CN106834343A - Sucrose synthase is in the regulation and control developmental application of fruit - Google Patents

Sucrose synthase is in the regulation and control developmental application of fruit Download PDF

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CN106834343A
CN106834343A CN201710092423.4A CN201710092423A CN106834343A CN 106834343 A CN106834343 A CN 106834343A CN 201710092423 A CN201710092423 A CN 201710092423A CN 106834343 A CN106834343 A CN 106834343A
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plant
fruit
sucrose synthase
sucrose
sequence
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CN106834343B (en
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张振贤
范敬伟
李翔
王虹云
眭晓蕾
李欣
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China Agricultural University
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Abstract

The invention discloses a kind of sucrose synthase in the regulation and control developmental application of fruit.It is to import in purpose plant the encoding gene of sucrose synthase the invention provides a kind of method for cultivating genetically modified plants, obtains genetically modified plants；The genetically modified plants fruit yield is more than the purpose plant.The encoding gene of the sucrose synthase concretely sucrose synthase gene CsSus4.Present invention finds the key effect that sucrose synthase gene CsSus4 is played in cucumber fruits growth course, can be with the strong development so as to promote fruit of regulation warehouse.Efficient volume increase of this excellent function for improveing cucumber has directive significance.

Description

Sucrose synthase is in the regulation and control developmental application of fruit
Technical field
The present invention relates to a kind of sucrose synthase in the regulation and control developmental application of fruit.
Background technology
Cucumber is one of important vegetables that countries in the world are generally cultivated, and is also one of big vegetable crop of China five.With it is most Number higher plants are identical, and the photosynthesis of cucumber is occurred mainly in the chlorenchymas such as blade, fruit and the stem of maturation, and we claim Be " source ".And for root, flower and seed etc. the non-green tissue without photosynthetic capacity, we term it " storehouse ".Sucrose is Green plants carries out photosynthetic primary product, is also photosynthate prevailing traffic form, and sucrose is by bast to storehouse device Official career is defeated, is grow supply carbon source and the energy of cell；Sucrose Metabolism is in development of plants, stress response and yield composition mistake Played an important role in journey, it mainly by generate it is a series of sugar as fuel metabolins and synthesis compound (including Protein, cellulose and starch) and adjust expression, transcription factor and other genes of microRNA as signaling molecule and swash The reciprocation of element, oxidative stress and defence signal.Pass through bast in the form of polysaccharide from the carbohydrate of source organ's synthesis Transport and store or absorb in each storehouse organ in portion.Storehouse organ is reached, least a portion of disaccharides or polysaccharide can be by storehouse organs Directly absorb, most of polysaccharide hydrolysis are absorbed by monose by plant.It is related to Sucrose Metabolism main in plant There are three kinds of enzymes：1st, Sucrose Phosphate Synthase (Sucrose Phosphate Synthase；SPS, EC 2.3.1.14), catalysis is anti- Should：Fructose -6-P+UDPG → sucrose -6-P+UDP, is the key enzyme of catalysing sucrose synthesis.2nd, invertase (Invertase； Inv, EC2.1.1.26), catalytic reaction：Sucrose+H2O → glucose+fructose, the key enzyme in sucrose decomposition process.3、 Sucrose synthase (Sucrose Synthase；SUS, E.C.2.4.1.13), catalysis is a reversible reaction：Sucrose+UDP ← → UDPG+ fructose.Invertase and sucrose synthase are all decomposing sucroses in plant glycometabolism, in regulation source and sink relation process In play an important role.The product of sucrose synthase decomposing sucrose is UDPG, for cellulosic electrode provides substrate.Sucrose is closed A key enzyme during being Sucrose Metabolism into enzyme, the enzyme is studied in many plants, and is considered as in plant Played an important role in internal energy metabolism, the transfer of sucrose, cyto-architectural shape are directly participated in plant cell Into and carbohydrate accumulation.But the function of the process that sucrose synthase is developed in cucumber fruits is not clear.
The content of the invention
It is an object of the invention to provide a kind of sucrose synthase in the regulation and control developmental application of fruit.
It is by the encoding gene of sucrose synthase the invention provides a kind of method (method first) for cultivating genetically modified plants Import in purpose plant, obtain genetically modified plants；The genetically modified plants meet at least one of following (a1)-(a5) proterties：
(a1) fruit yield is more than the purpose plant；
(a2) fruit length is more than the purpose plant；
(a3) fruit rugosity (diameter) is more than the purpose plant；
(a4) fruit weight is more than the purpose plant；
(a5) male flower is more than the purpose plant.
In methods described first, the encoding gene of the sucrose synthase can import purpose and plant by recombinant expression carrier Thing.The recombinant expression carrier can be by Ti-plasmids, Ri plasmids, plant viral vector, directly delivered DNA, microinjection, electricity Lead, the conventional biology methods such as agriculture bacillus mediated are transformed into purpose plant.
The recombinant expression carrier of the gene can be contained with existing plant expression vector construction.The plant expression vector Including double base agrobacterium vector with the carrier that can be used for plant micropellet bombardment etc..Use the gene constructed recombinant expression carrier When, any enhanced, composing type, organizing specific type or inducible promoter can be added before its transcription initiation nucleotides, They can be used alone or are used in combination with other plant promoters；Additionally, using the gene constructed recombinant expression carrier When, enhancer, including translational enhancer or transcriptional enhancer are it is also possible to use, these enhancer regions can be ATG initiation codons Son or neighboring region initiation codon etc., but must be identical with the reading frame of coded sequence, to ensure that the correct of whole sequence turns over Translate.The source of the translation control signal and initiation codon is extensive, can be natural, or synthesis.Turn over Translate initiation region and can come from transcription initiation region or structural gene.For the ease of being reflected to transgenic plant cells or plant It is fixed and screen, plant expression vector used can be processed, be such as added in expression in plant can produce color change enzyme or The gene of luminophor, resistant antibiotic marker or anti-chemical reagent marker gene etc..
The recombinant expression carrier concretely inserts the sequence of sequence table in the MCS of carrier pBI 121 The recombinant plasmid that double chain DNA molecule shown in 2 is obtained.
The recombinant expression carrier is concretely by the small fragment between XbaI the and SmaI restriction enzyme sites of carrier pBI 121 The recombinant plasmid for replacing the DNA molecular shown in the sequence 2 for sequence table to obtain.
The present invention also application of the protection methods described first in plant breeding.
The plant breeding is in order to seed selection fruit yield is high and/or fruit length big and/or fruit rugosity (diameter) is big And/or the big plant of male flower.
The present invention also protects a kind of method (method second) for cultivating genetically modified plants, is to suppress Sucrose synthesis in purpose plant The expression of the encoding gene of enzyme, obtains genetically modified plants；The genetically modified plants meet at least one of following (b1)-(b5) property Shape：
(b1) fruit yield is less than the purpose plant；
(b2) fruit length is less than the purpose plant；
(b3) fruit rugosity (diameter) is less than the purpose plant；
(b4) fruit weight is less than the purpose plant；
(b5) male flower is less than the purpose plant.
In methods described second, described " suppressing the expression of the encoding gene of sucrose synthase in purpose plant " is by interference What carrier was realized.
The interference carrier can be by Ti-plasmids, Ri plasmids, plant viral vector, directly delivered DNA, microinjection, electricity Lead, the conventional biology methods such as agriculture bacillus mediated are transformed into purpose plant.
The interference carrier concretely recombinant expression carrier containing interference fragment.
The interference fragment includes section first and section second.The section first and the section second are reverse complementary sequence. The sequence of the section first is as shown in the sequence 3 of sequence table.The interference fragment is concretely shown in the sequence 4 of sequence table.
The interference carrier concretely inserts the institute of sequence 4 of sequence table in the MCS of carrier pFGC1008 The recombinant expression carrier that the double chain DNA molecule for showing is obtained.
The interference carrier concretely takes the small fragment between AscI the and SpeI restriction enzyme sites of carrier pFGC1008 On behalf of sequence table sequence 4 shown in the recombinant expression carrier that obtains of DNA molecular.
The present invention also application of the protection methods described second in plant breeding.
The plant breeding is for seed selection low fruit yields and/or fruit length is small and/or fruit rugosity (diameter) is small And/or the small plant of male flower.
In any of the above methods described, the purpose plant concretely dicotyledon.The dicotyledon can be Cucurbitaceous plant.The cucurbitaceous plant can be cucumber plants.The cucumber plants concretely cucumber, such as Xintai City is close Thorn cucumber.
The present invention also protects the application of sucrose synthase, is at least one in following (e1)-(e16)：
(e1) regulation and control fruit development；
(e2) regulation and control plant storehouse is strong；
(e3) sucrose synthase activity in regulation and control plant organ；
(e4) sucrose synthase activity in regulation and control fruit；
(e5) sucrose synthase activity in regulation and control plant male flower；
(e6) sucrose synthase catalysing sucrose is converted into the activity of fructose in regulation and control plant organ；
(e7) sucrose synthase catalysing sucrose is converted into the activity of fructose in regulation and control fruit；
(e8) sucrose synthase catalysing sucrose is converted into the activity of fructose in regulation and control plant male flower；
(e9) fruit development is promoted；
(e10) plant storehouse is improved strong；
(e11) sucrose synthase activity in plant organ is promoted；
(e12) sucrose synthase activity in fruit is promoted；
(e13) sucrose synthase activity in plant male flower is promoted；
(e14) sucrose synthase catalysing sucrose is converted into the activity of fructose in promotion plant organ；
(e15) sucrose synthase catalysing sucrose is converted into the activity of fructose in promotion fruit；
(e16) sucrose synthase catalysing sucrose is converted into the activity of fructose in promotion plant male flower.
The plant concretely dicotyledon.The dicotyledon can be cucurbitaceous plant.The Curcurbitaceae is planted Thing can be cucumber plants.The cucumber plants concretely cucumber, the close thorn cucumber in such as Xintai City.
The present invention also protects a kind of specific DNA molecular, including section first and section second.The section first and the section second It is reverse complementary sequence.The sequence of the section first is as shown in the sequence 3 of sequence table.The specific DNA molecular such as sequence table Shown in sequence 4.
The present invention also protects a kind of interference carrier, and specific DNA molecular importing expression vector is obtained.
The interference carrier concretely inserts the institute of sequence 4 of sequence table in the MCS of carrier pFGC1008 The recombinant expression carrier that the double chain DNA molecule for showing is obtained.
The interference carrier concretely takes the small fragment between AscI the and SpeI restriction enzyme sites of carrier pFGC1008 On behalf of sequence table sequence 4 shown in the recombinant expression carrier that obtains of DNA molecular.
The present invention also protects the application of the specific DNA molecular or the interference carrier in genetically modified plants are cultivated；Institute State genetically modified plants and meet at least one of following (f1)-(f5) proterties：
(f1) fruit yield is less than the plant that sets out；
(f2) fruit length is less than the plant that sets out；
(f3) fruit rugosity (diameter) is less than the plant that sets out；
(f4) fruit weight is less than the plant that sets out；
(f5) male flower is less than the plant that sets out.
The plant concretely dicotyledon that sets out.The dicotyledon can be cucurbitaceous plant.The cucurbit Section plant can be cucumber plants.The cucumber plants concretely cucumber, the close thorn cucumber in such as Xintai City.
The concretely sucrose synthase CsSUS4 of sucrose synthase described in any of the above.
The sucrose synthase CsSUS4, is following (c1) or (c2) available from cucumber：
(c1) protein that the amino acid sequence shown in sequence in sequence table 1 is constituted；
(c2) by the amino acid sequence of sequence 1 is by the substitution of one or several amino acid residues and/or missing and/or adds Plus and with identical function as derived from sequence 1 protein.
In order that the sucrose synthase CsSUS4 in (c1) is easy to purify and is detected, can be in as sequence table shown in sequence 1 Amino acid sequence composition protein amino terminal or the upper label as shown in table 1 of carboxyl terminal connection.
The sequence of the label of table 1
Sucrose synthase CsSUS4 in above-mentioned (c2) can be artificial synthesized, also can first synthesize its encoding gene, then given birth to Thing expression is obtained.The encoding gene of the sucrose synthase CsSUS4 in above-mentioned (c2) can be by by shown in sequence in sequence table 2 Lack the codon of one or several amino acid residues in DNA sequence dna, and/or carry out the missense of one or several base-pairs and dash forward Become, and/or obtained in the coded sequence that its 5 ' end and/or 3 ' ends connect the label shown in table 1.
The encoding gene of sucrose synthase described in any of the above concretely CsSUS4 genes.
The CsSUS4 genes are any described DNA molecular in following (d1)-(d3)：
(d1) DNA molecular of the code area as shown in sequence 2 in sequence table；
(d2) the DNA sequence dna hybridization for being limited with (d1) under strict conditions and the DNA molecular of encoding sucrose synzyme；
(d3) DNA sequence dna limited with (d1) or (d2) has the DNA of more than 90% homology and encoding sucrose synzyme Molecule.
Above-mentioned stringent condition can be that with 0.1 × SSPE (or 0.1 × SSC), the solution of 0.1%SDS is miscellaneous in DNA or RNA Hand over and hybridize at 65 DEG C in testing and wash film.
Present invention finds the key work that sucrose synthase gene CsSUS4 is played in cucumber fruits growth course With can be with the strong development so as to promote fruit of regulation warehouse.Efficient volume increase of this excellent function for improveing cucumber has finger Lead meaning.
Brief description of the drawings
Fig. 1 is overexpression transgenic line PCR qualification results.
Fig. 2 is interference of transgene strain PCR qualification results.
Fig. 3 is cucumber different tissues position and different times CsSUS4 gene expression amounts and enzyme activity assay result.
Fig. 4 is enzyme activity assay result in transfer-gen plant fruit phenotype and fruit.
Fig. 5 is enzyme activity assay result in transfer-gen plant male flower phenotype and male flower.
Specific embodiment
Following embodiment facilitates a better understanding of the present invention, but does not limit the present invention.Experiment in following embodiments Method, unless otherwise specified, is conventional method.Test material used in following embodiments, unless otherwise specified, is certainly What routine biochemistry reagent shop was commercially available.Quantitative test in following examples, is respectively provided with three repetitions and tests, and as a result makes even Average.
The close thorn cucumber in Xintai City：Bibliography：Cheng J, Wang Z, Yao F, et al.Down-Regulating CsHT1, a Cucumber Pollen-Specific Hexose Transporter, Inhibits Pollen Germination, Tube Growth, and Seed Development. [J] .Plant Physiology, 2015,168 (2)：635-47.；The public can obtain from China Agricultural University.
Carrier pBI 121：Cheng J, Wang Z, Yao F, et al.Down-Regulating CsHT1, a Cucumber Pollen-Specific Hexose Transporter, Inhibits Pollen Germination, Tube Growth, and Seed Development. [J] .Plant Physiology, 2015,168 (2)：635-47.；The public can be with Obtained from China Agricultural University.
Carrier pFGC1008：Cheng J, Wang Z, Yao F, et al.Down-Regulating CsHT1, a Cucumber Pollen-Specific Hexose Transporter, Inhibits Pollen Germination, Tube Growth, and Seed Development. [J] .Plant Physiology, 2015,168 (2)：635-47.；The public can be with Obtained from China Agricultural University.
Agrobacterium LBA4404：Cheng J, Wang Z, Yao F, et al.Down-Regulating CsHT1, a Cucumber Pollen-Specific Hexose Transporter, Inhibits Pollen Germination, Tube Growth, and Seed Development. [J] .Plant Physiology, 2015,168 (2)：635-47.；The public can be with Obtained from China Agricultural University.
MS solid mediums：PH 5.7- are adjusted in MS culture medium 4.43g, sucrose 30g, plant gel 2.5g, moisturizing to 1L 5.8。
MS differential mediums：MS culture medium 4.43g, sucrose 30g, plant gel 2.5g, 6-BA 0.5mg, ABA 1mg, mend Water is to 1L, pH 5.7-5.8.
MS culture mediums：West Beijing Mei Jie Science and Technology Ltd.s, article No.：M519.
3,5- dinitrosalicylic acid solutions：To addition 6.3g 3,5- dinitrosalicylic acids in the 262mL 2M NaOH aqueous solution Powder, then mixes with hydrothermal solutions of the 500mL containing 185g sodium potassium tartrate tetrahydrates, is stirred after adding 5g crystalline phenols and 5g sodium sulfites Dissolving is mixed, adds distilled water to be settled to 1000mL after cooling, brown bottle is used after preserving a week.
The preparation method of crude enzyme liquid is in following examples：1g testing samples are taken in the mortar of ice bath, adds 4mL enzymes to carry (solvent is pH7.5,50mM HEPES-NaOH buffer solutions to take liquid；MgCl containing 10mM2、1mM EDTA、2.5mM DTT、0.1g/ 100mlBSA and volumn concentration are 0.05% TritonX-100) homogenate is ground to form, 13000rpm centrifugation 20min take Clearly.Supernatant is fitted into bag filter, (TritonX-100 is not contained, the concentration of remaining each solute is zyme extract in dialyzate Dialysis 16h in the ten of middle concentration/), takes the liquid in dialyzate, as crude enzyme liquid, for enzyme assay.
The detection method of sucrose synthase activity is in following examples：Prepare 700 μ L reaction systems, 30 DEG C of reactions 30min；To adding 1500 μ L 3,5- dinitrosalicylic acid solution terminating reactions in reaction system；By reaction system boiling water bath 10min, determines OD after cooling540nmValue.Enzyme activity catalysing sucrose point within the unit interval with the crude enzyme liquid in unit fruit fresh weight Solution produces a μm ol numbers for fructose to represent.Unit is：μmol·h-1·g-1FW (FW refers to sample fresh weight).
Reaction system (700 μ L)：Solvent is 80mM Mes buffer solutions (pH 5.5)；Sodium fluoride containing 5mM, 100mM sucrose, 5mM UDP, 500 μ L crude enzyme liquids.
Control reaction system (700 μ L)：Solvent is 80mM Mes buffer solutions (pH 5.5)；Sodium fluoride containing 5mM, 100mM sugarcanes Sugar, 500 μ L crude enzyme liquids.
The detection method of acidic conversion enzymatic activity or alkaline invertase activity is in following examples：Prepare reaction system, By 37 DEG C of reaction system, water-bath 60min；To addition 1mL 3,5- dinitrosalicylic acid solution terminating reactions in reaction system；Will Flowing water cools down rapidly (4000rpm centrifugations 10min takes supernatant if having precipitation) after 100 DEG C of reaction system boils 5min, determines OD520nmValue.
Acid invertase reaction system：0.6ml0.1mol/L acetic acid second-acid sodium (pH4.8), 0.2ml0.1mol/L sucrose, 0.2ml crude enzyme liquids.
Acid invertase control reaction system：0.6ml0.1mol/L acetic acid second-acid sodium (pH4.8), 0.2ml distilled water, 0.2ml crude enzyme liquids.
Alkaline invertase reaction system：0.6ml0.1mol/L KH2PO4- 0.1mol/L sodium citrates (pH 7.2), 0.2ml0.1mol/L sucrose, 0.2ml crude enzyme liquids.
Alkaline invertase control reaction system：0.6ml0.1mol/L KH2PO4- 0.1mol/L sodium citrates (pH 7.2), 0.2ml distilled water, 0.2ml crude enzyme liquids.
The amount and enzymatic activity of its generation reduced sugar are calculated according to reduced sugar (glucose) standard curve.Enzyme activity unit is：μ mol·h-1·g-1FW (FW refers to sample fresh weight).
The acquisition of embodiment 1, sucrose synthase protein and its encoding gene
Sequence analysis, section interception and functional verification are carried out to each kind Cucumber germplasm, is sent out from the close thorn cucumber in Xintai City A kind of existing sucrose synthase protein, is named as CsSUS4 albumen, as shown in the sequence 1 of sequence table.CsSUS4 eggs will be encoded White unnamed gene CsSUS4 genes, as shown in the sequence 2 of sequence table.
Embodiment 2, cucumber different tissues position different times CsSUS4 gene expression amounts and enzyme activity assay
1st, the following material in Xintai City's close thorn cucumber following period is taken respectively：
Space expression is sampled：Root (R), stem (S) on same cucumber plant, climax leaves (ML), spire (YL), male flower (FF) With female flower (MF).
Temporal expressions are sampled：Flower a few days ago fruit (- 2DAF), bloom the same day fruit (0DAF), spend after three days fruits (3DAF), spend rear six days fruits (6DAF) and spend rear nine days fruits (commodity mellow fruit, 9DAF).
Every part of sample takes 3 biology and repeats above.
The total serum IgE of above-mentioned material is extracted, and synthesizes the first chain cDNA, CsSUS4 genes are detected using the method for qRT-PCR Expression (with TUA genes as reference gene), the primer pair constituted using primers F P1 and primer RP1 detects CsSUS4 bases The expression of cause, the primer pair constituted using primers F P2 and primer RP2 detects the expression of TUA genes.
FP1：5’-CATTTTTCTTGTCAATTTACTGCTG-3’；
RP1：5’-GCCCGACACGAAACGAC-3’；
FP2：5’-ACGCTGTTGGTGGTGGTAC-3’；
RP2：5’-GAGAGGGGTAAACAGTGAATC-3’.
2nd, the following material in Xintai City's close thorn cucumber following period is taken respectively：Flower a few days ago fruit (- 2DAF), bloom the same day fruit Real (0DAF), spend after three days fruits (3DAF), spend after six days fruits (6DAF) and nine days fruits after spending (commodity mellow fruit, 9DAF).Sucrose synthase activity, acidic conversion enzymatic activity and alkaline invertase activity in above-mentioned material are detected respectively.
Every part of sample takes 3 biology and repeats above.
Result is as shown in Figure 3.Result shows that CsSUS4 genes have expression at the different tissues position of cucumber, wherein Express higher in the organ of storehouse, including male flower, female flower, root and fruit.And the expression quantity of CsSUS4 genes is with the development of fruit Gradually rise, sucrose synthase activity and alkaline invertase activity are also constantly raised with fruit development, the activity of acid invertase Keep constant.
The acquisition of embodiment 3, transfer-gen plant
First, the structure of over-express vector
1st, the total serum IgE of the close thorn cucumber in Xintai City is extracted, and reverse transcription is cDNA, with cDNA as template, using SS4F and SA4R The primer pair of composition enters performing PCR amplification, reclaims pcr amplification product.
SS4F：5’-GCTCTAGAATGGCTTCTTTGGTGGTAAATCATCATAACGGT-3’；
SA4R：5’-TCCCCCGGGTTACTTTTGAATCCGAGATTGGGTGCGCTT-3’。
In SS4F and SA4R, underscore marks XbaI and SmaI restriction enzyme sites respectively.
2nd, with the pcr amplification product of restriction enzyme XbaI and SmaI double digestion step 1, digestion products are reclaimed.
3rd, with restriction enzyme XbaI and SmaI double digestion carrier pBI 121, the carrier framework of about 14750bp is reclaimed.
4th, the carrier framework of the digestion products of step 2 and step 3 is connected, obtains over-express vector pBI 121- CsSUS4.According to sequencing result, structure is carried out to over-express vector pBI 121-CsSUS4 and is described as follows：By carrier pBI 121 XbaI and SmaI restriction enzyme sites between small fragment substitution in order to sequence table sequence 2 shown in DNA molecular.Over-express vector PBI 121-CsSUS4 parts original papers schematic diagram is as shown in Figure 4 A.
2nd, the structure of interference carrier
1st, extract Xintai City it is close thorn cucumber total serum IgE, and reverse transcription be cDNA, with cDNA as template, using+S4iF and+ The primer pair of S4iR compositions enters performing PCR amplification, reclaims pcr amplification product.
+S4iF：5’-AGGCGCGCCTCTTTGGTGGTAAATCATCATAACG-3’；
+S4iR：5’-GCATTTAAATATATTCCCAACATCCTGGTTCTG-3’。
In+S4iF and+S4iF, underscore marks AscI and SwaI restriction enzyme sites respectively.
2nd, with the pcr amplification product of restriction enzyme A scI and SwaI double digestion step 1, digestion products are reclaimed.
3rd, with restriction enzyme A scI and SwaI double digestion carrier pFGC1008, the carrier framework of about 10907bp is reclaimed.
4th, the carrier framework of the digestion products of step 2 and step 3 is connected, obtains recombinant vector pFGC1008-RNAi-1. According to sequencing result, structure is carried out to recombinant vector pFGC1008-RNAi-1 and is described as follows：By the AscI of carrier pFGC1008 and Small fragment between SwaI restriction enzyme sites replaces the DNA molecular shown in the sequence 3 for sequence table.
5th, extract Xintai City it is close thorn cucumber total serum IgE, and reverse transcription be cDNA, with cDNA as template, using-S4iF and- The primer pair of S4iR compositions enters performing PCR amplification, reclaims pcr amplification product.
-S4iF：5’-GACTAGTTCTTTGGTGGTAAATCATCATAACG-3’；
-S4iR：5’-CGGGATCCATATTCCCAACATCCTGGTTCTG-3’。
In-S4iF and-S4iF, underscore marks SpeI and BamHI restriction enzyme sites respectively.
6th, with the pcr amplification product of restriction enzyme SpeI and BamHI double digestion step 5, digestion products are reclaimed.
7th, the recombinant vector pFGC1008-RNAi-1 obtained with restriction enzyme SpeI and BamHI step 4, reclaims about The carrier framework of 11190bp.
8th, the carrier framework of the digestion products of step 6 and step 7 is connected, obtains interference carrier pFGC1008-RNAi- CsSUS4.According to sequencing result, structure is carried out to interference carrier pFGC1008-RNAi-CsSUS4 and is described as follows：By recombinant vector Small fragment between AscI the and SpeI restriction enzyme sites of pFGC1008 replaces the DNA molecular shown in the sequence 4 for sequence table.It is dry Disturb carrier pFGC1008-RNAi-CsSUS4 parts original papers schematic diagram as shown in Figure 4 A.
3rd, the acquisition of overexpression transfer-gen plant
1st, the over-express vector pBI 121-CsSUS4 for obtaining step one import Agrobacterium LBA4404, obtain recombinating agriculture Bacillus.
2nd, the recombinational agrobacterium that step 1 is obtained is taken, is seeded to containing 25 μ g/ml rifampins and 100 μ g/ml kanamycins In YEB fluid nutrient mediums, 120rpm, 28 DEG C of cultures to bacterium solution OD600nmReach 0.6-0.8；Bacterium solution 5000rpm centrifugations 5min is received Collection bacterial sediment, after bacterial sediment is washed into 2 times with 1/2MS fluid nutrient mediums, is resuspended in 1/2MS fluid nutrient mediums, Adjustment bacterium solution OD600nmIt is 0.2.
3rd, the close thorn cucumber seeds in Xintai City are taken, 28 DEG C of constant temperature light cultures to seed is sprouted 2 days in being seeded in MS solid mediums After carry out subsequent experimental.
4th, the cucumber tender cotyledon of children after step 3 is sprouted 2 days is taken, every cotyledon is uniformly divided into 2 pieces after removal growing point, only Retain near the part of growing point half.
5th, the cucumber cotyledons after step 4 is processed are immersed in the Agrobacterium bacterium solution that step 2 is obtained and infect 15 minutes, then Unnecessary bacterium solution is blotted with the filter paper of sterilizing, cucumber cotyledons are seeded in MS differential mediums, after the culture 2 days of 28 DEG C of dark condition Harvest explant.
The 6th, the explant that step 5 is obtained is seeded to the MS containing 50mg/L kanamycins and 500mg/L carbenicillins On differential medium, 25 DEG C, 2000LX intensities of illumination (14h illumination/10h dark) culture, whne resistant budses it is long to 1cm when, by it Root induction in MS solid mediums of the immigration containing 100mg/L kanamycins and 200mg/L carbenicillins is cut, T is obtained0 For overexpression transfer-gen plant, after plant to be planted root system development is good, immigration is filled in the flowerpot of sterile soil, greenhouse Routine Management.T0 For plant selfing, T is obtained1For plant.T1For plant selfing, T is obtained2For plant.
7th, the T of some transgenic lines that step 6 is obtained is taken2Identified for plant.Take plant seedling leaf to be measured Simultaneously reverse transcription is cDNA to total serum IgE, with cDNA as template, enters performing PCR using primer 35S2-F and 35S2-R and identifies.
35S2-F：5’-TGGTTAGAGAGGCTTACGCAGCAGGTC-3’；
35S2-R：5’-CCATCTTTGGGACCACTGTCGGCA-3’.
If for a certain T0For plant, the T of its sampling Detection2PCR qualification results for plant are the positive, the T0Generation Plant and its self progeny are an overexpression transgenic line for homozygosis.
It is the negative control of template that setting uses close thorn cucumber (wild type) cDNA in Xintai City.
It is the positive control of template that setting uses over-express vector pBI 121-CsSUS4 DNA.
Qualification result is as shown in Figure 1.
4th, the acquisition of interference of transgene plant
Interference carrier pFGC 1008-RNAi-CsSUS4 are obtained instead of over-express vector pBI 121- using step 2 CsSUS4, is operated according to step 3, obtains T0For interference of transgene plant, after plant to be planted root system development is good, immigration is filled In the flowerpot of sterile soil, greenhouse Routine Management.T0For plant selfing, T is obtained1For plant.T1For plant selfing, T is obtained2Dai Zhi Strain.Using the method in step 3 to the T of interference of transgene strain2Identified for plant.
It is the negative control of template that setting uses close thorn cucumber (wild type) cDNA in Xintai City.
It is the positive control of template that setting uses interference carrier pFGC1008-RNAi-CsSUS4 DNA.
Qualification result is as shown in Figure 2.
5th, the acquisition of empty carrier plant is turned
1st, over-express vector pBI 121-CsSUS4 are replaced using carrier pBI 121, is operated according to step 3, obtained Overexpression non-transgenic control lines, after plant to be planted root system development is good, immigration is filled in the flowerpot of sterile soil, greenhouse Routine Management.
2nd, over-express vector pBI 121-CsSUS4 are replaced using carrier pFGC1008, is operated according to step 3, obtained To interference of transgene adjoining tree, after plant to be planted root system development is good, immigration is filled in the flowerpot of sterile soil, greenhouse Routine Management.
The Phenotypic Observation of embodiment 4, transfer-gen plant
Plant to be measured is：Xintai City it is close thorn cucumber (WT), embodiment 3 build overexpression transgenic line (OE6-10 and OE17-2 T)2For plant, the T of interference of transgene strain (Ri54-12 and Ri37-5)2For plant, the strain of overexpression non-transgenic control The T of system2The T of strain is compareed for plant and interference of transgene2For plant.
1st, taking plant unification to be measured carries out daily field management.Remove root melon, be listed on the day of second melon is bloomed, The melon that Post flowering grows into 9 days is taken, the statistics of single fruit weight, single fruit length and single fruit rugosity is carried out, and using the side in embodiment 2 The expression of the CsSUS4 genes in method detection fruit, and detect fruit sucrose synthase activity.
Result is as shown in Figure 4.Result shows, compared with the close thorn cucumber (WT) in wild type Xintai City, CsSUS4 genes it is relative Expression quantity raises in overexpression transgenic line (OE6-10 and OE17-2) fruit, interference of transgene strain (Ri54-12 and Ri37-5) (Fig. 4 B) is lowered in fruit；Compared with the close thorn cucumber in wild type Xintai City, the activity of sucrose synthase is in interference of transgene Raised in strain (Ri54-12 and Ri37-5) fruit, lowered in interference of transgene strain (Ri54-12 and Ri37-5) fruit (Fig. 4 C)；The fruit of interference of transgene strain (CsSUS4-Ri) is significantly less than wild type；And overexpression transgenic line (CsSUS4-OE) fruit is significantly greater than wild type (Fig. 4 D)；Overexpression transgenic line (OE6-10 and OE17-2) fruit and open country Raw type is compared, and no matter fruit length, rugosity and weight are dramatically increased, interference of transgene strain (Ri54-12 and Ri37-5) fruit Compared with WT strain, no matter fruit length, rugosity and weight are substantially reduced (Fig. 4 E, F, G).Overexpression non-transgenic control is planted The statistics of strain and interference of transgene adjoining tree is with wild type without significant difference.
2nd, taking plant unification to be measured carries out daily field management.The phenotype of male flower, each strain are counted in male flower full-bloom stage 50 weight of male flower are at least counted, the average single flower weight of the hemp nettle of each strain 5,6 hemp nettles, 7 hemp nettles and 8 hemp nettles are counted respectively, The quantitative proportion shared by the hemp nettle of each strain 5,6 hemp nettles, 7 hemp nettles and 8 hemp nettles, Sucrose synthesis enzyme activity in detection male flower are counted respectively Property.
Result is as shown in Figure 5.Result shows, Sucrose synthesis in overexpression transgenic line (OE6-10 and OE17-2) male flower Activity ratio's wild type (WT) is high；Sucrose synthase activity is than wild in interference of transgene strain (Ri54-12 and Ri37-5) male flower Raw type is low (Fig. 5 A)；Compared with wild type male flower, overexpression transgenic line (OE6-10 and OE17-2) male flower is substantially done greatly Disturb transgenic line (Ri54-12 and Ri37-5) male flower substantially small (Fig. 5 B)；Overexpression transgenic line (OE6-10 and OE17- 2), in interference of transgene strain (Ri54-12 and Ri37-5) and wild type 5 hemp nettles, 6 hemp nettles, 7 hemp nettles and 8 hemp nettles average list There were significant differences (Fig. 5 C) for flower weight；Occurs the ratio of 7 hemp nettles and 8 hemp nettles in overexpression transgenic line (OE6-10 and OE17-2) Example is higher than wild type, occur in interference of transgene strain (Ri54-12 and Ri37-5) ratio of 7 hemp nettles and 8 hemp nettles with it is wild Type is quite (Fig. 5 D).The statistics of overexpression non-transgenic control lines and interference of transgene adjoining tree is with wild type without notable Difference.
<110>China Agricultural University
<120>Sucrose synthase is in the regulation and control developmental application of fruit
<160> 4
<210> 1
<211> 834
<212> PRT
<213>Cucumber（Cucumis sativus L.）
<400> 1
Met Ala Ser Leu Val Val Asn His His Asn Gly Glu Ser Ile Gly Asp
1 5 10 15
Gly Ile Val Glu Ala Leu Lys Gln Asn His Asn Tyr Met Lys Arg Cys
20 25 30
Phe Gly Lys Phe Val Glu Lys Gly Asn Arg Ser Leu Lys Lys Lys Glu
35 40 45
Leu Met Glu Glu Met Glu Leu Val Ile Asp Asp Lys Ile Glu Arg Asn
50 55 60
Arg Val Met Glu Gly Val Leu Gly His Met Leu Thr Ser Thr Gln Val
65 70 75 80
Ala Ile Val Ile Pro Pro Tyr Val Ala Phe Ala Ile Arg Pro Glu Pro
85 90 95
Gly Cys Trp Glu Tyr Val Lys Val Ser Ser Leu Asp Leu Ser Leu Gln
100 105 110
Ser Leu Thr Ser Thr Glu Phe Leu Lys Leu Lys Glu Met Ile Tyr Asp
115 120 125
Glu Glu Trp Ala Asn Asp Glu Asn Ala Leu Glu Val Asp Phe Gly Ala
130 135 140
Ile Glu Phe Thr Thr Pro Arg Leu Ser Leu Pro Ser Ser Ile Gly Asp
145 150 155 160
Gly Leu Ser Tyr Thr Thr Lys Phe Leu Thr Ser Lys Leu Ser Gly Lys
165 170 175
Ser Glu Asn Leu Gln Pro Leu Val Asp Tyr Leu Leu Ser Leu Asp Tyr
180 185 190
Gln Gly Glu Lys Leu Met Ile Asn Glu Thr Leu Ser Thr Ala Ser Lys
195 200 205
Leu Gln Met Thr Leu Ile Leu Ala Asp Ile Phe Leu Ser Val Leu Pro
210 215 220
Pro Asp Thr Pro Tyr Asp Asp Phe His Leu Lys Phe Lys Gln Trp Gly
225 230 235 240
Phe Glu Arg Gly Trp Gly Asp Cys Ala Gly Arg Val Lys Glu Thr Ile
245 250 255
Arg Cys Leu Ser Glu Ile Phe Gln Ala Tyr Asp Pro Ile Gln Met Glu
260 265 270
Lys Phe Phe Ser Arg Leu Pro Thr Ile Phe Asn Val Val Ile Leu Ser
275 280 285
Pro His Gly Tyr Phe Gly Gln Ala Gly Val Leu Gly Leu Pro Asp Thr
290 295 300
Gly Gly Gln Val Val Tyr Ile Leu Asp Gln Val Lys Ala Met Glu Glu
305 310 315 320
Glu Leu Leu Leu Arg Ile Lys Gln Gln Gly Leu Asn Phe Lys Pro Gln
325 330 335
Ile Ile Ile Ile Thr Arg Leu Ile Pro Asp Ala Lys Gly Thr Lys Cys
340 345 350
Asn Gln Glu Ile Glu Pro Val Ile Gly Thr Thr Tyr Ser Lys Ile Val
355 360 365
Arg Val Pro Phe Lys Thr Glu Asn Gly Thr Leu His Arg Trp Val Ser
370 375 380
Arg Phe Asp Ile Tyr Pro Tyr Leu Glu Lys Phe Ala Gln Asp Ala Ser
385 390 395 400
Asp Lys Ile Leu Glu Leu Met Glu Ala Lys Pro Asp Leu Ile Ile Gly
405 410 415
Asn Tyr Thr Asp Gly Asn Leu Val Ala Ser Leu Met Ala Ser Arg Leu
420 425 430
Gly Val Thr Gln Gly Thr Ile Ala His Ala Leu Glu Lys Thr Lys Tyr
435 440 445
Glu Asp Ser Asp Leu Lys Trp Lys Glu Leu Asp Ser Lys Tyr His Phe
450 455 460
Ser Cys Gln Phe Thr Ala Asp Ile Leu Ala Met Asn Ala Thr Asp Phe
465 470 475 480
Val Ile Ala Ser Thr Phe Gln Glu Ile Ala Gly Ser Lys Glu Lys Pro
485 490 495
Gly Gln Tyr Glu Ser His Glu Ala Phe Thr Leu Pro Gly Leu Cys Arg
500 505 510
Phe Val Ser Gly Ile Asn Val Phe Asp Pro Lys Phe Asn Ile Ala Ala
515 520 525
Pro Gly Ala Asp Gln Ser Val Tyr Phe Pro Tyr Thr Thr Lys Glu Leu
530 535 540
Arg Phe Ala Ser Phe Gln Pro Ala Ile Glu Glu Leu Leu Phe Ser Lys
545 550 555 560
Val Glu Asn Asp Glu His Ile Gly Tyr Leu Ala Asp Arg Lys Lys Pro
565 570 575
Ile Ile Phe Ser Met Ala Arg Leu Asp Val Val Lys Asn Ile Thr Gly
580 585 590
Leu Val Glu Trp Phe Gly Lys Asn Glu Lys Leu Arg Asn Leu Val Asn
595 600 605
Leu Val Val Val Gly Gly Phe Phe Asp Pro Ser Lys Ser Lys Asp Arg
610 615 620
Glu Glu Met Ala Glu Ile Arg Lys Met His Glu Leu Ile Asp Lys Tyr
625 630 635 640
Gln Leu Lys Gly Gln Ile Arg Trp Ile Ala Ala Gln Thr Asp Arg Arg
645 650 655
Arg Asn Gly Glu Leu Tyr Arg Cys Ile Ala Asp Thr Lys Gly Ala Phe
660 665 670
Val Gln Pro Ala Leu Tyr Glu Ala Phe Gly Leu Thr Val Ile Glu Ala
675 680 685
Met Asn Cys Gly Leu Pro Thr Phe Ala Thr Asn Gln Gly Gly Pro Ala
690 695 700
Glu Ile Ile Val Asp Gly Val Ser Gly Phe Gln Ile Asp Pro Asn Asn
705 710 715 720
Gly Thr Glu Ser Ser Gln Lys Ile Ala Asn Phe Phe Glu Lys Cys Lys
725 730 735
Asn Asp Pro Thr Tyr Trp Asn Glu Ile Ser Asn His Gly Leu Gln Arg
740 745 750
Ile Asn Glu Cys Tyr Thr Trp Lys Ile Tyr Ala Lys Lys Val Leu Asn
755 760 765
Met Gly Ser Thr Tyr Ser Phe Trp Lys Gln Val Asn Lys Asn Gln Lys
770 775 780
Gln Ala Lys Asp Arg Tyr Ile Gln Met Phe Tyr Asn Leu Leu Phe Lys
785 790 795 800
Asn Leu Val Lys Asn Val Pro Ile Val Val His Glu Asp Ser His Pro
805 810 815
Glu Asn Pro Arg Leu Pro Gln Val Ser Lys Arg Thr Gln Ser Arg Ile
820 825 830
Gln Lys
<210> 2
<211> 2505
<212> DNA
<213>Cucumber（Cucumis sativus L.）
<400> 2
atggcttctt tggtggtaaa tcatcataac ggtgagtcga ttggtgatgg aattgtagag 60
gcattgaagc aaaaccataa ttatatgaag agatgctttg gtaagtttgt agagaaaggg 120
aataggagct tgaagaagaa ggaattaatg gaggaaatgg agcttgttat agatgataag 180
atagaaagga atagggttat ggagggtgtc cttggtcata tgttgacttc aactcaggtg 240
gcgattgtga ttccaccata tgttgcattt gcaataagac cagaaccagg atgttgggaa 300
tatgttaaag ttagttctct tgatctttca ctccaatctc tcacttcaac tgaatttctc 360
aaattgaagg agatgattta cgacgaagaa tgggcaaatg atgaaaatgc tttggaagtg 420
gatttcggag caattgaatt tacaactcct cggttaagcc ttccatcttc aattggagat 480
ggacttagtt atactacaaa gtttctcacc tcaaagctga gtgggaagtc ggagaattta 540
caacctcttg tagattactt gttgtccctt gattatcaag gagagaaact tatgatcaat 600
gaaactctaa gcacagcttc caaactccaa atgacattaa ttcttgctga tattttcctc 660
tctgttcttc ctccagacac cccatatgat gacttccatc tcaaattcaa gcaatgggga 720
tttgagagag gatggggaga ttgtgctgga agagtaaagg aaacaataag atgtttatct 780
gaaatattcc aagcctatga tccaatccaa atggagaagt tcttcagcag gcttcctact 840
attttcaatg ttgtcatttt gtctcctcat ggatactttg gccaagctgg tgttcttggt 900
ttgcccgaca ccggaggtca ggttgtgtac atactcgatc aagttaaagc tatggaagaa 960
gaacttctgc tcagaattaa gcaacaaggc ctcaatttca agcctcaaat tattattatt 1020
acaagactta ttccagatgc gaaggggact aagtgcaacc aagaaataga acctgtcatt 1080
gggactactt actccaagat tgttagggtg cccttcaaga ctgaaaatgg cacccttcat 1140
cgttgggttt ctcgtttcga catttatcct tatctcgaga aatttgcaca agatgcgagc 1200
gacaaaatat tagagctcat ggaagcaaag ccagatctaa tcattggaaa ctacacagat 1260
ggaaatcttg tggcatctct catggccagc agattaggag taacccaagg aactattgca 1320
catgccttgg agaagacaaa gtatgaagat tcagatctta aatggaagga attggactcc 1380
aagtaccatt tttcttgtca atttactgct gatattcttg ctatgaatgc gactgatttt 1440
gtcatcgcaa gcactttcca agagattgca ggaagcaaag aaaagccagg ccaatatgaa 1500
agccacgagg catttacgct tccaggactg tgtcgtttcg tgtcgggcat caatgtgttt 1560
gatcctaagt tcaacattgc agcaccaggg gctgatcagt ctgtctattt tccttacacg 1620
accaaagaac ttcgatttgc atcgtttcaa cctgccattg aagaacttct ttttagcaaa 1680
gttgagaacg atgagcatat aggatatctg gccgatagga aaaagccgat catcttttca 1740
atggcacggc ttgatgttgt caagaacatt accgggttgg tcgaatggtt tgggaagaat 1800
gagaagctga gaaatttggt gaatcttgtt gtggttggag gattttttga tccttccaaa 1860
tcaaaggaca gagaagaaat ggcagagata agaaagatgc atgaattaat tgacaaatac 1920
caactcaaag gtcagatcag gtggatagca gcacagactg atcgccgtcg taatggagaa 1980
ctctaccgct gcattgctga cacaaaagga gcctttgtgc agcctgctct ctatgaagct 2040
tttggtctca cagtcattga ggcaatgaat tgtggtctac caacctttgc tacgaaccaa 2100
gggggtccag ctgagatcat tgttgatggg gtctctggct tccaaattga tcccaacaat 2160
ggcactgaat caagccaaaa gattgctaac ttttttgaga aatgcaagaa tgatccaacc 2220
tactggaacg aaatttcgaa tcacggtctt caacgtatca atgaatgtta cacatggaaa 2280
atctatgcaa aaaaggtgct aaatatggga agcacttaca gtttttggaa gcaagtgaac 2340
aaaaaccaaa agcaggcaaa ggacagatac atccaaatgt tctacaatct actttttaag 2400
aacttggtga aaaacgtgcc aatcgtggtc cacgaagatt cacatccaga aaatccacgg 2460
ttaccacaag tctcgaagcg cacccaatct cggattcaaa agtaa 2505
<210> 3
<211> 297
<212> DNA
<213>Cucumber（Cucumis sativus L.）
<400> 3
tctttggtgg taaatcatca taacggtgag tcgattggtg atggaattgt agaggcattg 60
aagcaaaacc ataattatat gaagagatgc tttggtaagt ttgtagagaa agggaatagg 120
agcttgaaga agaaggaatt aatggaggaa atggagcttg ttatagatga taagatagaa 180
aggaataggg ttatggaggg tgtccttggt catatgttga cttcaactca ggtggcgatt 240
gtgattccac catatgttgc atttgcaata agaccagaac caggatgttg ggaatat 297
<210> 4
<211> 975
<212> DNA
<213>Artificial sequence
<220>
<223>
<400> 4
tctttggtgg taaatcatca taacggtgag tcgattggtg atggaattgt agaggcattg 60
aagcaaaacc ataattatat gaagagatgc tttggtaagt ttgtagagaa agggaatagg 120
agcttgaaga agaaggaatt aatggaggaa atggagcttg ttatagatga taagatagaa 180
aggaataggg ttatggaggg tgtccttggt catatgttga cttcaactca ggtggcgatt 240
gtgattccac catatgttgc atttgcaata agaccagaac caggatgttg ggaatatatt 300
taaatcccca gatgaacatg gcatcgtggt gattgatgaa actgctgctg tcggctttaa 360
cctctcttta ggcattggtt tcgaagcggg caacaagccg aaagaactgt acagcgaaga 420
ggcagtcaac ggggaaactc agcaagcgca cttacaggcg attaaagagc tgatagcgcg 480
tgacaaaaac cacccaagcg tggtgatgtg gagtattgcc aacgaaccgg atacccgtcc 540
gcaaggtgca cgggaatatt tcgcgccact ggcggaagca acgcgtaaac tcgacccgac 600
gcgtccgatc acctgcgtca atgtaatgtt ctgcgacgct cacaccgata ccatcagcga 660
tctctttgat ggggatccat attcccaaca tcctggttct ggtcttattg caaatgcaac 720
atatggtgga atcacaatcg ccacctgagt tgaagtcaac atatgaccaa ggacaccctc 780
cataacccta ttcctttcta tcttatcatc tataacaagc tccatttcct ccattaattc 840
cttcttcttc aagctcctat tccctttctc tacaaactta ccaaagcatc tcttcatata 900
attatggttt tgcttcaatg cctctacaat tccatcacca atcgactcac cgttatgatg 960
atttaccacc aaaga 975

Claims (10)

1. a kind of method for cultivating genetically modified plants, is to import in purpose plant the encoding gene of sucrose synthase, is turned Gene plant；The genetically modified plants meet at least one of following (a1)-(a5) proterties：
(a1) fruit yield is more than the purpose plant；
(a2) fruit length is more than the purpose plant；
(a3) fruit rugosity is more than the purpose plant；
(a4) fruit weight is more than the purpose plant；
(a5) male flower is more than the purpose plant.
2. a kind of method for cultivating genetically modified plants, is the expression of the encoding gene for suppressing sucrose synthase in purpose plant, is obtained To genetically modified plants；The genetically modified plants meet at least one of following (b1)-(b5) proterties：
(b1) fruit yield is less than the purpose plant；
(b2) fruit length is less than the purpose plant；
(b3) fruit rugosity is less than the purpose plant；
(b4) fruit weight is less than the purpose plant；
(b5) male flower is less than the purpose plant.
3. method as claimed in claim 1 or 2, it is characterised in that：The sucrose synthase is following (c1) or (c2)：
(c1) protein that the amino acid sequence shown in sequence in sequence table 1 is constituted；
(c2) by the amino acid sequence of sequence 1 by the substitution of one or several amino acid residues and/or missing and/or addition and With identical function as derived from sequence 1 protein.
4. the method as described in claim 1-3 is any, it is characterised in that：The encoding gene of the sucrose synthase is as follows (d1) any described DNA molecular in-(d3)：
(d1) DNA molecular of the code area as shown in sequence 2 in sequence table；
(d2) the DNA sequence dna hybridization for being limited with (d1) under strict conditions and the DNA molecular of encoding sucrose synzyme；
(d3) DNA sequence dna limited with (d1) or (d2) has the DNA molecular of more than 90% homology and encoding sucrose synzyme.
5. the method as described in claim 1-4 is any, it is characterised in that：The purpose plant is cucumber.
6. application of any described methods of claim 1-5 in plant breeding.
7. the application of sucrose synthase, is at least one in following (e1)-(e16)：
(e1) regulation and control fruit development；
(e2) regulation and control plant storehouse is strong；
(e3) sucrose synthase activity in regulation and control plant organ；
(e4) sucrose synthase activity in regulation and control fruit；
(e5) sucrose synthase activity in regulation and control plant male flower；
(e6) sucrose synthase catalysing sucrose is converted into the activity of fructose in regulation and control plant organ；
(e7) sucrose synthase catalysing sucrose is converted into the activity of fructose in regulation and control fruit；
(e8) sucrose synthase catalysing sucrose is converted into the activity of fructose in regulation and control plant male flower；
(e9) fruit development is promoted；
(e10) plant storehouse is improved strong；
(e11) sucrose synthase activity in plant organ is promoted；
(e12) sucrose synthase activity in fruit is promoted；
(e13) sucrose synthase activity in plant male flower is promoted；
(e14) sucrose synthase catalysing sucrose is converted into the activity of fructose in promotion plant organ；
(e15) sucrose synthase catalysing sucrose is converted into the activity of fructose in promotion fruit；
(e16) sucrose synthase catalysing sucrose is converted into the activity of fructose in promotion plant male flower.
8. a kind of specific DNA molecular, including section first and section second；The section first and the section second are reverse complemental sequence Row；The sequence of the section first is as shown in the sequence 3 of sequence table.
9. a kind of interference carrier, is the recombinant expression carrier containing specific DNA molecular described in claim 8.
10. the specific DNA molecular described in claim 8, or, the interference carrier described in claim 9 is cultivating genetically modified plants In application；The genetically modified plants meet at least one of following (f1)-(f5) proterties：
(f1) fruit yield is less than the plant that sets out；
(f2) fruit length is less than the plant that sets out；
(f3) fruit rugosity is less than the plant that sets out；
(f4) fruit weight is less than the plant that sets out；
(f5) male flower is less than the plant that sets out.
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* Cited by examiner, † Cited by third party
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CN110894221A (en) * 2019-12-12 2020-03-20 中国科学院植物研究所 Strawberry maturation associated transcription factor gene FaNAC2 and application thereof

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