CN114410663A - Application of corn ZmDek701 gene in regulation and control of plant grain quality and mutant thereof - Google Patents

Abstract

The invention relates to the technical field of botany and molecular biology, and particularly discloses application of a corn ZmDek701 gene in regulation and control of plant grain quality and a mutant thereof. The invention discovers that the ZmDek701 gene of the corn can regulate and control the quality of plant seeds, and the specific regulation and control of the quality of the plant seeds refers to any one of the following: (1) reducing the total starch content and/or starch particle size in the grain; (2) increasing the total protein content in the grains; (3) reducing the kernel. The invention also provides a corn ZmDek701 gene mutant, and the nucleotide sequence of the mutant is shown in SEQ ID NO. 2. The method not only can provide scientific basis for analyzing the development mechanism of grain formation, but also can provide theoretical guidance for genetic improvement and yield improvement of the corn.

Description

Application of corn ZmDek701 gene in regulation and control of plant grain quality and mutant thereof

Technical Field

The invention relates to the technical field of botany and molecular biology, in particular to application of a corn ZmDek701 gene in regulation and control of plant grain quality and a mutant thereof.

Background

Corn originates from central and south america, is one of important grain, feed, energy and industrial crops, and has large annual demand of seeds, high yield value and large proportion of the industrial yield value of crop species. Seeds are the major part of the harvest of cereal crops, and the development of seeds and the maturity of seeds are not only closely related to the harvest yield of the crop, but also directly affect the vigor of the seeds.

In the beginning of the 21 st century, the first crystal structures of bacterial RNA polymerase and yeast RNA polymerase II (pol II) were obtained by X-ray diffraction and their basic structures were analyzed, while the analysis of the crystal structures of yeast RNA polymerase I (pol I) took more than 10 years, and the structures of yeast RNA polymerase III (pol III) were obtained thanks to the low-temperature electron microscopy.

All RNA polymerases are multi-subunit combinations, and bacterial DNA-dependent RNA polymerases have five core subunits (β', β, ω, α; two α molecules in the core enzyme) which share homology among RNA polymerases from archaea and eukaryotes, but archaea RNA polymerases and eukaryotic organisms Pol I, II and III are more complex. In addition to these five core subunits, eukaryotes Pol I, Pol II, and Pol III share five additional subunits, forming a 10 subunit catalytic core component. The core component has a typical crab claw shape, which surrounds a central cleft, in which DNA is occluded, and has two channels, one for the entry of substrate NTPs and the other for the export of RNA products. Two tweezers, stabilizing the DNA at the downstream end and controlling the opening and closing of the cleft. In order for transcription to occur, the enzyme must maintain a transcription bubble bound to the isolated DNA strand, facilitate nucleotide incorporation, translocate along the template, stabilize the DNA: RNA strand, and finally allow the DNA strand to reanneal. These are achieved by many conserved elements of the active site, including fork loops, rudders, trigger loops, and bridge helixes, among others.

With the exception of the core component, all eukaryotic RNA polymerases share two additional, more distantly related subunits, which constitute the peripheral component of the polymerase. In Pol I and Pol III, the core region is further modified to heterodimer (Pol I and Pol III) and heterotrimer (Pol III) complexes. Thus, Pol I and Pol III comprise 14 and 17 subunits, respectively, compared to 12 subunits of Pol II enzyme. Relative to the core element, which is critical for transcriptional elongation, the peripheral components are involved in the initiation, termination and cleavage of transcription of RNA, mainly through protein-protein interactions or direct contact with nucleic acids.

Saccharomyces cerevisiae RNA polymerase II is relatively well defined because its 12 subunit genes RPB1-RPB12 have been cloned and characterized. Most of the molecular mass of RNA polymerase consists of the two largest subunits, RPB1 and RPB 2. These two subunits are homologous to the β' and β subunits of E.coli RNA polymerase and are believed to have similar functions. Similar to β', RPB1 appears to be involved in DNA binding, while both β and RPB2 appear to be involved in RNA catalysis. RPB3 has some sequence similarity to the e.coli alpha subunit, and, like alpha, appears to play a role in subunit assembly. RPB4-RPB12, also plays a key role in transcription, as most are essential for cell survival and are evolutionarily conserved, of these 9 small subunits, 5 RPB5, RPB6, RPB8, RPB10, and RPB12 are assembled into all 3 eukaryotic RNA polymerases. The remaining four subunits, RPB4, RPB7, RPB9, and RPB11, are uniquely expressed in RNA polymerase II. Three of RPB7, RPB9, and RPB11 have sequence similarity with subunits of other types of yeast RNA polymerases, RPB7 is related to the C25 subunit of RNA polymerase III; RPB9 is associated with the a12.2 subunit of RNA polymerase I; RPBll is associated with the AC19 subunit in RNA polymerases I and III.

Arabidopsis genome sequencing revealed the expected genes for Pol I, II and III catalytic subunits, but unexpectedly found two atypical largest subunit genes and two atypical second largest subunit genes. Furthermore, the five shared subunits of Pol I, II and III are normally encoded by a single gene in yeast and mammals, i.e. RPB5, RPB6, RPB8, RPB10 and RPB12, in arabidopsis by a multigene family, as are Pol II-specific subunits RPB3, RPB4, RPB7 and RPB 9.

In all organisms, the flow of genetic information is a two-step process: first, DNA is transcribed into RNA, which is then used as a template for protein synthesis during translation. In bacteria, archaea and eukaryotes, transcription is performed by multi-subunit RNA polymerases that share a core conserved structure. RNA polymerase takes DNA as a template and catalyzes the high-precision polymerization of RNA of an NTP building block through the assistance of transcription factors in the transcription initiation, extension and termination stages. The complexity of this highly dynamic process is represented by the complex network of protein-protein and protein-nucleic acid interactions in the transcription complex, and the large conformational changes that occur to RNA polymerase throughout the transcription cycle.

RNA polymerase activity was first discovered by Weiss and Gladstone in 1959, and when added to rat liver nuclear extracts, all 4 NTPs were integrated into RNA. Roeder and Rutter are the first to describe the transcription of eukaryotic genomes not one but three different enzymes Pol I, II and III.

Bacteria and archaea use a single type of RNA polymerase, while the genome of eukaryotes is transcribed by at least three specialized RNA polymerases that are specialized for different genes. Pol I transcribes ribosomal RNA precursors of mature 25/28S, 18S and 5.8S rRNAs, Pol II is responsible for transcription of messenger RNA (mrna) and many non-coding RNAs, while Pol III synthesizes small structural RNAs such as transfer RNA (trna), intra-splice U6 small nuclear RNA (snrna), ribosomal 5S rRNA and 7sl RNA. Higher plants have added two additional nuclear RNA polymerases, Pol IV and Pol V, which are used exclusively for sirna-mediated DNA methylation and gene silencing. The subunit composition of RNA polymerase is organized into three functional domains: catalytic core, assembly platform and ancillary specialized functions. Some common shared subunits are important for RNA polymerase function, such as transcription efficiency, enzyme stability, nuclear localization, or coordination and regulation of rRNA, mRNA, and tRNA synthesis.

The research on the action mechanism of key genes in the corn kernel development process not only can provide scientific basis for analyzing the development mechanism of kernel formation, but also can provide theoretical guidance for the genetic improvement and yield improvement of corn. Therefore, it is necessary to research.

Disclosure of Invention

The invention aims to provide an RPB10 subunit protein of coding DNA-directed RNA polymerase, the coding gene is ZmDek701, and the coding protein can regulate and control the quality of plant (corn) grains through verification.

Specifically, the technical scheme of the invention is as follows:

in a first aspect, the invention provides an application of a corn ZmDek701 gene or a biological material containing the same in regulating and controlling plant grain quality.

Specifically, the regulation and control of plant grain quality refers to any one of the following:

(1) reducing the total starch content and/or starch particle size in the grain;

(2) increasing the total protein content in the grains;

(3) reducing the kernel.

Particularly, the volume and the weight of the grains can be reduced.

In a second aspect, the use of the maize ZmDek701 gene, or a biological material comprising the same, in any one of:

(1) breeding plants with low total starch content and/or high total protein content in grains;

(2) preparing a plant having a grain with a low total starch content and/or a high total protein content;

(3) identifying or screening plants or germplasm resources thereof with low total starch content and/or high total protein content in grains.

Specifically, the corn ZmDek701 gene or a biological material containing the same can improve the content of prolamin and non-prolamin in plant grains.

In the invention, the biological material is an expression cassette, a vector, a host cell or a recombinant bacterium.

In the present invention, the plant is a cereal crop; preferably wheat, rice, maize, soybean or sorghum. More preferably corn.

In the invention, the wheat is wheat, barley, oat, rye, highland barley and the like, and the rice is indica rice, japonica rice, glutinous rice and the like.

In a third aspect, the invention provides a method for changing the quality of plant grains, which controls the expression of a ZmDek701 gene of a plant through a transgenic, crossing, backcrossing, selfing or asexual propagation method.

Preferably, the transgenosis comprises the step of silencing expression or reducing expression quantity of a ZmDek701 gene by using a DNA homologous recombination technology, a Cre/Loxp technology and a Crispr/Gas9 technology to obtain a transgenic plant line.

In a fourth aspect, the invention provides a corn ZmDek701 gene mutant, and the nucleotide sequence of the mutant is shown as SEQ ID NO. 2.

In a fifth aspect, the invention provides any one of the following applications of the maize ZmDek701 gene mutant:

(1) the application of the composition in reducing the total starch content and/or the starch grain size in corn grains;

(2) application in increasing the total protein content in corn grains;

(3) application in reducing corn kernels;

(4) the application in breeding corn with low total starch content and/or high total protein content in grains;

(5) use in the preparation of corn having a low total starch content and/or a high total protein content in the kernel;

(6) the application in identifying or screening the corn with low total starch content and/or high total protein content or germplasm resources thereof.

In a sixth aspect, the invention provides a method for preparing corn with low total starch content and/or high total protein content of grains, and particularly relates to a method for expressing the corn ZmDek701 gene mutant in the corn.

The invention has the beneficial effects that: the invention finds that the ZmDek701 gene of the corn can regulate and control the quality of plant (corn) grains, and particularly, the invention finds that the ZmDek701 gene of the corn can regulate and control the content of total starch in the plant (corn) grains and/or improve the content of total protein in the plant (corn) grains. The invention also discloses a ZmDek701 gene mutant. The discovery not only can provide scientific basis for analyzing the development mechanism of grain formation, but also can provide theoretical guidance for genetic improvement and yield improvement of the corn.

Drawings

FIG. 1 is a view of dek701(ZmDek701) mutant grain phenotype observation under different genetic backgrounds of the invention.

FIG. 2 is a section view of dek701 mutant grains according to the present invention;

wherein FIG. 2A is a comparison of dek701 and WT (wild-type) mature seed cross section;

FIG. 2B is a comparison of dek701 and paraffin sections of WT whole grain;

FIG. 2C is a partial enlarged comparison of dek701 and paraffin sections of WT whole kernel;

in the figure, En represents endosperm, Em represents embryo, EC represents endosperm cells, SC represents scutellum, LP represents leaf primordium, SAM represents apical (shoot tip) meristem, and RAM represents root tip meristem;

the white/black (scale) line segments in the figure represent: fig. 2A Bar 2.5 mm; fig. 2B Bar 1 mm; fig. 2C Bars 500 μm.

FIG. 3 is a diagram showing the result of the present BSR-Seq locating ZmDek701 gene on chromosome 8.

FIG. 4 is a fine mapping of the ZmDek701 gene of the present invention. Wherein, FIG. 4A is a fine mapping of ZmDek701 gene, and FIG. 4B is a semi-quantitative PCR method for analyzing the expression of ZmDek701 gene in WT and dek 701; GAPDH is the internal control.

FIG. 5 shows the measurement results of the hundred grain weight of Wild Type (WT) and dek701 grains, and the total starch and protein content of endosperm;

wherein, fig. 5A is a grain weight measurement result;

FIG. 5B shows the measurement results of total starch content in endosperm;

fig. 5C is a measurement of endosperm protein content, wherein dek represents dek701 grain;

in the figure, denotes T test, P value is less than 0.001; and represents T test, and the P value is less than 0.01.

FIG. 6 is an electrophoretogram of total protein of endosperm of Wild Type (WT) and dek701 according to the present invention.

FIG. 7 is a scanning electron microscope image of starch granules in the center region of endosperm of Wild Type (WT) and dek701 according to the present invention; wherein PM represents proteosome, and SG represents starch granule.

Detailed Description

Preferred embodiments of the present invention will be described in detail with reference to the following examples. It is to be understood that the following examples are given for illustrative purposes only and are not intended to limit the scope of the present invention. Various modifications and alterations of this invention will become apparent to those skilled in the art without departing from the spirit and scope of this invention.

The experimental procedures used in the following examples are all conventional procedures unless otherwise specified. Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.

Example 1 discovery procedure of the invention

Dek701 mutant grain phenotype observation under different genetic backgrounds:

the invention was originally developed by finding dek701-ref (dek701) mutant (ZmDek701 gene mutant maize line) in the field and crossing this mutant with B73, ZHENG58(Z58), Chang7-2 and Mo17 to produce F₁And (5) plant growing. Then selfing to obtain F₂The population, found to show normal and small phenotype in different backgrounds, indicates that the mutation site can be stably inherited and is not influenced by the genetic background (see figure 1).

Cross-sectional observation of mature seeds (see FIG. 2) revealed that dek701 seeds were significantly smaller than WT (wild type B73), but both embryo and endosperm tissues were clearly visible, indicating that the organogenesis was essentially normal (see FIG. 2A). Paraffin sections of the entire grain from 16DAP (16 days of pollination) showed that dek701 had delayed embryo and endosperm development relative to WT (see fig. 2B), but the embryos retained the ability to differentiate, forming typical embryo structures including the scutellum, leaf primordia, shoot apical meristem and root apical meristem (see fig. 2C).

Localization of the ZmDek701 gene:

for cloning candidate genes, F hybridized with B73 was taken₂And (3) separating seeds for generation, namely 15 seeds of the normal seeds and 15 seeds of the mutant seeds respectively, mixing pools respectively, extracting RNA, and positioning the candidate genes on chromosome 8 through BSR-Seq (see figure 3).

Sequence analysis of the ZmDek701 gene (see fig. 4):

7680 mutant grains are collected from an F2 generation population of dek701 multiplied by B73, 3840 mutant grains are collected from an F2 generation population of dek701 multiplied by Z58, single grain DNA is extracted, fine positioning is carried out by utilizing polymorphism markers in an initial positioning interval, the interval is narrowed to 22kb, and 4 coding protein genes are contained (see figure 4A). Then, by genome re-sequencing, a large difference occurred in the first 218bp of the gene sequence of dek701, compared to the B73 reference genome sequence (nucleotide sequence see https:// www.maizegdb.org /) (V3). The sequence of the unmutated ZmDek701 gene is shown in SEQ ID NO. 1. The sequence of the ZmDek701 gene after mutation is shown as SEQ ID NO. 2.

The differential gene fragment resulted in incomplete ZmDek701 gene, missing the 5' UTR region and the first exon region of the gene, resulting in transcript deletion (see fig. 4B). The expression of the ZmDek701 gene in WT and dek701 was analyzed using a semi-quantitative PCR method, and in the homozygous mutant, no transcript of the ZmDek701 gene was detected (see fig. 4B).

Since dek701 kernels had serious developmental defects, their hundredth weight, total starch content and protein content were also measured in this example (see FIG. 5). Referring specifically to fig. 5A, 5B, 5C, dek701 had a hundred weight of only 48.16% of wild type. The endosperm component is changed in dek701, and the total starch content is obviously reduced by 13.99% compared with the wild type. The total protein amount in mature dek701 endosperm was significantly increased by 17.45%, prolamin by 20.03%, and non-prolamin by a slight increase of 15.71%. It is shown that dek701 mutation has great influence on the accumulation of nutrients in the endosperm of corn kernels.

Specific hundred particle weight test data are shown in table 1.

TABLE 1

The specific total starch content, total protein content, prolamin, non-prolamin test modes were as follows:

in order to accurately measure the content difference between Total starch and Total protein, the invention adopts a kit (Total starch measuring Total static assay kit (Megazyme, K-TSTA-100A); BCA kit (Total gold, DQ111)) to measure the Total protein content, 50 grains are taken from three groups (wild type, WT) of normal materials (wild type, WT) and ground in liquid nitrogen, then vacuum-dried, then 100mg is weighed for starch measurement, 50mg is weighed for protein measurement), and three groups (50 grains are taken from each group of samples, ground in liquid nitrogen, vacuum-dried, then 100mg is weighed for starch measurement, and 50mg is weighed for protein measurement) of mutant materials (dek), and the results are shown in figure 5B and figure 5C.

Total starch content was determined using the Total starch assay kit (Megazyme, K-TSTA-100A), as directed by the instructions, in which the self-prepared solutions were as follows:

1. sodium acetate buffer (1.2M, pH 3.8)

69.6mL of glacial acetic acid was adjusted to pH 3.8 with 4M NaOH and the volume of deionized water was adjusted to 1L. Can be stored at room temperature for 12 months.

2. Sodium acetate buffer (100mM, calcium chloride 5mM, pH 5.0)

5.8mL of glacial acetic acid and 0.74g of calcium chloride dihydrate, wherein the volume of the deionized water is constant to 1L after the glacial acetic acid and the calcium chloride dihydrate are completely dissolved, the pH value is adjusted to 5 by using 1M NaOH, and the mixture can be stored for 6 months at 4 ℃.

3. Potassium hydroxide solution (2M)

112.2g KOH was weighed and dissolved in 900mL deionized water, and the volume was 1L after stirring and dissolving completely. Storing at room temperature, and storing in a sealed container.

The specific method for extracting total protein, alcohol soluble protein and non-alcohol soluble protein comprises the following steps:

1. weighing 50mg of the drained powder, filling the powder into an EP tube, adding 1mL of petroleum ether, uniformly mixing by oscillation, and incubating for 1h by oscillation at 4 ℃ to achieve the purpose of degreasing.

2.13200rpm for 12min, discard the supernatant (pipette off the supernatant), and pump the pellet on a freeze-pump dryer.

3. Sodium borate solution (1 mL), 2% mercaptoethanol (20. mu.L, to which the sample to be electrophoresed must be added) was added and incubated overnight at 37 ℃ in a shaker.

4.13200rpm for 12min, and aspirating all supernatant, i.e., total protein, into a new EP tube.

5. And (3) sucking 300 mu L of total protein, adding 700 mu L of absolute ethyl alcohol, uniformly mixing at room temperature, and then incubating for 2h in a shaking table.

Centrifuging at 6.13200rpm for 12min, sucking all the supernatant into a new EP tube, and vacuum-drying to obtain prolamin (zein).

7. Washing the rest precipitate with 70% ethanol twice, centrifuging at 13200rpm for 5min, air drying in a fume hood, and dissolving with 200 μ L IPG buffer to obtain non-prolamin.

8. Total protein, prolamin and non-prolamin were separately vacuum dried on a freeze-pump dryer and tested by dissolving in 200. mu.L IPG buffer.

Specific total starch test data are shown in table 2.

TABLE 2

Specific total protein, prolamin, non-prolamin test data are shown in table 3.

TABLE 3

This example performed electrophoretic testing of Wild Type (WT) and dek701 endosperm total protein, and 19kD α -prolamin, 22kD α -prolamin and 27kD α -prolamin were relatively enriched in the electropherogram of total protein at dek701 (see FIG. 6).

This example also showed scanning electron microscopy of starch granules from the central region of endosperm in Wild Type (WT) and dek 701. Electron microscopy of the mature endosperm showed that the starch grain of the dek701 endosperm was smaller than that of the wild type (see FIG. 7).

Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those 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> institute of crop science of Chinese academy of agricultural sciences

Application of <120> corn ZmDek701 gene in regulation and control of plant grain quality and mutant thereof

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caagtaccca acgcgtcgtg attcgggtac accgcttgct agactacccg actaccagcc 300

gcgtctcgag catgactgcg acttagggtt ttggtggtga cgacgttcct tagggttttg 360

atttttcgtt aggtttgttc ttatgttgag aatctcgggg ggagagagac attaccgtga 420

aaagtgagtt gaactaaggt gcgtttgttt ctcgctattt caggttattg ggaacaagtg 480

ggatgtctac ctcgatctcc tccaggctga ctactcagaa gggtaaaacc tcgtgtcttt 540

tgttcacaag ccttatattg ctgttttcta actgcatctc gagtggtgtt tctgttgcat 600

cgtgctatga ggtaaagata tatgctactc ctggataact gaacacaact gcacaagccg 660

tcaagccaaa aggataaatt taccatgctt cttctcccta tatagtatat ataaataaat 720

atattcattt attagttctg tcaattcagc tctttcctaa aatttgttgt gaaacctggc 780

atacgagatc atttgtacat tatcttcagt gtggtagctc atgttaattg aaacacttca 840

gtaatctttt attataaaat tcggactcta ggctgtcaag aatgacactg caatatgctg 900

tatatctcgt tacggacaaa gataaccaat agagtgtatt gatctgtagt taatcaataa 960

gagcttaggt acactgattc agcagcagta gcagtgtgtc attgactata cgcctgacaa 1020

tgattaacag tttactgacc agaggtcact gatctgtagg tggatttctt tgtacagaat 1080

taagagattt gtagcctata agccttattt gtatgctttc aaattgtcat cgtgtggcct 1140

ccctacacct acatgttatc ttagttcaga ttagctaaga tatatatgtc cgtaacttca 1200

ttttgtttct ttttgtgccc ttctaggatc tcatgcttaa ccttctatca tatttataca 1260

tcctgaacgt tgatagttga aattaatcat tacataatgc tattttatta ctcatggcaa 1320

cgaaattaca gaagtttgtg aaacttaaac catccttatc ctgcaaccaa ctactctcat 1380

ccttacttaa ggcccagggt tcagatgagg tggcaaccac tgcactaaaa caccaaaacg 1440

aaccctgagt tcagatgata tgatcctaca taaaggaaca tgtttaacag tacatagaag 1500

acacatttgg ggcctgtttg tgagtgaagt aattttgtag tttctaggcg ataccatggt 1560

ttttagtaat accatagcat tttttgccat caggtgtttg gttgcattct tgaaaactat 1620

gttttcaaaa ttatgatatt ctagatagta gataccatgg tatttttaga gtattggaaa 1680

caccactcta acccaaagtt ttcatggcat gactagcttt tgcctaatat catggtttac 1740

aatattgtat ccaaacaatg tttctcaaaa tcatggtatt ttcagagctg accaaaacta 1800

cggtattgtc atgacaattg tgaagtatag tattgtaaaa ccatggtttt gaaaaacatc 1860

gtagccaaac atgctcttga ttgtgagccc aagaaactaa atccattaaa caagtggaca 1920

cctatcatgg gtgcagtggg ctaggccata taggctgggt cagttagtat taaaggttca 1980

ttagggtttt ggttagagat aagatgagct atcttgcttt gggggtcaag taaggctcta 2040

gcgatacaag gaggggtatt gtatcaatta aatcaagcaa gaagaaatag ctataaaagg 2100

ctgtctccag ctgattttgc ttatggtcgt gcaaaacatt gttttgcact atagactgca 2160

ctgtttacag agtgaagttt aaaatagaag atgagatggg agataggatg aggctgtctt 2220

cggtggatcc tataaaatag gagacacgga actgtagatg acactttttg acattgtctg 2280

cagagtagag tttgaaatag gggatgggat agggaatgtg atagaggatc tattggagat 2340

gacctaagtt gctagagaca gcctaaggaa aggtgatctt tcaaatatca ataaatcaag 2400

gaagaaatag atctattatc ccagtcccct tctgtcccta cccttaaggt ggctgtctga 2460

tgtcgggctt cgtaagttgg ggccatccag atcaaaatcc ataagatctt actatatgga 2520

aatgtacctc catgtctgca attcttcata tatgtttatt tgtgattttt cctccaacat 2580

gctggggagt ggggagctac atatcatcat atgaagagcc agccttattc tctgacgtga 2640

gggcagataa gatgggataa tccttttgag cctgctcata ctccaaaact gcatctcctc 2700

actaattagc gatagcactt ctgctaggtt aggagaccct aaagcacact ggttgcggtg 2760

gttacaaatc gaccatgctc ctaagagatg gagttgagcc ccttcctaac ttgtccatct 2820

actgtttcgt ttgcactatt gccaccaatc attaaaggaa aactcatgtc aacaataatc 2880

taaatgtact agtgtattat gattttgtgc tatgtgttgc agttggtaaa gattttgacc 2940

tactgagttc cctgagcata caagggctcc aagtccttgt tactgattga attgtatttt 3000

ctgtcaagta tcgttaaaaa ttatctgtat ttttctgtgc tttctattgc cagggatgct 3060

ctggatgctt tggaattgtt ccgctactgc tgcaggcgaa tgctcatgac acatgttgac 3120

ctcattgaga agttgctcaa ctacaacagt aagttggttt ttgagatgtt attcacgttg 3180

tgattttcat ttatggctgc accacttata gatgtatcaa tctttatttt cagccctaga 3240

gaagaccgag acaagttaag cgagcacatc atgctccaaa aacactactg tttggcagta 3300

tcatagagct taggtagtat gttgtctctt ctctatcagt agaaatgggt tgtgagtgtg 3360

aacatgcttt tgctcatgag cattcaagga cttttcatgc gacagtgctt gttgccacct 3420

atgcaacgat gttggatatt ttaagtgata taattgctaa aagaaagcta atgtatttgt 3480

gtgaattggg tcgataatat ttcgttgaca gatgtgcgtt tttggttgga ctacatttag 3540

tgatgcatgc tggtacatat tttattattg tattctataa ctctaatcgt catttctgat 3600

acataggaga tattgctgtc tctatatccg aagtattaat attattcaat cgcatgcggc 3660

cgttctggta gaggttctaa tgatgtccag caggcgatgg cgttacctgc cagtaccaga 3720

caaccaaaca tcaatcaacg aaagagactt gaaagggcaa tatcgagagt ttaccttgag 3780

actgcctttg ttgcctcgcc aactcatcgt agcattgccg tctggtaagg tatcagtccg 3840

tggccagaaa aaataggcaa tggtggaata cattatgctg ctttggttca gccacaaaag 3900

aggcaacgtt aatgggaata tctaccctag gtatctgtta catgcaatga tcagcgggtt 3960

ttgtttggtt gcaacggagg tgaacggcgt ggcattgaat cacgactagt ggggagcggt 4020

aatgattttt tcactccagg tggtgaagag ccgacatcga ggtgccaaac ctttatgtcg 4080

atgtgtcttg gggaagatta accttttatc catagagtaa cttttatttg tt 4132

Claims (10)

1. The corn ZmDek701 gene or a biological material containing the same is applied to the regulation and control of the quality of plant grains.

2. The use of claim 1, wherein said modulating plant grain quality refers to any of:

(1) reducing the total starch content and/or starch particle size in the grain;

(2) increasing the total protein content in the grains;

(3) reducing the kernel.

3. The application of the corn ZmDek701 gene or the biological material containing the same in any one of the following items:

(1) breeding plants with low total starch content and/or high total protein content in grains;

(2) preparing a plant having a grain with a low total starch content and/or a high total protein content;

(3) identifying or screening plants or germplasm resources thereof with low total starch content and/or high total protein content in grains.

4. The use according to any one of claims 1 to 3, wherein the biological material is an expression cassette, a vector, a host cell or a recombinant bacterium.

5. Use according to any one of claims 1 to 4, wherein the plant is a cereal crop; preferably wheat, rice, maize, soybean or sorghum; more preferably corn.

6. The method for changing the quality of plant grains is characterized in that the expression of the ZmDek701 gene of the plant is controlled by a transgenic, hybridization, backcross, selfing or asexual propagation method.

7. The method of claim 6, wherein the transgene comprises silent expression or reduced expression of ZmDek701 gene by using DNA homologous recombination technology, Cre/Loxp technology and Crispr/Gas9 technology to obtain a transgenic plant line.

8. The corn ZmDek701 gene mutant is characterized in that the nucleotide sequence is shown as SEQ ID NO. 2.

9. The corn ZmDek701 gene mutant of claim 8, which is applied to any one of the following applications:

(1) the application of the composition in reducing the total starch content and/or the starch grain size in corn grains;

(2) application in increasing the total protein content in corn grains;

(3) application in reducing corn kernels;

(4) the application in breeding corn with low total starch content and/or high total protein content in grains;

(5) use in the preparation of corn having a low total starch content and/or a high total protein content in the kernel;

(6) the application in identifying or screening the corn with low total starch content and/or high total protein content or germplasm resources thereof.

10. A method for preparing corn with low total starch content and/or high total protein content, which is characterized in that the corn is made to express the corn ZmDek701 gene mutant of claim 8.

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