CN114540379A - Gene participating in synthesis of peach fruit fragrance type aromatic substances and application thereof - Google Patents
Gene participating in synthesis of peach fruit fragrance type aromatic substances and application thereof Download PDFInfo
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Abstract
The invention provides a gene participating in synthesis of peach fruit fragrance type aromatic substances and application thereof. The expression of the gene provided by the invention is gradually increased in the process of peach fruit ripening, and is positively correlated with the content change of fruit-flavor gamma-decalactone, delta-decalactone and hexyl acetate; the expression level of the gene in different peach varieties is positively correlated with the content of lactone and hexyl acetate aromatic substances in fruits; over-expression of the gene in peach fruits can promote accumulation of gamma-decalactone, delta-decalactone and hexyl acetate, and silencing of the gene reduces the content of gamma-decalactone, delta-decalactone and hexyl acetate. The overexpression of the gene can promote the synthesis of the fruity aromatic substance ester and lactone, and provides a target gene for the improvement of fruit quality.
Description
Technical Field
The invention relates to the technical field of plant molecular biotechnology and genetic engineering, and relates to a gene participating in synthesis of a peach fruit fragrance type aromatic substance and application thereof.
Background
Peach (Prunus persica) is a peach genus of Rosaceae family, originally produced in China, rich in germplasm resources, and the cultivation area and yield of peach in China are the top of the world. The aroma can strengthen the sensory memory of the consumers to the fruits and is a key factor influencing the flavor quality of the fruits. Consumers have recently complained of a lighter peach fruit flavor, with loss of aroma being the primary cause.
Esters such as acetic acid hexyl ester, acetic acid-E-2-hexenyl ester and acetic acid-Z-3-hexenyl ester, and lactones such as gamma-decalactone and delta-decalactone are the material bases for forming the fruity fragrance quality of mature peach fruits. Sensory evaluation analysis shows that the content of ester and lactone aromatic substances is positively correlated with the preference of consumers and gradually accumulates along with the ripening of peach fruits. Alcohol Acyltransferase (AAT) involved in the synthesis of ester aromatic substances from peach fruits has been identified and located at the end of the synthetic pathway. In addition to being involved in ester synthesis, peach PpAAT1 also catalyzes the attachment of the hydroxyl group of 4-hydroxydecanoyl CoA to the acyl group to form γ -decalactone. In addition, the portal enzyme PpACX1 (acyl CoA oxidase) of the fatty acid beta-oxidation pathway is also involved in the synthesis of peach fruit lactones.
The esters and lactones take fatty acid as precursor substances, and the synthesis of fruity aromatic substances such as peach fruit esters and lactones can be influenced by regulating and controlling the component content of the fatty acid. Genes involved in fatty acid synthesis and transformation include fatty acid desaturase FAD, fatty acid lyase LIP, and the like. Currently, LIP is found in tomato fruits to be involved in the synthesis of aldehydes and alcohols aromatics, and the fatty acid desaturase FaFAD1, which controls the synthesis of gamma-decalactone, is identified in strawberry fruits. However, the genes that control the synthesis of ester and lactone aromatic substances by affecting the fatty acid content of precursor substances have not been identified. Considering the important role of ester and lactone aromatic substances in the formation of aromatic quality of peach fruits and other fruits during the ripening period, the identification of genes capable of simultaneously regulating and controlling the synthesis of the aromatic substances has important application value in the improvement of fruit quality.
Disclosure of Invention
The invention aims to provide a gene participating in synthesis of peach fruit fragrance type aromatic substances, which is a gene for simultaneously regulating and controlling synthesis of peach fruit ester and lactone aromatic substances. In order to achieve the above object, the present invention mainly provides the following technical solutions:
the nucleotide sequence of the gene participating in the synthesis of the peach fruit fragrance type aromatic substance provided by the invention is shown in SEQ ID NO. 1.
The amino acid sequence of the protein coded by the gene provided by the invention is shown in SEQ ID NO. 2.
The invention also provides a recombinant vector TRV2-PpFAD2 containing the gene in the scheme.
The invention also provides a recombinant microorganism containing the recombinant vector of the scheme.
The invention also aims to provide application of the gene in promoting or inhibiting synthesis of peach fruit lactone aromatic substances, in particular application in promoting synthesis of peach fruit gamma-decalactone and delta-decalactone aromatic substances.
The invention further aims to provide application of the gene in promoting or inhibiting synthesis of peach fruit ester aromatic substances, in particular application in promoting synthesis of peach fruit hexyl acetate aromatic substances.
The 4 th purpose of the invention is to provide the application of the gene in peach variety breeding and aroma quality improvement.
The invention has the beneficial effects that:
the expression of the gene provided by the invention is gradually increased in the process of peach fruit ripening, and is positively correlated with the content change of fruit-flavor gamma-decalactone, delta-decalactone and hexyl acetate; the expression level of the gene in different peach varieties is positively correlated with the content of lactone and hexyl acetate aromatic substances in fruits; over-expression of the gene in peach fruits can promote accumulation of gamma-decalactone, delta-decalactone and hexyl acetate, and silencing of the gene reduces the content of gamma-decalactone, delta-decalactone and hexyl acetate. The overexpression of the gene can promote the synthesis of the fruity aromatic substance ester and lactone, and provides a target gene for the improvement of fruit quality.
Drawings
FIG. 1: the expression of PpFAD2 in the peach fruit development process is positively correlated with the content of lactone and ester.
FIG. 2: the contents of lactone and ester in peach fruits and the expression of PpFAD2 are regulated and controlled by treating ethylene and 1-MCP.
FIG. 3: the correlation between the lactone and ester contents of different varieties of peach fruits and the PpFAD2 gene expression.
FIG. 4: transient over-expression of PpFAD2 in peach fruits promotes synthesis of lactones and ester aromatics.
FIG. 5: VIGS silencing PpFAD2 in peach fruits inhibits synthesis of lactone and ester aromatic substances.
Detailed Description
The gene PpFAD2 involved in the synthesis of peach fruit fruity lactone and ester aromatic substance and the application thereof provided by the present invention will be described in detail with reference to the following specific examples and the accompanying drawings, but the examples do not limit the scope of the present invention.
Example 1: PpFAD2 gene expression and lactone content positive correlation in peach fruit development process
(I) Experimental method
1. Peach fruit material
'Hujing honeydew' (Prunus persica L. Batsch cv. Hujingmiu) fruits collected from 5 different growth and development stages of Fenghua City peach institute in Zhejiang province: 34 days after flowering (young fruit period), 71 days after flowering (expansion period), 94 days after flowering (color change period), 108 days after flowering (mature period), 111 days after flowering (mature fruit is placed at 20 ℃ for 3 days after harvest), and 114 days after flowering (mature fruit is placed at 20 ℃ for 6 days after harvest). Sample collection when transported to the laboratory in the day, fruits with consistent size and maturity and free from plant diseases, insect pests and mechanical injuries are selected, 3 biological replicates are set at each sampling time point, and 5 fruits are replicated at each time point. The pulp tissue was frozen in liquid nitrogen and stored at-80 ℃.
2. Analysis of lactone and ester aromatic substance content
Grinding peach pulp sample with liquid nitrogen, weighing 5g, adding 3mL 200mM EDTA solution 3mL 20% CaCl2The solution and 20. mu.L of internal standard 2-octanol (0.8mg/mL) were sealed and mixed uniformly, and after 30min of constant temperature equilibration, 65 μm polydimethylsiloxane and a divinylbenzene (PDMS-DVB) extraction head (Supelco Co.) were used for 30min of solid phase microextraction. The extraction head is injected in a GC-MS (Agilent 7890-5975)Desorbing in the mouth for 5min, and purifying with DB-WAX capillary chromatography column (0.25mm, 30m, 0.25 μm, J)&W Scientific) were isolated. The temperature raising program is from 40 ℃ to 3 ℃ per min-1At a rate of 100 ℃ and then at 5 ℃ min-1The rate increased to 245 ℃. At a rate of 1.0 mL/min-1Helium is taken as carrier gas, the MS ion source temperature is 230 ℃, an electron bombardment ionization mode is adopted, the electron energy is 70eV, and the mass spectrum scanning range is 35-350 m/z. The mass spectrum library NIST-8(NIST/EPA/NIH) and Retention Index (RI) are adopted for substance identification, and the concentration of the substance is calculated by an internal standard area normalization method.
RNA extraction and transcriptome analysis
Freezing 1g of the ground sample by liquid nitrogen, adding 4mL of CTAB/beta-mercaptoethanol extract preheated at 65 ℃, carrying out vortex mixing, and carrying out water bath at 65 ℃ for 5 min; 4mL of chloroform was added: extracting isoamyl alcohol (24: 1) and fully mixing in a vortex manner; centrifuging at 15 ℃ and 10000rpm for 10min, sucking the supernatant into a new centrifugal tube, and re-extracting once; sucking the supernatant into a new centrifuge tube, adding 1/4 volumes of 10mol/L LiCl, and standing at 4 ℃ for 12 h; centrifuging at 4 deg.C and 10000rpm for 20min, removing supernatant, adding 400 μ L SSTE preheated at 65 deg.C into the precipitate to dissolve the precipitate; then 400 μ L chloroform was added: extracting isoamyl alcohol (24: 1) and mixing in a vortex mode; transferring to a 1.5mL centrifuge tube, centrifuging at 20 ℃ and 10000rpm for 10min, sucking supernatant into a new centrifuge tube, adding 2 times volume of-20 ℃ precooled absolute ethyl alcohol, turning upside down, mixing uniformly, and standing at-80 ℃ for 30 min; centrifuging at 4 deg.C and 10000rpm for 25min, removing supernatant, centrifuging for a short time, sucking out residual liquid, and air drying the precipitate in a fume hood; the precipitate was dissolved with 20. mu.L of DEPC water to obtain a total RNA sample. Total RNA residual genomic DNA was removed using TURBO DNase (Ambion) kit. Gel electrophoresis was performed to check RNA integrity. Baimaike's company was entrusted to perform RNA-seq transcriptome sequencing with the test platform HiSeq-2000.
(II) results of the experiment
Gamma-decalactone and delta-decalactone are main lactone aroma substances in the peach fruits of the 'lake Jing nectar', the content of the main lactone aroma substances is not detected in the fruits before the color conversion period (94 days), and the main lactone aroma substances are gradually accumulated after the color conversion and in the fruit after-ripening process; hexyl acetate is an ester aromatic substance synthesized by taking linoleic acid as a substrate in peach fruits, the content of the hexyl acetate also approximately increases along with the ripening of the fruits, and the hexyl acetate has a content peak in the color conversion period. The expression level of the PpFAD2 gene is lower in the prophase of peach fruit development, and rapidly increases after a color transition period, and is obviously and positively correlated with the content change of aroma substances of gamma-decalactone (R ═ 0.90) and delta-decalactone (R ═ 0.93), and also positively correlated with the content change of hexyl acetate (R ═ 0.67) (figure 1).
Example 2: ethylene and 1-MCP treatment influence peach fruit lactone content and PpFAD2 expression
(I) Experimental method
1. Experimental Material
The fruits of 'lake Jing Honey dew' (94DAB) before the color-changing period were divided into 3 groups. Ethylene treatment group: sealing treatment is carried out on 100 mu L/L of ethylene at room temperature for 12 h; 1-MCP treatment group: sealing and processing the 5 mu L/L ethylene signal transduction inhibitor 1-MCP for 12h at room temperature; control group: the fruits were not treated and sealed at room temperature for 12 h. The treated fruits are placed at the room temperature of 25 ℃ and stored for 24h, 60h and 96h, then frozen and sampled by liquid nitrogen respectively, and stored at the temperature of-80 ℃ for subsequent analysis. Each set of experiments set 3 biological replicates, each replicate 5 fruits.
2. Analysis of lactone and ester aromatic substance content
The contents of lactone and ester-type aromatic substances were analyzed in accordance with example 1. The results of the experiment are shown in FIG. 2.
RN A extraction and transcriptome sequencing analysis
Extraction of total RNA by CTAB method: refer to example 1. Baimaike corporation was entrusted with RNA-seq transcriptome sequencing with the test platform HiSeq-2000. The results of the experiment are shown in FIG. 2.
(II) results of the experiment
Compared with a control fruit, the ethylene treatment can remarkably induce the synthesis of gamma-decalactone, delta-decalactone and hexyl acetate, and the 1-MCP can remarkably inhibit the synthesis of the gamma-decalactone, the delta-decalactone and the hexyl acetate; gene expression analysis showed that ethylene treatment significantly induced the expression of PpFAD2, while 1-MCP treatment significantly inhibited the expression of PpFAD2 gene (fig. 2). The results show that the expression of PpFAD2 and the synthesis of gamma-decalactone, delta-decalactone and hexyl acetate are all regulated and controlled by ethylene.
Example 3: expression analysis of lactone and PpFAD2 of different varieties of peach fruits
(I) Experimental method
1. Experimental Material
The method comprises the steps of collecting 127 different varieties of peach fruits from a peach resource garden of an agricultural academy of agricultural sciences in Jiangsu province, wherein 3 biological replicates are arranged for each variety, and 5 fruits are replicated for each variety. The pulp tissue is frozen by liquid nitrogen and sampled, and preserved at-80 ℃ for subsequent analysis of aroma substances and gene expression.
2. Analysis of lactone and ester aromatic substance content
The contents of lactone and ester-type aromatic substances were analyzed in accordance with example 1. The results of the experiment are shown in FIG. 3.
RNA extraction and transcriptome sequencing analysis
Extraction of total RNA by CTAB method: refer to example 1. Baimaike corporation was entrusted with RNA-seq transcriptome sequencing with the test platform HiSeq-2000. The results of the experiment are shown in FIG. 3.
(II) results of the experiment
The content of lactones and ester aromatic substances in different varieties of peach fruits is greatly different, and 127 parts of peach fruits are divided into the following parts according to the difference of the total amount of gamma-decalactone and delta-decalactone: low lactone (<100ng/g), medium lactone (>100ng/g and <400ng/g) and high lactone (>400ng/g)3 groups. Analyzing the expression of the PpFAD2 gene in the fruits of the 3 groups, wherein the result shows that the average expression level of the PpFAD2 gene in the peach fruits of the low lactone group is lower, and the average expression level of the PpFAD2 gene in the peach fruits of the high lactone group is higher; peach fruit varieties are divided into 3 groups according to the content of hexyl acetate: low ester (<30ng/g), medium ester (>30ng/g and <100ng/g) and high ester (>100 ng/g). The PpFAD2 gene expression in the 3 peach groups also showed a trend from low to high. Analysis on the correlation between the gene expression level of PpFAD2 and the substance content shows that the gene expression level of PpFAD2 is in positive correlation with the lactone content (R is 0.61, P is less than 0.01) and the hexyl acetate content (R is 0.46, P is less than 0.01) (FIG. 3). The results show that the expression of the PpFAD2 gene can influence the content of lactone and ester aromatic substances in different peach fruits.
Example 4: peach fruit transient overexpression PpFAD2 promotes synthesis of peach fruit lactone and ester aromatic substances
(I) Experimental method
cDNA Synthesis and PpFAD2 Gene expression analysis
The total RNA of peach fruit was 1.0. mu.g, and after removing genomic DNA using TAKARA kit, cDNA was synthesized by reverse transcription according to the protocol. PpFAD2 gene expression is analyzed by QPCR, peach PpTEF2 is used as an internal reference gene, and PpFAD2 primer sequences are SEQ ID NO.3 and SEQ ID NO. 4. The QPCR reaction system included 10. mu.L of Ssofast EvaGreen Supermix (Bio-Rad), 1. mu.L of each of the upstream and downstream primers (10. mu.M), 2. mu.L of cDNA, and 6. mu. L H2O. The PCR procedure was: 3min at 95 ℃; 10s at 95 ℃, 30s at 60 ℃ and 45 cycles; 10s at 95 ℃; the fluorescence signal was read every 0.5 ℃ rise from 65 ℃ to 95 ℃. The used instrument is a Bio-Rad CFX96 real-time fluorescence quantitative PCR instrument, and QPCR primer specificity is verified by melting point curve analysis and QPCR product sequencing. The results of the experiment are shown in FIG. 4.
Construction of SK-PpFAD2 recombinant vector and transformation of Agrobacterium
And amplifying the PpFAD2 gene reference sequence in the peach genome database by using cDNA as a template and using primer pairs SEQ ID NO.5 and SEQ ID NO.6 to obtain the full-length SEQ ID NO.1 of PpFAD 2. The PCR reaction system is 50 mu L, and the components are respectively as follows: mu.L of PrimeSTAR Max Premix (2X) enzyme (TAKARA), 2. mu.L of each of upstream and downstream primers (10. mu.M), 1. mu.L of peach fruit cDNA, 20. mu. L H2And O. And connecting the PCR product with a pGreen II 002962-SK vector cut by restriction enzymes BamH I and Sal I by using a homologous recombination method. The ligation product was added to 20ul DH5 alpha to be competent, placed on ice for 30min, heat shocked at 42 ℃ for 90s, transformed into DH5 alpha, and positive colonies were picked for sequencing verification.
3. The SK-PpFAD2 vector, pTRV1 vector and TRV2-PpFAD2 vector with correct sequences are respectively transferred into agrobacterium GV 3101:pSomu p by an electric shock transformation method, and positive clones are respectively picked and stored. Agrobacterium was transformed with unloaded SK and TRV2 in the same manner and used as a negative control.
Peach fruit homologous transient overexpression
Culturing Agrobacterium GV3101 containing SK-PpFAD2 vector on solid culture medium containing Kan (50mg/L) and Get (25mg/L) at 28 deg.C for 2d, selecting single clone strain in 5mL LB containing Kan and Get, culturing overnight, transferring to 500mL LB (Kan 50mg/L + Get 25mg/L), culturingTo OD600 of 0.8 to 1.0. Centrifuging at 4 deg.C and 5000g for 10min to collect bacteria. With equal volume of MES permeate (10mM MES, 10mM MgCl)2150mM acetosyringone, 0.04% TritonRX-100, pH 5.6), and standing at room temperature for 2 h. Agrobacterium containing the empty SK vector was prepared in the same manner as a control.
Taking peach fruit after color transition period, injecting bacterial liquid containing target gene while injecting control with the belly line as boundary. 3 wells were injected per side, 300 μ L of bacterial suspension per well. Each fruit is a repeat, and 20 repeats are respectively arranged in overexpression and gene silencing groups. And (3) placing the injected fruits in an incubator at 25 ℃, taking a pulp sample with the diameter of 1cm near an injection hole after 8-10 days, and freezing the pulp sample by using liquid nitrogen for subsequent gene expression and lactone content determination. The results of the experiment are shown in FIG. 4.
(II) results of the experiment
After the PpFAD2 gene was transiently overexpressed in peach fruits, the expression level of the PpFAD2 gene was increased to 2.5 times that of the control, while the content of γ -decalactone in the fruits was increased to 3.6 times that of the control fruits, the content of δ -decalactone was increased to 2.6 times that of the control fruits, and the content of hexyl acetate was increased to 3.8 times that of the control fruits (fig. 4). The results show that the overexpression of the PpFAD2 gene in the peach fruits can promote the synthesis of lactones and ester aromatic substances.
Example 5: peach fruit VIGS (Virus-mediated Gene silencing) PpFAD2 inhibits lactone and ester synthesis
(I) Experimental method
cDNA Synthesis and PpFAD2 Gene expression analysis
The method for synthesizing peach fruit cDNA and the method for analyzing PpFAD2 gene expression were as in example 4.
Construction of VIGS recombinant vector and Agrobacterium transformation
The cDNA is taken as a template, and a primer pair SEQ ID NO.7 and SEQ ID NO.8 are utilized to amplify to obtain a sequence of 400bp of the CDs region of PpFAD2, which is shown as SEQ ID NO. 9. The PCR product was ligated with pTRV2 vector digested with restriction enzymes Xho I and BamH I by homologous recombination. The ligation product was added to 20. mu.L of DH 5. alpha. to be competent, incubated on ice for 30min, heat-shocked at 42 ℃ for 90s, transformed into DH 5. alpha. and positive colonies were picked for sequencing verification.
The pTRV2-PpFAD2 vector and the helper virus vector pTRV1 with the sequences confirmed to be correct are respectively transferred into Agrobacterium GV3101:: pSo μ p by an electric shock transformation method, and positive clones are respectively picked and stored. Agrobacterium was transformed with the empty pTRV2 in the same manner and used as a negative control.
1. Peach fruit homologous transient VIGS gene silencing
Agrobacterium harboring pTRV1, unloaded pTRV2, pTRV2-PpFAD2 plasmids were streaked on 3-Resistant (RGK) LB medium (25mg/L Rif +25mg/L Get +50mg/L Kan), respectively, and cultured at 28 ℃ for 2-3 d. Single colonies were picked and shaken with LB liquid medium containing 3 antibody (10mg/L Rif +25mg/L Gen +50mg/L Kan) at 200r/min for 16-18h at 28 ℃. Adding the bacterial liquid into an induction culture medium (containing 10mg/L Rif, 25mg/L Get, 50mg/L Kan and 200 mu M acetosyringone) according to a ratio of 1:25, and shaking at 28 ℃ for 16-24h to enable the OD600 value of the bacterial liquid to be 0.5-0.8. The cells were collected, centrifuged at 5000r/min for 10min and resuspended in the same volume of MES suspension. The cells were again centrifuged and collected and resuspended in 1/2 volumes of MES permeate. Acetosyringone (final concentration 400. mu.M) was added to the suspension containing pTRV1 plasmid and left at room temperature for 3-5 h. The pTRV 1-containing bacterial suspension was mixed with pTRV2-PpFAD2 and pTRV2 (as negative control) in equal volumes, and used for peach fruit injection. Peach fruit injection infection method referring to example 4, the experimental results are shown in fig. 5.
(II) results of the experiment
PpFAD2 gene in peach fruit is silenced by VIGS, the expression level of the PpFAD2 gene is reduced to 1/2 of a control fruit, and simultaneously compared with the control fruit, the content of gamma-decalactone, the content of delta-decalactone and the content of hexyl acetate in the fruit with gene silencing are reduced by 3.6 times, 1 time and 1.5 times respectively (figure 5). The results prove that the silencing of PpFAD2 gene expression can inhibit the synthesis of lactone and ester of peach fruit.
Sequence listing
<110> Zhejiang university
<120> a gene participating in synthesis of peach fruit fragrance type aromatic substances and application thereof
<160> 9
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1149
<212> DNA
<213> peach (Prunus persica)
<400> 1
atgggtgccg gtggaagaat gtccgcaccc cctacccgta agaacgctga aaccgacaac 60
cccaagcgag tgccatactc aaaacctcca ttcagtctcg gccagatcaa gaaagccatc 120
ccacctcatt gctttcagcg ctctgttatc cgctccttct cctatgtctt ttatgacctt 180
accattgcct ttctcctgta ctacattgct tgcaattaca tccagcctct ctctcaacct 240
ttctctttct tggcgtggcc agtttactgg tatgttcagg gctgtgtcct gactggtgtt 300
tgggtgatag cgcatgagtg tggtcaccat gcttttagtg attatcaatg gctggatgac 360
acagtcggtc taatcctcca ctctgccctc cttgtgcctt acttttcttg gaagtatagc 420
catcgccgtc accactctaa cacaggttcc attgagcgag atgaggtctt tgtccccaag 480
cagaagtctg ctattggatg gtactccaaa tatctaaaca acccaccggg caggttcctt 540
acactcacca tccaactcac tctaggctgg cctctgtatc tggctttcaa tgtttcaggg 600
aggcactatg aaggctttgc ttgccactac catccatatg gccccatctt ctccgatcgt 660
gaacgattgc agattttcct ttctgatgcc ggtgttcttg cagtcgtata tgggctatac 720
cgtctagctg tggcgaaagg gcttgcttgg gttgtatgct attatggagg acctttgatg 780
gtggtgaatg gatttttggt gctgatcaca tacttgcagc acacacaccc ttcattgcca 840
cactacgatt cctcagaatg ggactggttg agaggagctt tggcaacagt tgacagagat 900
tacggcatcc tgaacaaggt tttccataac atcacagaca ctcatgttgc tcaccatttg 960
ttctcaacca tgccacatta ccatgcaatg gaggccacca aagctatcaa gccgattttg 1020
ggcgattact atcagttgga caggacgccg gttttcaagg caatgttcag ggaggcaaag 1080
gagtgtattt atgttgagcg agatgagggt gacaagaaag gtgtcttctg gtacaataat 1140
aagttataa 1149
<210> 2
<211> 382
<212> PRT
<213> peach (Prunus persica)
<400> 2
Met Gly Ala Gly Gly Arg Met Ser Ala Pro Pro Thr Arg Lys Asn Ala
1 5 10 15
Glu Thr Asp Asn Pro Lys Arg Val Pro Tyr Ser Lys Pro Pro Phe Ser
20 25 30
Leu Gly Gln Ile Lys Lys Ala Ile Pro Pro His Cys Phe Gln Arg Ser
35 40 45
Val Ile Arg Ser Phe Ser Tyr Val Phe Tyr Asp Leu Thr Ile Ala Phe
50 55 60
Leu Leu Tyr Tyr Ile Ala Cys Asn Tyr Ile Gln Pro Leu Ser Gln Pro
65 70 75 80
Phe Ser Phe Leu Ala Trp Pro Val Tyr Trp Tyr Val Gln Gly Cys Val
85 90 95
Leu Thr Gly Val Trp Val Ile Ala His Glu Cys Gly His His Ala Phe
100 105 110
Ser Asp Tyr Gln Trp Leu Asp Asp Thr Val Gly Leu Ile Leu His Ser
115 120 125
Ala Leu Leu Val Pro Tyr Phe Ser Trp Lys Tyr Ser His Arg Arg His
130 135 140
His Ser Asn Thr Gly Ser Ile Glu Arg Asp Glu Val Phe Val Pro Lys
145 150 155 160
Gln Lys Ser Ala Ile Gly Trp Tyr Ser Lys Tyr Leu Asn Asn Pro Pro
165 170 175
Gly Arg Phe Leu Thr Leu Thr Ile Gln Leu Thr Leu Gly Trp Pro Leu
180 185 190
Tyr Leu Ala Phe Asn Val Ser Gly Arg His Tyr Glu Gly Phe Ala Cys
195 200 205
His Tyr His Pro Tyr Gly Pro Ile Phe Ser Asp Arg Glu Arg Leu Gln
210 215 220
Ile Phe Leu Ser Asp Ala Gly Val Leu Ala Val Val Tyr Gly Leu Tyr
225 230 235 240
Arg Leu Ala Val Ala Lys Gly Leu Ala Trp Val Val Cys Tyr Tyr Gly
245 250 255
Gly Pro Leu Met Val Val Asn Gly Phe Leu Val Leu Ile Thr Tyr Leu
260 265 270
Gln His Thr His Pro Ser Leu Pro His Tyr Asp Ser Ser Glu Trp Asp
275 280 285
Trp Leu Arg Gly Ala Leu Ala Thr Val Asp Arg Asp Tyr Gly Ile Leu
290 295 300
Asn Lys Val Phe His Asn Ile Thr Asp Thr His Val Ala His His Leu
305 310 315 320
Phe Ser Thr Met Pro His Tyr His Ala Met Glu Ala Thr Lys Ala Ile
325 330 335
Lys Pro Ile Leu Gly Asp Tyr Tyr Gln Leu Asp Arg Thr Pro Val Phe
340 345 350
Lys Ala Met Phe Arg Glu Ala Lys Glu Cys Ile Tyr Val Glu Arg Asp
355 360 365
Glu Gly Asp Lys Lys Gly Val Phe Trp Tyr Asn Asn Lys Leu
370 375 380
<210> 3
<211> 20
<212> DNA
<213> Artificial sequence (Unknown)
<400> 3
cggttttcaa ggcaatgttc 20
<210> 4
<211> 20
<212> DNA
<213> Artificial sequence (Unknown)
<400> 4
<210> 5
<211> 36
<212> DNA
<213> Artificial sequence (Unknown)
<400> 5
agaactagtg gatccatggg tgccggtgga agaatg 36
<210> 6
<211> 37
<212> DNA
<213> Artificial sequence (Unknown)
<400> 6
cccctcgagg tcgacttata acttattatt gtaccag 37
<210> 7
<211> 35
<212> DNA
<213> Artificial sequence (Unknown)
<400> 7
tgcccgggcc tcgagggtac tccaaatatc taaac 35
<210> 8
<211> 35
<212> DNA
<213> Artificial sequence (Unknown)
<400> 8
cctccatggg gatcctctct gtcaactgtt gccaa 35
<210> 9
<211> 400
<212> DNA
<213> peach (Prunus persica)
<400> 9
ggtactccaa atatctaaac aacccaccgg gcaggttcct tacactcacc atccaactca 60
ctctaggctg gcctctgtat ctggctttca atgtttcagg gaggcactat gaaggctttg 120
cttgccacta ccatccatat ggccccatct tctccgatcg tgaacgattg cagattttcc 180
tttctgatgc cggtgttctt gcagtcgtat atgggctata ccgtctagct gtggcgaaag 240
ggcttgcttg ggttgtatgc tattatggag gacctttgat ggtggtgaat ggatttttgg 300
tgctgatcac atacttgcag cacacacacc cttcattgcc acactacgat tcctcagaat 360
gggactggtt gagaggagct ttggcaacag ttgacagaga 400
Claims (7)
1. A gene participating in the synthesis of peach fruit fragrance type aromatic substances is characterized in that the nucleotide sequence of the gene is shown as SEQ ID NO. 1.
2. The gene of claim 1, wherein the amino acid sequence of the protein encoded by the gene is shown in SEQ ID No. 2.
3. A recombinant vector TRV2-PpFAD2 containing the gene of claim 1.
4. A recombinant microorganism comprising the recombinant vector of claim 3.
5. The application of the gene of claim 1 in promoting or inhibiting the synthesis of peach fruit lactone aromatic substances.
6. The application of the gene of claim 1 in promoting or inhibiting the synthesis of ester aromatic substances of peach fruits.
7. The application of the gene of claim 1 in peach variety breeding and aroma quality improvement.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210246733.8A CN114540379B (en) | 2022-03-14 | 2022-03-14 | Gene participating in synthesis of peach fruit fragrance type aromatic substances and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210246733.8A CN114540379B (en) | 2022-03-14 | 2022-03-14 | Gene participating in synthesis of peach fruit fragrance type aromatic substances and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114540379A true CN114540379A (en) | 2022-05-27 |
| CN114540379B CN114540379B (en) | 2023-07-21 |
Family
ID=81664429
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210246733.8A Active CN114540379B (en) | 2022-03-14 | 2022-03-14 | Gene participating in synthesis of peach fruit fragrance type aromatic substances and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114540379B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105400805A (en) * | 2015-12-24 | 2016-03-16 | 浙江大学 | Gene participating in peach fruit fatty acid forming and application thereof |
| CN109913469A (en) * | 2019-03-25 | 2019-06-21 | 浙江大学 | It is a kind of participate in peach esters aromatic substance synthesis regulation transcription factor PpNAC1 and its application |
-
2022
- 2022-03-14 CN CN202210246733.8A patent/CN114540379B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105400805A (en) * | 2015-12-24 | 2016-03-16 | 浙江大学 | Gene participating in peach fruit fatty acid forming and application thereof |
| CN109913469A (en) * | 2019-03-25 | 2019-06-21 | 浙江大学 | It is a kind of participate in peach esters aromatic substance synthesis regulation transcription factor PpNAC1 and its application |
Non-Patent Citations (1)
| Title |
|---|
| PRUNUS等: "PREDICTED: Prunus persica delta(12)-fatty-acid desaturase FAD2 (LOC18769718),mRNA" * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114540379B (en) | 2023-07-21 |
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