CN101643742A - Gene for regulating fruit maturity and quality and encoding product and application thereof - Google Patents
Gene for regulating fruit maturity and quality and encoding product and application thereof Download PDFInfo
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Abstract
The invention relates to a gene for regulating fruit maturity and quality and an encoding product and an application thereof and discloses a gene MaGAD1 which is related to the maturation process of banana fruits after harvest and is one of nucleotide sequences as follows: 1) the DNA sequence in the <400>1 term in the sequence table; 2) protein in the <400>1 term with the amino acid residue sequence in the <400>2 term or the derived protein with the same activity of the amino acid residue sequence in the <400>2 term by replacing, deleting or adding one or more of amino acid residues to the amino acid residue sequence in pseudo column. The gene can be separated and inhibited by gamma-aminobutyric acid (GABA) and aminoxyacetic acid (AOAA) so as to regulate the changes of the maturity, hardness, total soluble sugar, starch content and the like of the banana fruits after harvest. The application of the gene has an important theoretical and practical significance for building a new technology for regulating the maturation process of banana fruits after harvest and culturing good varieties.
Description
[technical field]
(glutamate decarboxylase gene, the MaGAD1) acquisition of sequence, expression analysis and external regulate gene expression are regulated banana maturation and quality to the present invention relates to the banana glutamic acid decarboxylase gene.
[background technology]
(Glutamate decarboxylase GAD) extensively is present in protokaryon and the eukaryote L-Glutamic decarboxylase, is an intracellular enzyme, mainly is present in the tenuigenin, combines the catalysis glutamic acid rotating with the pyridoxal phosphate cofactor and turns to GABA and CO
2The C-terminal of GAD has special and calmodulin (CaM) bonded zone in plant, and can combine with CaM increases enzyme activity (Chen et al.1994).Plant when the many different stimulation of reply, such as, physical abuse, cold stimulation, thermal stimulus, histanoxia, plant hormone stimulation etc., GAD is by intracellular H
+Or Ca
2+The increase of level and activating, GABA can a large amount of accumulation.GABA is accumulated in (the Bown et al.1997 that plays an important role in the physiological processs such as PH regulation and control, N storage, development of plants, fruit maturation, plant defense; Gallegoet al.1995).
The GAD gene is cloned in plants such as Arabidopis thaliana, petunia, paddy rice, tomato, tobacco, the result shows that this gene may be the multigene family coding, and be prevalent in each tissue of plant, it is expressed to change and present differential expression in different physiological process such as development of plants, external stimulus and fruit maturation.GAD expression of gene and fruit maturation and ethene biosynthesizing also have relation, Kathiresan etc. (Kathiresan 1997) have studied the relation that GAD and ethene produce in the Sunflower Receptacle, external source GABA handled the Sunflower Receptacle cotyledon 12 hours, caused the ethene production rate to increase by 14 times.Its regulation process is, external source GABA has promoted synthesizing of endogenous GA BA, thereby promoted the accumulation of ACC synthetic enzyme mRNA and ACC content, and the increase of acc oxidase mRNA and external acc oxidase vigor, and proved that GABA is not the Ethylene Biosynthesis precursor, promptly GABA participates in the ethene biosynthesizing by pathways metabolism.Infer that external source GABA promotes endogenous GA BA synthetic process to be: GABA has at first promoted the GAD expression of gene, has increased the GAD enzymic activity, thereby has promoted synthesizing of endogenous GA BA.GABA promotes that the ethene biosynthesizing mainly is the enhancing by inducing the ACC synthetic enzyme to transcribe.Gallego etc. (Gallego et al.1995) obtain a GAD cDNA total length from tamato fruit cDNA library, tissue specific expression analysis revealed, this gene expression amount in tissues such as root, stem, leaf is very low, and expression amount is higher in fruit.Fruit is adopted after ripening different steps expression analysis and is shown that this gene expression amount in normal mature and rin mutant is the highest when all fruit for the first time colour-change takes place after adopting; This gene descends after expression amount reaches peak value immediately in the normal mature fruit, and this genetic expression maintains higher level always in the rin mutant.The differential expression of (adopting back 10d) gene when we (Jinet al.2008) utilizes cDNA microarray and RT-PCR analysis and research banana to adopt back ethene biosynthesizing startup, showing the obvious up-regulated expression of banana glutamic acid decarboxylase gene, is of expression amount maximum in 16 up-regulated expression genes.Sum up above-mentioned result of study: in ethene biosynthesizing and ripening of fruits, GAD catalysis glutamic acid rotating turns to GABA, and GABA may participate in the organic acid degraded by pathways metabolism, the PH regulation and control, thus the metabolism of N causes ripe relevant physiological change; GABA also may be as ethene biosynthesizing in the signaling molecule regulating fruit ripening process.Therefore, the development and use of GAD gene will help to understand in depth more banana and adopt after ripening mechanism, set up the cheap fruit post-harvest fresh-keeping novel method of safety more.
[summary of the invention]
Adopt after ripening mechanism in order to further investigate banana, set up the cheap effectively fruit new fresh keeping technique of safety, improve the commodity value of fruit, separation is with the fruit maturation genes involved and identify that their function is very necessary, by to these genoid functional study and regulation and control, to adopt the back quality to improving fruit, cultivation storage endurance new variety are significant.
Technical scheme provided by the invention is: make up banana and adopt back 0 day, 2 days inhibition reduction libraries, obtain a differential expression L-Glutamic decarboxylase homologous cDNA sequence higher, RACE technology this full length gene sequence (MaGAD1) that increases with the GAD dna homolog.Fluorescence quantifying PCR method research induces maturation and 1-MCP (1-methyl cyclopropene) to suppress under the sophisticated condition at normal mature, ethene, and MaGAD1 adopts the back expression in different stage of maturity at banana.By expression external evoked, that suppress MaGAD1, research fruit maturation characteristic and quality.
1. banana is adopted and was suppressed subtracted library structure and analysis in back 2 days:
Adopt hot borate method method to extract banana and adopted back 0 day and 2 days total RNA of fruit, electrophoresis detection RNA quality, store in after quality examination finishes-76 ℃ standby.PCR-Select according to Clontech company
TMCDNA SubstractionKit, Advantage 2 PCR Kit product operation instructions are reduced hybridization.Suppress reduction hybridization product through being connected with pGEM-T Easy Vecter, Transformed E .coli DH5 α makes up and suppresses subtracted library.Obtain library independent cloning order-checking, analyze.
2. banana glutamic acid decarboxylase gene full length sequence is cloned:
Gene fragment that obtains according to SSH and the comparison result of NCBI as can be known, this gene fragment 3 ' end has contained the TAA terminator codon.With one 5 of Primer Premier 5 design '-the RACE primer, the banana cDNA library that makes up with this research department is a template, pcr amplification 5 ' terminal sequence.Full length sequence with extension increasing sequence and existing 3 ' terminal sequence splicing acquisition gene.
3. fluorescent quantitation research MaGAD1 and fruit are adopted the relation of after ripening:
Adopt that the back banana sets up that ethene is handled, 1-MCP handles separately and 3 groups of normal matures, extract the total RNA reverse transcription of the fruit cDNA in different stage of maturity in 3 groups, in the fluorescent quantitation research 3 under the different treatment condition MaGAD1 express and sophisticated relation.
4.MaGAD1 inductor γ-An Jidingsuan (GABA) and inhibitor AOAA (AOAA) are handled fruit, study the influence of external regulate gene expression to fruit maturation and quality:
Banana after adopting is handled respectively with γ-An Jidingsuan (GABA) and AOAA (AOAA) according to finite concentration, observe and handle the fruit maturation variation, measure the relevant physical signs of fruit maturation, analyze expression by adopting the external regulation and control in back MaGAD1 the influence of fruit maturation and quality.
[interpretation of result]
1. successfully make up banana and adopt the inhibition reduction library of back 0 day and 2 days: subtract the PCR product with difference, make up the plasmid vector library with the T/A cloning, subtracted library comprises that 312 whites clone, and it is full clear to clone.The application PCR method is screened 312 clones of subtracted library, obtains 302 recons, inserts fragment and concentrates between the 300bp-500bp, meets the requirement that suppresses the reduction library.
2. successfully clone banana glutamic acid decarboxylase gene full length sequence: to 302 reorganization sequencing analysis in the inhibition reduction library that obtains, there are 2 to have higher homology, further analyze and show that these two banana glutamic acid decarboxylase gene fragments belong to same gene with the sub-glutamic acid decarboxylase gene of other biological.According to banana glutamic acid decarboxylase gene fragment design 5 ' the end PCR primer that obtains, clone's 5 ' terminal sequence, with utilize the information biology software analysis behind original sequence assembly, has complete open-reading frames, be a complete genome called after MaGAD1, the complete open reading frame (sequence 1) that comprises a 1500bp, the polypeptide (sequence 2) of 499 amino-acid residues of coding.With paddy rice (NM_001068533), barley (AK248209), Arabidopis thaliana (NM_121739) and corn (AY103733) higher consistence is arranged, be respectively 82%, 80%, 80%, 78%.GenBank searches and does not appear in the newspapers.
3.MaGAD1 adopt the back by the ethene abduction delivering at banana, and may be adjusted to ripe ethene biosynthesizing: induce under the sophisticated condition at normal mature and ethene, MaGAD1 is significantly induced when ripe ethene biosynthesizing starts, its expression amount reaches maximum value fast, and has induced subsequently that rolling up of ethene makes it reach peak value; Under 1-MCP suppressed maturation condition, the burst size of endogenous ethylene and the relative expression quantity of MaGAD1 were adopted the back storage at banana and are maintained lower level (Fig. 1) period always.So the expression of MaGAD1 not only had been subjected to ethene to induce but also promote the biosynthesizing of ethene, adopt back ethene biosynthesizing with banana and fruit maturation closely related.
4.MaGAD1 inductor and inhibitor significantly promote and suppress this expression of gene, and then influence the ripening process and the quality of fruit: result of study proves, GABA and AOAA can induce and suppress the banana maturation respectively, show that by physiologic analyses this influence has changed fruit color, hardness, aromatoising substance, solubility total reducing sugar and starch content etc.Confirmation can be regulated fruit quality (Fig. 2, Fig. 3) to the regulation and control that MaGAD1 expresses.Studies show that further MaGAD1 regulating fruit maturation realizes by regulation and control MaACS1 expression of gene, and MaACS1 to be banana adopt starts ripe Ethylene Biosynthesis key gene in the after ripening process.Therefore, we infer that banana MaGAD1 is that banana is adopted ripe Ethylene Biosynthesis upstream regulation person in the after ripening process, and it forms by the ripening process and the quality of the expression control fruit of regulation and control MaACS1.
Description of drawings:
Under Fig. 1: 1-MCP processing and the ethene treatment condition, utilize quantitative fluorescent PCR to analyze MaGAD1 adopts the back different times at banana expression.
Fig. 2: GABA and AOAA handle the influence of banana being adopted after ripening
Fig. 3: GABA and AOAA handle adopting the influence of back banana hardness, starch, solubility total reducing sugar.
[embodiment]
The present invention is described in detail below in conjunction with embodiment:
1, material source: Brazilian banana (M.AAA Group Cavendish) fruit, pick up from China tropic Agriculture Academy Sciences tropic Biotechnology Research Institute experiment base.
2, method:
(1) banana is adopted and suppressed subtracted library structure and analysis in back 2 days: the hot borate method with reference to Ching-Yi Wan and Thea A.Wilkins (1994) extracts banana RNA.According to PCR-Select
TMCDNA SubstractionKit, Advantage 2 PCR Kit product operation instructions were carried out 0 day and 2 days fruit RNA suppress reduction hybridization.Suppress reduction hybridization product through be connected Transformed E .coli DH5 α with pGEM-T Easy Vecter.Adopt the method for PCR that recon is identified, PCR is provided by the primer that adopts test kit to provide:
Nested PCR primer 1:5 '-TCGAGCGGCCGCCCGGGCAGGT-3 '
Nested PCR primer 2 R:5 '-AGCGTGGTCGCGGCCGAGGT-3 '
With plasmid to be measured is template, in the enterprising performing PCR reaction of Biometra type PCR instrument.Reaction conditions is 94 ℃ of sex change 5.0 minutes; 94 ℃ 45 seconds, 68 ℃ 1.0 minutes, 72 ℃ 1.0 minutes, 35 circulations of increasing; 72 ℃ were extended 7.0 minutes.Get 5.0 μ l PCR reaction solutions then and carry out 1.2% agarose gel electrophoresis, electrophoresis was observed recon down in ultraviolet lamp and is inserted clip size after 30 minutes under the 5v/cm constant-pressure conditions.To be inserted with the segmental recon order-checking of purpose, analyze.
(2) banana glutamic acid decarboxylase gene full length sequence clone: gene fragment that obtains according to SSH and the comparison result of NCBI as can be known, this gene fragment 3 ' end has contained the TAA terminator codon.With one 5 of Primer Premier 5 design '-the RACE primer:
5 '-TATGCGAACCCGAAACTCCCAAAG-3 ', it is synthetic to transfer to Shanghai biotechnology company limited.The banana cDNA library that makes up with my research department is a template, and another primer of PCR is the joint primer in cDNA library: ptr5 ': 5 '-CTCCGAGATCTGGACGAGC-3 '; In the 0.2mL centrifuge tube, add successively:
Storehouse, cDNA library liquid 1.0 μ L
10 * Buffer (contains 2.5mM Mg
2+) 2.5 μ L
dNTP(dA/G/C/TTP：10mMeach) 0.5μL
ptr5’(10pM) 1.0μL
BR1695’-RACE(10pM) 1.0μL
Taq polysaccharase (5u/ μ L) 0.3 μ L
ddH
2O 18.7μL
Cumulative volume 25.0 μ L
Carry out the PCR reaction with following PCR reaction conditions:
The result one section 1042bp that increases contains the ATG initiator codon.According to suppress sequence and 5 in the reduction library '-2 primers of sequence nucleotide sequence splicing back design that RACE obtained clone this gene cDNA total length from banana cDNA library.PCR and sequencing result show that this gene cDNA ORF is 1500bp.
(3) fluorescent quantitation research MaGAD1 and fruit are adopted the relation of after ripening:
Extract ethene processing, 1-MCP processing and adopt the template of the total RNA reverse transcription of the fruit cDNA in back normal mature different ripening stages of banana as quantitative fluorescent PCR.
With conserved domain analysis software MaGAD1, the Ma-actin12 gene conservative structure of NCBI, guarantee that the amplified fragments of designed primer is positioned at non-conserved regions; According to the design of primers principle of quantitative fluorescent PCR, design primers then with primer premier 5.
MaGAD1 upstream primer: 5 '-GTCGTCCCCAAGAAGAGCGTCAA-3 ';
MaGAD1 downstream primer: 5 ' ATCGCCAAACCATAACTAAACCACA-3 '.
The Ma-actin1 upstream primer: 5 '-CGAGGCTCAATCAAAGA-3 '
The Ma-actin1 downstream primer: 5 '-ACCAGCAAGGTCCAAAC-3 '
On the Mx3000P of Stratagene instrument, carry out quantitative fluorescent PCR.In the PCR of 0.2mL reaction tubes, add (TAKARA) (TAKARA) each 0.75 μ L of a pair of primer of 0.5 μ L, 5 μ M of 12.5 μ L, Rox reference Dye II (50 *) of SYBR Premix Ex Taq (2 *), cDNA sample 1 μ L, water complements to 25 μ L then.Each sample should be used for the amplifying target genes internal control gene Ma-actin1 that increases again, and three repetitions are all done in the amplification of each gene.According to 94 ℃ of pre-sex change 3min, 94 ℃ of sex change 7s, 55 ℃ of annealing 15s, 72 ℃ are extended 20s, and totally 40 round-robin response procedures increase (response procedures of four genes is consistent), and extend the phase acquisition fluorescent signal in each round-robin.Reaction is done 94 ℃-55 ℃ melt curve analysis analysis after finishing.
Fluorescent quantitative PCR result proves, MaGAD1 expresses and is subjected to that ethene is induced, 1-MCP suppresses, and adopts at banana to play a part the biosynthesizing of regulation and control ethene, fruit maturation in the after ripening process.
(4) MaGAD1 inductor GABA and inhibitor AOAA handle the influence of fruit to ripening stage and quality: the fruit of gathering is cut into single any of several broadleaf plants refer to, carry out surface sterilization 10min with 0.1% clorox, simultaneously the dried flower at fruit top is erased, place and dry an evening, the more consistent banana of picked at random ripening degree is divided into 3 processing: normal mature, GABA handle, AOAA handles.30-40 fruit chosen in each processing, and per 3 fruits are put in the crisper.The fruit of normal mature places under 22 ℃ of constant temperatures ripe.
Observe different treatment banana is adopted the after ripening influence in period, measure banana hardness, solubility total reducing sugar under the different treatment condition.The fruit starch content.The result shows that GABA handles and obviously can shorten the time that reaches each ripening degree, the promotion maturation; AOAA handles obviously can prolong the time that reaches each ripening degree, delays maturation.And adopt physiological change regulating fruit ripening processs such as back banana ripening degree, hardness, solubility total reducing sugar and starch content by regulation and control.
SEQUENCE?LISTING
＜110〉Ministry of Agriculture of China tropic Agriculture Academy Sciences tropic Biotechnology Research Institute tropical crops biotechnology emphasis open laboratory Chinese Academy of Tropical Agricultural Sciences Haikou experiment centre
Gold, will is strong
Slowly, jasper
Recklessly, big
Liu, chrysanthemum China
Open, build refined
The merchant is color red
Tan, the light orchid
＜120〉gene and the coded product and the application of the ripe and quality of regulating fruit
<160>2
<170>PatentIn?version?3.3
<210>1
<211>1500
<212>DNA
＜213〉banana (Musa acuminata)
<220>
<221>gene
<222>(1)..(1500)
<400>1
atgactctct?cggcggtagc?atcggatgcc?gatgattcgg?tcgcttatac?attcgcttcg 60
cgatacgttc?gcgaggctct?tccccggttc?aggataccgg?agcagtcgat?ccccaaggat 120
gcggcgtacc?agatcatcaa?cgacgagctg?atgctcgacg?ggaacccgcg?gttgaatctg 180
gcgtcgttcg?tgacgacgtg?gatggagccg?gagtgcgatc?gcctcatcat?ggcggccgtc 240
aacaagaact?acgtcgacat?ggacgagtac?cccgtcacca?ccgagctcca?gaatcgctgc 300
gtaaatatga?tagcccacct?tttcaatgcc?ccaattgggg?aagacgaaac?ggctgttgga 360
gttggaaotg?tgggttcctc?agaagcaatc?atgcttgcag?gacttgcatt?caagaggaaa 420
tggcagaaca?aaagaaaggc?agaggagaag?ccttacgaca?aacccaacat?tgttaccggt 480
gcaaatgttc?aggtttgctg?ggagaaattt?gcaaggtatt?ttgaagttga?actgaaagaa 540
gtgaagttga?aagagggata?ttatgtaatg?gatcctgcca?aggcagtaga?aatggttgat 600
gagaatacga?tatgtgttgc?tgccatcttg?ggttcaactc?tcactggaga?gtttgaagat 660
gttaagcttc?taaatgatct?cctgacagaa?aaaaaccaag?aaactgggtg?ggacacaccc 720
atacatgtcg?atgctgcaag?tgggggattt?atagcgcctt?tcctctatcc?tgaactagag 780
tgggacttcc?ggctccctca?ggtgaagagc?ataaatgtca?gcggccacaa?atatggcttg 840
gtttatgctg?gagtaggttg?ggttgtctgg?aggaacaaag?aggatttacc?tgaagagctt 900
attttccata?ttaattatct?tggtgcagat?caacctacct?tcactctcaa?cttttctaaa 960
ggatctagtc?agatcattgc?acagtactac?cagtttatac?gcttgggctt?tgagggtttc 1020
agaaatatca?tggaaaactg?tatggacaat?gccaaggtgt?tgaaggcggg?catcgagggg 1080
acagagacat?tcgacattgt?gtccaaggat?gtgggtgtgc?cgctcgtggc?attctcactc 1140
aaggacagcg?gcaagtacac?ggtcttcgac?gtatcggaga?ccttgaggag?gttcggatgg 1200
atcgttcccg?cgtacaccat?gcctgccgac?gcagagcatg?ttgcggtgct?ccgcgtggtg 1260
atcagggagg?acttcagtcg?gagtctggcg?gagcgcctcg?tgacggacat?caagaaggtg 1320
ttgacagact?tggaaaaccg?gtggacgaaa?gccaccatga?tcacccatgt?caaagccgag 1380
gagaacccgg?acggcggcgg?cgccgtcgtc?cccaagaaga?gcgtcaagca?gacgcacgag 1440
gagatcgcca?gatactggaa?gcgattggtg?gatcgtaaga?agaccagcgg?cgtttgctga 1500
<210>2
<211>499
<212>PRT
＜213〉banana (Musa acuminata)
<220>
<221>PRT
<222>(1)..(499)
<400>2
Met?Thr?Leu?Ser?Ala?Val?Ala?Ser?Asp?Ala?Asp?Asp?Ser?Val?Ala?Tyr
1 5 10 15
Thr?Phe?Ala?Ser?Arg?Tyr?Val?Arg?Glu?Ala?Leu?Pro?Arg?Phe?Arg?Ile
20 25 30
Pro?Glu?Gln?Ser?Ile?Pro?Lys?Asp?Ala?Ala?Tyr?Gln?Ile?Ile?Asn?Asp
35 40 45
Glu?Leu?Met?Leu?Asp?Gly?Asn?Pro?Arg?Leu?Asn?Leu?Ala?Ser?Phe?Val
50 55 60
Thr?Thr?Trp?Met?Glu?Pro?Glu?Cys?Asp?Arg?Leu?Ile?Met?Ala?Ala?Val
65 70 75 80
Asn?Lys?Asn?Tyr?Val?Asp?Met?Asp?Glu?Tyr?Pro?Val?Thr?Thr?Glu?Leu
85 90 95
Gln?Asn?Arg?Cys?Val?Asn?Met?Ile?Ala?His?Leu?Phe?Asn?Ala?Pro?Ile
100 105 110
Gly?Glu?Asp?Glu?Thr?Ala?Val?Gly?Val?Gly?Thr?Val?Gly?Ser?Ser?Glu
115 120 125
Ala?Ile?Met?Leu?Ala?Gly?Leu?Ala?Phe?Lys?Arg?Lys?Trp?Gln?Asn?Lys
130 135 140
Arg?Lys?Ala?Glu?Glu?Lys?Pro?Tyr?Asp?Lys?Pro?Asn?Ile?Val?Thr?Gly
145 150 155 160
Ala?Asn?Val?Gln?Val?Cys?Trp?Glu?Lys?Phe?Ala?Arg?Tyr?phe?Glu?Val
165 170 175
Glu?Leu?Lys?Glu?Val?Lys?Leu?Lys?Glu?Gly?Tyr?Tyr?Val?Met?Asp?Pro
180 185 190
Ala?Lys?Ala?Val?Glu?Met?Val?Asp?Glu?Asn?Thr?Ile?Cys?Val?Ala?Ala
195 200 205
Ile?Leu?Gly?Ser?Thr?Leu?Thr?Gly?Glu?Phe?Glu?Asp?Val?Lys?Leu?Leu
210 215 220
Asn?Asp?Leu?Leu?Thr?Glu?Lys?Asn?Gln?Glu?Thr?Gly?Trp?Asp?Thr?Pro
225 230 235 240
Ile?His?Val?Asp?Ala?Ala?Ser?Gly?Gly?Phe?Ile?Ala?Pro?Phe?Leu?Tyr
245 250 255
Pro?Glu?Leu?Glu?Trp?Asp?Phe?Arg?Leu?Pro?Gln?Val?Lys?Ser?Ile?Asn
260 265 270
Val?Ser?Gly?His?Lys?Tyr?Gly?Leu?Val?Tyr?Ala?Gly?Val?Gly?Trp?Val
275 280 285
Val?Trp?Arg?Asn?Lys?Glu?Asp?Leu?Pro?Glu?Glu?Leu?Ile?Phe?His?Ile
290 295 300
Asn?Tyr?Leu?Gly?Ala?Asp?Gln?pro?Thr?Phe?Thr?Leu?Asn?Phe?Ser?Lys
305 310 315 320
Gly?Ser?Ser?Gln?Ile?Ile?Ala?Gln?Tyr?Tyr?Gln?phe?Ile?Arg?Leu?Gly
325 330 335
Phe?Glu?Gly?Phe?Arg?Asn?Ile?Met?Glu?Asn?Cys?Met?Asp?Asn?Ala?Lys
340 345 350
Val?Leu?Lys?Ala?Gly?Ile?Glu?Gly?Thr?Glu?Thr?Phe?Asp?Ile?Val?Ser
355 360 365
Lys?Asp?Val?Gly?Val?Pro?Leu?Val?Ala?Phe?Ser?Leu?Lys?Asp?Ser?Gly
370 375 380
Lys?Tyr?Thr?Val?Phe?Asp?Val?Ser?Glu?Thr?Leu?Arg?Arg?Phe?Gly?Trp
385 390 395 400
Ile?Val?Pro?Ala?Tyr?Thr?Met?Pro?Ala?Asp?Ala?Glu?His?Val?Ala?Val
405 410 415
Leu?Arg?Val?Val?Ile?Arg?Glu?Asp?Phe?Ser?Arg?Ser?Leu?Ala?Glu?Arg
420 425 430
Leu?Val?Thr?Asp?Ile?Lys?Lys?Val?Leu?Thr?Asp?Leu?Glu?Asn?Arg?Trp
435 440 445
Thr?Lys?Ala?Thr?Met?Ile?Thr?His?Val?Lys?Ala?Glu?Glu?Asn?Pro?Asp
450 455 460
Gly?Gly?Gly?Ala?Val?Val?Pro?Lys?Lys?Ser?Val?Lys?Gln?Thr?His?Glu
465 470 475 480
Glu?Ile?Ala?Arg?Tyr?Trp?Lys?Arg?Leu?Val?Asp?Arg?Lys?Lys?Thr?Ser
485 490 495
Gly?Val?Cys
Claims (3)
1. gene and the coded product and the application of the ripe and quality of regulating fruit is characterized in that having sequence table＜sequence under 400〉1.
2. sequence table＜400〉protein that sequence under 1 limits, have sequence table＜400〉2 down amino acid residue sequence or the amino acid residue sequence of sequence passed through replacement, disappearance or the interpolation of one or several amino-acid residue and have and sequence table＜400 amino acid residue sequence under 2 is identical active by sequence deutero-protein.
3. γ-An Jidingsuan (GABA) and AOAA (AOAA) are handled banana and are adopted the back fruit, can induce and suppress the coded genetic expression of sequence in the claim 1, and then influence the variation of hardness, solubility total reducing sugar and the starch content etc. of ripening of fruits, fruit, effectively regulating fruit is adopted after ripening and quality.
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| CN110387430A (en) * | 2018-04-20 | 2019-10-29 | 中国农业大学 | A method of based on apple transcription factor ERF4 the 225th amino acid residue category forecasting Apple hardness |
| CN111793641A (en) * | 2020-07-20 | 2020-10-20 | 中国农业科学院郑州果树研究所 | Application of sweet cherry PavSS or PavSPS gene in regulation and control of fruit coloring or fruit ripening and softening |
| CN114600712A (en) * | 2022-02-28 | 2022-06-10 | 浙江大学 | Method for promoting tomato fruit ripening and improving tomato fruit quality |
| CN114651653A (en) * | 2022-02-21 | 2022-06-24 | 西北农林科技大学 | Amino acid application method for improving water utilization rate and fruit quality of apple dwarf rootstock |
| CN117487821A (en) * | 2023-12-29 | 2024-02-02 | 中国热带农业科学院三亚研究院 | Malil 6 gene for Brazilian banana ethylene signal transduction and application thereof |
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2009
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