CN113817755A - Rice long-grain gene LOG1 and application thereof - Google Patents
Rice long-grain gene LOG1 and application thereof Download PDFInfo
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Abstract
The invention belongs to the field of plant genetic engineering, and particularly relates to a separation clone and functional verification of a gene LOG1 for controlling rice grain development, and application thereof in rice production practice. The invention discloses a rice long-grain gene LOG1 and a protein coded by the same, and also discloses an application of the rice long-grain gene LOG 1: the grain length and the grain width of rice grains are increased, so that the rice yield is improved.
Description
Technical Field
The invention belongs to the field of plant genetic engineering, and particularly relates to a separation clone and functional verification of a gene LOG1 for controlling rice grain development, and application thereof in rice production practice.
Background
Rice is an important food crop for human beings, and more than one third of the world population is cultivated. As the population increases year by year, it is predicted that the world food production must increase by 70% by 2050 to meet the increasing population demand for food. In high-yield breeding of rice, thousand kernel weight is one of three factors that determine yield, and grain shape is an important agronomic trait that determines thousand kernel weight (Zhang Jian et al, 2020). Wherein, the indexes reflecting the particle shape mainly comprise the particle length, the particle width, the particle thickness and the length-width ratio. These indicators are not only closely related to the rice yield, but also affect the appearance quality of rice, and determine the final commodity value. To date, a number of genes controlling grain shape have been cloned, which lays an important foundation for increasing rice yield and improving rice quality (Liuxi et al, 2018).
The grain length, the grain width, the grain thickness and the length-width ratio of the rice are quantitative traits controlled by multiple genes, most related Quantitative Trait Loci (QTL) of the rice mainly regulate grain shape by influencing the division of glume cells, and a molecular regulation mechanism of the grain shape is preliminarily disclosed. GS3 encodes a transmembrane protein containing 3 domains, of which the OSR domain is an essential element affecting grain length, while the VWFC domain has an inhibitory effect on the OSR domain (Mao et al, 2010). After the GS3 function is lost, grains are lengthened, narrowed and the grain weight is increased; after overexpression of GS3, grains become small, plants are dwarfed, and growth and development are inhibited, which indicates that GS3 is a negative regulatory factor of grain length. Compared with the short-grain variety, the long-grain variety has nonsense mutation of the gene in the second exon, and the protein translation is terminated early. Genome association analysis of rice cultivars showed that this nonsense mutation located in the second exon of GS3 is responsible for the difference in grain size in the natural population of rice (Fan et al, 2009). qGL3/GL3.1 is located on chromosome 3 and encodes a Kelch repeat domain phosphatase protein. qGL3/GL3.1 mainly changes glume length by changing glume cell number, and also influences rice filling rate, so that the size and yield of rice grains are changed, and the rice grain length regulating factor is a negative regulating factor of rice grain length (Zhang et al, 2012; Qi et al, 2012). In rice variety N411, a D364E rare allelic variation in the second Kelch domain of the osppcl 1 protein occurred, resulting in a long grain phenotype (Zhang et al, 2012). GL6 is located on chromosome 6 and encodes an AT-rich and zinc-binding protein that positively regulates grain length and grain weight. The GL6 mutation causes the shortening of grains; it over-expressed kernel becomes long (Wang et al, 2019). GW2 is located on chromosome 2, is a major QTL for grain width and grain weight, encodes a RING protein with E3 ubiquitin ligase activity, which is degraded by anchoring its substrate to the proteasome, thereby negatively regulating cell division. GW2 increases glume size mainly by increasing glume cell number, and also increases rice filling rate, thereby changing grain size and yield (Song et al, 2007). Besides affecting the development of grains, the GW2 gene also regulates the development of the whole plant. Therefore, it belongs to the pleiotropic gene as the GS3 gene. The GS5 gene is located on chromosome 5 and encodes a serine carboxypeptidase that positively regulates rice grain width and grain weight (Li et al, 2011). The GS5 gene influences the width of grains by influencing the number and the size of cells, and the increase of the expression quantity of the GS5 gene promotes glume cell division, so that the grain width is increased, and the opposite is true. Studies have shown that both broad and narrow varieties encode the complete GS5 protein, and thus the difference in grain width may be caused by the difference in the amount of expression of the gene. GS9 is located in chromosome 9, encodes an unknown domain protein, and is used as a transcription activator to regulate the size and appearance quality of grains. Loss of function of GS9 results in a narrowing of the kernel, which overexpresses kernel widening (Zhao et al, 2018). Plant hormones and small molecular RNA are used as important regulating factors for plant growth and are also involved in the development and regulation of grain shape. TGW6 hydrolyzes indole acetic acid-glucose to free indole acetic acid and glucose, and affects the transformation of endosperm from the syncretic to the cellularizing stage by controlling indole acetic acid supply, thereby limiting cell number and kernel length. SG1 encodes a protein of unknown function, involved in the synthesis and signal transduction of Brassinosteroids (BR). Overexpression of SG1 resulted in short grain and dwarf phenotypes, SG1 reduced the response to BR, inhibiting the length of grain by reducing cell proliferation (Nakagawa et al, 2012). BG1 encodes an unknown functional protein involved in auxin transport, is a positive regulator of auxin response and transport, is a plant specific regulator of organ size, and enhances polar transport capacity of auxin over-expressed plants, thereby positively regulating grain size (Liu et al, 2015). GS2/GL2 is regulated by OsmiR396, a rare dominant mutation occurs at a target site of the OsmiR396, shearing of the OsmiR396 on the target site is influenced, the expression quantity of the gene is obviously increased, and therefore division and growth of cells are promoted, and a long-grained phenotype is generated (Che et al, 2015; Duan et al, 2015; Hu et al, 2015).
Rice (Oryza sativa) is a basic food for nearly half of the global population to live, and the cultivation of high-yield and high-quality rice varieties is still urgent.
Disclosure of Invention
The invention aims to solve the technical problem of providing a rice long-grain gene LOG1 and application thereof in rice breeding.
In order to solve the technical problems, the invention provides a rice long grain gene LOG1, wherein the nucleotide sequence of the gene LOG1 is shown as SEQ ID NO: 2, the preparation method is as follows.
The invention also provides the protein coded by the rice long-grain gene LOG1, and the amino acid sequence of the protein is shown as SEQ ID NO: 4, the method is described in the specification.
The invention also provides a recombinant vector and a transformant containing the gene.
The invention also provides the application of the rice long-grain mutant gene LOG 1: increasing the grain size of gramineous plants and increasing the yield.
The improvement of the application of the rice long-grain gene LOG1 of the invention is as follows: the gramineous plants are rice, and the grain length and the grain width of rice grains are increased, so that the rice yield is increased.
The invention also provides a method for regulating and controlling the grain development of the gramineous plants, which comprises the following steps: the method comprises the steps of transforming gramineous plant (such as rice) cells by using the gene LOG1, and then culturing the transformed gramineous plant cells into plants, wherein the grains of the plants are enlarged, and the thousand kernel weight of the plants is increased.
The improvement of the method for regulating and controlling the development of the gramineous plant kernel comprises the following steps: the grain length and the grain width of the grains are increased.
The invention also provides a plasmid containing the gene, and engineering bacteria or host cells containing the gene or the vector.
The engineered bacteria and host cells are understood to be those used by the skilled person in the transgenic process. However, with the development of science and technology, the selection of the engineering bacteria and the host cells may be changed, or in the application field of non-transgenic purpose, the utilization of the vector and the engineering bacteria is also related, but the invention is within the protection scope as long as the gene or the vector of the invention is contained.
Further, the invention also provides a host cell, which contains the gene sequence and is an Escherichia coli cell, an Agrobacterium cell or a plant cell.
Another object of the present invention is to provide the use of the above gene for transgenic crop improvement.
The preparation of the transgenic rice is a conventional technical means in the field, the invention is not limited, and the technical scheme of utilizing the gene to perform rice transgenosis is within the protection scope of the invention.
The specific technical steps for realizing the invention are as follows:
isolation and genetic analysis of mutant LOG 1:
the rice long-grain mutant log1 is from EMS mutant library of indica rice variety Shuhui 527(SH 527). The mutant is proved to be controlled by a recessive single gene through a positive and negative cross experiment with a wild type (SH 527).
Comparison of Lo 1 with wild type spikelets
Log1 kernels were observed during maturity. The log1 mutant can obviously increase the grain length and the grain width, and obviously increase the rice yield (figure 1).
Third, map-based cloning of LOG1 Gene
In order to separate LOG1 gene, the invention firstly constructs a positioning group, uses japonica rice Nipponbare (NIP) as recurrent parent, and backcrosses with LOG1 to separate group BC2F2And preliminarily positioning the LOG1 locus on the 7 th chromosome between the R35 marker and the R17 marker by using a plurality of molecular markers through a map-based cloning method. By analyzing the sequence between these two markers, a new polymorphic marker was developed to finely map the LOG1 gene to a region of about 36kb between the markers S53 and S99, and the DNA sequence of LOC _ Os07g01820 gene of the LOG1 mutant was found to have base substitutions compared with SH527 by sequencing alignment analysis (FIG. 2). Blast analysis found that the LOG1 gene (LOC _ Os07g01820) was allelic with the previously reported DEP/OsMADS 15. The log1 mutant site is inconsistent with the previously reported mutant site of DEP/OsMADS15 allele, and the mutant DNA sequence in log1 has base substitution to result in amino acid change, and the mutant amino acid is different from that of DEP/OsMADS15 allele. LOG1 gene expression analysis indicated expression predominantly in the ear, consistent with its phenotype (fig. 3).
The rice long-grain mutant log1 of the invention has obvious phenotype difference with the existing dep/osmads15 mutant. The method specifically comprises the following steps: the dep/osmads15 mutant is mainly characterized by palea (i.e., the palea is present to be thinned and the palea degeneration is reduced), whereas the log1 mutant of the present invention mainly shows that the kernel is obviously lengthened and widened, the thousand kernel weight is increased and the palea is normal.
Therefore, the cloning and the deep functional analysis of the LOG1 gene are helpful for deeply clarifying the genetic regulation mechanism of rice grain development; namely, the invention has important practical and theoretical significance for further improvement of the yield of rice and molecular design breeding.
In conclusion, the cloned gene LOG1 for controlling long grain and the protein coded by the gene have important production and application values in improving the rice yield.
Drawings
The following describes embodiments of the present invention in further detail with reference to the accompanying drawings.
FIG. 1 is a comparison of grain phenotypes of wild type (SH527) and log1 mutants;
in fig. 1:
a is the length comparison of SH527 and log1 mutant grains;
b is the width comparison of SH527 and log1 mutant grains;
c is the length statistical data of SH527 and log1 mutant grains;
d is the statistic data of the widths of SH527 and log1 mutant grains;
e is statistic data of thousand kernel weight of SH527 and log1 mutant grains;
FIG. 2 shows the phenotype of transgenic rice tested for gene localization and functional complementation;
in fig. 2:
a is the location of the LOG1 gene;
b is LOG1 gene encoding cinnamyl alcohol dehydrogenase, the mutation of which results in premature termination of protein translation;
c is a genetic complementation experiment; wt represents grain of wild type plants, log1 represents grain of mutant plants, com represents grain of complementary plants.
FIG. 3 is the expression of the LOG1 gene;
0.5cm represents a 0.5cm long ear, 0.5-2 cm represents a 0.5-2 cm long ear, 2-5 cm represents a 2-5 cm long ear, R represents a root, C represents a stem, and L represents a leaf.
Detailed Description
The following examples illustrate the invention in detail: the present embodiment is implemented on the premise of the technical solution of the present invention, and a detailed implementation manner and a specific operation process are given, but the protection scope of the present invention is not limited to the following embodiments.
Example 1: rice material
Rice (Oryza sativa L.) mutant log1(long grain 1), original wild type material indica rice variety shuhui 527(SH527) (fig. 1). log1 was derived from the EMS mutant pool of the indica variety shuhui 527(SH 527).
Example 2:
1. analyzing and locating populations
Backcross segregation population BC formed by combining japonica rice variety NIP as recurrent parent with log12F2And from BC2F25628 individuals with long grain phenotype are selected from the group as the positioning group.
2. DNA extraction
Approximately 1g of leaves were taken from each plant during heading to extract total DNA. The method comprises the following specific steps:
firstly, 1g of rice leaves are weighed and ground into powder by liquid nitrogen, then 600 mul of DNA extraction buffer solution prepared by CTAB solution (2% (m/V) CTAB, 100mmol/L Tris-Cl, 20mmol/L EDTA, 1.4mol/L NaCl, pH8.0) is added, and water bath is carried out at 65 ℃ for 40 minutes. Then 600. mu.l of chloroform/isoamyl alcohol (24:1 by volume) was added thereto and mixed well. Centrifuge at 10,000rpm for 5 minutes and transfer the supernatant to a new centrifuge tube.
And secondly, adding 2/3-1 times of volume of precooled (to 4 ℃) ethanol into the supernatant obtained after centrifugation in the step I, and gently mixing the mixture until DNA precipitates. Centrifuged at 12,000rpm for 10 minutes and the supernatant was decanted.
③ washing the DNA precipitate obtained in the previous step with 500. mu.l of 75% (volume concentration) ethanol.
Fourthly, the washed DNA is dried and dissolved in 500 mul of pure water.
Fifthly, detecting the integrity of the DNA by using 1 percent agarose gel electrophoresis. The intact DNA was used for PCR amplification and the incomplete DNA was re-extracted until the intact DNA was obtained.
Example 3:
1. preliminary mapping of LOG1 Gene
Backcross segregation population BC formed by combining japonica rice variety NIP as recurrent parent with log12F2188 long-grain plants are randomly selected from the group, polymorphism analysis is carried out on a small population, and PCR amplification is carried out according to known reaction conditions for linkage analysis according to primers uniformly distributed on 12 chromosomes in a laboratory. The PCR amplification conditions were as follows: the total PCR reaction was 10. mu.l: wherein 100 ng/. mu.l rice genome DNA 1. mu.l, 10 XPCR Buffer 1. mu.l, 2mM dNTP 1. mu.l, 10uM primer 2. mu.l, 5U/. mu.l rTaq 0.05. mu.l, ddH2O4.95. mu.l. The PCR amplification conditions are specifically: pre-denaturation at 94 ℃ for 5 min; denaturation at 94 ℃ for 30 seconds, annealing at 55 ℃ for 30 seconds, extension at 72 ℃ for 30 seconds, 35 cycles, separation by 4% agarose gel electrophoresis and staining with Gelred nucleic acid dye, detection of polymorphisms in PCR products, and preliminary mapping of the LOG1 gene between the R35 and R17 markers on chromosome 7 (fig. 2).
2. Fine localization of LOG1 Gene
Segregating populations BC using backcrosses assembled from NIP and log12F2The remaining 5440 recessive individuals continue to design molecular markers based on the initial localization, and finally precisely localize the LOG1 gene in the region of about 36kb between the markers S53 and S99. The sequence of the coding region of LOC _ Os07g01820 gene of the log1 mutant was found to undergo base substitutions compared to SH527 by sequencing alignment analysis, resulting in amino acid substitutions (FIG. 2). Blast analysis found that the LOG1 gene (LOC _ Os07g01820) was allelic with the previously reported DEP/OsMADS 15. The log1 mutant site is inconsistent with the mutant site of DEP/OsMADS15 allele reported previously, and the base substitution of the mutant coding region sequence in log1 results in amino acid substitution, and the mutant amino acid is different from the mutant amino acid of DEP/OsMADS15 allele.
The primer sequences are shown in table 1:
TABLE 1 location marker sequence of LOG1 Gene
| Name of label | Front primer (5 '-3') | Rear primer (5 '-3') | Fragment size |
| R35 | TATGTCCTTAGCATGGAAGACC | CGTTGTTGATCATCTGGTACG | 122bp |
| R17 | CGTAGGTGATATATTGATCC | GTTCAAATTTTAACTAGCCA | 142bp |
| S53 | CACATGGGTCCCACGCTGAG | CACCTCAGACCATTAGCAC | 374bp |
| S99 | CTGCGGATATAGGATGAGTTC | CACGCCACTGTCGACGTTTG | 655bp |
3. Gene prediction and comparative analysis:
from the fine localization results, a total of 4 candidate genes were found in this interval, as predicted by the Rice Genome Annotation Project http:// Rice. Candidate genes were amplified from the LOG1 mutant and wild-type SH527 genomes respectively by PCR for sequencing analysis by designing sequencing primers. The DNA sequence of LOC _ Os07g01820 gene of the log1 mutant underwent base substitution, resulting in amino acid change. Blast analysis found that the LOG1 gene (LOC _ Os07g01820) was allelic with the previously reported DEP/OsMADS 15. The log1 mutation site was not consistent with the previously reported mutation site for the DEP/OsMADS15 allele. The mutant DNA sequence in log1 underwent base substitutions resulting in amino acid changes.
The results are verified repeatedly for 3 times respectively, the same result is obtained, and the accuracy of the result is proved. Based on the gene annotation information (NCBI), the LOG1 gene was predicted to encode a MADS-box gene. Expression pattern analysis showed that the LOG1 gene was predominantly expressed in young ears, with reduced expression as ear length increased (fig. 3).
The PCR reaction system is as follows: 2. mu.l of 100 ng/. mu.l rice genomic DNA,
GXL Buffer 10. mu.l, 2.5mM dNTP 4. mu.l, 10. mu.M primer 4. mu.l,
GXL DNA Polymerase 4μl，ddH2o26. mu.l, 50. mu.l in total. The PCR amplification conditions were: pre-denaturation at 94 ℃ for 2 min; denaturation at 98 ℃ for 10 seconds, annealing at 60 ℃ for 15 seconds, extension at 68 ℃ for 70 seconds, 30 cycles, gel cutting and recovery after 1% agarose gel electrophoresis, transferring to escherichia coli, and selecting positive monoclonal sequencing. The LOG1 gene sequencing primers were as follows: LOG1CX-1F: TAGCTTGTAGCAGCTAAGGTTAGGTC and LOG1CX-1R: CGCTAGGATCGATACCAATGACTTC.
The nucleotide sequence of the rice long-grain gene LOG1 is shown as SEQ ID NO: 2, the amino acid sequence of the encoded protein is shown as SEQ ID NO: 4 is shown in the specification; SEQ ID NO: 4 is provided with a terminator.
The nucleotide sequence of original wild type material indica rice variety Shuhui 527(SH527) is SEQ ID NO: 1, the amino acid sequence of the protein coded by SH527 is shown as SEQ ID NO: 3, the process is carried out; SEQ ID NO: and a terminator is arranged at the tail end of the 3.
Description of the drawings: SEQ ID NO: 1 includes exon, intron; SEQ ID NO: 2 comprises 5-terminal exon and intron.
Example 4, plant transformation:
the DNA sequence of SH527 LOG1 (containing SEQ ID NO: 1) was amplified using homologous recombination and PCR techniques. And recovering and purifying DNA fragments after electrophoresis, connecting products with the correct sizes of the recovered and purified fragments to a vector after enzyme digestion for escherichia coli transformation, and selecting positive monoclonal sequencing. Obtaining a correct transformation vector pCA1301-LOG1, and transforming the rice mutant LOG1 callus in an Agrobacterium tumefaciens (Agrobacterium tumefaciens) strain LBA4404 by an electric shock method. The callus induced by the mutant seeds is cultured for 3 weeks by an induction culture medium, and the vigorous growing callus is selected to be used as a transformation receptor. The rice calli were infected with LBA4404 strain containing recombinant plasmid vector, cultured in dark at 25 ℃ for 3 days, and then cultured on the screening medium containing 300mg/LG 418. Selection of resistant calli was cultured on pre-differentiation medium containing 250mg/L G418 for about 10 days. The pre-differentiated calli were transferred to differentiation medium and cultured under light at 25 ℃. Obtaining resistant transgenic plants in about two months, and transplanting the resistant transgenic plants to a field for continuous growth. The plants were phenotypically identified and observed at maturity and found to recover from log1 grain size (fig. 2), with the data shown in table 2 below. It was shown that the present invention obtained transgenic rice that restored the normal phenotype of the log1 mutant (FIG. 2).
TABLE 2
| Length of grain | Grain width | Thousand seed weight | |
| wt | 10.52±0.21 | 2.81±0.13 | 33.56±0.39 |
| log1 mutant | 13.06±0.30 | 3.03±0.12 | 39.12±0.56 |
| Resistant transgenic plants | 10.81±0.15 | 2.78±0.17 | 33.71±0.42 |
Finally, it is also noted that the above-mentioned lists merely illustrate a few specific embodiments of the invention. It is obvious that the invention is not limited to the above embodiments, but that many variations are possible. All modifications which can be derived or suggested by a person skilled in the art from the disclosure of the present invention are to be considered within the scope of the invention.
Sequence listing
<110> institute of Rice research in China
<120> rice long-grain gene LOG1 and application thereof
<160> 4
<170> SIPOSequenceListing 1.0
<210> 1
<211> 5880
<212> DNA
<213> Rice (Oryza sativa L.)
<400> 1
atggggcggg ggaaggtgca gctgaagcgg atagagaaca agatcaacag gcaggtgacg 60
ttctccaaga ggaggaatgg attgctgaag aaggcgcacg agatctccgt cctctgcgac 120
gccgaggtcg ccgccatcgt cttctccccc aagggcaagc tctacgagta cgccactgac 180
tccaggtaca cctatagcct gatcgattcc ttgccatttc tggagcacct gagcccgcct 240
gagatggatg aactcaataa ttctcctagc tttttgccgt ttctggccgg ccgcttccat 300
ggatttcagc tgctgctctc tgaatttgcc atgtttggtt tgtgcatctg gttgcatgtg 360
tgctacagct gatgatttta tgcttcacta gtgctgcaac acatgcaaca cagtccaatc 420
caagctagct cagctactcc aggaggagga gtcagctgca tgcctcctca tttcgggaaa 480
tcatctctct ctctttctat tttttccctt tcctttcttg tttccccttt ttctttcctg 540
ctagcttgct tctctctcta caccctgcat accctttttg ggattgatgg ccggttttaa 600
ttaatttccc ctcctttctt tagttaatta ctaccctact actaattcat ctcctcccga 660
tcctctcaat ttctcatcat atggagtact actaaaatat atatatgtag tacgcacttg 720
gtgtggccgg tctttgttaa catcatgtgc ttgcttagat tcaaggggat tatatatata 780
agaggatcta ctactaagtg tactttgtga ttacatcagc tatctatgta gctgatgatc 840
ttgctttggt cttttgctga tccattgagc tagtgctaat tctcatcacc cttccatttg 900
atgtactgca aatagttttt tattcattcg cttcttctcc ctcttcccca tgtgatttga 960
ttgctagctc cttcctgact tttatttggg ttgatttcgt cacatcttcc tctcaaatta 1020
atctcatgcg cgtgcctttg ttctccacct gtcaagaaat taattaaaga atatatcaca 1080
cgaattacga ggagtccacc accagtgtca tgcactcctc cctattgttc acagtcacag 1140
ctgatgcatg caaaacatct tctgtttctc aagtaattaa ttcccctctt cttgtgaggc 1200
ttggcttaaa attactcaaa cagctgtttt tcttgcttat gcatcatctt cctcagatac 1260
caaaacgttg gagatggaca gatctggcct ttcctcttcc cttctctctc atcctcttct 1320
atgtttctgc ctcttcttat gatgaatctt tatttggtat tattcacgtt aagcatctct 1380
ttaaagagcc ctagatcaaa tttgatctat ctcttttggt ttaacttcat ggccagttgc 1440
ttgtgtgttt tggtctagaa cgaccacgta cgcgagatat ttccttttct gctcagctga 1500
ttaattaata atgatgggat aattcactaa ttattctgga gttagatcaa gagatgtact 1560
taaacatcct gtgacacgat gacttcaaaa aataatcttg ctctctgtaa aatcacataa 1620
tagctagcca tcgcgcgtgt ttcagaaaaa taatttatct tgcttgctag cttttgctca 1680
aaatgcgttt ctgacgtcta ggtaaagagg acttttacgg tcgtctgtgt tgtactggaa 1740
cagtgccctg cacactcccc agcgtcaaga tcagtctaag gcgcttcatt tctagctcag 1800
aaatcttcac ttatttgatc ttaattatag atcttcctga gatagatcta agggtgttcc 1860
agtaaaacag aagaagtcaa gttcttcagg atcctcgtga tataattaaa ctgtagtctc 1920
acaatcttac tgatcatcaa ggagtctgta aacacctata cgtacggcta taggattatg 1980
gggaggttgg tctaaaataa aggtgttaaa ttacttgcaa ttatcaagat gtgcaaatca 2040
tcacccagta attcaatggt gctattatat atagtctgtt ttgaacagtt ccactggttt 2100
tgttagggtc cttgctgatg gctgtttaac ggtgctgtta tatataatct tgattatttt 2160
ttttgctaaa ttttcctaag caacccactt caaattttgg aatgggtgaa ttcttatgtt 2220
cacaccctca tatcacatgt gtcatgagga gctttttttt cgtcacacag tcccatacaa 2280
cgtccctgtt ttcttgaaga taagataatt catgcatgca cttgaatata accgagtgtt 2340
ttcatgtgca tgctttttat ttggctgtgt cgagtgttta gacagcaaaa gagctcttgt 2400
gcatacaaat gccgatatct atatattagt agtcttaatc cacatttcag ctttaattgc 2460
ttggaatagt ttggtaggca ttcctagctt cagaccattt aaatatactt cctaaacgtt 2520
gtcatgagaa tgctgaatgc aaagtattta catatatagt ggactaattt cagattctgc 2580
tcatgatggt tttccctgca ggatggacaa aatccttgaa cgttatgagc gctattcata 2640
tgctgaaaag gctcttattt cagctgaatc cgagagtgag gtaaaacaca gtaggttgta 2700
tgatatataa cttttacatg aagggaatat tattgttcct tttctattgc attagcattt 2760
caatcatatc atttccttgt ggccttttaa atcacatgtt aacaaagcag tttttctgaa 2820
ttctatcgtt ctacttgcca aaactagatg ttggtaacct tctgttttat actttttaac 2880
ttcaaacctc gtgtcttttg cactctggca tatttcttaa aatttctact tattgggata 2940
tcatgcagga gagtcctcat tcccttcatt taatcccctc aaattttatt tattatggtc 3000
atacaattaa tgggccacat gcatggaagt cgatcgccag cttgcgatca gcatcctatc 3060
attaatgtta tgatctatac taattcttct gtaaataaat taaaattgca ctagttttta 3120
actaaatgct actattctat ttttttatcc atactccgtt tgagtggttt cttttgtgtt 3180
tcatgaagca atataatatg cttaagaaaa catagaaaca tgcacaatac aataatagcg 3240
ctggtagaat ccggttggag accaaatgtt tcgatattct aagtgcatat ttatatatac 3300
taagtaaaac tgtaatcata aacatgcata tggactgtga tcgatgaaat gtttcttcat 3360
tttttttttc aagataactc ttccccagtt gacaacatgc actgcatcca gaagtactca 3420
tggtatttgt tttcagtact gtttgatgag taaaactttg gtaagtgtta aaaaatgatg 3480
ataagtgcaa atatttgttc ctttaccact tccaccttct tggagtaaag aaaaaaatgt 3540
gtgtggtttg gcaaagttgt tgcatatatc acggcattag tgtgcggata atgcgaataa 3600
atgcatcaat aagcactagg agtaagttaa ttagttctct accttaattg aaaagttcat 3660
gtacatttaa aaaaattaaa catgcctctt tttatcattt gttgaatttc tacatttatc 3720
aagacagcaa aggagttctc atctactaat aaattaattt aaggagctat aactccctta 3780
attttatcaa atattcacat acacggttgc atttgaggat tttctatata ctccactaaa 3840
tgaggatgca tttaggttct ttataatatt ttatttccat cgtttcatca tctgtttatg 3900
tttacagtaa tttagtaata gttctctaat tttgacagta atggttggct cacagaaaac 3960
atcaaccaac catatatatt ggtttatact gacaaagagt ttttttttgt acagggaaat 4020
tggtgccatg aatacaggaa acttaaggca aagattgaga ccatacaaaa atgtcacaag 4080
taatcgatat atatatccaa tttctataat aatgttaaat aaatttcgaa ctaattgcac 4140
taattattat ttttaaacca atttaaaaga gaacttccgg tattgactgc tgaagatgac 4200
gtgaaactaa cttatgtatc ttgtgtagac acctcatggg agaggatcta gaatccctga 4260
atctcaaaga actccaacag ctagagcagc agctggagag ttcattgaag cacataatat 4320
caagaaaggt agtatttttt tggatagtgg aatcttttta tttgaactcc cgacctctac 4380
acaccaaatg tacagaaaaa aaggagcaaa ttattaaaag aaacgtagta atatatttag 4440
catgttctgt ttcaaactgg actgaattga aataattcat gcatgcgcat gcctgagtta 4500
cataaatata tatccaaatg tcttgttgtc actgctactt tgaaacaaat gttctttttt 4560
ttttcctttt tgagaaaata tatgagtaaa tattttcagc acattctgga ggaacaatta 4620
ccaaatataa actttttagc acattctgga gtacaattac caaatatgac ggactaacta 4680
gaacaatatg tttgtgtttg cagagccacc ttatgcttga gtccatttcc gagctgcaga 4740
aaaaggtgac caattgaact gcactactgt gtatgcatgt atatatgaat cgtgcatgca 4800
aaccgttgtt agctagtagt tagttatagt ttactcttgg ttagtctcaa ctgatagtgc 4860
agttttcttg catgagtgca ggagaggtca ctgcaggagg agaacaaggc tctgcagaag 4920
gaagtaagtt gtcgaaaata tagtatttac tgctgcttat attaattaat tcattctgct 4980
tgactagcca atggaggcac taaacataat taattcatgc gcattggttt tgatcaattg 5040
atttatcttg ttgacaattt tccaatctat ccatgggttt aattactagt tttgtcaatg 5100
gaggcattag agataattaa ttcatatgcg cactcgaaag tttgaaattg ctgtagttta 5160
cttgttgggt ggatcatatc gccaataatt ttttccccca cagttgaccg caaatgagaa 5220
gtaattttct ttgccttcta gacttgacgg ctattttttt agcgtgttgg caattgaaca 5280
tatgaccttg ggttaaaacc acacatcttt tatcattgcg ctatcaaata catctcgata 5340
aatttgtccc ttactagcgg tttatgtggt ttccttggtg gtcggtttgt gcctcttaga 5400
aaaaaatcgt gatgtttgta gtgtgcgcat gcagctggtg gagaggcaga agaatgtgag 5460
gggccagcag caagtagggc agtgggacca aacccaggtc caggcccagg cccaagccca 5520
accccaagcc cagacaagct cctcctcctc ctccatgctg agggatcagc aggcacttct 5580
tccaccacaa aatatctggt acgtatatgt ctgcattcat gtctcatcag ttgctattca 5640
gttataaatt aatctgttac tgcactaatt aagatcgatc acacccttgt taagttctaa 5700
gtctctaatt aattaactgc atgtatataa ttatgcacat ctgtgcatag tagctacccg 5760
ccggtgatga tgggcgagag aaatgatgcg gcggcggcgg cggcggtggc ggcgcagggc 5820
caggtgcaac tccgcatcgg aggtcttccg ccatggatgc tgagccacct caatgcttaa 5880
<210> 2
<211> 5880
<212> DNA
<213> Rice (Oryza sativa L.)
<400> 2
atggggcggg ggaaggtgca gctgaagcgg atagagaaca agatcaacag gcaggtgacg 60
ttctccaaga ggaggaatgg attgctgaag aaggcgcacg agatctccgt cctctgcgac 120
gccgaggtcg ccgccatcgt cttctccccc aagggcaagc tctacgagta cgccactgac 180
tccacgtaca cctatagcct gatcgattcc ttgccatttc tggagcacct gagcccgcct 240
gagatggatg aactcaataa ttctcctagc tttttgccgt ttctggccgg ccgcttccat 300
ggatttcagc tgctgctctc tgaatttgcc atgtttggtt tgtgcatctg gttgcatgtg 360
tgctacagct gatgatttta tgcttcacta gtgctgcaac acatgcaaca cagtccaatc 420
caagctagct cagctactcc aggaggagga gtcagctgca tgcctcctca tttcgggaaa 480
tcatctctct ctctttctat tttttccctt tcctttcttg tttccccttt ttctttcctg 540
ctagcttgct tctctctcta caccctgcat accctttttg ggattgatgg ccggttttaa 600
ttaatttccc ctcctttctt tagttaatta ctaccctact actaattcat ctcctcccga 660
tcctctcaat ttctcatcat atggagtact actaaaatat atatatgtag tacgcacttg 720
gtgtggccgg tctttgttaa catcatgtgc ttgcttagat tcaaggggat tatatatata 780
agaggatcta ctactaagtg tactttgtga ttacatcagc tatctatgta gctgatgatc 840
ttgctttggt cttttgctga tccattgagc tagtgctaat tctcatcacc cttccatttg 900
atgtactgca aatagttttt tattcattcg cttcttctcc ctcttcccca tgtgatttga 960
ttgctagctc cttcctgact tttatttggg ttgatttcgt cacatcttcc tctcaaatta 1020
atctcatgcg cgtgcctttg ttctccacct gtcaagaaat taattaaaga atatatcaca 1080
cgaattacga ggagtccacc accagtgtca tgcactcctc cctattgttc acagtcacag 1140
ctgatgcatg caaaacatct tctgtttctc aagtaattaa ttcccctctt cttgtgaggc 1200
ttggcttaaa attactcaaa cagctgtttt tcttgcttat gcatcatctt cctcagatac 1260
caaaacgttg gagatggaca gatctggcct ttcctcttcc cttctctctc atcctcttct 1320
atgtttctgc ctcttcttat gatgaatctt tatttggtat tattcacgtt aagcatctct 1380
ttaaagagcc ctagatcaaa tttgatctat ctcttttggt ttaacttcat ggccagttgc 1440
ttgtgtgttt tggtctagaa cgaccacgta cgcgagatat ttccttttct gctcagctga 1500
ttaattaata atgatgggat aattcactaa ttattctgga gttagatcaa gagatgtact 1560
taaacatcct gtgacacgat gacttcaaaa aataatcttg ctctctgtaa aatcacataa 1620
tagctagcca tcgcgcgtgt ttcagaaaaa taatttatct tgcttgctag cttttgctca 1680
aaatgcgttt ctgacgtcta ggtaaagagg acttttacgg tcgtctgtgt tgtactggaa 1740
cagtgccctg cacactcccc agcgtcaaga tcagtctaag gcgcttcatt tctagctcag 1800
aaatcttcac ttatttgatc ttaattatag atcttcctga gatagatcta agggtgttcc 1860
agtaaaacag aagaagtcaa gttcttcagg atcctcgtga tataattaaa ctgtagtctc 1920
acaatcttac tgatcatcaa ggagtctgta aacacctata cgtacggcta taggattatg 1980
gggaggttgg tctaaaataa aggtgttaaa ttacttgcaa ttatcaagat gtgcaaatca 2040
tcacccagta attcaatggt gctattatat atagtctgtt ttgaacagtt ccactggttt 2100
tgttagggtc cttgctgatg gctgtttaac ggtgctgtta tatataatct tgattatttt 2160
ttttgctaaa ttttcctaag caacccactt caaattttgg aatgggtgaa ttcttatgtt 2220
cacaccctca tatcacatgt gtcatgagga gctttttttt cgtcacacag tcccatacaa 2280
cgtccctgtt ttcttgaaga taagataatt catgcatgca cttgaatata accgagtgtt 2340
ttcatgtgca tgctttttat ttggctgtgt cgagtgttta gacagcaaaa gagctcttgt 2400
gcatacaaat gccgatatct atatattagt agtcttaatc cacatttcag ctttaattgc 2460
ttggaatagt ttggtaggca ttcctagctt cagaccattt aaatatactt cctaaacgtt 2520
gtcatgagaa tgctgaatgc aaagtattta catatatagt ggactaattt cagattctgc 2580
tcatgatggt tttccctgca ggatggacaa aatccttgaa cgttatgagc gctattcata 2640
tgctgaaaag gctcttattt cagctgaatc cgagagtgag gtaaaacaca gtaggttgta 2700
tgatatataa cttttacatg aagggaatat tattgttcct tttctattgc attagcattt 2760
caatcatatc atttccttgt ggccttttaa atcacatgtt aacaaagcag tttttctgaa 2820
ttctatcgtt ctacttgcca aaactagatg ttggtaacct tctgttttat actttttaac 2880
ttcaaacctc gtgtcttttg cactctggca tatttcttaa aatttctact tattgggata 2940
tcatgcagga gagtcctcat tcccttcatt taatcccctc aaattttatt tattatggtc 3000
atacaattaa tgggccacat gcatggaagt cgatcgccag cttgcgatca gcatcctatc 3060
attaatgtta tgatctatac taattcttct gtaaataaat taaaattgca ctagttttta 3120
actaaatgct actattctat ttttttatcc atactccgtt tgagtggttt cttttgtgtt 3180
tcatgaagca atataatatg cttaagaaaa catagaaaca tgcacaatac aataatagcg 3240
ctggtagaat ccggttggag accaaatgtt tcgatattct aagtgcatat ttatatatac 3300
taagtaaaac tgtaatcata aacatgcata tggactgtga tcgatgaaat gtttcttcat 3360
tttttttttc aagataactc ttccccagtt gacaacatgc actgcatcca gaagtactca 3420
tggtatttgt tttcagtact gtttgatgag taaaactttg gtaagtgtta aaaaatgatg 3480
ataagtgcaa atatttgttc ctttaccact tccaccttct tggagtaaag aaaaaaatgt 3540
gtgtggtttg gcaaagttgt tgcatatatc acggcattag tgtgcggata atgcgaataa 3600
atgcatcaat aagcactagg agtaagttaa ttagttctct accttaattg aaaagttcat 3660
gtacatttaa aaaaattaaa catgcctctt tttatcattt gttgaatttc tacatttatc 3720
aagacagcaa aggagttctc atctactaat aaattaattt aaggagctat aactccctta 3780
attttatcaa atattcacat acacggttgc atttgaggat tttctatata ctccactaaa 3840
tgaggatgca tttaggttct ttataatatt ttatttccat cgtttcatca tctgtttatg 3900
tttacagtaa tttagtaata gttctctaat tttgacagta atggttggct cacagaaaac 3960
atcaaccaac catatatatt ggtttatact gacaaagagt ttttttttgt acagggaaat 4020
tggtgccatg aatacaggaa acttaaggca aagattgaga ccatacaaaa atgtcacaag 4080
taatcgatat atatatccaa tttctataat aatgttaaat aaatttcgaa ctaattgcac 4140
taattattat ttttaaacca atttaaaaga gaacttccgg tattgactgc tgaagatgac 4200
gtgaaactaa cttatgtatc ttgtgtagac acctcatggg agaggatcta gaatccctga 4260
atctcaaaga actccaacag ctagagcagc agctggagag ttcattgaag cacataatat 4320
caagaaaggt agtatttttt tggatagtgg aatcttttta tttgaactcc cgacctctac 4380
acaccaaatg tacagaaaaa aaggagcaaa ttattaaaag aaacgtagta atatatttag 4440
catgttctgt ttcaaactgg actgaattga aataattcat gcatgcgcat gcctgagtta 4500
cataaatata tatccaaatg tcttgttgtc actgctactt tgaaacaaat gttctttttt 4560
ttttcctttt tgagaaaata tatgagtaaa tattttcagc acattctgga ggaacaatta 4620
ccaaatataa actttttagc acattctgga gtacaattac caaatatgac ggactaacta 4680
gaacaatatg tttgtgtttg cagagccacc ttatgcttga gtccatttcc gagctgcaga 4740
aaaaggtgac caattgaact gcactactgt gtatgcatgt atatatgaat cgtgcatgca 4800
aaccgttgtt agctagtagt tagttatagt ttactcttgg ttagtctcaa ctgatagtgc 4860
agttttcttg catgagtgca ggagaggtca ctgcaggagg agaacaaggc tctgcagaag 4920
gaagtaagtt gtcgaaaata tagtatttac tgctgcttat attaattaat tcattctgct 4980
tgactagcca atggaggcac taaacataat taattcatgc gcattggttt tgatcaattg 5040
atttatcttg ttgacaattt tccaatctat ccatgggttt aattactagt tttgtcaatg 5100
gaggcattag agataattaa ttcatatgcg cactcgaaag tttgaaattg ctgtagttta 5160
cttgttgggt ggatcatatc gccaataatt ttttccccca cagttgaccg caaatgagaa 5220
gtaattttct ttgccttcta gacttgacgg ctattttttt agcgtgttgg caattgaaca 5280
tatgaccttg ggttaaaacc acacatcttt tatcattgcg ctatcaaata catctcgata 5340
aatttgtccc ttactagcgg tttatgtggt ttccttggtg gtcggtttgt gcctcttaga 5400
aaaaaatcgt gatgtttgta gtgtgcgcat gcagctggtg gagaggcaga agaatgtgag 5460
gggccagcag caagtagggc agtgggacca aacccaggtc caggcccagg cccaagccca 5520
accccaagcc cagacaagct cctcctcctc ctccatgctg agggatcagc aggcacttct 5580
tccaccacaa aatatctggt acgtatatgt ctgcattcat gtctcatcag ttgctattca 5640
gttataaatt aatctgttac tgcactaatt aagatcgatc acacccttgt taagttctaa 5700
gtctctaatt aattaactgc atgtatataa ttatgcacat ctgtgcatag tagctacccg 5760
ccggtgatga tgggcgagag aaatgatgcg gcggcggcgg cggcggtggc ggcgcagggc 5820
caggtgcaac tccgcatcgg aggtcttccg ccatggatgc tgagccacct caatgcttaa 5880
<210> 3
<211> 296
<212> PRT
<213> Rice (Oryza sativa L.)
<400> 3
Met Gly Arg Gly Lys Val Gln Leu Lys Arg Ile Glu Asn Lys Ile Asn
1 5 10 15
Arg Gln Val Thr Phe Ser Lys Arg Arg Asn Gly Leu Leu Lys Lys Ala
20 25 30
His Glu Ile Ser Val Leu Cys Asp Ala Glu Val Ala Ala Ile Val Phe
35 40 45
Ser Pro Lys Gly Lys Leu Tyr Glu Tyr Ala Thr Asp Ser Arg Met Asp
50 55 60
Lys Ile Leu Glu Arg Tyr Glu Arg Tyr Ser Tyr Ala Glu Lys Ala Leu
65 70 75 80
Ile Ser Ala Glu Ser Glu Ser Glu Ile Thr Leu Pro Gln Leu Thr Thr
85 90 95
Cys Thr Ala Ser Arg Ser Thr His Gly Ile Cys Phe Gln Tyr Cys Leu
100 105 110
Met Ser Lys Thr Leu Gly Asn Trp Cys His Glu Tyr Arg Lys Leu Lys
115 120 125
Ala Lys Ile Glu Thr Ile Gln Lys Cys His Lys His Leu Met Gly Glu
130 135 140
Asp Leu Glu Ser Leu Asn Leu Lys Glu Leu Gln Gln Leu Glu Gln Gln
145 150 155 160
Leu Glu Ser Ser Leu Lys His Ile Ile Ser Arg Lys Ser His Leu Met
165 170 175
Leu Glu Ser Ile Ser Glu Leu Gln Lys Lys Glu Arg Ser Leu Gln Glu
180 185 190
Glu Asn Lys Ala Leu Gln Lys Glu Leu Val Glu Arg Gln Lys Asn Val
195 200 205
Arg Gly Gln Gln Gln Val Gly Gln Trp Asp Gln Thr Gln Val Gln Ala
210 215 220
Gln Ala Gln Ala Gln Pro Gln Ala Gln Thr Ser Ser Ser Ser Ser Ser
225 230 235 240
Met Leu Arg Asp Gln Gln Ala Leu Leu Pro Pro Gln Asn Ile Cys Tyr
245 250 255
Pro Pro Val Met Met Gly Glu Arg Asn Asp Ala Ala Ala Ala Ala Ala
260 265 270
Val Ala Ala Gln Gly Gln Val Gln Leu Arg Ile Gly Gly Leu Pro Pro
275 280 285
Trp Met Leu Ser His Leu Asn Ala
290 295
<210> 4
<211> 296
<212> PRT
<213> Rice (Oryza sativa L.)
<400> 4
Met Gly Arg Gly Lys Val Gln Leu Lys Arg Ile Glu Asn Lys Ile Asn
1 5 10 15
Arg Gln Val Thr Phe Ser Lys Arg Arg Asn Gly Leu Leu Lys Lys Ala
20 25 30
His Glu Ile Ser Val Leu Cys Asp Ala Glu Val Ala Ala Ile Val Phe
35 40 45
Ser Pro Lys Gly Lys Leu Tyr Glu Tyr Ala Thr Asp Ser Thr Met Asp
50 55 60
Lys Ile Leu Glu Arg Tyr Glu Arg Tyr Ser Tyr Ala Glu Lys Ala Leu
65 70 75 80
Ile Ser Ala Glu Ser Glu Ser Glu Ile Thr Leu Pro Gln Leu Thr Thr
85 90 95
Cys Thr Ala Ser Arg Ser Thr His Gly Ile Cys Phe Gln Tyr Cys Leu
100 105 110
Met Ser Lys Thr Leu Gly Asn Trp Cys His Glu Tyr Arg Lys Leu Lys
115 120 125
Ala Lys Ile Glu Thr Ile Gln Lys Cys His Lys His Leu Met Gly Glu
130 135 140
Asp Leu Glu Ser Leu Asn Leu Lys Glu Leu Gln Gln Leu Glu Gln Gln
145 150 155 160
Leu Glu Ser Ser Leu Lys His Ile Ile Ser Arg Lys Ser His Leu Met
165 170 175
Leu Glu Ser Ile Ser Glu Leu Gln Lys Lys Glu Arg Ser Leu Gln Glu
180 185 190
Glu Asn Lys Ala Leu Gln Lys Glu Leu Val Glu Arg Gln Lys Asn Val
195 200 205
Arg Gly Gln Gln Gln Val Gly Gln Trp Asp Gln Thr Gln Val Gln Ala
210 215 220
Gln Ala Gln Ala Gln Pro Gln Ala Gln Thr Ser Ser Ser Ser Ser Ser
225 230 235 240
Met Leu Arg Asp Gln Gln Ala Leu Leu Pro Pro Gln Asn Ile Cys Tyr
245 250 255
Pro Pro Val Met Met Gly Glu Arg Asn Asp Ala Ala Ala Ala Ala Ala
260 265 270
Val Ala Ala Gln Gly Gln Val Gln Leu Arg Ile Gly Gly Leu Pro Pro
275 280 285
Trp Met Leu Ser His Leu Asn Ala
290 295
Claims (8)
1. Rice long grain gene LOG1, characterized by: the nucleotide sequence of the gene LOG1 is shown as SEQ ID NO: 2, the preparation method is as follows.
2. The rice long grain gene LOG 1-encoding protein of claim 1, which is characterized in that: the amino acid sequence of the protein is shown as SEQ ID NO: 4, the method is described in the specification.
3. A recombinant vector comprising the gene of claim 1.
4. A transformant containing the gene according to claim 1.
5. The application of the rice long-grain gene LOG1 is characterized in that: increasing the grain size of gramineous plants and increasing the yield.
6. The use of the rice long grain gene LOG1 as claimed in claim 5, wherein: the gramineous plants are rice, and the grain length and the grain width of rice grains are increased, so that the rice yield is increased.
7. A method for regulating and controlling the development of gramineous plant kernels is characterized in that: comprises transforming a graminaceous plant cell with the gene LG1 as described in claim 1, and then cultivating the transformed graminaceous plant cell into a plant, wherein the grain size of the plant is increased and the thousand kernel weight is increased.
8. The method of modulating grain development in a graminaceous plant according to claim 7, wherein said step of modulating grain development comprises: the grain length and the grain width of the grains are increased.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111101492.XA CN113817755B (en) | 2021-09-18 | 2021-09-18 | Rice long-grain gene LOG1 and application thereof |
| ZA2022/03310A ZA202203310B (en) | 2021-09-18 | 2022-03-22 | Rice long grain gene log1 and use thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111101492.XA CN113817755B (en) | 2021-09-18 | 2021-09-18 | Rice long-grain gene LOG1 and application thereof |
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| Publication Number | Publication Date |
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| CN113817755A true CN113817755A (en) | 2021-12-21 |
| CN113817755B CN113817755B (en) | 2023-03-31 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN202111101492.XA Active CN113817755B (en) | 2021-09-18 | 2021-09-18 | Rice long-grain gene LOG1 and application thereof |
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| ZA (1) | ZA202203310B (en) |
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| WO2013060136A1 (en) * | 2011-10-24 | 2013-05-02 | 南京农业大学 | Cloning and application of semi-dominant gene qgl3 capable of controlling grain length and grain weight of rice kernel |
| WO2015143972A1 (en) * | 2014-03-26 | 2015-10-01 | 湖南杂交水稻研究中心 | Rice panicle traits regulatory gene pt2 and application thereof |
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2021
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| CN113817755B (en) | 2023-03-31 |
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