CN112458102B

CN112458102B - Peach heat shock transcription factor PpHF 5 and application thereof

Info

Publication number: CN112458102B
Application number: CN202011542666.1A
Authority: CN
Inventors: 谭彬; 冯建灿; 连晓东; 郑先波; 严柳; 栗焕楠; 程钧; 王伟; 王小贝; 叶霞; 李继东; 张郎郎; 李志谦; 张海朋
Original assignee: Henan Agricultural University
Current assignee: Henan Agricultural University
Priority date: 2020-12-21
Filing date: 2020-12-21
Publication date: 2022-03-15
Anticipated expiration: 2040-12-21
Also published as: CN112458102A

Abstract

The invention discloses a peach heat shock transcription factor PpHF 5 and application thereof, and the invention verifies the function of a target gene by cloning PpHF 5 in a peach heat shock transcription factor HSF gene family and finally transferring the gene into arabidopsis thaliana by an agrobacterium-mediated method, thereby being beneficial to explaining a molecular mechanism related to the dwarfing aspect of peach trees from the molecular mechanism and having important significance for dwarfing germplasm cultivation by utilizing a molecular means.

Description

Peach heat shock transcription factor PpHF 5 and application thereof

Technical Field

The invention belongs to the technical field of biology, and particularly relates to a peach heat shock transcription factor PpHSF5 and application thereof.

Background

At present, more and more researches on dwarfing breeding in various species are carried out, and a few achievements are obtained, particularly, the dwarfing breeding of rice in China gradually moves to maturity. There are two mechanisms of dwarfing in rice, one is quality trait inheritance, which is controlled by a single gene, and the other is quantitative trait inheritance, which is controlled by multiple genes. Wheat is one of the most important grain crops in China and the world, and the plant height of wheat belongs to quantitative trait inheritance in heredity, is influenced by various factors, is influenced by environmental effects except genetic background and dwarfing genes. In the study of wheat dwarf stem, the plant height is not only controlled by major gene, but also modified by some modifying genes.

Through high-temperature pretreatment, the plants can obtain certain heat resistance. As an important class of molecular chaperones, the accumulation of Heat Shock Proteins (HSPs) plays a critical role in this process. The expression of heat shock protein coding genes is regulated and controlled by heat shock transcription factors (HSF) at the transcription level, and at present, the research on the heat shock transcription factors is more, and a large number of HSFs are found in animals and plants, which indicates that the HSF genes can play different roles in different stresses or development. Most of the HSF genes are heat-induced, and the expression of some genes is enhanced by calcium chloride stress. Therefore, the study of heat resistance of the plant is mainly related at present, and the plant can activate the expression of heat shock genes under heat stress, belongs to transcription regulation genes and regulates the growth and development of the plant.

In plants, HSF is mainly divided into three groups, namely, A-type HSF, B-type HSF and C-type HSF. A large number of researches show that only the A-type HSF has a transcription activation function and can independently complete the regulation of gene expression, and a large number of B-type and C-type members have no transcription activation function. From the conclusions of these studies, it can be seen that HSF in different HSF classes or different plants has different functions, which is also of interest in studying HSF function in different plants. At present, researches on HSF functions of different plants in different categories are mainly focused on heat resistance of the plants, but researches on the HSF functions and plant dwarfing are rarely reported, and only researches at present show that overexpression of AtHsfB4 (Arabidopsis thaliana type B HSF) genes can influence early root development of the plants, the root surfaces of transgenic plants are rough, cells fall off, but the functions of causing the plant dwarfing are not reported.

Disclosure of Invention

An object of the present invention is to provide peach (Prunus persica L.) heat shock transcription factor PpHF 5 and its encoded protein.

The second purpose of the invention is to provide a recombinant vector of the peach heat shock transcription factor PpHSF 5.

The invention also aims to provide the application of the peach heat shock transcription factor PpHSF5, the protein coded by the peach heat shock transcription factor PpHSF5 and the recombinant vector in plant dwarfing.

In order to achieve the purpose, the invention adopts the following technical scheme:

the invention provides a peach heat shock transcription factor PpHF 5, the sequence of which is shown in SEQ ID NO. 1.

The invention also provides a protein coded by the peach heat shock transcription factor PpHF 5, and the amino acid sequence of the protein is shown as SEQ ID NO. 2.

The invention also provides a recombinant vector containing the peach heat shock transcription factor PpHF 5. Preferably, the recombinant vector is an agrobacterium expression vector.

The invention also provides application of the peach heat shock transcription factor PpHF 5, the protein coded by the peach heat shock transcription factor PpHF 5 and the recombinant vector of the peach heat shock transcription factor PpHF 5 in plant dwarfing, and transgenic plants are obtained by using the genes or the proteins, wherein the plants include but are not limited to Arabidopsis thaliana.

Specifically, the step of obtaining the transgenic arabidopsis thaliana comprises the following steps:

a: constructing a recombinant vector containing the peach heat shock transcription factor PpHF 5;

b: transforming the constructed recombinant vector into an arabidopsis tissue or cell;

c: and culturing and screening to obtain transgenic arabidopsis plants.

Wherein, the SEQ ID NO.1 in the sequence has 1170bp, the amino acid sequence of the codified protein is shown as SEQ ID NO. 2 in the sequence table, and the total 389 amino acids are obtained.

The term "recombinant expression vector" in the present invention refers to a bacterial plasmid, phage, yeast plasmid, plant cell virus, mammalian cell virus or other vectors well known in the art. In general, any plasmid or vector can be used as long as it can replicate and is stable in the host. An important feature of expression vectors is that they generally contain an origin of replication, a promoter, a marker gene and translation control elements.

Vectors comprising the appropriate DNA sequences described above, together with appropriate promoter or control sequences, may be used to transform appropriate host cells to enable expression of the protein. Wherein, the host cell may be a prokaryotic cell, such as a bacterial cell; or lower eukaryotic cells, such as yeast cells; or higher eukaryotic cells, such as plant cells. Representative examples are: escherichia coli, Streptomyces, Agrobacterium; fungal cells such as yeast; plant cells, and the like.

It will be clear to one of ordinary skill in the art how to select appropriate vectors, promoters, enhancers and host cells.

Transformation of a host cell with recombinant DNA can be carried out using conventional techniques well known to those skilled in the art. The transformed plant may be transformed by methods such as Agrobacterium transformation or particle gun transformation, for example, spray method, leaf disk method, rice immature embryo transformation method, etc. The transformed plant cells, tissues or organs can be regenerated into plants by conventional methods.

The invention also provides the application of the gene or the protein in obtaining transgenic plants.

In one embodiment of the present invention, the polynucleotide is cloned into an appropriate vector by a conventional method, and the recombinant vector with the exogenous gene is introduced into a plant cell to make the plant cell express the peach heat shock transcription factor PpHSF 5. Plants overexpressing the polypeptide can be obtained by regenerating the plant cells into plants.

In the present invention, there is no particular limitation on the plant suitable for use in the present invention, as long as it is suitable for carrying out a gene transformation operation, such as various crops, flowering plants, forestry plants, or the like. The plant may be, for example (without limitation): dicotyledonous, monocotyledonous, or gymnosperm. More specifically, the plants include (but are not limited to): wheat, barley, rye, rice, corn, sorghum, sugar beet, apple, pear, plum, peach, apricot, cherry, strawberry, raspberry, blackberry, bean, lentil, pea, soybean, rape, mustard, poppy, olea, sunflower, coconut, castor oil plant, cocoa bean, peanut, gourd, cucumber, watermelon, cotton, flax, hemp, jute, citrus, lemon, grapefruit, spinach, garland, asparagus, cabbage, chinese cabbage, pakchair, carrot, onion, potato, tomato, green pepper, avocado, cinnamon, camphor, tobacco, nut, coffee, eggplant, sugarcane, tea, pepper, grape tree, oyster hemp, banana, natural rubber tree, ornamental plants and the like.

As a preferred mode, the "plant" includes but is not limited to: plants of Solanaceae, Brassicaceae, Rosaceae, and Vitaceae. For example, the term "plant" includes, but is not limited to: tobacco and tomato of the solanaceae family; arabidopsis thaliana of Brassicaceae, apple of Rosaceae, and strawberry; grapes of the family Vitaceae, and the like.

The invention has the following advantages:

1. the invention verifies the function of the target gene by cloning PpHF 5 in the peach heat shock transcription factor HSF gene family and finally transferring the gene into arabidopsis thaliana by an agrobacterium-mediated method, is favorable for explaining a molecular mechanism related to the dwarfing aspect of peach trees from a molecular mechanism, and has important significance for dwarfing germplasm cultivation by utilizing a molecular means.

2. The invention can also separate the complete cDNA of the heat-activated transcription factor PpHF 5 gene from the peach, connect the complete cDNA to the plant expression vector, and transform the plant by using the agrobacterium infection method to obtain the transgenic plant, the root system of which is obviously shortened, the growth is retarded, the plant is smaller, the dwarfing character is shown, and the invention has very important meaning for obtaining the dwarfing plant, therefore, the gene can also be applied to the plant genetic improvement.

Drawings

FIG. 1 is a PCR amplification electrophoretogram of peach heat shock transcription factor PpHF 5;

in the figure, M of a left strip is DL 2000marker, and the size of a gene PpHF 5 gene fragment is 1170 bp.

FIG. 2 is an evolutionary analysis of peach PpHF 5 and Arabidopsis AtHSF;

FIG. 3 is the expression of peach PpHF 5 in different tissues of peach # 14';

FIG. 4 is identification of transgenic Arabidopsis positive plants of peach PpHF 5;

in the figure, A: transgenic plants; p: plasmid control; WT: wild type arabidopsis control; m: DL 2000 Marker; h₂O: blank control;

FIG. 5 is the analysis of the relative expression of the positive plants of transgenic Arabidopsis thaliana of peach PpHSF 5;

FIG. 6 is root phenotype observation and analysis of variance after one week of transgenic Arabidopsis of peach PpHSF 5;

FIG. 7 is a two-week aerial phenotype observation and analysis of variance of transgenic Arabidopsis thaliana from peach PpHSF 5;

FIG. 8 is a three-week aerial phenotype observation and analysis of variance of transgenic Arabidopsis thaliana from peach PpHSF 5;

FIG. 9 is phenotypic observations and analysis of variance of three weeks underground portions of peach PpHSF5 transgenic Arabidopsis;

FIG. 10 is temperature treatment of peach PpHF 5 transgenic Arabidopsis.

Detailed Description

The present invention will be described in detail below with reference to specific examples. These embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art.

In the following description and in the claims, the terms "include" and "comprise" are used in an open-ended fashion, and thus should be interpreted to mean "include, but not limited to. The description which follows is a preferred embodiment of the invention, but is made for the purpose of illustrating the general principles of the invention and not for the purpose of limiting the scope of the invention. The scope of the present invention is defined by the appended claims.

Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art. The test methods in the following examples are conventional methods unless otherwise specified. The reagents and materials used are commercially available, unless otherwise specified.

The 'Zhongshitao No. 14' is a new variety of semi-dwarf nectarine cultivated by hybridization of Quejia Queensis of Zhengzhou fruit tree institute of Chinese academy of agricultural sciences, the variety is already on the market and can be obtained in the market through a purchase way, and the experimental material adopted by the invention is provided by the national peach and grape improvement center of Zhengzhou fruit tree institute of Chinese academy of agricultural sciences.

Example one

1. Isolation of Gene PpHF 5

The total RNA of the leaf of the Chinese nectarine No. 14 is extracted by using a column type plant total RNA extraction and purification Kit (Shanghai Biotechnology), and reverse transcription is carried out by using a FastQuant RT Kit (with gDNase) reverse transcription Kit produced by Tiangen Biochemical technology (Beijing) Co., Ltd to obtain single-stranded cDNA, the single-stranded cDNA is used as a template, the following sequence is used as a primer, PCR amplification is carried out to obtain the full-length sequence of the gene PpHF 5, and a PCR amplification electrophoresis chart is shown in figure 1. The full-length sequence of the gene PpHF 5 is shown as SEQ ID NO.1 in the sequence table and has 1170bp in total, the amino acid sequence of the codified protein is shown as SEQ ID NO. 2 in the sequence table and has 389 amino acids in total. The amino acid sequence alignment result of peach PpHF 5 and Arabidopsis AtHSFB-4 is shown in FIG. 2. The results in FIG. 2 show that PpHF 5, i.e., PpHF 5, in peach has an amino acid sequence similarity of 51.81% (no high homology) with AtHSFB-4.

The primer sequence is as follows:

positive PpHF 5-F: 5'-ATGGCTCTGATGATGGACAATTG-3', respectively;

reverse PpHF 5-R: 5'-CTAACATGCGGAGGGAGGCAT-3' are provided.

The PCR amplification conditions were: reacting at 98 ℃ for 10s, at 55 ℃ for 15s and at 72 ℃ for 40s, and performing 35 cycles; storing at 4 ℃.

2. Expression analysis of Gene PpHF 5

Fluorescent quantitative PCR operation steps: total RNA of stem tip, flower, embryo, tender leaf, mature leaf and fruit of 'Zhonghua peach No. 14' is extracted by a column type plant total RNA extraction and purification Kit (Shanghai Biotechnology), reverse transcription is carried out by a FastQuant RT Kit (with gDNase) reverse transcription Kit produced by Tiangen Biotechnology (Beijing) Co., Ltd to obtain single-chain cDNA, amplification reaction is carried out by a fluorescence quantitative Kit SYBR Select Master Mix (Applied Biosystems, Mardrid, CA, USA), and qRT-PCR Detection is carried out by ABIPRISM 7500FAST Sequence Detection System (Applied Biosystems, Mardrid, CA, USA) fluorescence quantification.

The fluorescent quantitative PCR reaction system was 20. mu.l, including 200ng cDNA (1. mu.L), SYBR Select Master Mix (10. mu.L), 0.5. mu. mol. L^-1Upstream and downstream primers (1. mu.L each) and RNase free water (7. mu.L).

Reaction procedure: pre-denaturation at 95 ℃ for 2 min; denaturation at 95 ℃ for 30s, annealing at 60 ℃ for 30s, and extension at 72 ℃ for 30s for 42 cycles. Each sample was replicated 3 times. Design of quantitative primers Using Primer Premier 5.0 software

The sequence of the quantitative primer is as follows:

positive Q PpHF 5-F: 5'-CTTGGCGGAGGACAATGAGA-3', respectively;

reverse Q PpHF 5-R: 5'-AGAAGCAAAGAAGAAGGGTAGGA-3' are provided.

And detecting the expression condition of the PpHF 5 gene in different tissues of No. 14 nectarine by adopting qRT-PCR. As can be seen from FIG. 3, PpHSF5 is expressed in the shoot tip, young leaf, mature leaf, flower, embryo and fruit of the annual shoot of "Zhongyou No. 14", but the total expression level is low. The expression level was highest in young leaves, followed by shoot tips, and lowest in embryos and mature leaves.

3. Functional identification test of PpHF 5 Gene

To investigate whether the gene PpHSF5 can cause dwarfing of peach trees, its function was analyzed and identified by transgenic Arabidopsis thaliana.

3.1 construction of recombinant vectors

The PpHF 5 gene amplified from the oil 14 of the ` medium oil was ligated to the over-expression vector pSAK-277 using a one-step cloning kit (Vazyme, N.Nanjing Novozam Biotech Co., Ltd.) by overnight ligation at 16 ℃ using T4 ligase from NEB. The primers are as follows:

G-PpHSF5-F：

5'-CCGCTCGAGATGGCTCTGATGATGGACAATTG-3'；

G-PpHSF5-R：5'-TGCTCTAGACTAACATGCGGAGGGAGGCAT-3'。

and detecting positive clones. The positive clones were then sent to Shanghai Biotechnology GmbH for sequencing.

3.2 screening of transgenic Arabidopsis Positive strains

As a high-efficiency mature peach genetic transformation system is not established in peaches, the model plant Arabidopsis is adopted for functional verification of the PpHSF5 gene. The sequenced pSAK-277-PpHF 5 plasmid is extracted and transferred into agrobacterium GV3101 by a liquid nitrogen freeze-thaw method. Adjusting the concentration OD of the agrobacterium liquid₆₀₀When the concentration is up to 0.8, wild arabidopsis thaliana plants are infected by a flower dipping method, and T is harvested₀After seed generation, resistant plants were selected on MS medium containing kanamycin. After the seedlings grow into seedlings, extracting DNA, obtaining positive transgenic plants through PCR detection, and collecting T₁And (5) seed generation. T is₁After sowing seeds, use T₂Transgenic Arabidopsis thaliana was observed to differ from the wild type. The following experiments all utilize T₂Generation and progeny homozygous lines.

PCR molecular identification positive plant primer:

positive PpHF 5-F: 5'-ATGGCTCTGATGATGGACAATTG-3', respectively;

reverse PpHF 5-R: 5'-CTAACATGCGGAGGGAGGCAT-3' are provided.

For confirmation of fragment insertion.

Two transgenic lines of PpHSF5 gene (PpHSF5-1, PpHSF5-2) were obtained as a representative line for comparison with the wild type as shown in FIG. 4.

3.3 analysis of expression levels of wild type Arabidopsis and transgenic Arabidopsis positive strains

Fluorescent quantitative PCR operation steps: total RNA of Arabidopsis thaliana leaves is extracted by adopting a polysaccharide polyphenol RNA extraction Kit SK8662 (Life, Shanghai), reverse transcription is carried out by utilizing a FastQuant RT Kit (with gDNase) reverse transcription Kit produced by Tiangen Biotechnology (Beijing) Co., Ltd to obtain single-chain cDNA, amplification reaction is carried out by utilizing a fluorescence quantitative Kit SYBR Select Master Mix (Applied Biosystems, Mardrid, CA, USA), and qRT-PCR Detection is carried out by utilizing an ABI PRISM 7500FAST Sequence Detection System (Applied Biosystems, Mardrid, CA, USA) fluorescence quantitative instrument.

The sequence of the quantitative primer is as follows:

positive Q PpHF 5-F: 5'-CTTGGCGGAGGACAATGAGA-3', respectively;

reverse Q PpHF 5-R: 5'-AGAAGCAAAGAAGAAGGGTAGGA-3' are provided.

Compared with the wild type, the expression level of two PpHF 5 gene strains (PpHF 5-1 and PpHF 5-2) obtained in the way of figure 5 is 4376.3 times that of the strain PpHF 5-1 and 1698.3 times that of the strain PpHF 5-2.

EXAMPLE Effect of the Secondary Gene PpHSF5 in transgenic Arabidopsis

2.1 Effect of the Gene PpHSF5 on root growth of transgenic Arabidopsis

Seeding T₂The generation transgene and the wild arabidopsis thaliana grow on an MS culture medium, after two cotyledons grow, the arabidopsis thaliana is transplanted into a square culture dish with the length of 10 multiplied by 10cm, and the root length is observed one week after the transplanting. As shown in FIG. 6, T transformed with PpHSF5, as compared with wild type Arabidopsis thaliana₂The root line of the generation transgenic line is shorter, the root length of the PpHF 5-1 line is 43.2 percent of that of the wild type line, and the root length of the PpHF 5-2 line is 49.4 percent of that of the wild type line.

2.2 Effect of the Gene PpHSF5 on the growth of aerial parts of transgenic Arabidopsis

Seeding T₂The generation transgene and the wild arabidopsis thaliana grow on an MS culture medium, and are transplanted into a matrix after two cotyledons grow, and phenotype observation and variance analysis are carried out when the two cotyledons grow to two weeks. The transgenic lines PpHF 5-1 and PpHF 5-2 have rosette leaf length and width significantly lower than those of the wild type line, but there is no significant difference in rosette leaf number, as shown in FIG. 7. When the plants grow to three weeks, compared with wild type arabidopsis thaliana, the transgenic lines PpHF 5-1 and PpHF 5-2 show that the plants are dwarf in whole, the average plant height of PpHF 5-1 is 16.83cm, the average plant height of PpHF 5-2 is 19.37cm, the average plant height of WT is 26.77cm, and the plant height of the transgenic lines is obviously lower than that of WT; meanwhile, the rosette leaf length and leaf width of the transgenic line are obviously lower than those of the wild type plant, but no obvious difference exists in the number of rosette leaves, as shown in figure 8.

2.3 Effect of the Gene PpHSF5 on the growth of the underground portion of transgenic Arabidopsis

Seeding T₂And (3) growing the generation transgenosis and the wild type arabidopsis thaliana on an MS culture medium, transplanting the arabidopsis thaliana into a matrix after two cotyledons grow out, stripping a root system from the matrix after three weeks, photographing for observation, and scanning by using a root system scanner. As shown in FIG. 9, the total root length of both lines of transgenic Arabidopsis was significantly lower than that of wild type Arabidopsis, and the total root length of PpHSF5-1 was 219.34cm, about 54% of wild type lines; the total volume of the two lines of transgenic arabidopsis is also significantly lower than that of wild-type arabidopsis, which is about 20% less than WT; meanwhile, the root tip number and the bifurcation number of the root system of the transgenic arabidopsis are obviously lower than those of wild type strains. The results of root scanning show that transgenic arabidopsis root lines grown under normal conditions are significantly shorter than wild type.

2.4 response of Gene PpHSF5 to temperature

Seeding T₂The generation transgene and the wild type arabidopsis are grown on the same MS culture medium, three times of biological repetition are carried out, the arabidopsis plants at the age of 5 days are placed in an incubator at 46 ℃ for treatment for 30min, then are taken out and placed under normal conditions for culture, and the growth vigor and the germination rate of the arabidopsis are observed and recorded. As shown in FIG. 9, after high temperature treatment for 1d, the germination rate of two transgenic lines is as high as 93.3% or more, while that of WT is only 8.3%; after the high-temperature treatment for 3d, the germination rates of the two transgenic lines reach 100 percent, and the germination rate of WT is improved to 50 percent; after high temperature treatment for 5 days, two transgenic lines grow two cotyledons, while the germination rate of WT is 70%, and two cotyledons with weak growth vigor appear. From FIG. 10, after the seeds of Arabidopsis were treated at high temperature, the germination rate of wild Arabidopsis was significantly lower than that of the transgenic lines, and the transgenic lines showed high temperature resistance.

Although the invention has been described in detail above with reference to a general description and specific examples, it will be apparent to one skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.

Sequence listing

<110> Henan university of agriculture

<120> peach heat shock transcription factor PpHF 5 and application thereof

<130> 2020

<160> 2

<170> SIPOSequenceListing 1.0

<210> 1

<211> 1170

<212> DNA

<213> Prunus persica L.

<400> 1

atggctctga tgatggacaa ttgtgagggc atattgctct ccctcgactc gcacaagtcg 60

gtgccggctc ccttcctgac caaaacgtac cagctcgtgg atgatccggc caccgaccac 120

atcgtctcgt ggggcgaaga cgacgccacc ttcgtcgttt ggcgccctcc cgagttcgcc 180

cgcgacctcc tccccaacta cttcaagcac aacaacttct ccagcttcgt ccgccagctc 240

aacacctacg gttttaggaa gattgtaccg gacagatggg agtttgccaa tgagttcttc 300

aagaaaggag agaagcattt gctctgtgag atccatagaa gaaagacagc tcagcctcat 360

caggtgggtt tcagccacca ccaccaccac caccacaact cgcattcgcc actcggcatc 420

aacggccacc atcatccgag cttcttcccc ttccctagtc gtggcagcat ctccccctcc 480

gactcggacg agccgcccaa ctggtgtgac tcggactcac cacctctccc atcaccaacc 540

ggaggtatta acaatcacaa caacaacaat aataatttta tgagtattaa taatgcgtcg 600

gtgacgggct tggcggagga caatgagagg ctgcggcgga gcaactccat gctgatgtca 660

gagctagccc acatgagaaa actctacaac gacatcatct actttgttca gaatcatgtc 720

aagcctgtgg ctccaagcaa ttcctaccct tcttctttgc ttctctgtaa ccctcctccg 780

aattccatgg ctcctgctgc tactgctact aagcctagta ataatttcaa ccagcttctt 840

gggtactatc cagctcctgc tacaaatgct aagcaaaccc ctcacatgtc tacgacgacc 900

caccatcatg ttatgaactc ttccagcccg agcaacacca cgtccaagag cagctcagtg 960

acgattctgg aagaccagca acaacccagt agcaatgggt gcaaaaatac caagctgttt 1020

ggggtgccgc tgcttcactc gaagaagcgg ttgcacccgg aggagtatgg ctcgaaccat 1080

gggaccagca tgatggaggc cagcaaggct cgtctgattt tggaaaaaga tgacttaggt 1140

ctccatctca tgcctccctc cgcatgttag 1170

<210> 2

<211> 389

<212> PRT

<213> Prunus persica L.

<400> 2

Met Ala Leu Met Met Asp Asn Cys Glu Gly Ile Leu Leu Ser Leu Asp

1 5 10 15

Ser His Lys Ser Val Pro Ala Pro Phe Leu Thr Lys Thr Tyr Gln Leu

20 25 30

Val Asp Asp Pro Ala Thr Asp His Ile Val Ser Trp Gly Glu Asp Asp

35 40 45

Ala Thr Phe Val Val Trp Arg Pro Pro Glu Phe Ala Arg Asp Leu Leu

50 55 60

Pro Asn Tyr Phe Lys His Asn Asn Phe Ser Ser Phe Val Arg Gln Leu

65 70 75 80

Asn Thr Tyr Gly Phe Arg Lys Ile Val Pro Asp Arg Trp Glu Phe Ala

85 90 95

Asn Glu Phe Phe Lys Lys Gly Glu Lys His Leu Leu Cys Glu Ile His

100 105 110

Arg Arg Lys Thr Ala Gln Pro His Gln Val Gly Phe Ser His His His

115 120 125

His His His His Asn Ser His Ser Pro Leu Gly Ile Asn Gly His His

130 135 140

His Pro Ser Phe Phe Pro Phe Pro Ser Arg Gly Ser Ile Ser Pro Ser

145 150 155 160

Asp Ser Asp Glu Pro Pro Asn Trp Cys Asp Ser Asp Ser Pro Pro Leu

165 170 175

Pro Ser Pro Thr Gly Gly Ile Asn Asn His Asn Asn Asn Asn Asn Asn

180 185 190

Phe Met Ser Ile Asn Asn Ala Ser Val Thr Gly Leu Ala Glu Asp Asn

195 200 205

Glu Arg Leu Arg Arg Ser Asn Ser Met Leu Met Ser Glu Leu Ala His

210 215 220

Met Arg Lys Leu Tyr Asn Asp Ile Ile Tyr Phe Val Gln Asn His Val

225 230 235 240

Lys Pro Val Ala Pro Ser Asn Ser Tyr Pro Ser Ser Leu Leu Leu Cys

245 250 255

Asn Pro Pro Pro Asn Ser Met Ala Pro Ala Ala Thr Ala Thr Lys Pro

260 265 270

Ser Asn Asn Phe Asn Gln Leu Leu Gly Tyr Tyr Pro Ala Pro Ala Thr

275 280 285

Asn Ala Lys Gln Thr Pro His Met Ser Thr Thr Thr His His His Val

290 295 300

Met Asn Ser Ser Ser Pro Ser Asn Thr Thr Ser Lys Ser Ser Ser Val

305 310 315 320

Thr Ile Leu Glu Asp Gln Gln Gln Pro Ser Ser Asn Gly Cys Lys Asn

325 330 335

Thr Lys Leu Phe Gly Val Pro Leu Leu His Ser Lys Lys Arg Leu His

340 345 350

Pro Glu Glu Tyr Gly Ser Asn His Gly Thr Ser Met Met Glu Ala Ser

355 360 365

Lys Ala Arg Leu Ile Leu Glu Lys Asp Asp Leu Gly Leu His Leu Met

370 375 380

Pro Pro Ser Ala Cys

385

Claims

1. Peach heat shock transcription factorPpHSF5Use in dwarf plants of a heat shock transcription factor of peachPpHSF5The DNA sequence of (1) is shown in SEQ ID NO.

2. The use of claim 1, wherein the peach heat shock transcription factorPpHSF5The amino acid sequence of the coded protein is shown as SEQ ID NO. 2.

3. Use according to claim 1, wherein the gene or protein is used for obtaining transgenic plants.

4. The use according to claim 1, wherein the plant is peach, arabidopsis.