CN114957417A - Protein related to pollen development and application of coding gene thereof - Google Patents
Protein related to pollen development and application of coding gene thereof Download PDFInfo
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- CN114957417A CN114957417A CN202110195290.XA CN202110195290A CN114957417A CN 114957417 A CN114957417 A CN 114957417A CN 202110195290 A CN202110195290 A CN 202110195290A CN 114957417 A CN114957417 A CN 114957417A
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Abstract
The invention discloses a protein related to pollen development and application of a coding gene thereof. The invention provides application of a SlPP2C5 protein in regulation and control of plant pollen formation. The invention also provides application of the nucleic acid molecule for encoding the SlPP2C5 protein in preparation of transgenic plants with increased pollen aberration rate. The invention also provides application of the nucleic acid molecule for encoding the SlPP2C5 protein in preparation of male-sterile transgenic plants. The inventor of the invention finds that the overexpression of the SlPP2C5 gene in tomatoes can cause abnormal tomato flower development and increase pollen aberration rate. Therefore, the SlPP2C5 protein is related to the tomato flower development process and pollen development. The SlPP2C5 protein and the coding gene thereof play an important role in high-quality breeding of tomatoes.
Description
Technical Field
The invention belongs to the field of plant genetic engineering, and particularly relates to a protein related to pollen development and application of a coding gene thereof.
Background
The growth and development of higher plants are divided into two stages of vegetative growth and reproductive growth, when a specific period is reached, the plants are switched from vegetative growth to reproductive growth, and the process of normal flowering is an important node for reproductive growth of the plants, so that flowering is considered to be a central link in the process of plant ontogeny. The flower organ of the plant plays an important role in the aspects of people appreciation, diet, fruit and vegetable, crop high yield and the like, so that the research on the plant flowering process and the regulation and control mechanism of flower development has a good significance in theory and practical application.
Flower development and flowering of plants are a complex and highly programmed process, which is subject to the co-action of both external and internal genetic mechanisms, in which various plant hormones and their regulatory genes are involved.
Disclosure of Invention
The invention aims to provide a protein related to pollen development and application of a coding gene thereof.
The invention provides application of a SlPP2C5 protein in regulation and control of plant pollen formation.
The invention also provides application of the SlPP2C5 protein in regulation and control of plant pollen development; .
The invention also provides application of the SlPP2C5 protein in regulation and control of plant flower development.
The invention also provides application of the nucleic acid molecule for encoding the SlPP2C5 protein in preparation of transgenic plants with increased pollen aberration rate.
The invention also provides application of the nucleic acid molecule for encoding the SlPP2C5 protein in preparation of male-sterile transgenic plants.
The invention also provides a method for cultivating the transgenic plant, which comprises the following steps: and (3) introducing a nucleic acid molecule for coding the SlPP2C5 protein into a receptor plant to obtain a transgenic plant with an increased pollen aberration rate.
The invention also provides a plant breeding method, which comprises the following steps: the content and/or activity of SlPP2C5 protein in the target plant is increased, so that the pollen aberration rate of the plant is increased.
The invention also provides a method for cultivating transgenic plants, which comprises the following steps: and (3) introducing a nucleic acid molecule for encoding the SlPP2C5 protein into a receptor plant to obtain a male-sterile transgenic plant.
The invention also provides a plant breeding method, which comprises the following steps: and (3) increasing the content and/or activity of the SlPP2C5 protein in the target plant, so that the plant is male sterile.
The SlPP2C5 protein is obtained from tomato (Solanum lycopersicum) and is (a1) or (a2) or (a3) or (a4) as follows:
(a1) protein shown in a sequence 1 in a sequence table;
(a2) a protein which is obtained by substituting and/or deleting and/or adding one or more amino acid residues to the protein shown in the sequence 1 of the sequence table, is related to the formation of plant pollen and/or the development of pollen and is derived from the protein;
(a3) a fusion protein obtained by attaching a tag to the N-terminus or/and the C-terminus of the protein of (a 1);
(a4) a protein derived from tomato and having 98% or more identity to (a1) and associated with plant pollen formation and/or pollen development.
The labels are specifically shown in table 1.
TABLE 1 sequences of tags
| Label (R) | Residue of | Sequence of |
| Poly-Arg | 5-6 (typically 5) | RRRRR |
| Poly-His | 2-10 (generally 6) | HHHHHH |
| FLAG | 8 | DYKDDDDK |
| Strep-tag II | 8 | WSHPQFEK |
| c- |
10 | EQKLISEEDL |
| HA | 9 | YPYDVPDYA |
The nucleic acid molecule encoding the SlPP2C5 protein is a DNA molecule of (b1) or (b2) or (b3) or (b4) as follows:
(b1) the coding region is shown as the DNA molecule shown by the 167 th and 1609 th nucleotides in the sequence 2 of the sequence table;
(b2) a DNA molecule shown in a sequence 2 of a sequence table;
(b3) a DNA molecule which hybridizes under stringent conditions to the DNA molecule defined in (b1) or (b2) and encodes the protein;
(4) a DNA molecule derived from tomato and having at least 98% homology with the DNA molecule defined in (b1) or (b2) and encoding said protein.
The stringent conditions are hybridization and washing of the membrane 2 times 5min at 68 ℃ in a solution of 2 XSSC, 0.1% SDS and 2 times 15min at 68 ℃ in a solution of 0.5 XSSC, 0.1% SDS.
The introduction of a nucleic acid molecule encoding a SlPP2C5 protein into a recipient plant as described above may specifically be the introduction of a recombinant plasmid having a nucleic acid molecule encoding a SlPP2C5 protein into a recipient plant. Specifically, the recombinant plasmid can be obtained by inserting a double-stranded DNA molecule shown by the 167-1609 th nucleotide in the sequence 2 of the sequence table into a multiple cloning site (for example, between XbaI and SacI enzyme cutting sites) of the pRI101-AN vector.
Any of the above plants can be a dicot.
Any of the above plants may be a solanaceous plant.
Any of the above plants may be a plant of the genus Lycopersicon.
Any of the above plants may be a tomato, e.g., tomato Micro-Tom.
The inventor of the invention finds that the tomato flower dysplasia and pollen teratogenesis rate can be increased by over-expressing the SlPP2C5 gene in the tomato. Therefore, the SlPP2C5 protein is related to the tomato flower development process and pollen development. The SlPP2C5 protein and the coding gene thereof play an important role in high-quality breeding of tomatoes.
Drawings
FIG. 1 shows the expression change of SlPP2C5 gene in the tomato fruit development and maturation process.
FIG. 2 is a photograph of pollen taken under an electron microscope.
FIG. 3 shows the pollen teratogenicity rate.
Detailed Description
The present invention is described in further detail below with reference to specific embodiments, which are given for the purpose of illustration only and are not intended to limit the scope of the invention. The examples provided below serve as a guide for further modifications by a person skilled in the art and do not constitute a limitation of the invention in any way.
The experimental procedures in the following examples, unless otherwise indicated, are conventional and are carried out according to the techniques or conditions described in the literature in the field or according to the instructions of the products. Materials, reagents and the like used in the following examples are commercially available unless otherwise specified. Unless otherwise stated, the quantitative tests in the following examples were carried out in triplicate, and the results were averaged. Tomato Micro-Tom, a conventional variety of tomato, also known as wild type tomato, is denoted by WT. Tomato peel refers to the portion of the fruit excluding the exocarp, pectin and seeds.
A new protein is found from tomato, and is shown as a sequence 1 in a sequence table and named as SlPP2C5 protein. The gene encoding the SlPP2C5 protein was named SlPP2C5 gene. In the tomato cDNA, the SlPP2C5 gene is shown as a sequence 2 in a sequence table (an open reading frame is shown as a 167-1609 site in the sequence 2).
Example 1 expression Change of SlPP2C5 Gene in tomato fruit development and maturation Process
According to the fruit setting days after the full-bloom stage of the tomato plant, the fruit size and the color change condition, the fruit development and maturation process is divided into a young fruit stage (IG), a green mature stage (MG), a color breaking stage (B), a color turning stage (T) and a red mature stage (RR).
The roots, stems, leaves, flowers of tomato Micro-Tom plants and the fruit peel of fruits in each period are frozen and preserved by liquid nitrogen. Taking a frozen sample of liquid nitrogen, extracting total RNA, and performing reverse transcription to obtain cDNA. And detecting the expression level of the SlPP2C5 gene by real-time quantitative qPCR (the internal reference gene is SlSAND gene) by taking the cDNA as a template.
The primers used for detecting the SlPP2C5 gene were as follows:
SlPP2C5-qF (upstream primer): GTGTATTTGGCGTTCTTGCAATGTC, respectively;
SlPP2C5-qR (downstream primer): CAGGCAGAGGGTTAGTCCCGTTC are provided.
Reaction system: 2 × SYBR Premix Ex Taq 10 μ L, upstream primer (10mM)0.5 μ L, downstream primer (10mM)0.5 μ L, cDNA 1.5 μ L, ddH 2 O 7.5μL。
The results are shown in FIG. 1.
Example 2 obtaining of transgenic tomato with SlPP2C5 Gene and testing of fruit-related traits
Construction of recombinant expression vector
Inserting the double-stranded DNA molecule shown by the 167-1609 th nucleotide in the sequence 2 of the sequence table between XbaI and SacI enzyme cutting sites of the pRI101-AN vector to obtain the recombinant plasmid. The recombinant plasmid was sequence verified.
Secondly, preparing transgenic tomato plants
1. Taking tomato Micro-Tom seeds, sterilizing with 75% ethanol water solution for 30s, then sterilizing with sodium hypochlorite water solution for 15min, and then washing with sterile water for 6-8 times.
2. And (3) taking the seeds which are subjected to the step 1, placing the seeds in a solid 1/2MS culture medium, and alternately culturing for 7-8 days at 25 ℃ in dark and light (at the moment, cotyledons are fully expanded, true leaves are not formed yet).
3. After the step 2 is finished, cutting off cotyledons, removing two ends, and cutting the middle part into a square with the side length of about 0.5cm, namely an explant; the explants were placed in preculture medium and cultured in the dark at 25 ℃ for 2 days.
Pre-culture medium: solid MS medium containing 2.5 mg/L6-BA and 0.2mg/L IAA.
4. And (3) after the step 3 is finished, placing the explant in an infection solution for infection for 5min, then taking out the explant, sucking the residual bacterial liquid, then placing the explant in a co-culture medium, and carrying out dark culture for 2 days at 25 ℃.
Immersing a dye solution: introducing the recombinant plasmid obtained in the step one into agrobacterium LBA4404 to obtain recombinant agrobacterium; resuspending the recombinant Agrobacterium with liquid MS medium containing 100mg/L AS to OD 600nm And (5) obtaining the staining solution.
Co-culture medium: solid MS medium containing 2.5 mg/L6-BA, 0.2mg/L IAA and 100mg/L AS.
5. After the step 4 is completed, the explants are transferred to a sterilization culture medium, and are cultured in the dark at 25 ℃ for 2-3 days, and then are cultured in the dark at 25 ℃ for 3-4 days alternately.
A sterilization culture medium: solid MS culture medium containing 2.5 mg/L6-BA, 0.2mg/L IAA, 200mg/L Carb and 300mg/L Cef.
6. After step 5 is completed, the explants are transferred to a screening medium and cultured alternately in light and dark at 25 ℃ for 30-45 days (subcultured every 15 days) to obtain plants.
Screening a culture medium: solid MS culture medium containing 2.5 mg/L6-BA, 0.2mg/L IAA, 200mg/L Carb, 300mg/L Cef and 4mg/L Hyg.
7. And (6) after the step 6 is finished, transferring the plants to a rooting culture medium, and performing light-dark alternate culture at 25 ℃ to obtain rooted plants, namely T0 generation regeneration plants.
Rooting culture medium: 1/2MS solid medium containing 0.2mg/L IAA and 200mg/L Carb.
8. Transgenic plants were selected from regenerated plants of T0 generation.
The method for screening transgenic plants comprises the following steps: and when the plant grows to 2-4cm long, taking the plant leaves for PCR identification, and if a specific amplification product is obtained, the regenerated plant is a positive plant for PCR identification, namely a transgenic plant.
The primers used for PCR identification were as follows:
35S-F:GCAAGACCCTTCCTCTATATAAGG；
SlPP2C5-R：ACTTTTGCTTTTGAACTTCCTGTG。
9. selfing the T0 transgenic plants to obtain progeny plants, namely T1 transgenic plants; transgenic plants were selected from T1 generation plants (same procedure as in step 8).
10. Selfing the T1 transgenic plants to obtain progeny plants, namely T2 plants; transgenic plants were selected from T2 generation plants (same procedure as in step 8).
For a certain T1 generation transgenic plant, if the T2 generation plants obtained by selfing are all transgenic plants, the T1 generation plants are homozygous transgenic plants, and the generation after selfing is homozygous transgenic lines. Taking 2 transgenic lines (namely a SlPP2C5-OE-2 line and a SlPP2C5-OE-10 line) to carry out the fourth step.
Thirdly, preparing empty carrier tomato plants
And (5) replacing the recombinant plasmid with the pRI101-AN vector, and performing operation according to the second step to obtain a transgenic empty vector strain.
Fourth, fruit quality analysis
Test plants: t3 plants of a SlPP2C5-OE-2 strain, T3 plants of a SlPP2C5-OE-10 strain, wild type tomato plants and T3 plants of a transgenic empty vector strain.
Normally culturing the tested plants under parallel conditions, taking pollen in full-bloom stage for electron microscope observation, and counting the pollen aberration rate.
The photograph of the pollen under an electron microscope is shown in FIG. 2. Pollen grains of wild-type tomato plants are in the shape of wheat grains and have three germination ditches. Pollen grains of the plants of the SlPP2C5-OE-2 line and the plants of the SlPP2C5-OE-10 line are shrunken and deformed, the germination ditches are shallow and the middle part is convex. Pollen tubes usually enter from the germination groove, and thus pollen drying out and germination groove malformation of transgenic plants result in reduced pollen viability. Compared with wild tomato plants, the pollen phenotype of the plants of the empty vector line has no significant difference.
Pollen teratogenicity is shown in FIG. 3 (at least 3 plants tested per line). Pollen aberration rate of wild type tomato plant was 10%. The pollen aberration rate of the empty vector line strain plants is 11%. The pollen distortion rates of the plant of the SlPP2C5-OE-2 line and the plant of the SlPP2C5-OE-10 line are 66.7 percent and 81.8 percent respectively, namely the pollen distortion rate of the transgenic plant is obviously higher than that of a wild plant.
The results show that the normal development of tomato flowers is influenced by over-expression of the SlPP2C5 gene, and the SlPP2C5 protein plays an important role in the development process of tomato pollen.
The present invention has been described in detail above. It will be apparent to those skilled in the art that the invention can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the invention and without undue experimentation. While the invention has been described with reference to specific embodiments, it will be appreciated that the invention can be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains. The use of some of the essential features is possible within the scope of the claims attached below.
Sequence listing
<120> protein related to pollen development and application of coding gene thereof
<130> GNCYX210768
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 480
<212> PRT
<213> Solanum lycopersicum
<400> 1
Met Lys Val Asp Val Gly Arg Val Pro Leu Leu Thr Leu Gly Glu Ser
1 5 10 15
Ser Gly Lys Cys Ser Leu Pro Gln Thr Val Leu Gly Ala Glu Asn Gly
20 25 30
Leu Ile Val Ser Asp Ser Ile Ile Gln Gly Ser Asp Glu Asp Glu Ile
35 40 45
Leu Ser Val Gly Glu Asp Pro Cys Gly Ile Asn Gly Glu Glu Leu Leu
50 55 60
Pro Leu Gly Ala Ser Leu Gln Leu Ser Leu Pro Ile Ala Val Glu Ile
65 70 75 80
Glu Gly Ile Asp Asn Gly Gln Ile Val Ala Lys Val Ile Ser Leu Glu
85 90 95
Glu Arg Ser Leu Asp Arg Lys Val Ser Asn Thr Ile Val Ala Leu Pro
100 105 110
Asp Asp Glu Ile Thr Ser Gly Pro Thr Leu Lys Ala Ser Val Val Ala
115 120 125
Leu Pro Leu Pro Ser Glu Lys Glu Pro Val Lys Glu Ser Val Lys Ser
130 135 140
Val Phe Glu Leu Glu Cys Val Pro Leu Trp Gly Ser Val Ser Ile Cys
145 150 155 160
Gly Lys Arg Pro Glu Met Glu Asp Ala Leu Val Val Val Pro Asn Phe
165 170 175
Met Lys Ile Pro Ile Lys Met Phe Ile Gly Asp Arg Val Ile Asp Gly
180 185 190
Leu Ser Gln Ser Leu Ser His Leu Thr Ser His Phe Tyr Gly Val Tyr
195 200 205
Asp Gly His Gly Gly Ser Gln Val Ala Asp Tyr Cys Arg Lys Arg Val
210 215 220
His Leu Ala Leu Val Glu Glu Leu Lys Leu Pro Lys His Asp Leu Val
225 230 235 240
Asp Gly Ser Val Arg Asp Thr Arg Gln Val Gln Trp Glu Lys Val Phe
245 250 255
Thr Asn Cys Phe Leu Lys Val Asp Asp Glu Val Gly Gly Lys Val Ile
260 265 270
Asp Leu Cys Asp Asp Asn Ile Asn Ala Ser Ser Cys Thr Ser Glu Pro
275 280 285
Ile Ala Pro Glu Thr Val Gly Ser Thr Ala Val Val Ala Val Ile Cys
290 295 300
Ser Ser His Ile Ile Val Ala Asn Cys Gly Asp Ser Arg Ala Val Leu
305 310 315 320
Tyr Arg Gly Lys Glu Ala Val Ala Leu Ser Ile Asp His Lys Pro Ser
325 330 335
Arg Glu Asp Glu Tyr Ala Arg Ile Glu Ala Ser Gly Gly Lys Val Ile
340 345 350
Gln Trp Asn Gly His Arg Val Phe Gly Val Leu Ala Met Ser Arg Ser
355 360 365
Ile Gly Asp Arg Tyr Leu Lys Pro Trp Ile Ile Pro Glu Pro Glu Val
370 375 380
Met Phe Val Pro Arg Ala Arg Glu Asp Glu Cys Leu Val Leu Ala Ser
385 390 395 400
Asp Gly Leu Trp Asp Val Met Thr Asn Glu Glu Ala Cys Glu Met Ala
405 410 415
Arg Arg Arg Ile Leu Leu Trp His Lys Lys Asn Gly Thr Asn Pro Leu
420 425 430
Pro Glu Arg Gly Gln Gly Val Asp Leu Ala Ala Gln Ala Ala Ala Glu
435 440 445
Tyr Leu Ser Ser Met Ala Leu Gln Lys Gly Ser Lys Asp Asn Ile Ser
450 455 460
Val Ile Val Val Asp Leu Lys Ala His Arg Lys Phe Lys Ser Lys Ser
465 470 475 480
<210> 2
<211> 1919
<212> DNA
<213> Solanum lycopersicum
<400> 2
aagttgtatt tttgttctct ctcatgtgta tcacaaagaa aaaaagaaaa taaaatcttg 60
taaaggggct gtagctgcaa ggtgttttgg taaaagggtt tttgttgatc ctacatcttt 120
gttgcatgct gagtcaaata ccagttggga tggaaaaagc aacggtatga aagttgatgt 180
tggtagagtt cccttgttga ccctaggaga aagctctgga aaatgtagtc tgccgcagac 240
tgtattggga gctgaaaatg gcctgattgt tagcgatagc atcattcagg gaagtgatga 300
agatgagatt ttatctgttg gagaggatcc atgtggaatt aatggcgagg agttgttgcc 360
actgggcgct agcttgcagt tgagcttgcc aattgctgtt gaaattgagg gtattgacaa 420
tggacaaata gttgccaagg tcataagttt ggaagaaagg agtttagata gaaaggttag 480
taataccata gttgctcttc cagatgatga aattactagt ggccctacac ttaaggcatc 540
tgtagtggcc cttccattgc ccagtgagaa ggagcctgtc aaagaaagtg tcaagagtgt 600
gtttgaattg gaatgtgtgc cactctgggg ttctgtatct atctgtggaa agagaccgga 660
gatggaggat gctcttgtgg ttgttcctaa tttcatgaaa attcctatca agatgtttat 720
tggtgatcgt gtaattgatg gactaagtca aagtttgagt cacctgacat ctcatttcta 780
tggagtatat gatggtcatg gaggatctca ggttgcggat tattgccgta aacgtgttca 840
tctagcatta gttgaggaat taaaacttcc caaacatgat ttggtggatg gaagtgtaag 900
ggatacccgg caggtgcagt gggagaaggt ttttactaat tgctttctca aggttgatga 960
tgaagttgga ggaaaggtca tagatctctg tgatgacaac attaatgcct ctagctgcac 1020
ctctgagcct atagctccag aaactgttgg gtccaccgca gttgtagcgg tgatttgttc 1080
atctcatatt atagttgcta actgtgggga ttcaagagca gtcctttatc gtggcaaaga 1140
agcagtggca ttgtcaatcg atcacaaacc aagcagagaa gatgagtatg ccagaattga 1200
agcatctggt ggtaaggtca ttcagtggaa tggacatcgt gtatttggcg ttcttgcaat 1260
gtcaagatct attggtgaca gatatttgaa accatggata atacctgaac cagaagttat 1320
gtttgtacca cgtgctagag aagatgaatg cctagtttta gccagtgacg gtttgtggga 1380
tgtgatgacg aatgaagaag cttgtgaaat ggctagacgg cgaattctgc tgtggcacaa 1440
aaagaacggg actaaccctc tgcctgaaag gggccaggga gtggatcttg ctgcacaagc 1500
agcagcggag tatctttcat cgatggctct tcagaaaggc agcaaagaca atatatccgt 1560
gattgtggtg gaccttaaag ctcacaggaa gttcaaaagc aaaagttaga gatgacatgt 1620
tcactacatt tggtttagta taaatctgta cacggcgacg ggtataaatc tcattattac 1680
ataactcagt ccattaattt ttcctatggg cttaaggtgt gtgtatgaga atagtatttt 1740
agcaatgtat tatagaagaa aaaacagttg acaaacgacg tttatccaaa ttttttggtg 1800
tttgttgcgc cagaaaatgg ctatgtaaat tgagcatgtt gtagcaaata tcagaaatgc 1860
aatttcttac ttcttccgtt cgagtatatg ttataaaagc taagccacaa ttggttact 1919
Claims (10)
- The application of the SlPP2C5 protein in regulation and control of plant pollen formation;the SlPP2C5 protein is (a1) or (a2) or (a3) or (a4) as follows:(a1) protein shown in a sequence 1 in a sequence table;(a2) a protein which is obtained by substituting and/or deleting and/or adding one or more amino acid residues to the protein shown in the sequence 1 of the sequence table, is related to the formation of plant pollen and/or the development of pollen and is derived from the protein;(a3) a fusion protein obtained by attaching a tag to the N-terminus or/and the C-terminus of the protein of (a 1);(a4) a protein derived from tomato and having 98% or more identity to (a1) and associated with plant pollen formation and/or pollen development.
- 2, application of SlPP2C5 protein in regulation and control of plant pollen development; the SlPP2C5 protein is the SlPP2C5 protein as claimed in claim 1.
- The application of the SlPP2C5 protein in regulating and controlling the development of plant flowers; the SlPP2C5 protein is the SlPP2C5 protein as claimed in claim 1.
- 4. The application of nucleic acid molecules for coding SlPP2C5 protein in preparing transgenic plants with increased pollen aberration rate; the SlPP2C5 protein is the SlPP2C5 protein as claimed in claim 1.
- 5. The application of nucleic acid molecules for coding SlPP2C5 protein in preparing male sterile transgenic plants; the SlPP2C5 protein is the SlPP2C5 protein as claimed in claim 1.
- 6. Use according to claim 4 or 5, characterized in that:the nucleic acid molecule encoding the SlPP2C5 protein is a DNA molecule of (b1) or (b2) or (b3) or (b4) as follows:(b1) the coding region is shown as the DNA molecule shown by the 167 th and 1609 th nucleotides in the sequence 2 of the sequence table;(b2) a DNA molecule shown in a sequence 2 of a sequence table;(b3) a DNA molecule which hybridizes under stringent conditions to the DNA molecule defined in (b1) or (b2) and encodes the protein;(4) a DNA molecule derived from tomato and having at least 98% homology with the DNA molecule defined in (b1) or (b2) and encoding said protein.
- 7. A method of breeding a transgenic plant comprising the steps of: introducing a nucleic acid molecule for coding the SlPP2C5 protein into a receptor plant to obtain a transgenic plant with an increased pollen aberration rate; the SlPP2C5 protein is the SlPP2C5 protein as claimed in claim 1.
- 8. A method of plant breeding comprising the steps of: increasing the content and/or activity of SlPP2C5 protein in a target plant, thereby increasing the pollen teratogenesis of the plant; the SlPP2C5 protein is the SlPP2C5 protein as claimed in claim 1.
- 9. A method of growing a transgenic plant comprising the steps of: introducing a nucleic acid molecule encoding an SlPP2C5 protein into a receptor plant to obtain a male-sterile transgenic plant; the SlPP2C5 protein is the SlPP2C5 protein as claimed in claim 1.
- 10. A method of plant breeding comprising the steps of: increasing the content and/or activity of SlPP2C5 protein in a target plant, thereby making the plant male sterile; the SlPP2C5 protein is the SlPP2C5 protein as claimed in claim 1.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20010106331A (en) * | 2001-07-30 | 2001-11-29 | 박찬구 | Nucleic acid molecule encoding the catalytic subunit of a protein phosphatase 2A that regulates flowering time in plants |
| CN102149818A (en) * | 2008-08-15 | 2011-08-10 | 纳幕尔杜邦公司 | Plants with altered root architecture, related constructs and methods involving genes encoding protein phophatase 2C (PP2C) polypeptides and homologs thereof |
| US20140007298A1 (en) * | 2011-02-07 | 2014-01-02 | Pioneer Hi Bred International Inc | Plants with altered root architecture, related constructs and methods involving genes encoding protein phophatase 2c (pp2c) polypeptides and homologs thereof |
| CN109423492A (en) * | 2017-08-21 | 2019-03-05 | 中国科学院遗传与发育生物学研究所 | Application of the SlTOE1 gene in regulation tomato flowering time and yield |
-
2021
- 2021-02-20 CN CN202110195290.XA patent/CN114957417B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20010106331A (en) * | 2001-07-30 | 2001-11-29 | 박찬구 | Nucleic acid molecule encoding the catalytic subunit of a protein phosphatase 2A that regulates flowering time in plants |
| CN102149818A (en) * | 2008-08-15 | 2011-08-10 | 纳幕尔杜邦公司 | Plants with altered root architecture, related constructs and methods involving genes encoding protein phophatase 2C (PP2C) polypeptides and homologs thereof |
| US20140007298A1 (en) * | 2011-02-07 | 2014-01-02 | Pioneer Hi Bred International Inc | Plants with altered root architecture, related constructs and methods involving genes encoding protein phophatase 2c (pp2c) polypeptides and homologs thereof |
| CN109423492A (en) * | 2017-08-21 | 2019-03-05 | 中国科学院遗传与发育生物学研究所 | Application of the SlTOE1 gene in regulation tomato flowering time and yield |
Non-Patent Citations (2)
| Title |
|---|
| YUSHU ZHANG等: "Suppressing Type 2C Protein Phosphatases Alters Fruit Ripening and the Stress Response in Tomato" * |
| 姜丽等: "关于ABA信号转导核心组份PP2C的研究进展分析" * |
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