the decline of coarse-fibred content.
In the method for above-mentioned raising plant prod quality, described raising plant prod quality can be the quality improving described plant prod by promoting the maturation of described plant prod.In one embodiment of the invention, VdAL uses the maturation promoting cotton production, and before the frost of cotton, flower output increases, and then improves cotton quality.
In the method for above-mentioned raising plant prod quality, described plant can be dicotyledons or monocotyledons.
In the method for above-mentioned raising plant prod quality, described dicotyledons can be following a), b), c) or d):
A) cucurbitaceous plant;
B) watermelon;
C) Malvaceae plant;
D) cotton.
For solving the problems of the technologies described above, present invention also offers described biotechnological formulation and improving the application in plant prod quality.
Above-mentioned biotechnological formulation is improving in the application in plant prod quality, and described raising plant prod quality can be the content that improves described plant prod nutritive ingredient and/or improves the content of described plant prod medium trace element and/or improve the palatability of described plant prod.Described plant prod nutritive ingredient specifically can be vitamins C, soluble sugar and/or protein.Described trace element specifically can be Cu, Fe, K, Mn and/or Zn.Described palatability may be embodied in dry rate and/or the decline of coarse-fibred content.
Above-mentioned biotechnological formulation is improving in the application in plant prod quality, and described raising plant prod quality can be the quality improving described plant prod by promoting the maturation of described plant prod.In one embodiment of the invention, VdAL uses the maturation promoting cotton production, and before the frost of cotton, flower output increases, and then improves cotton quality.
Above-mentioned biotechnological formulation is improving in the application in plant prod quality, and described plant can be dicotyledons or monocotyledons.
Above-mentioned biotechnological formulation is improving in the application in plant prod quality, described dicotyledons can be following a), b), c) or d):
A) cucurbitaceous plant;
B) watermelon;
C) Malvaceae plant;
D) cotton.
In the present invention, can there is different results because of the difference of VdAL or described biotechnological formulation concentration to the effect of different plant in VdAL or described biotechnological formulation.When using VdAL or described biotechnological formulation, the concentration of VdAL or described biotechnological formulation can according to crop or its kind and the object (content as object plant prod Middle nutrition material as described in improving) using VdAL or described biotechnological formulation regulate VdAL or as described in the concentration of biotechnological formulation.
In an embodiment of the present invention, described biotechnological formulation dilutes the content that 2000 times of liquid obtained can improve watermelon Middle nutrition material and trace element.The liquid that described biotechnological formulation dilution 3000-5000 doubly obtains can improve the ginning outturn of cotton.
Experiment proves, is that Lynn Vidali peace biotechnological formulation (VDLA) of activeconstituents can improve plant prod quality, also can promotes fruit maturation in the present invention with VdAL:
1, after the process of Lynn Vidali peace, in watermelon, main nutrient composition and micronutrient levels increase, and palatability improves.After the process of Lynn Vidali peace, Vc content significantly improves, and improves 50.41%; Soluble sugar content improves 8.88%, and in dry-matter, protein content improves 14.2%, and in fresh watermelon, protein content improves 0.62%, and dry rate declines 12.4%, and coarse-fibred content has a small amount of decline.After the process of Lynn Vidali peace, the quality of Ca, Cu, Fe, K, Mg, Mn, Na, P and Zn of often liter of fresh watermelon all has increase, and increasing amount is respectively 12.75%, 16.54%, 19.21%, 16.46%, 1.05%, 20.45%, 2.13%, 10.02%, 19.73%; In fresh watermelon, the mass content of Cu, Fe, K, Mn and Zn all has increase, and increasing amount is respectively 2.09%, 19.09%, 1.88%, 5.72% and 5.07%.
2, after the process of Lynn Vidali peace, before cotton frost, flower output and ginning outturn all increase.With No. 30, the VDLA process Shandong of dilution 3000 times, flower output increased 19.48% before frost; During VDLA process with dilution 5000 times, flower output increased 19.06% before flower output increased 21.25%, 616 frost before the frost of No. 30, Shandong, flower output increased 16.37% before the frost of No. 25, Shandong.During VDLA process with dilution 3000 times, ginning outturn increment rate is 0.24%; During VDLA process with dilution 5000 times, ginning outturn increment rate is 0.49%.
Experiment proves, protein VdAL of the present invention and biotechnological formulation thereof can improve plant prod quality.
Accompanying drawing explanation
Fig. 1 is the western-blot detected result of VdAL during the sight of Lynn Vidali Anwai and Lynn Vidali are pacified.Wherein, A is that Lynn Vidali Anwai is seen; B is the western-blot detected result of VdAL in Lynn Vidali peace, and blower fan represents the comparatively fine powder entering air outlet position in sample drying process, and king-tower represents the normal powder in sample drying process.
Fig. 2 is that Lynn Vidali peace can promote nourishing and growing of watermelon.Wherein, CK represents untreated watermelon, and VdAL represents the watermelon of Lynn Vidali peace process.
Fig. 3 is that Lynn Vidali peace can promote cucumber setting and expand.Wherein, CK represents untreated cucumber, the cucumber of the VdAL process of 300 expression dilutions 300 times, the cucumber of the VdAL process of 600 expression dilutions 600 times, the cucumber of the VdAL process of 900 expression dilutions 900 times.
Fig. 4 is the growing way of the cucumber of different treatment.Wherein, CK represents untreated cucumber, the cucumber of the VdAL process of 300 expression dilutions 300 times.
Fig. 5 is the old and feeble situation of the cucumber of different treatment.Wherein, CK represents untreated cucumber, the cucumber of the VdAL process of 2000 × expression dilution 2000 times.
Fig. 6 is the setting situation of the cherry tomato of different treatment.Wherein, CK represents untreated cherry tomato, the cherry tomato of the VdAL process of 2000 × expression dilution 2000 times.
Fig. 7 is the strawberry of different treatment.Wherein, CK represents untreated strawberry, the strawberry of the VdAL process of 2000 × expression dilution 2000 times.
Fig. 8 is the muskmelon of different treatment.Wherein, CK represents untreated muskmelon, the muskmelon of the VdAL process of 2000 × expression dilution 2000 times.
Fig. 9 is the pimento of different treatment.Wherein, CK represents untreated pimento, the pimento of the VdAL process of 1000 × expression dilution 1000 times.
Figure 10 is the germination of the soybean seeds of different treatment.Wherein, CK represents untreated soybean seeds, the soybean seeds of the VdAL process of 1000 expression dilutions 1000 times, the soybean seeds of the VdAL process of 3000 expression dilutions 3000 times, the soybean seeds of the VdAL process of 5000 expression dilutions 5000 times, the soybean seeds of the VdAL process of 10000 expression dilutions 10000 times, the soybean seeds of the VdAL process of 20000 expression dilutions 20000 times.
Figure 11 is the Soybean Seedlings Growth situation of different treatment.Wherein, CK represents untreated soybean seedling, the soybean seedling of the VdAL process of 5000 × expression dilution 5000 times.
Embodiment
Below in conjunction with embodiment, the present invention is further described in detail, the embodiment provided only in order to illustrate the present invention, instead of in order to limit the scope of the invention.
Experimental technique in following embodiment, if no special instructions, is ordinary method.
Material used in following embodiment, reagent etc., if no special instructions, all can obtain from commercial channels.
E. coli jm109 in following embodiment is Beijing Suo Laibao Science and Technology Ltd. product, and catalog number is C1300.
Carrier pET42a (+) in following embodiment is Bi Chenglan bio tech ltd, Beijing product, and catalog number is S18-16.
New land in following embodiment early No. 58 be Xinjiang brocade cotton seed industry Science and Technology Co., Ltd. product, No. 30, Shandong, 616, to grind cotton No. 24 be Shandong Nong Xingzhong industry limited liability company product for No. 25, Shandong and Shandong.
Variety of watermelon gold Lido in following embodiment is accumulate virtue commerce and trade company limited product in Shouguang.
Middle wheat 816 in following embodiment is Beijing Long Shengyuan development in science and technology company limited product.
Tomato variety Fuchs in following embodiment is Zhong Ye company limited of Jinpeng of Shouguang City product.
Magnificent green grass or young crops stalk dish F1 in following embodiment is sky, Hongqiao in Shanghai dragon species industry company limited product.
Assorted No. 1 of cucumber variety Tianjin in following embodiment is Shandong Nong Xingzhong industry company limited product.
Red No. 2 of cherry tomato kind capital in following embodiment is Beijing North Nong Lvheng development in science and technology company limited product.
Strawberry cultivars Feng Xiang in following embodiment is Beijing North Nong Lvheng development in science and technology company limited product.
The precious fragrant melon variety of super kingfisher in following embodiment is Jinan Rui Mao agricultural science and technology development corporation, Ltd. product.
Sweet Pepper Varieties in following embodiment farsighted excellent 816 is Jinan Rui Mao agricultural science and technology development corporation, Ltd. product.
Soybean varieties China soybean king in following embodiment is Jinan Rui Mao agricultural science and technology development corporation, Ltd. product.
The preparation of embodiment 1, Lynn Vidali peace
Lynn Vidali peace is developed by China Agricultural University, by Huimin Hu Hao bio tech ltd, Shandong Development and Production.
1, the structure of recombinant vectors and recombinant bacterium
The DNA molecular shown in the Nucleotide of 1-894 position of sequence 2, i.e. VdAL gene in artificial synthesized sequence table.Sequence between NdeI and the KpnI recognition site of carrier pET42a (+) is replaced with the DNA molecular (i.e. VdAL gene) shown in the Nucleotide of 1-894 position of sequence 2 in sequence table, keep other sequences of pET42a (+) constant, obtain recombinant vectors, by the recombinant vectors called after pET42a-VdAL obtained.Protein VdAL shown in sequence 1 in recombinant vectors pET42a-VdAL expressed sequence table.
Wherein, sequence 2 is made up of 894 Nucleotide, the aminoacid sequence shown in encoding sequence 1.
PET42a-VdAL is imported in agrobacterium tumefaciens e. coli jm109, obtain recombinant bacterium, by the recombinant bacterium called after JM109-pET42a-VdAL obtained, the protein shown in JM109-pET42a-VdAL expressed sequence 1.
2, the preparation of Lynn Vidali peace
Being carried out by JM109-pET42a-VdAL fermenting under 37 DEG C of conditions to OD value is 0.6 obtain pre-fermentation broth, and in pre-fermentation broth, add IPTG, the concentration making IPTG is 1mM, obtains induced liquid, and induced liquid is obtained fermented liquid in 6 hours at 25 DEG C of condition bottom fermentations.Fermented liquid is carried out centrifugal, abandons supernatant liquor, by broken for the bacterial sediment obtained, then carry out spraying dry at 150 DEG C and obtain dry powder formulations (in Fig. 1 A), its called after Lynn Vidali is pacified.
The method of western-blot is utilized to detect the VdAL in Lynn Vidali peace, primary antibodie is VdAL antibody (VdAL antibody be the polyclonal antibody that immunogen prepares for the protein shown in sequence 2), result shows, containing VdAL (in Fig. 1 B) in Lynn Vidali peace, in Lynn Vidali peace, the content of VdAL is 7.28mg/g.
With clear water, Lynn Vidali peace is dissolved, obtain Lynn Vidali peace concentration respectively and be respectively 10g/3kg, 10g/6kg, 10g/9kg, 1g/kg, 1g/2kg, 1g/2.5kg, 1g/3kg, 1g/4kg, 1g/5kg, 1g/7.5kg, the liquid of 1g/10kg and 1g/20kg, by these liquid respectively called after dilute the VdAL of 300 times, dilute the VdAL of 600 times, dilute the VdAL of 900 times, dilute the VdAL of 1000 times, dilute the VdAL of 2000 times, dilute the VdAL of 2500 times, dilute the VdAL of 3000 times, dilute the VdAL of 4000 times, dilute the VdAL of 5000 times, dilute the VdAL of 7500 times, dilute the VdAL of 10000 times and the VdAL of dilution 20000 times.
Embodiment 2, Lynn Vidali peace can promote the growth of cotton
1, Lynn Vidali peace can promote the germination of cotton seeds
New land early No. 58 seeds 24 hours are soaked respectively with the VdAL of the VdAL of the dilution 5000 times of embodiment 1, dilution 7500 times and the VdAL of dilution 10000 times and clear water, germination test is carried out in large flowerpot, temperature is 25 DEG C, sowing the 8th day, and statistics Cotton Seed rate.Often kind of process 100 seeds, experiment in triplicate.
Result shows, dilute the VdAL of 5000 times, after diluting the VdAL of 7500 times and the VdAL of dilution 10000 times and clear water process cotton seeds, the average germination percentage of cotton is respectively 74.00%, 75.50%, 71.00% and 70.23%, dilute the VdAL of 5000 times, after diluting the VdAL of 7500 times and the dilution VdAL process cotton seeds of 10000 times, the average germination percentage of cotton is respectively 1.05 times of the average germination percentage of cotton after clear water process cotton seeds, 1.08 doubly, 1.01 doubly, show, Lynn Vidali peace can improve the percentage of germination of cotton seeds, the ability that Lynn Vidali peace improves Cotton Seed rate changes with the change of Lynn Vidali peace concentration.
2, Lynn Vidali peace can improve the output of cotton
Experiment one:
Field experimental design is as follows: experiment adopts randomized block design, 3 iterons are set, each iteron arranges nine treatment zones at random, be respectively VdAL process 1 district, No. 30, Shandong, VdAL process 2 district, No. 30, Shandong, control treatment district, No. 30, Shandong, 616VdAL process 1 district, 616VdAL process 2 district, 616 control treatment districts, VdAL process 1 district, No. 25, Shandong, VdAL process 2 district, No. 25, Shandong and control treatment district, No. 25, Shandong, the area of each treatment zone is 0.2 mu.
In Shandong, seedling stage of No. 30, the Shandong in No. 30 VdAL process 1 districts and Hua Ling process early stage respectively in the following manner: adopt the mode of top spray (machinery sprays) cotton leaf to be sprayed to the VdAL of the dilution 3000 times of embodiment 1, every mu sprays 45-50kg, obtains No. 30, the Shandong of the VdAL process of dilution 3000 times.Seedling stage and Hua Ling respectively spray 1 time early stage; Spray the time in the morning or at dusk, preferably avoid the high temperature period at high noon; Forbid when spraying to mix with medicine and fertilizer; If rain in 2 hours after spraying, needs fill spray and execute once.
According to the treatment process in No. 30 VdAL process 1 districts, Shandong, the VdAL VdAL of dilution 3000 times being replaced with dilution 5000 times processes VdAL process 2 district, No. 30, Shandong, and other steps are all constant, obtain No. 30, the Shandong of the VdAL process of dilution 5000 times.
According to the treatment process in No. 30 VdAL process 1 districts, Shandong, the VdAL of dilution 3000 times is replaced with clear water and process control treatment district, No. 30, Shandong, other steps are all constant, obtain No. 30, untreated Shandong.
According to the treatment process in No. 30 VdAL process 1 districts, Shandong, process 616VdAL process 1 district and VdAL process 1 district, No. 25, Shandong respectively, other steps are all constant, obtain 616 of the VdAL process of dilution 3000 times respectively, No. 25, Shandong.
According to the treatment process in No. 30 VdAL process 1 districts, Shandong, the VdAL of dilution 3000 times is replaced with the VdAL of dilution 5000 times, process 616VdAL process 2 district and VdAL process 2 district, No. 25, Shandong respectively, other steps are all constant, obtain 616 of the VdAL process of dilution 5000 times respectively, No. 25, Shandong.
According to the treatment process in No. 30 VdAL process 1 districts, Shandong, the VdAL of dilution 3000 times is replaced with clear water, process 616 control treatment districts and control treatment district, No. 25, Shandong respectively, other steps are all constant, obtain untreated 616, No. 25, Shandong respectively.
Add up respectively No. 30, the Shandong of the VdAL process of dilution 3000 times, No. 30, the Shandong of VdAL process of dilution 5000 times, No. 30, untreated Shandong, dilution 3000 times 616 of VdAL process, the VdAL process of dilution 5000 times 616, untreated 616, No. 25, the Shandong of VdAL process of dilution 3000 times, No. 25, the Shandong of the VdAL process of dilution 5000 times and No. 25, untreated Shandong frost front spend output with, whitely spend output and ultimate production afterwards, result is as shown in table 1.
The mean yield of the cotton of table 1, different treatment
Note: process 1 represents the process that the VdAL of dilution 3000 times carries out, process 2 represents the process that the VdAL of dilution 5000 times carries out, and CK represents untreated (namely with the process that clear water carries out).
Result shows, and the Lynn Vidali peace of proper concn can improve the output of cotton: with No. 30, the VdAL process Shandong of dilution 3000 times, flower output increased 19.48% before frost, and ultimate production improves 17.04%; During VdAL process with dilution 5000 times, flower output increased 21.25% before the frost of No. 30, Shandong, ultimate production improves 20.00%, flower output increased 19.06% before 616 frosts, ultimate production improves 19.69%, and flower output increased 16.37% before the frost of No. 25, Shandong, ultimate production improves 17.60%.
Experiment two:
Field experimental design is as follows: experiment adopts randomized block design, 3 iterons are set, each iteron arranges four treatment zones at random, and be respectively VdAL process 1 district, VdAL process 2 district, VdAL process 3 district and control treatment district, the area of each treatment zone is 10 mu.
The seedling stage and the Hua Ling that grind cotton No. 24 in the Shandong in VdAL process 1 district process early stage respectively in the following manner: adopt the mode of top spray (machinery sprays) cotton leaf to be sprayed to the VdAL of the dilution 3000 times of embodiment 1, every mu sprays 45-50kg, locomotive shunting speed control is at speed per hour 6 km, and the Shandong obtaining the VdAL process of dilution 3000 times grinds cotton No. 24.Seedling stage and Hua Ling respectively spray 1 time early stage; Spray the time in the morning or at dusk, preferably avoid the high temperature period at high noon; Forbid when spraying to mix with medicine and fertilizer; If rain in 2 hours after spraying, needs fill spray and execute once.
According to the treatment process in VdAL process 1 district, the VdAL VdAL of dilution 3000 times being replaced with dilution 5000 times processes VdAL process 2 district, and other steps are all constant, and the Shandong obtaining the VdAL process of dilution 5000 times grinds cotton No. 24.
According to the treatment process in VdAL process 1 district, the VdAL VdAL of dilution 3000 times being replaced with dilution 10000 times processes VdAL process 3 district, and other steps are all constant, and the Shandong obtaining the VdAL process of dilution 10000 times grinds cotton No. 24.
According to the treatment process in VdAL process 1 district, the VdAL of dilution 3000 times is replaced with clear water and processes control treatment district, other steps are all constant, obtain untreated Shandong and grind cotton No. 24.
The Shandong adding up each process grind cotton No. 24 every mu of plant number, single-strain bell-forming number, the total bell number of mu, Single boll weight, ginning outturn and per mu yield, the mean value of projects and increment are compared with the control as shown in table 2 and table 3.
Mean value and the increment of cotton No. 24 projects are ground in the Shandong of table 2, each process
Note: process 1 represents the process that the VdAL of dilution 3000 times carries out, process 2 represents the process that the VdAL of dilution 5000 times carries out, and process 3 represents the process that the VdAL of dilution 10000 times carries out, and CK represents untreated (namely with the process that clear water carries out).
Mean value and the increment of cotton No. 24 projects are ground in the Shandong of table 3, each process
Note: process 1 represents the process that the VdAL of dilution 3000 times carries out, process 2 represents the process that the VdAL of dilution 5000 times carries out, and process 3 represents the process that the VdAL of dilution 10000 times carries out, and CK represents untreated (namely with the process that clear water carries out).
Result shows, the Lynn Vidali of proper concn pacifies every mu of plant number, single-strain bell-forming number, the total bell number of mu, Single boll weight, ginning outturn and the per mu yield that can improve cotton: when using the VdAL process of dilution 3000 times, the total bell number of single-strain bell-forming number, mu, ginning outturn and per mu yield all have increase, and increment rate is respectively 4.5%, 1.98%, 0.24% and 1.76%; During VdAL process with dilution 5000 times, every mu of plant number, single-strain bell-forming number, the total bell number of mu, Single boll weight, ginning outturn and per mu yield all have increase, and increment rate is respectively 1.4%, 3.41%, 4.41%, 1.9%, 0.49% and 5.22%; During VdAL process with dilution 10000 times, the increasing amount of projects is high with the VdAL process of dilution 3000 times with when diluting the VdAL process of 5000 times for the increasing amount of projects.Show, the Lynn Vidali peace using proper concn can improve the quality and yield of cotton.
Embodiment 3, Lynn Vidali peace can improve the quality and yield of watermelon
Field experimental design is as follows: experiment adopts randomized block design, and arrange 3 iterons, each iteron arranges 2 treatment zones at random, is respectively VdLA treatment zone and control treatment district, and the area of each treatment zone is 0.2 mu.
Variety of watermelon gold Lido in VdLA treatment zone processes in the following manner: adopt the mode of top spray (machinery sprays) water melon leaf to be sprayed to the VdLA of the dilution 2000 times of embodiment 1, every mu sprays 30kg, obtains the watermelon of the VdAL process of dilution 2000 times.Spray the Seedling Stage from watermelon, and spray 1 time again after spraying 15 days in first time, spray 2 times altogether; Spray the time in the morning or at dusk, preferably avoid the high temperature period at high noon; Preferably do not mix with medicine and fertilizer when spraying; If rain in 2 hours after spraying, needs fill spray and execute once.
According to the treatment process of VdAL treatment zone, the VdAL of dilution 2000 times is replaced with clear water and processes control treatment district, other steps are all constant, obtain untreated watermelon.
Found that, Lynn Vidali peace to watermelon nourish and grow and setting has promoter action (Fig. 2).The yield of watermelon in statistics different treatment district, found that, diluting per plant number after the VdAL process watermelon of 2000 times is 1.43 ± 0.5, and output is 5200.2 ± 500kg/ mu, and the per plant number of untreated watermelon is 1.21 ± 0.5, output is 3898.2 ± 500kg/ mu.Compared with untreated watermelon, the per plant number of watermelon of Lynn Vidali peace process improves 18.2%, output increased 33.4%.
The content (table 5) of the water content of different treatment watermelon, dry rate, Vc content, soluble sugar content, titratable acid content, soluble solid content, melon skin thickness, protein content, crude fiber content (table 4) and trace elements of Ca, Cu, Fe, K, Mg, Mn, Na, P and Zn is measured respectively in the watermelon ripening stage.Wherein, water content and dry rate (in watermelon, dry-matter accounts for the per-cent of watermelon fresh weight) all measure after watermelon drying, Vc content is according to 2, 6-dichloroindophenol volumetry measures, soluble sugar content measures according to anthrone colorimetry, titratable acid content measures according to colorimetry, soluble solid content, protein content measures according to spectrophotometric metering method, crude fiber content detects robust fibre method according to Wen Defa and measures, trace elements of Ca, Cu, Fe, K, Mg, Mn, Na, the content of P and Zn all utilizes microelement analyzer to measure.
The average content of table 4, each index of different treatment watermelon
| Index | Untreated | VdLA process |
| Water content (%) | 90.22 | 91.43 |
| Dry rate (%) | 9.78 | 8.57 |
| Vc(mg/100g) | 12.1 | 18.2 |
| Soluble sugar (%) | 6.83 | 7.44 |
| Titratable acid (%) | 0.09 | 0.09 |
| Soluble solid (%) | 9.1 | 9.1 |
| Melon skin thick (cm) | 0.97 | 1.23 |
| Protein (%DW) | 5.54 | 6.33 |
| Protein (%FW) | 0.54 | 0.54 |
| Robust fibre (%DW) | 2.06 | 2.02 |
| Robust fibre (%FW) | 0.2 | 0.17 |
The average content of table 5, each trace element of different treatment watermelon
| Numbering | Index | Untreated | VdAL process |
| 1 | Water ratio (%) | 90.22 | 91.43 |
| 2 | Ca(mg/L) | 20.13 | 22.70 |
| 3 | Ca content (mg/100gFw) | 9.83 | 9.71 |
| 4 | Cu(mg/L) | 0.13 | 0.15 |
| 5 | Cu content (mg/100gFw) | 0.06 | 0.06 |
| 6 | Fe(mg/L) | 0.50 | 0.60 |
| 7 | Fe content (mg/100gFw) | 2.50 | 2.98 |
| 8 | K(mg/L) | 158.00 | 184.00 |
| 9 | K content (mg/100gFw) | 77.19 | 78.64 |
| 10 | Mg(mg/L) | 22.27 | 22.50 |
| 11 | Mg content (mg/100gFw) | 10.88 | 9.63 |
| 12 | Mn(mg/L) | 0.10 | 0.12 |
| 13 | Mn content (mg/100gFw) | 0.05 | 0.05 |
| 14 | Na(mg/L) | 7.20 | 7.35 |
| 15 | Na content (mg/100gFw) | 3.52 | 3.14 |
| 16 | P(mg/L) | 18.30 | 20.13 |
| 17 | P content (mg/100gFw) | 8.93 | 8.65 |
| 18 | Zn(mg/L) | 0.25 | 0.30 |
| 19 | Zn content (mg/100gFw) | 0.12 | 0.13 |
Note: 3, in 5,7,9,11,13,15,17 and 19 row, the content of each trace element all refers to the quality of each trace element in often liter of fresh watermelon, and in 2,4,6,8,10,12,14,16 and 18 row, the content of each trace element all refers to the content of each trace element in fresh watermelon.
Result shows, and after the process of Lynn Vidali peace, the water content of watermelon slightly increases, and adds 1.34%; Vc content significantly improves, and improves 50.41%; Soluble sugar content improves 8.88%, in dry-matter, protein content improves 14.2%, and in fresh watermelon, protein content improves 0.62%, and the content of titratable acid and soluble solid does not all change, and dry rate declines 12.4%, coarse-fibred content has a small amount of decline.Show, after the process of Lynn Vidali peace, the main nutrient composition content in watermelon increases; And acidity is not significantly increased, dry rate and coarse-fibred content all have decline, improve the palatability of watermelon.After the process of Lynn Vidali peace, the quality of Ca, Cu, Fe, K, Mg, Mn, Na, P and Zn of often liter of fresh watermelon all has increase, and increasing amount is respectively 12.75%, 16.54%, 19.21%, 16.46%, 1.05%, 20.45%, 2.13%, 10.02%, 19.73%; In fresh watermelon, the mass content of Cu, Fe, K, Mn and Zn all has increase, and increasing amount is respectively 2.09%, 19.09%, 1.88%, 5.72% and 5.07%.Show, Lynn Vidali pacifies the content that can significantly improve to human body healthy trace elements with household Cu, Fe, K, Mn and Zn in watermelon.More than test proof, Lynn Vidali peace can improve the quality of watermelon.
Embodiment 4, Lynn Vidali peace can promote the growth of wheat, improve the output of wheat
1, Lynn Vidali peace can promote the germination of wheat and nourish and grow
Experiment one:
Middle wheat 816 seed is soaked seed 12 hours respectively in the VdAL of the VdAL of the VdAL of the dilution 1000 times of embodiment 1, dilution 2000 times, dilution 5000 times, the VdAL of dilution 10000 times and clear water, then vernalization 96 hours at 25 DEG C in germination box (being covered with the aseptic filter paper that two-layer sterilized water soaks in germination box), statistics wheat seed germinating rate (wheat seed germinating standard is: the long half being or exceeding seed length of seed bud, and root length is or exceedes seed length).Often kind of process 100 seeds, experiment in triplicate.
Result shows, the VdAL of contrast and dilution 1000 times, dilute the VdAL of 2000 times, the VdAL diluting 5000 times, the percentage of germination that dilutes the VdAL of 10000 times and the wheat seed of clear water process is respectively 95.31 ± 1%, 83.26 ± 1%, 92.51 ± 1%, 97.67 ± 1% and 99.50 ± 1%, after diluting the VdAL of 1000 times, the VdAL of dilution 2000 times, the VdAL of dilution 5000 times, the dilution VdAL process wheat seed of 10000 times, percentage of germination improves-12.64% ,-0.00%, 2.48% and 4.40% respectively.Show, the Lynn Vidali peace of suitable concentration can promote the germination of wheat seed.
Experiment two:
Middle wheat 816 seed is soaked seed 24 hours respectively in the VdAL of the dilution 5000 times of embodiment 1, the VdAL of dilution 10000 times and clear water, then be seeded in soil, sowing is designated as sowing the 1st day the same day, wheat seedling was taken out from soil in the 3rd day in sowing, weigh wheat seedling fresh weight.
Result shows, 0.128 ± 0.022g, 0.141 ± 0.022g and 0.111 ± 0.022g is respectively with VdAL, the dilution VdAL of 10000 times of dilution 5000 times and the single-strain fresh weight of the wheat of clear water process, after the VdAL diluting 5000 times and the VdAL diluting 10000 times soaks seed, wheat single-strain fresh weight improves 15.32% and 27.03% respectively, and the single-strain fresh weight diluted after the VdAL seed soaking of 10000 times is significantly higher than the wheat single-strain fresh weight after Seed soaking.Show, Lynn Vidali peace can promote nourishing and growing of wheat.
2, Lynn Vidali peace can improve the output of wheat
Field experimental design is as follows: experiment adopts randomized block design, 3 iterons are set, each iteron arranges five treatment zones at random, and be respectively VdAL process 1 district, VdAL process 2 district, VdAL process 3 district, VdAL process 4 district and control treatment district, the area of each treatment zone is 0.5 mu.
In VdAL process 1 district, the blooming stage of wheat 816 carries out wheat process 1 time in the following manner: adopt the mode of top spray (artificial spraying) wheat leaf blade to be sprayed to the VdAL of the dilution 2500 times of embodiment 1, every mu sprays 45-50kg, obtains the wheat of the VdAL process of dilution 2500 times.Spray the time in the morning or at dusk, preferably avoid the high temperature period at high noon; Forbid when spraying to mix with medicine and fertilizer; If rain in 2 hours after spraying, needs fill spray and execute once.
According to the treatment process in VdAL process 1 district, the VdAL VdAL of dilution 2500 times being replaced with dilution 5000 times processes VdAL process 2 district, and other steps are all constant, obtains the wheat of the VdAL process of dilution 5000 times.
According to the treatment process in VdAL process 1 district, the VdAL VdAL of dilution 2500 times being replaced with dilution 10000 times processes VdAL process 3 district, and other steps are all constant, obtains the wheat of the VdAL process of dilution 10000 times.
According to the treatment process in VdAL process 1 district, the VdAL VdAL of dilution 2500 times being replaced with dilution 20000 times processes VdAL process 4 district, and other steps are all constant, obtains the wheat of the VdAL process of dilution 20000 times.
According to the treatment process in VdAL process 1 district, the VdAL of dilution 2500 times is replaced with clear water and processes control treatment district, other steps are all constant, obtain untreated wheat.
Add up the wheat yield of each process, result shows, after diluting the VdAL process wheat of 2500 times, the output of wheat does not improve, output is 8679kg/ha, dilute the VdAL of 5000 times, VdAL, the dilution VdAL of 20000 times of dilution 10000 times and the mean yield of the wheat of clear water process and be respectively 8942kg/ha, 9485kg/ha, 9231kg/ha and 8772kg/ha, after diluting the VdAL of 5000 times, the VdAL of dilution 10000 times, the dilution VdAL process wheat of 20000 times, output improves 1.94%, 8.13%, 5.23% respectively.
Add up the thousand seed weight of the wheat of different treatment, number of productive ear, grain number per spike and spike length, result is as shown in table 6.Result shows, and after diluting the VdAL process wheat of 5000 times, number of productive ear adds 1.5%, and grain number per spike adds 0.5%; Dilute the VdAL of 10000 times, number of productive ear adds 7%, and grain number per spike adds 3.5%; Dilute the VdAL of 20000 times, number of productive ear adds 2.3%, and grain number per spike adds 3.7%.
The mean value of the thousand seed weight of the wheat of table 6, different treatment, number of productive ear, grain number per spike and spike length
Note: process 1 represents the process that the VdAL of dilution 20000 times carries out, process 2 represents the process that the VdAL of dilution 10000 times carries out, process 3 represents the process that the VdAL of dilution 5000 times carries out, process 4 represents the process that the VdAL of dilution 2500 times carries out, and CK represents untreated (namely with the process that clear water carries out).
Embodiment 5, Lynn Vidali peace can increase tomato per plant number
Field experimental design is as follows: experiment adopts randomized block design, and arrange 3 iterons, each iteron arranges three treatment zones at random, is respectively VdAL process 1 district, VdAL process 2 district and control treatment district, and the area of each treatment zone is 0.1 mu.
The coming into bloom and spray VdAL in the following manner respectively on the 15th day after first time process of tomato variety Fuchs in VdAL process 1 district: adopt the mode of top spray (artificial spraying) tomato leaf to be sprayed to the VdAL of the dilution 1000 times of embodiment 1, every mu sprays the tomato that 45-50kg obtains the VdAL process of dilution 1000 times, sprays to be designated as the same day to spray latter 0th day by the 1st time.Spray the time in the morning or at dusk, preferably avoid the high temperature period at high noon; Forbid when spraying to mix with medicine and fertilizer; If rain in 2 hours after spraying, needs fill spray and execute once.
According to the treatment process in VdAL process 1 district, the VdAL VdAL of dilution 1000 times being replaced with dilution 3000 times processes VdAL process 2 district, and other steps are all constant, obtains the tomato of the VdAL process of dilution 3000 times.
According to the treatment process in VdAL process 1 district, the VdAL of dilution 1000 times is replaced with clear water and processes control treatment district, other steps are all constant, obtain untreated tomato.
The per plant number of the 20th day statistics tomato after spraying, result shows, after diluting the VdAL of 1000 times, the dilution VdAL of 3000 times and clear water process tomato, the average per plant number of tomato is respectively 34.9/strain, 40.4/strain and 33.3/strain, after the VdAL diluting 1000 times and the dilution VdAL process tomato of 3000 times, the per plant number of tomato improves 4.8% and 21.3% respectively.Show, Lynn Vidali peace can promote tomato result.
Embodiment 6, Lynn Vidali peace can promote the growth of blue or green stalk dish
Field experimental design is as follows: experiment adopts randomized block design, 3 iterons are set, each iteron arranges five treatment zones at random, and be respectively VdAL process 1 district, VdAL process 2 district, VdAL process 3 district, VdAL process 4 district and control treatment district, the area of each treatment zone is 0.2 mu.
Two leaf one heart stages of the magnificent hybridization blue or green stalk dish in VdAL process 1 district in the following manner VdAL spray 1 time: adopt the mode of top spray (artificial spraying) tomato leaf to be sprayed to the VdAL of the dilution 2000 times of embodiment 1, every mu sprays the green grass or young crops stalk dish that 45-50kg obtains the VdAL process of dilution 2000 times, Lynn Vidali peace is sprayed and be designated as process the same day the 0th day.Spray the time in the morning or at dusk, preferably avoid the high temperature period at high noon; Forbid when spraying to mix with medicine and fertilizer; If rain in 2 hours after spraying, needs fill spray and execute once.
According to the treatment process in VdAL process 1 district, the VdAL VdAL of dilution 2000 times being replaced with dilution 2500 times processes VdAL process 2 district, and other steps are all constant, obtains the green grass or young crops stalk dish of the VdAL process of dilution 2500 times.
According to the treatment process in VdAL process 1 district, the VdAL VdAL of dilution 2000 times being replaced with dilution 3000 times processes VdAL process 3 district, and other steps are all constant, obtains the green grass or young crops stalk dish of the VdAL process of dilution 3000 times.
According to the treatment process in VdAL process 1 district, the VdAL VdAL of dilution 2000 times being replaced with dilution 4000 times processes VdAL process 4 district, and other steps are all constant, obtains the green grass or young crops stalk dish of the VdAL process of dilution 4000 times.
According to the treatment process in VdAL process 1 district, the VdAL of dilution 2000 times is replaced with clear water and processes control treatment district, other steps are all constant, obtain untreated green grass or young crops stalk dish.
Process the 20th day, add up the fresh weight of 50 strain green grass or young crops stalk dishes of different treatment respectively, result shows, dilute the VdAL of 2000 times, dilute the VdAL of 2500 times, dilute the VdAL of 3000 times, after diluting the VdAL of 4000 times and clear water process, average 50 strains green grass or young crops stalk dish fresh weights are respectively 2.960kg, 3.270kg, 3.835kg, 3.895kg and 1.912kg, dilute the VdAL of 2000 times, dilute the VdAL of 2500 times, after the VdAL diluting 3000 times and the VdAL process green grass or young crops of diluting 4000 times obstruct dish, 50 strain green grass or young crops stalk dish fresh weights improve 54.8% respectively, 71.0%, 100.6% and 103.7%.Show, Lynn Vidali peace can promote the growth of blue or green stalk dish.
Embodiment 7, Lynn Vidali are positioned on the effect in other crops
1, Lynn Vidali peace can promote cucumber growth, improves output, delay senility
Field experimental design is as follows: experiment adopts randomized block design, 3 iterons are set, each iteron arranges five treatment zones at random, and be respectively VdAL process 1 district, VdAL process 2 district, VdAL process 3 district, VdAL process 4 district and control treatment district, the area of each treatment zone is 0.1 mu.
To mix in the cucumber variety Tianjin in VdAL process 1 district No. 1 come into bloom and spray VdAL in the following manner respectively on the 15th day after first time process: adopt the mode of top spray (artificial spraying) cucumber leaves to be sprayed to the VdAL of the dilution 300 times of embodiment 1, every mu sprays 45-50kg, obtain the cucumber of the VdAL process of dilution 300 times, the 1st time is sprayed Lynn Vidali peace and spray to be designated as the same day and spray the 0th day.Spray the time in the morning or at dusk, preferably avoid the high temperature period at high noon; Forbid when spraying to mix with medicine and fertilizer; If rain in 2 hours after spraying, needs fill spray and execute once.
According to the treatment process in VdAL process 1 district, the VdAL VdAL of dilution 300 times being replaced with dilution 600 times processes VdAL process 2 district, and other steps are all constant, obtains the cucumber of the VdAL process of dilution 600 times.
According to the treatment process in VdAL process 1 district, the VdAL VdAL of dilution 300 times being replaced with dilution 900 times processes VdAL process 3 district, and other steps are all constant, obtains the cucumber of the VdAL process of dilution 900 times.
According to the treatment process in VdAL process 1 district, the VdAL VdAL of dilution 300 times being replaced with dilution 2000 times processes VdAL process 4 district, and other steps are all constant, obtains the cucumber of the VdAL process of dilution 2000 times.
According to the treatment process in VdAL process 1 district, the VdAL of dilution 300 times is replaced with clear water and processes control treatment district, other steps are all constant, obtain untreated cucumber.
Spraying the 20th day, the result situation of statistics cucumber, found that, Lynn Vidali peace can improve cucumber percentage of fertile fruit, promote the expanding of cucumber, improve cucumber yield (Fig. 3), show, Lynn Vidali peace can promote nourishing and growing and reproductive growth of cucumber.Spraying the 20th day, observing the growing way of cucumber, found that, the growing way of the cucumber of Lynn Vidali peace process is better than the growing way (Fig. 4) of untreated cucumber, and show, Lynn Vidali peace can promote nourishing and growing of cucumber.Spraying the 45th day, observing the old and feeble situation of cucumber, found that, the old and feeble situation of untreated cucumber is than the old and feeble situation serious (Fig. 5) of the cucumber of Lynn Vidali peace process, and show, Lynn Vidali peace can delay the aging of cucumber.
2, Lynn Vidali peace can improve percentage of fertile fruit, the raising output of cherry tomato
Field experimental design is as follows: experiment adopts randomized block design, and arrange 3 iterons, each iteron arranges two treatment zones at random, is respectively VdAL treatment zone and control treatment district, and the area of each treatment zone is 1 mu.
In the cherry tomato kind capital of VdAL treatment zone, the flowering period of red No. 2, VdAL sprayed 1 time in the following manner: adopt the mode of top spray (artificial spraying) to the VdAL of the dilution 2000 times of cherry tomato blade spraying embodiment 1, every mu sprays 45-50kg, obtain the cherry tomato of the VdAL process of dilution 2000 times, Lynn Vidali peace is sprayed and was designated as process the same day the 0th day.Spray the time in the morning or at dusk, preferably avoid the high temperature period at high noon; Forbid when spraying to mix with medicine and fertilizer; If rain in 2 hours after spraying, needs fill spray and execute once.
According to the treatment process of VdAL treatment zone, the VdAL of dilution 2000 times is replaced with clear water and processes control treatment district, other steps are all constant, obtain untreated cherry tomato.
At process 7-10 days, observe the result situation of cherry tomato, found that, Lynn Vidali peace can improve the percentage of fertile fruit of cherry tomato, improves cherry tomato output (Fig. 6), shows, Lynn Vidali peace can promote the growth of cherry tomato.
3, Lynn Vidali peace can promote the growth of strawberry
Field experimental design is as follows: experiment adopts randomized block design, and arrange 3 iterons, each iteron arranges two treatment zones at random, is respectively VdAL treatment zone and control treatment district, and the area of each treatment zone is 1 mu.
Strawberry cultivars in VdAL treatment zone be Feng Xiang phase in flowering period in the following manner VdAL spray 1 time: adopt the mode of top spray (artificial spraying) From Strawberry Leaves to be sprayed to the VdAL of the dilution 2000 times of embodiment 1, every mu sprays 45-50kg, obtain the strawberry of the VdAL process of dilution 2000 times, Lynn Vidali peace is sprayed and was designated as process the same day the 0th day.Spray the time in the morning or at dusk, preferably avoid the high temperature period at high noon; Forbid when spraying to mix with medicine and fertilizer; If rain in 2 hours after spraying, needs fill spray and execute once.
According to the treatment process of VdAL treatment zone, the VdAL of dilution 2000 times is replaced with clear water and processes control treatment district, other steps are all constant, obtain untreated strawberry.
At process 7-10 days, observe the growing state of strawberry, found that, Lynn Vidali peace can promote nourish and grow (Fig. 7) of strawberry.
4, Lynn Vidali peace can promote the growth of muskmelon
Field experimental design is as follows: experiment adopts randomized block design, and arrange 3 iterons, each iteron arranges two treatment zones at random, is respectively VdAL treatment zone and control treatment district, and the area of each treatment zone is 1 mu.
The precious fragrant melon variety of the super kingfisher of VdAL treatment zone seedling stage in the following manner VdAL spray 1 time: adopt top spray (artificial spraying) mode to the VdAL of the dilution 2000 times of muskmelon blade spraying embodiment 1, every mu sprays 45-50kg, obtain the muskmelon of the VdAL process of dilution 2000 times, Lynn Vidali peace is sprayed and was designated as process the same day the 0th day.Spray the time in the morning or at dusk, preferably avoid the high temperature period at high noon; Forbid when spraying to mix with medicine and fertilizer; If rain in 2 hours after spraying, needs fill spray and execute once.
According to the treatment process of VdAL treatment zone, the VdAL of dilution 2000 times is replaced with clear water and processes control treatment district, other steps are all constant, obtain untreated muskmelon.
Process the 30th day, observe the growing state of muskmelon, found that, Lynn Vidali peace can promote nourish and grow (Fig. 8) of muskmelon.
5, Lynn Vidali peace can promote the growth of pimento
Field experimental design is as follows: experiment adopts randomized block design, and arrange 3 iterons, each iteron arranges two treatment zones at random, is respectively VdAL treatment zone and control treatment district, and the area of each treatment zone is 1 mu.
The Sweet Pepper Varieties of VdAL treatment zone farsighted flowering period of excellent 816 in the following manner VdAL spray 1 time: adopt the mode of top spray (artificial spraying) to the VdAL of the dilution 1000 times of pimento blade spraying embodiment 1, every mu sprays 45-50kg, obtain the pimento of the VdAL process of dilution 1000 times, Lynn Vidali peace is sprayed and was designated as process the same day the 0th day.Spray the time in the morning or at dusk, preferably avoid the high temperature period at high noon; Forbid when spraying to mix with medicine and fertilizer; If rain in 2 hours after spraying, needs fill spray and execute once.
According to the treatment process of VdAL treatment zone, the VdAL of dilution 1000 times is replaced with clear water and processes control treatment district, other steps are all constant, obtain untreated pimento.
At process 7-10 days, observe the growing state of pimento, found that, after spraying Lynn Vidali peace, pimento growing way is healthy and strong, blade is dark green (Fig. 9), shows that Lynn Vidali peace can promote nourishing and growing of pimento.
6, Lynn Vidali peace can promote the emergence and growth of soybean
Soybean varieties Chinese soybean king seed is soaked seed 24 hours respectively in the VdAL of the VdAL of the VdAL of the VdAL of the dilution 1000 times of embodiment 1, dilution 3000 times, dilution 5000 times, dilution 10000 times, the VdAL of dilution 20000 times and clear water, then vernalization 72 hours at 25 DEG C in pallet (being covered with the aseptic filter paper that two-layer sterilized water soaks in pallet), statistics soybean seeds percentage of germination.Often kind of process 100 seeds, experiment in triplicate.
Result display (Figure 10), dilute 3000 times VdAL and after diluting the VdAL soybean seeds of 5000 times, percentage of germination is improved.Show, Lynn Vidali peace can promote the germination of soybean seeds.
Experiment two:
Soybean varieties China soybean king seed is soaked seed 24 hours respectively in the VdAL and clear water of the dilution 5000 times of embodiment 1, then be seeded in soil, sowing was designated as sowing the 1st day the same day, soybean seedling was taken out from soil in 20th day in sowing, weigh soybean seedling fresh weight (Figure 11).
Result shows, and is respectively 1.59 ± 0.3g and 1.36 ± 0.3g with the single-strain fresh weight of the VdAL of dilution 5000 times and the soybean of clear water process, and after diluting the VdAL seed soaking of 5000 times, soybean per plant fresh weight improves 16.91%; 1.36 ± 0.3g and 1.06 ± 0.3g is respectively with the over-ground part single-strain fresh weight of the VdAL of dilution 5000 times and the soybean of clear water process, after diluting the VdAL seed soaking of 5000 times, soybeans upper part single-strain fresh weight improves 28.30%, is significantly higher than the over-ground part single-strain fresh weight of the soybean of clear water process.Show, Lynn Vidali peace can promote nourishing and growing of soybean.
Claims (10)
1. protein is improving the application in plant prod quality; Described protein is following A1) or A2) or A3):
A1) aminoacid sequence is the protein of sequence 1;
A2) in the aminoacid sequence of sequence 1 through replacement and/or disappearance and/or add that one or several amino-acid residue obtains have identical function by A1) derivative protein;
A3) at A1) or N end A2) or/and C end connects the fused protein that obtains of label.
2. the biomaterial relevant to protein described in claim 1 is improving the application in plant prod quality;
Described biomaterial is following B1) to B20) in any one:
B1) nucleic acid molecule of protein described in coding claim 1;
B2) containing B1) expression cassette of described nucleic acid molecule;
B3) containing B1) recombinant vectors of described nucleic acid molecule;
B4) containing B2) recombinant vectors of described expression cassette;
B5) containing B1) recombinant microorganism of described nucleic acid molecule;
B6) containing B2) recombinant microorganism of described expression cassette;
B7) containing B3) recombinant microorganism of described recombinant vectors;
B8) containing B4) recombinant microorganism of described recombinant vectors;
B9) containing B1) the transgenic plant cells system of described nucleic acid molecule;
B10) containing B2) the transgenic plant cells system of described expression cassette;
B11) containing B3) the transgenic plant cells system of described recombinant vectors;
B12) containing B4) the transgenic plant cells system of described recombinant vectors;
B13) containing B1) Transgenic plant tissue of described nucleic acid molecule;
B14) containing B2) Transgenic plant tissue of described expression cassette;
B15) containing B3) Transgenic plant tissue of described recombinant vectors;
B16) containing B4) Transgenic plant tissue of described recombinant vectors;
B17) containing B1) the transgenic plant organ of described nucleic acid molecule;
B18) containing B2) the transgenic plant organ of described expression cassette;
B19) containing B3) the transgenic plant organ of described recombinant vectors;
B20) containing B4) the transgenic plant organ of described recombinant vectors.
3. application according to claim 2, is characterized in that: B1) described nucleic acid molecule is following b1) b2) or b3) gene:
B1) nucleotide sequence is cDNA molecule or the DNA molecular of sequence 2 in sequence table;
B2) nucleotide sequence and b1) limited has more than 75% or 75% identity, and the cDNA molecule of protein described in coding claim 1 or genomic DNA molecule;
B3) under strict conditions with b1) nucleotide sequence hybridization that limits, and the cDNA molecule of protein described in coding claim 1 or genomic DNA molecule.
4. improve a method for plant prod quality, comprise and biomaterial or biotechnological formulation described in protein described in claim 1, Claims 2 or 3 are used to object plant; The activeconstituents of described biotechnological formulation is biomaterial described in protein described in claim 1 or Claims 2 or 3.
5. method according to claim 4, it is characterized in that: described biotechnological formulation is according to method 1) preparation, described method 1) be: arbitrary described recombinant microorganism B5 in cultivation Claims 2 or 3)-B8), the encoding gene of described protein is expressed, obtains described biotechnological formulation.
6. the method according to claim 4 or 5, is characterized in that: described method 1) be following 11) and 12):
11) cultivate B5 in Claims 2 or 3)-B8) in arbitrary described recombinant microorganism, described encoding gene is expressed, obtains the recombinant microorganism culture of expressing described protein;
12) thalline in broken described microorganisms cultures, obtains described biotechnological formulation.
7. in claim 4-5, arbitrary described biotechnological formulation is improving the application in plant prod quality.
8. application according to arbitrary described method in the arbitrary described application of claim 1-3, claim 4-6 or claim 7, is characterized in that: described raising plant prod quality is following H1 and/or H2:
H1, improve the content of described plant prod nutritive ingredient and/or improve the content of described plant prod medium trace element;
H2, improve the quality of described plant prod by promoting the maturation of described plant prod.
9. arbitrary described method in the application according to claim 1,2,3,7 or 8 or claim 4-6, is characterized in that: described plant is dicotyledons or monocotyledons.
10. application according to claim 9 or method, is characterized in that: described dicotyledons be following a), b), c) or d):
A) cucurbitaceous plant;
B) watermelon;
C) Malvaceae plant;
D) cotton.
Priority Applications (6)
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| CN201510691295.6A CN105198967B (en) | 2015-10-22 | 2015-10-22 | Protein VdAL is improving the application in plant product quality |
| PCT/CN2016/088074 WO2017067214A1 (en) | 2015-10-22 | 2016-07-01 | Applications of protein vdal in improving output, product quality and drought resistance of plant and in improving fruit coloring of plant |
| US15/769,746 US10806145B2 (en) | 2015-10-22 | 2016-07-01 | Applications of protein VdAL in improving output, product quality and drought resistance of plant and in improving fruit coloring of plant |
| CA3002881A CA3002881A1 (en) | 2015-10-22 | 2016-07-01 | Applications of protein vdal in improving output, product quality and drought resistance of plant and in improving fruit coloring of plant |
| EP16856654.5A EP3351555A4 (en) | 2015-10-22 | 2016-07-01 | Applications of protein vdal in improving output, product quality and drought resistance of plant and in improving fruit coloring of plant |
| CL2018001026A CL2018001026A1 (en) | 2015-10-22 | 2018-04-20 | Applications of the protein vdal in the improvement of the yield, quality of the product and resistance to the drought of plants and in the promotion of the coloration of the fruits of the plant |
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| WO2017067214A1 (en) * | 2015-10-22 | 2017-04-27 | 中国农业大学 | Applications of protein vdal in improving output, product quality and drought resistance of plant and in improving fruit coloring of plant |
| EP3243830A4 (en) * | 2015-01-06 | 2018-07-18 | China Agricultural University | Protein associated with disease resistance and encoding gene thereof, and use thereof in regulation of plant disease resistance |
| CN115413678A (en) * | 2022-09-20 | 2022-12-02 | 中捷四方生物科技股份有限公司 | Wettable powder containing verticillium dahliae activator protein and application |
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| CN104558130A (en) * | 2015-01-06 | 2015-04-29 | 中国农业大学 | Resistance-related protein and encoding gene thereof, and application of resistance-related protein and encoding gene for regulating and controlling plant disease resistance |
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| CN104558130A (en) * | 2015-01-06 | 2015-04-29 | 中国农业大学 | Resistance-related protein and encoding gene thereof, and application of resistance-related protein and encoding gene for regulating and controlling plant disease resistance |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3243830A4 (en) * | 2015-01-06 | 2018-07-18 | China Agricultural University | Protein associated with disease resistance and encoding gene thereof, and use thereof in regulation of plant disease resistance |
| US11254715B2 (en) | 2015-01-06 | 2022-02-22 | Pherobio Technology Co., Ltd. | Protein associated with disease resistance and encoding gene thereof, and use thereof in regulation of plant disease resistance |
| WO2017067214A1 (en) * | 2015-10-22 | 2017-04-27 | 中国农业大学 | Applications of protein vdal in improving output, product quality and drought resistance of plant and in improving fruit coloring of plant |
| EP3351555A4 (en) * | 2015-10-22 | 2019-08-21 | Pherobio Technology Co., Ltd | Applications of protein vdal in improving output, product quality and drought resistance of plant and in improving fruit coloring of plant |
| US10806145B2 (en) | 2015-10-22 | 2020-10-20 | Pherobio Technology Co., Ltd | Applications of protein VdAL in improving output, product quality and drought resistance of plant and in improving fruit coloring of plant |
| CN115413678A (en) * | 2022-09-20 | 2022-12-02 | 中捷四方生物科技股份有限公司 | Wettable powder containing verticillium dahliae activator protein and application |
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