CN110904131B - Cotton GhGlu19 gene and application thereof in improving cotton yield - Google Patents

Cotton GhGlu19 gene and application thereof in improving cotton yield Download PDF

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CN110904131B
CN110904131B CN201911281742.5A CN201911281742A CN110904131B CN 110904131 B CN110904131 B CN 110904131B CN 201911281742 A CN201911281742 A CN 201911281742A CN 110904131 B CN110904131 B CN 110904131B
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郭旺珍
王海棠
周雪松
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Nanjing Agricultural University
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Abstract

The invention discloses a cotton GhGlu19 gene and application thereof in improving cotton yield, belonging to the field of biotechnology application. The invention relates to a GhGlu19 gene which codes beta-1, 3-glucanase. The invention provides a full-length ORF nucleotide sequence and an amino acid sequence of GhGlu19 in genome A subgroup and genome D subgroup in an allotetraploid upland cotton genetic standard line TM-1. The gene is used as a target gene to construct an antisense plant expression vector to create a transgenic cotton material, and the result shows that the inhibition of the gene expression can obviously increase the weight of bolls, the number of seeds per boll and the effective number of bolls of a single plant, improve the seed dressing, enhance the seed vigor and the like, and further obviously improve the cotton yield.

Description

Cotton GhGlu19 gene and application thereof in improving cotton yield
Technical Field
The invention belongs to the field of biotechnology application, and particularly relates to a cotton GhGLU19 gene and application thereof in improving cotton yield. Cloning the full-length ORF of the GhGLU19 gene by utilizing a PCR (polymerase chain reaction) technology and integrating the full-length ORF of the GhGLU19 gene on a pBI121 binary vector in an antisense direction to construct a gene antisense expression vector; vectors were introduced into cotton recipient W0 lines by agrobacterium-mediated methods. And obtaining a stable transgenic homozygous plant through PCR identification and qRT-PCR expression detection analysis on transgenic progeny plants. Through field character investigation for two consecutive years, it is determined that the inhibition of the expression of the GhGLU19 gene can increase the yield of cotton, more specifically, the weight of bolls, the number of seeds per boll and the effective number of bolls per plant can be increased, the seed dressing can be improved, the seed vigor can be enhanced, and the like.
Background
Beta-1, 3-glucanase (E.C. 3.2.1.39) hydrolyzes beta-1, 3-glucan (callose), which is widely present in viruses, bacteria, fungi) and seed plants. In plants, β -1, 3-glucanase is a conserved large gene family with 51 members in Arabidopsis, 44 members in Theobroma, 43 genes in Vitis vinifera, and 67 members in diploid Raymond cotton. The beta-1, 3-glucanase genes all have an N-terminal signal peptide sequence and a glycosyl hydrolyzed 17 core domain, and the C-terminal structure generally comprises a Carbohydrate-Binding module 43 (CBM43) domain and a C-terminal hydrophobic sequence. According to the protein structure classification, the Arabidopsis thaliana 51 β -1, 3-glucanase genes can be classified into 5 types (Type 1-Type 5) (Doxey et al, 2007). The protein structure with Type 3 is considered to be the ancestral β -1, 3-glucanase, whose function is related to cell division. During long-term evolution, certain members of the family lose the C-terminal domain, mature proteins are secreted into the cell wall, and new functions such as antifungal have evolved (Doxey et al, 2007). Beta-1, 3-glucanases play an important role in plant defense against pathogens (Leubner-Metzger and Mens 1999), cell division, pollen germination and pollen tube elongation (Worrall et al, 1992 Wan et al, 2011), plasmodesmata signaling (Levy et al, 2007a, levy et al, 2007 b), cold stress response (Hincha et al, 1997), seed germination and maturation (Leubner-Metzger and Mens 2000), among others.
Callose plays an important role in the development of pollen. During the development of male gametes, the synthesis and degradation of callose are strictly regulated. At the very beginning of meiosis, callose is deposited between the primary wall and the plasma membrane of pollen mother cells to prevent binding and fusion between pollen mother cells and surrounding diploid cells (Waterkeyn 1962). At the end of meiosis, upon formation of tetrads, the callose wall breaks, releasing microspores (Wan et al, 2011).
The time of pollen mother cell callose wall rupture is strictly controlled, and callose layer degradation in advance or prevention of callose layer degradation can affect the release of tetrads, thereby causing male sterility. The β -1, 3-glucanase gene is expressed in tobacco pollen mother cells in advance by a tapetum-specific promoter, and the callus layer is degraded in advance, releasing immature microspores, resulting in male sterility of tobacco (Worrall et al, 1992). The beta-1, 3-glucanase gene expressed in the rice floral organs is silenced by an RNA interference technology, the callose layer is not degraded when the tetrad structure is formed, and the microspores are adhered together and are not released. Microspores with no fertility are released later (Wan et al, 2011). Beta-1, 3-glucanase genes are expressed or silenced in advance in a tapetum, the degradation of a callus layer is advanced or hindered, and microspores without fertility are released, so that male sterility is realized, and a new idea is provided for molecular breeding of male sterility.
Cotton is an important economic crop, and the yield is guaranteed for planting benefit. Therefore, improving yield has always been an important goal in cotton breeding. The main methods and approaches for Chinese high-yield breeding are as follows: foreign introduction is used for replacing the original low-yield inferior varieties, the hybrid vigor is utilized to cultivate high-yield hybrid varieties and molecular marker-assisted breeding. Although the development and utilization of molecular markers associated with yield and the release of genome-wide sequences provide a good basis for molecular design and breeding of cotton, few genes have been explored for improvement of yield traits and little has been known about the molecular mechanisms of yield-related constitutive factors (well known, et al, 2016). The yield-related constitutive factors of cotton are: total number of plants per unit area, number of bolls per plant, boll weight and clothes score. Through planting tests of several main cultivars in Xinjiang northern Xinjiang over the last two decades, the yield and the change conditions of the constitutive factors are analyzed, and the conclusion is obtained: the weight per bell tends to increase in quality during variety changeovers (von Yang et al, 2016). Therefore, the search and cloning of genes related to the regulation and control of yield traits, especially genes for regulating and controlling the change of the boll weight have important significance for breeding high-yield cotton varieties by creating germplasm resource materials for improving the yield.
Disclosure of Invention
The invention aims to provide a method for improving cotton yield by inhibiting the expression of a cotton GhGLU19 gene. Cloning the full-length ORF of the GhGLU19 gene by utilizing a PCR technology, integrating the full-length ORF of the GhGLU19 gene on a pBI121 binary vector in an antisense direction, and constructing an antisense expression vector of the gene; vectors were introduced into cotton recipient W0 lines by agrobacterium-mediated methods. And obtaining a stable transgenic homozygous plant through PCR identification and qRT-PCR expression detection analysis on transgenic progeny plants. Through field character investigation for two consecutive years, it is determined that the inhibition of the expression of the GhGLU19 gene can increase the yield of cotton, more specifically, the weight of bolls, the number of seeds per boll and the effective number of bolls per plant can be increased, the seed dressing can be improved, the seed vigor can be enhanced, and the like.
Another objective of the invention is to provide a cotton GhGLU19 gene and a full-length cDNA ORF nucleotide sequence of the gene in Gossypium hirsutum TM-1A subgroup (GhGLU 19A) and Gossypium hirsutum D subgroup (GhGLU 19D). The gene has the amino acid sequence coded in A subgroup and D subgroup in upland cotton.
The invention also aims to provide a novel germplasm material with obviously increased cotton yield and application thereof in breeding and production of new cotton varieties.
The purpose of the invention is realized by the following technical scheme:
the application of the cotton GhGLU19 gene in improving the cotton yield and cultivating new cotton germplasm. The application is to improve the cotton yield by inhibiting the expression quantity of the GhGLU19 gene of cotton.
A method for improving cotton yield by inhibiting the expression of cotton GhGLU19 gene.
The expression quantity for inhibiting the cotton GhGLU19 gene is that a plant expression vector pBI121 is utilized to integrate the GhGLU19 gene on a vector in an antisense direction to construct the gene antisense expression vector, and the constructed antisense expression vector is transferred into cotton by an agrobacterium-mediated method to inhibit the expression level of the GhGLU19 gene. Preferably, the GhGLU19 gene coding sequence is introduced into PBI121 vector skeleton in an antisense direction to construct the gene antisense expression vector, and the constructed antisense expression vector is transformed into cotton by utilizing an agrobacterium-mediated method to obtain a transgenic cotton material for reducing the expression of the GhGLU19 gene. More preferably, the method for reducing the expression level of the GhGLU19 gene is to introduce the nucleotide sequence shown in SEQ ID NO.1 or SEQ ID NO.2 into a PBI121 vector skeleton in an antisense direction to construct the gene antisense expression vector. The vector is transformed into an agrobacterium strain LBA4404, cotton is transformed by an agrobacterium-mediated method, a transgenic cotton material for reducing GhGLU19 gene expression is obtained through cotton callus screening induction, somatic embryo differentiation and regeneration processes, and the obtained transgenic pure line for inhibiting the GhGLU19 expression is subjected to field growth and development index determination for two years continuously, so that the effects of inhibiting the GhGLU19 expression on increasing the boll weight, the number of seeds per boll and the effective number of bolls of a single plant, improving the seed dressing, enhancing the seed vigor and the like are proved, and the vector has important application value for creating new high-yield cotton varieties.
The plant expression vector is transferred into cotton, namely the hypocotyl stem segment of the aseptic cotton seedling is transformed, callus is induced, somatic embryo is obtained, and the regenerated seedling is obtained. The constitutive promoter carried by the plant expression vector is the CaMV35S promoter.
The improvement of the cotton yield is the improvement of the cotton yield characters, and the concrete expression is as follows: (1) increasing the ring weight; (2) increasing the number of seeds per bell; (3) increasing the effective ring number of the single plant; (4) increasing the clothes score; and (5) enhancing the seed vigor.
The cotton yield can be improved by inhibiting the expression of the GhGLU19 gene, wherein the GhGLU19 gene is any one of (1) to (3):
(1) The gene is in upland cotton TM-1A subgroup (GhGLU 19A), and the nucleotide sequence of the full-length cDNA ORF is shown as SEQ ID NO. 1;
(2) The gene is in a Gekko cotton TM-1D subgroup (GhGLU 19D), and the nucleotide sequence of the full-length cDNA ORF is shown as SEQ ID NO. 2;
(3) Nucleotide sequences with homology of more than 98 percent and identical or similar functions with the nucleotide sequences of the genes in (1) or (2).
The protein coded by the GhGLU19 gene has an amino acid sequence shown in SEQ ID NO. 3 and an amino acid sequence shown in SEQ ID NO. 4, wherein the protein coded by the nucleotide sequence shown in SEQ ID NO.1 and the protein coded by the nucleotide sequence shown in SEQ ID NO. 2.
A recombinant vector, an expression cassette, a transgenic cell line or a recombinant bacterium containing the GhGLU19 gene.
The recombinant vector, the expression cassette, the transgenic cell line or the recombinant strain are applied to the improvement of the yield of cotton and the cultivation of new cotton germplasm.
The invention reduces the expression quantity of the GhGLU19 gene of cotton by utilizing the plant genetic engineering technology, obtains a new germplasm material with increased cotton yield and applies the new germplasm material to production.
The invention has the advantages that:
the invention utilizes plant gene engineering technology to create transgenic material for inhibiting GhGLU19 gene expression, and carries out agronomic character investigation on the transgenic material to discover that: the important yield indexes such as the ring-back weight and the like are remarkably improved, and excellent materials are provided for researching molecular mechanisms for forming yield key characters and breeding application.
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FIG. 1 overexpression vector and antisense vector;
wherein A is an excessive expression vector pBI121-35S: : ghGLU19, B is antisense vector pBI121-35S: : antisenseGhGLU19.
FIG. 2 shows the results of PCR to prove the successful transformation of the vector;
wherein A is an excessive expression vector pBI121-35S: : the detection result of the GhGLU19 is M from left to right in a: marker molecular weight markers, +: plasmid control, -: receptor W0 control, 1-12 is overexpression vector transgenic plant; b is antisense vector pBI121-35S: : the detection result of the antisenseGhGLU19 is M: marker molecular weight markers, +: plasmid control, -: acceptor W0 control, 1-9 antisense vector transgenic plants.
FIG. 3 shows the real-time quantitative fluorescence detection results of the expression level of the GhGLU19 gene in different tissues of the transgenic line;
wherein A is the relative expression quantity of the GhGLU19 gene in roots of antisense vector strains (AS 19, AS87 and AS 105), transgenic receptor control (W0) and overexpression strains (OE 4, OE39, OE54 and OE 110); b is the relative expression quantity of the GhGLU19 gene in the stem; c is the relative expression level of the GhGLU19 gene in leaves.
The results show that the relative expression amount of the GhGLU19 gene in different tissues of the over-expression material is obviously increased, and the relative expression amount of the GhGLU19 gene in different tissues of the antisense material is obviously reduced.
FIG. 4 shows the results of seed germination tests;
wherein A is the top view of seedlings 7 days after seed germination of antisense vector lines (AS 19, AS87 and AS 105), transgenic recipient control (W0) and overexpression lines (OE 4, OE39, OE54 and OE 110); graph B shows a comparison of germination rates between different materials; FIG. C shows 7-day-old seedlings after germination of different materials; panels D-F are comparisons of the relative indices of germination vigor (hypocotyl length, cotyledon length and cotyledon width) for antisense, control and over-expressed material.
The results show that the seed germination rate of the antisense material is obviously improved compared with that of a control material and an over-expression material, and the germination potential index also tends to increase; the germination rate of the over-expressed material is obviously reduced compared with that of a contrast, the index value of the germination potential is also reduced, and the variation trend of each index is consistent with the expression quantity difference of the GhGLU19 gene in the material.
FIG. 5 comparison of yield-related growth and development indicators for transgenic material;
wherein, panels a and B are the plant heights of antisense vector strains (AS 19, AS87 and AS 105), transgenic acceptor control (W0) and overexpression strain (OE 4, OE39, OE54 and OE 110) materials in 2018 and 2019, respectively. The plant height of the antisense material is obviously increased compared with that of a control, and the plant heights of a plurality of over-expression strains are obviously reduced compared with that of the control; FIGS. C-D are the statistical results of the average effective bell per plant, showing that the cases of average effective bell per plant between materials are antisense strains > control > over-expressed strains; panel E shows comparison of cotton bolls at day 43 post-anthesis for antisense vector lines (AS 19, AS87 and AS 105), transgenic recipient control (W0) and over-expressed line (OE 4, OE39, OE54 and OE 110) material, where the bolls of the antisense material are significantly larger than the control and the bolls of the over-expressed line are smaller than the control. Panels F-G are comparisons of bell weights of antisense vector lines (AS 19, AS87, and AS 105), transgenic recipient control (W0), and overexpression line (OE 4, OE39, OE54, and OE 110) material. The ring weight of the antisense material is significantly higher than that of the control material and the excess material, and the ring weight of the over-expressed material is significantly reduced compared with that of the control material; FIG. H-I is a graph showing the statistical results of the number of seeds per bell for the antisense material, the number of seeds per bell for the control material being about 22-26 grains, and the number of seeds per bell for the over-expressed material being reduced by different amounts compared to the control material; FIGS. J-K are comparative results of the scores between different materials; the cotton fiber is an important economic benefit part of cotton, the lint is an important index, and from the graph J and the graph K, the expression level of GhGLU19 is changed through transgenosis, the lint ratio is not reduced, and the lint ratio tends to be increased in an antisense material.
Detailed Description
In order to make the technical problems, technical solutions and advantageous effects solved by the present invention more apparent, the present invention is further described in detail below with reference to the following embodiments.
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.
1. Creation of transgenic cotton material of GhGLU19 gene
PBI121 is a traditional plant binary expression vector, and the promoter of the PBI121 is a 35S promoter. Constructing an over-expression binary vector and an antisense binary vector of the 35S promoter on the basis, wherein the insertion position of the fragment is between 35S-P (promoter) and Nos-T (terminator); the reverse complementary sequence of the gene SEQ NO ID.1 sequence is used for constructing an antisense plant expression vector, and the specific process is as follows:
1. the cloning vector containing 1410bp target fragment (SEQ NO ID.1) is subjected to double enzyme digestion by XbaI and SacI, and a small fragment is recovered; the pBI121 expression vector was digested with XbaI and SacI, and the Gus gene was excised to recover a large fragment (about 13 kb). The overexpression vector (A in FIG. 1) was constructed by inserting the gene of interest into the pBI121 vector between XbaI and SacI cleavage sites using T4 ligase by a technique well known to those skilled in the art.
2. The cloning vector containing 1410bp target fragment (SEQ NO ID.1) is subjected to double enzyme digestion by XbaI and SacI, and a small fragment is recovered; the pBI121 expression vector was digested with XbaI and SacI, and the Gus gene was excised to recover a large fragment (about 13 kb). An antisense vector (B in FIG. 1) was constructed by inserting the gene of interest into the pBI121 vector in the antisense direction between the XbaI and SacI cleavage sites using T4 ligase by a technique known to those skilled in the art.
3. Transforming the constructed vector into agrobacterium; the specific operation steps are as follows: add 1ug of the constructed vector plasmid DNA into 200uL of competent cells, placed on ice for 30 minutes, frozen rapidly in liquid nitrogen for 5 minutes, thawed in water bath at 37 ℃ (5 minutes), added with 1ml of YEP medium, shake-cultured at 28 ℃ for 2-4 hours (180 rpm), centrifuged for 1 minute, suspended cells in 200uL of YEP medium, plated, and cultured at 28 ℃ for 2 days. Selecting a single clone, adding the single clone into 700uL LB containing 50mg/L Kan and 50mg/L Rif, culturing for 6 hours at 28 ℃, and taking 1uL bacterial liquid to carry out PCR positive identification, wherein a primer 1 and a primer 2 are used for detecting an over-expression vector, and a primer 3 and a primer 4 are used for detecting an antisense vector.
Primer 1:5'GCTCTAGAATGGCTTCCCAGGCGTTTTATC3'
Primer 2:5 'CGAGCTCTCTCAATGCCCTGTTGGAAACTC 3'
And (3) primer: 5 'CGAGCTCATGGCTCCCAGGCGTTTATC 3'
And (4) primer: 5'GCTCTAGATCAATGCCCTGTTGGAAACTC3'
4. Sucking 10uL of positive clone bacterial liquid into 50mL of LB containing 50mg/L kan and 50mg/L Rif, culturing for about 16 hours at 28 ℃, centrifuging for 10 minutes at 4000rpm after the OD value reaches 0.6, collecting the bacterial liquid, washing bacterial plaque twice by 20mL of LB, adjusting the OD value of the bacterial liquid to 0.3, infecting aseptic seedlings (seedling culture medium culture) of 7 days old for 15 minutes, sucking the bacterial liquid, culturing the stem sections in a co-culture medium for 2 days, subculturing in a screening callus induction medium for 3 weeks, subculturing for about 3 times, subculturing callus with the diameter of about 2cm in a callus proliferation medium for 2-4 times until the somatic embryos differentiate, subculturing the differentiated somatic embryos in a dark culture medium, after the seedlings grow to about 10cm, washing the seedling root culture medium by aseptic water, culturing for 1 week under the conditions of 25 ℃, 60% humidity and 16 hours illumination for 8 hours, and transplanting the seedlings into a substrate: vermiculite =1: the growth period was completed in 1 (w/w) pot.
The medium composition was as follows:
basic culture medium: macroelement mother liquor (50 ml. L) -1 ) + ferric salt mother liquor (5 ml. L) -1 ) + microelement mother liquor (5 ml. L) -1 ) + organic mother liquor (5 ml. L) -1 ) + glucose (30 g.L) -1 )+phytogel(3.0g·L -1 )+MgCl 2 (1.0g·L -1 )。
TABLE 1 stock solution formula Table for minimal Media (MSB)
Figure BDA0002316950380000071
Seedling culture medium: 1/2MS + plant gel (7 g.L) -1 ) The pH was adjusted to 5.8.
Co-culture medium: in the minimal medium, the ph was adjusted to 5.8.
Screening a callus induction culture medium: minimal medium +0.1 mg. L -1 2,4-D+0.1mg·L -1 KT+50mg·L -1 Kanamycin +500mg L -1 Timentin, ph adjusted to 5.8.
Callus proliferation medium: minimal medium +1.9 g.L -1 KNO 3 The ph value was adjusted to 5.8.
Differentiation medium: minimal medium does not contain NH 4 NO 3 +0.5g·L -1 Asparagine +1.0 g.L -1 Glutamine +1.9 g.L -1 KNO 3 The ph was adjusted to 6.0.
5, PCR verifies the positive of the transgenic material; extracting total DNA in plant leaves as a template, performing PCR amplification on the over-expressed material by using primers 5 and 6, and verifying whether the over-expressed material is transgenic; the antisense material was positively detected using primers 7 and 8. Follow-up testing was performed on each generation of transgenic material until a transgenic inbred was obtained (fig. 2).
Primer 5:5 'CACACAATCCCACTTCCTTCG3'
Primer 6:5 'ATTAGGAAGTGCGACGACCACAC 3'
Primer 7:5 'GAGGCCTATTCGGCTATGACTG3'
And (3) primer 8:5'TAGAAGGCGATGCGCTGCGA3'
Verifying the relative expression quantity of the GhGLU19 gene in a transgenic material and a control material by qRT-PCR; respectively extracting total RNA of leaves, roots and stems of an over-expression material, an anti-sense expression material and a control material, taking 1ug of RNA, carrying out reverse transcription to obtain cDNA, carrying out qRT-PCR amplification by taking the cDNA of 1uL 100ng/uL as a template, respectively amplifying specific sequences (amplification primers are a primer 9 and a primer 10) of a Histone3 gene of an internal reference gene and a GhGLU19 gene (amplification primers are a primer 11 and a primer 12), and calculating data by a 2^ delta Ct method to obtain the relative expression quantity of the GhGLU19 gene in the transgenic material and the control material. The results show that the GhGLU19 gene is up-regulated and expressed in different tissues (roots, stems and leaves) of the transgenic over-expression material, and the expression level of the GhGLU19 gene is obviously reduced in different tissues (roots, stems and leaves) of the antisense material (figure 3).
Primer 9:5 'GAAGCCTCCATCGATACCGTC 3'
Primer 10:5'CTACCACTACCATCATGG3'
Primer 11:5 'AGAGGGTAAGGCTGATTGTAGG'
Primer 12:5'CATATGTAGGACGTTGGGAGa3'
Transforming cotton hypocotyl by agrobacterium-mediated method, and obtaining transgenic regenerated seedling by callus culture and somatic cell differentiation regeneration. PCR identification and qRT-PCR verification prove that homologous genes of SEQ NO ID.1 and SEQ NO ID.2 sequences in the obtained 4 over-expression pure lines have stable and high expression; in 3 suppression expression clones, the homologous gene expression of SEQ NO id.1 and SEQ NO id.2 sequences was significantly reduced compared to the control.
2. Functional verification of GhGLU19 gene
Through field trait survey of the transgenic material obtained in the step one for two consecutive years (2018 and 2019), the germination and yield-related growth and development traits of the over-expression material and the antisense material are obviously changed. The specific results are as follows:
1. after harvested seeds are delinted by sulfuric acid, the seeds are aired for 2 days, 30 seeds are taken from each material and planted in paper cups with 40g of matrix and vermiculite mixed in equal ratio (volume ratio), 15 paper cups are used as one plate, and 500mL of water is added into the plate every other day. And after the germination data are stable, counting the germination rate of each material, and designing three times of repetition. The seed germination rate of the over-expressed material was significantly reduced compared to the control, while the germination rate of the antisense material was increased compared to the control (a and B in fig. 4); by measuring germination potential-related indexes such as hypocotyl length, cotyledon length, and cotyledon width of seedlings 7 days after germination, it was found that the germination potential of the excess material was reduced compared to the control, while the germination potential of the antisense material was higher than the control (C-F in FIG. 4).
2. Measuring the plant heights of all the plants in the two rows, and calculating the average value to obtain the average plant height of each pure material; the plant height of the over-expressed material was significantly reduced compared to the control, while the average plant height of the antisense material was significantly increased compared to the control (a-B in fig. 5).
3. In the boll opening period, counting the number of effective bolls (i.e. bolls with diameter larger than 2 cm) of each plant in two rows, and averaging to obtain the average effective boll number of each pure line material per plant (C-D in figure 5);
4. harvesting 25 cotton bolls at the middle-upper part after boll opening, picking seed cotton, weighing and dividing by 25 to obtain the weight of a single boll, repeating for three times, and calculating the average value to obtain the average boll weight; the results show that 4 excess materials had a significantly lower weight average bell than the control material, and 3 antisense materials had a higher weight average bell than the control material, all of which were shown to be very significant by statistical difference calculations (E-G in figure 5).
5. Respectively counting the number of seeds of 25 rings harvested in the step 4, dividing the number by 25 to obtain the number of seeds of each ring, repeating the steps for three times, and calculating an average value to obtain the average number of seeds of each ring; the number of seeds per bell in the over-expressed material was significantly reduced compared to the control, while the antisense material was significantly increased compared to the control (H-I in fig. 5).
6. Ginning 25 boll seed cotton harvested in the step 4, respectively weighing the weight of lint cotton and the weight of fiber, and according to a calculation formula of the lint: the ginning mark = ginned cotton weight/seed cotton weight is calculated by 100%, repeated for three times, and an average value is taken; the scores of all 3 antisense materials were higher than those of the control material, indicating that altering the expression of the GhGLU19 gene has potential application in increasing scores (J-K in FIG. 5).
7. Stable cotton quality is the basis for increased cotton yield. 25 cotton bolls of antisense vector strains (AS 19, AS87 and AS 105), transgenic acceptor control (W0) and overexpression strains (OE 4, OE39, OE54 and OE 110) planted in the field rows are harvested in 2018 and 2019 respectively, 20g of cotton fibers are weighed after cotton ginning and sent to a cotton quality detection center of Chinese academy of agricultural sciences for determination, and the determination indexes comprise Fiber Length (FL), specific strength (FS), micronaire value (FM), uniformity (FU) and elongation (FE) (the Fiber quality detection data results are shown in tables 2 and 3). The results show that there was no significant change in the fiber quality traits of the antisense vector lines (AS 19, AS87 and AS 105) and the overexpression vector lines (OE 4, OE39, OE54 and OE 110) compared to the recipient control (W0). GhGLU19 had no significant effect on fiber length, specific strength, micronaire value, etc.
TABLE 2 2018 fibre quality test results
Figure BDA0002316950380000091
TABLE 3 2019 fiber quality test results
Figure BDA0002316950380000092
The expression of germination and yield related traits in the transgenic material is integrated, the important function of the GhGLU19 in the growth and development of the germination and yield related traits of cotton is verified, and the change of the expression quantity of the GhGLU19 gene does not change the fiber quality, so that the purpose of regulating and controlling the germination and yield related traits of cotton can be realized by changing the expression quantity of the GhGLU19 gene, and the application prospect in the fields of genetic improvement of crops and the like is wide.
SEQ ID NO:1
Nucleotide sequence of GhGLU19 gene in upland cotton TM-1A subgroup (GhGLU 19A) ORF
ATGGCTTCCCAGGCGTTTTATCTTCTTGTTTCAATCATTGTTCTCTTATCCGCCATTGT TGTCTCAGGTTCGGGTTCGGTCGGGATTAACTACGGTCGTGTAGCAAACAACTTACC GTCACCGGAAAAAGTCGTGGAACTATTGAAATCACAAGGAATAAACAAAGTGAAGCTT TACGACACCGACGCGACGGTTTTGACGGCGTTAGCGGATTCCGGTATAACCGTGGTC GTCGCACTTCCTAATGAACTACTTTCATCCACCGCTGCTGATCAGTCGTTTGCAGACA ACTGGGTTGAAGCAAACATAACAAAGTTTTACCCAAAGACAAAAATCGAAGCCATAGC TGTCGGTAACGAAGTGTTTGTCGATCCGGCCAACACGACCAAATACCTCGTACCGGC GATGAAGAACATCCACGCTTCGTTAGTTAAATCCAAGCTTGATTCCGCCATTAAAATC TCTTCTCCGATAGCTTTCAGTGCTTTAAAAACCTCGTATCCTTCATCGGCCGGTTCGT TTAAGCCGGAATTGATTGAACCGGTGATTAAACCCATGTTGGATTTTTTGAAACAAAC CGGGTCTTACTTAATGGTTAATGCGTACCCGTTTTTTGCTTATTCGGCTAATTCGGATC AGATCTCACTTGATTATGCTTTGTTTAAGGATAACCCGGGTGTAGTAGATTCGGGTAA CGGGTTGAAATATTCTAGCCTTTTGGAAGCTCAAATCGACGCCGTTTTTGCAGCTATG TCGGCTATTAAATACGACGACGTAAAGATGGTTGTGACCGAAACGGGTTGGCCATCA ATGGGCGATGAAGATGAAATAGGTGCTAGTGAATCCAATGCGGCGTCGTATAATGGT AATTTAGTACGGAAAGTTTTGACCGGTAATGGGACCCCTTTAAGACCTCAAGACCCAC TTAACGTTTATTTATTTGCTTTATTTAATGAGAATAAAAAACCCGGTCCAACTTCCGAAA GGAATTACGGTTTATTTTACCCTAATGAACAAAAAGTATATAACATACCATTAACCAAA GAGGAGGCGAAAACCGGTCAGTCAACGCCGGTCAACAGCAACACGAGTCAGGTTCC GGTAGCCGGAGAAGTATCGAAGGCGAAAGTAGGTCAGACATGGTGCGTTGCAAACG GGAAAGCCGATGAGAAGAAGTTACAAGCGGCGTTGGATTATGCTTGCGGAGAGGGT AAGGCTGATTGTAGCCCTATTCAACCTGGCGCGACGTGTTATAATCCAAACACACTCG AAGCACATGCTTCGTACGCTTTCAATAGCTATTATCAGAAGAACACACGTGTGACTGG CACATGCGAGTTCGGTGGTGCGGCTTATGTGGTCTCCCAACGTCCTACATATGGGAG TTGTGAGTTTCCAACAGGGCATTGA
SEQ ID NO:2
Nucleotide sequence of GhGLU19 gene in Gossypium hirsutum TM-1D subgroup (GhGLU 19D) ORF
ATGGCTTCCCAGGCTTTTTATCTTATTGTTTCAATCATTGTTCTCTTATCCGCCATTGTT GTCTCAGGTTCGGGTTCGGTCGGGATTAACTACGGTCGTGTAGCAAACAACTTACCG TCACCGGAAAAAGTCGTGGAACTATTGAAATCACAGGGAATAAACAAAGTGAAGCTTT ACGATACCGACGCGACGGTTTTGACGGCGTTAGCGGATTCCGGTATAACCGTGGTC GTCGCACTTCCTAATGAACTACTTTCATCCATCGCCGCTGATCAGTCGTTTGCCGACA ACTGGGTTGAAGCAAACATAACAAAGTTTTATCCAAAGACAAAAATCGAAGCCATAGC TGTCGGTAACGAAGTGTTTGTCGATCCGGCCAACACGACCAAATACCTCGTACCGGC GATGAAGAACATCCACGCTTCGTTAGTTAAATCCAAGCTTGATTCCGCCATTAAAATC TCTTCTCCAATAGCTTTCAGTGCTTTGAAAACCTCGTACCCTTCATCGGCCGGTTCGT TTAAGCCGGAATTGATTGAACCGGTGATTAAACCCATGTTGGATTTTTTGAAACAAAC CGGGTCTTACTTAATGGTTAATGCGTACCCGTTTTTTGCTTATACGGCTAATTCGGAT CAAATCTCACTTGATTATGCTTTGTTTAAGGAAAACCCGGGTGTAGTGGATTCGGGTA ACGGGTTGAAATATTCTAGCCTTTTCGAAGCTCAAATCGACGCCGTTTTTGCAGCTAT GTCGGCTATTCAATACGACGACGTAAAGATGGTTGTGACCGAAACGGGTTGGCCTTC AATGGGAGATCAAGACGAAAAAGGTGCTAGTGAATCCAATGCGGCGTCGTATAATGG TAATTTAGTACGGAAAGTTTTGACTGGTAATGGGACCCCTTTAAGACCTCAAGACCCA CTTAACGTTTATTTATTTGCTTTATTTAATGAGAATCAAAAACCCGGTCCAACTTCCGA AAGGAATTACGGTTTATTTTACCCTAATGAACAAAAGGTATATACCATACCATTAACCA AAGAGGAGGCGAAAACCGGTGAGTCAACGCAGGTCAACACCAACACGAGTCTGGCT CCTGTAGCCGGAGAAGTATCGAAGGCAAAAGTAGGTCAGACGTGGTGCGTTGCGAA CGAGAAAGCCGATGAGAAGAAGTTACAAGCGGCGTTGGATTATGCTTGCGGAGAGG GTGAGGCTGATTGTAGCCCTATTCAACCTGGCGCGACGTGCTATAACCCAAACACAC TCGAAGCACATGCTTCATACGCTTTCAATAGTTATTATCAGAAGAACACACGTGCGAC TGGCACATGCGAGTTCGGTGGTGCTGCTTATGTGGTCACCCAACGTCCTACATATGG GAATTGTGAGTTTCCAACAGGGCATTGA
SEQ ID NO:3
The GhGLU19 gene has an amino acid sequence of a cotton TM-1A subgroup (GhGLU 19A) in upland cotton
MASQAFYLLVSIIVLLSAIVVSGSGSVGINYGRVANNLPSPEKVVELLKSQGINKVKLYDTDA TVLTALADSGITVVVALPNELLSSTAADQSFADNWVEANITKFYPKTKIEAIAVGNEVFVDP ANTTKYLVPAMKNIHASLVKSKLDSAIKISSPIAFSALKTSYPSSAGSFKPELIEPVIKPMLDF LKQTGSYLMVNAYPFFAYSANSDQISLDYALFKDNPGVVDSGNGLKYSSLLEAQIDAVFAA MSAIKYDDVKMVVTETGWPSMGDEDEIGASESNAASYNGNLVRKVLTGNGTPLRPQDPLN VYLFALFNENKKPGPTSERNYGLFYPNEQKVYNIPLTKEEAKTGQSTPVNSNTSQVPVAGE VSKAKVGQTWCVANGKADEKKLQAALDYACGEGKADCSPIQPGATCYNPNTLEAHASYA FNSYYQKNTRVTGTCEFGGAAYVVSQRPTYGSCEFPTGH
SEQ ID NO:4
The GhGLU19 gene has an amino acid sequence of a cotton TM-1D subgroup (GhGLU 19D) in upland cotton
MASQAFYLIVSIIVLLSAIVVSGSGSVGINYGRVANNLPSPEKVVELLKSQGINKVKLYDTDA TVLTALADSGITVVVALPNELLSSIAADQSFADNWVEANITKFYPKTKIEAIAVGNEVFVDPA NTTKYLVPAMKNIHASLVKSKLDSAIKISSPIAFSALKTSYPSSAGSFKPELIEPVIKPMLDFL KQTGSYLMVNAYPFFAYTANSDQISLDYALFKENPGVVDSGNGLKYSSLFEAQIDAVFAAM SAIQYDDVKMVVTETGWPSMGDQDEKGASESNAASYNGNLVRKVLTGNGTPLRPQDPLN VYLFALFNENQKPGPTSERNYGLFYPNEQKVYTIPLTKEEAKTGESTQVNTNTSLAPVAGEV SKAKVGQTWCVANEKADEKKLQAALDYACGEGEADCSPIQPGATCYNPNTLEAHASYAFN SYYQKNTRATGTCEFGGAAYVVTQRPTYGNCEFPTGH
Sequence listing
<110> Nanjing university of agriculture
<120> cotton GhGlu19 gene and application thereof in improving cotton yield
<160> 4
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1410
<212> DNA
<213> Cotton (Gossypium spp)
<400> 1
atggcttccc aggcgtttta tcttcttgtt tcaatcattg ttctcttatc cgccattgtt 60
gtctcaggtt cgggttcggt cgggattaac tacggtcgtg tagcaaacaa cttaccgtca 120
ccggaaaaag tcgtggaact attgaaatca caaggaataa acaaagtgaa gctttacgac 180
accgacgcga cggttttgac ggcgttagcg gattccggta taaccgtggt cgtcgcactt 240
cctaatgaac tactttcatc caccgctgct gatcagtcgt ttgcagacaa ctgggttgaa 300
gcaaacataa caaagtttta cccaaagaca aaaatcgaag ccatagctgt cggtaacgaa 360
gtgtttgtcg atccggccaa cacgaccaaa tacctcgtac cggcgatgaa gaacatccac 420
gcttcgttag ttaaatccaa gcttgattcc gccattaaaa tctcttctcc gatagctttc 480
agtgctttaa aaacctcgta tccttcatcg gccggttcgt ttaagccgga attgattgaa 540
ccggtgatta aacccatgtt ggattttttg aaacaaaccg ggtcttactt aatggttaat 600
gcgtacccgt tttttgctta ttcggctaat tcggatcaga tctcacttga ttatgctttg 660
tttaaggata acccgggtgt agtagattcg ggtaacgggt tgaaatattc tagccttttg 720
gaagctcaaa tcgacgccgt ttttgcagct atgtcggcta ttaaatacga cgacgtaaag 780
atggttgtga ccgaaacggg ttggccatca atgggcgatg aagatgaaat aggtgctagt 840
gaatccaatg cggcgtcgta taatggtaat ttagtacgga aagttttgac cggtaatggg 900
acccctttaa gacctcaaga cccacttaac gtttatttat ttgctttatt taatgagaat 960
aaaaaacccg gtccaacttc cgaaaggaat tacggtttat tttaccctaa tgaacaaaaa 1020
gtatataaca taccattaac caaagaggag gcgaaaaccg gtcagtcaac gccggtcaac 1080
agcaacacga gtcaggttcc ggtagccgga gaagtatcga aggcgaaagt aggtcagaca 1140
tggtgcgttg caaacgggaa agccgatgag aagaagttac aagcggcgtt ggattatgct 1200
tgcggagagg gtaaggctga ttgtagccct attcaacctg gcgcgacgtg ttataatcca 1260
aacacactcg aagcacatgc ttcgtacgct ttcaatagct attatcagaa gaacacacgt 1320
gtgactggca catgcgagtt cggtggtgcg gcttatgtgg tctcccaacg tcctacatat 1380
gggagttgtg agtttccaac agggcattga 1410
<210> 2
<211> 1410
<212> DNA
<213> Cotton (Gossypium spp)
<400> 2
atggcttccc aggcttttta tcttattgtt tcaatcattg ttctcttatc cgccattgtt 60
gtctcaggtt cgggttcggt cgggattaac tacggtcgtg tagcaaacaa cttaccgtca 120
ccggaaaaag tcgtggaact attgaaatca cagggaataa acaaagtgaa gctttacgat 180
accgacgcga cggttttgac ggcgttagcg gattccggta taaccgtggt cgtcgcactt 240
cctaatgaac tactttcatc catcgccgct gatcagtcgt ttgccgacaa ctgggttgaa 300
gcaaacataa caaagtttta tccaaagaca aaaatcgaag ccatagctgt cggtaacgaa 360
gtgtttgtcg atccggccaa cacgaccaaa tacctcgtac cggcgatgaa gaacatccac 420
gcttcgttag ttaaatccaa gcttgattcc gccattaaaa tctcttctcc aatagctttc 480
agtgctttga aaacctcgta cccttcatcg gccggttcgt ttaagccgga attgattgaa 540
ccggtgatta aacccatgtt ggattttttg aaacaaaccg ggtcttactt aatggttaat 600
gcgtacccgt tttttgctta tacggctaat tcggatcaaa tctcacttga ttatgctttg 660
tttaaggaaa acccgggtgt agtggattcg ggtaacgggt tgaaatattc tagccttttc 720
gaagctcaaa tcgacgccgt ttttgcagct atgtcggcta ttcaatacga cgacgtaaag 780
atggttgtga ccgaaacggg ttggccttca atgggagatc aagacgaaaa aggtgctagt 840
gaatccaatg cggcgtcgta taatggtaat ttagtacgga aagttttgac tggtaatggg 900
acccctttaa gacctcaaga cccacttaac gtttatttat ttgctttatt taatgagaat 960
caaaaacccg gtccaacttc cgaaaggaat tacggtttat tttaccctaa tgaacaaaag 1020
gtatatacca taccattaac caaagaggag gcgaaaaccg gtgagtcaac gcaggtcaac 1080
accaacacga gtctggctcc tgtagccgga gaagtatcga aggcaaaagt aggtcagacg 1140
tggtgcgttg cgaacgagaa agccgatgag aagaagttac aagcggcgtt ggattatgct 1200
tgcggagagg gtgaggctga ttgtagccct attcaacctg gcgcgacgtg ctataaccca 1260
aacacactcg aagcacatgc ttcatacgct ttcaatagtt attatcagaa gaacacacgt 1320
gcgactggca catgcgagtt cggtggtgct gcttatgtgg tcacccaacg tcctacatat 1380
gggaattgtg agtttccaac agggcattga 1410
<210> 3
<211> 469
<212> PRT
<213> Cotton (Gossypium spp)
<400> 3
Met Ala Ser Gln Ala Phe Tyr Leu Leu Val Ser Ile Ile Val Leu Leu
1 5 10 15
Ser Ala Ile Val Val Ser Gly Ser Gly Ser Val Gly Ile Asn Tyr Gly
20 25 30
Arg Val Ala Asn Asn Leu Pro Ser Pro Glu Lys Val Val Glu Leu Leu
35 40 45
Lys Ser Gln Gly Ile Asn Lys Val Lys Leu Tyr Asp Thr Asp Ala Thr
50 55 60
Val Leu Thr Ala Leu Ala Asp Ser Gly Ile Thr Val Val Val Ala Leu
65 70 75 80
Pro Asn Glu Leu Leu Ser Ser Thr Ala Ala Asp Gln Ser Phe Ala Asp
85 90 95
Asn Trp Val Glu Ala Asn Ile Thr Lys Phe Tyr Pro Lys Thr Lys Ile
100 105 110
Glu Ala Ile Ala Val Gly Asn Glu Val Phe Val Asp Pro Ala Asn Thr
115 120 125
Thr Lys Tyr Leu Val Pro Ala Met Lys Asn Ile His Ala Ser Leu Val
130 135 140
Lys Ser Lys Leu Asp Ser Ala Ile Lys Ile Ser Ser Pro Ile Ala Phe
145 150 155 160
Ser Ala Leu Lys Thr Ser Tyr Pro Ser Ser Ala Gly Ser Phe Lys Pro
165 170 175
Glu Leu Ile Glu Pro Val Ile Lys Pro Met Leu Asp Phe Leu Lys Gln
180 185 190
Thr Gly Ser Tyr Leu Met Val Asn Ala Tyr Pro Phe Phe Ala Tyr Ser
195 200 205
Ala Asn Ser Asp Gln Ile Ser Leu Asp Tyr Ala Leu Phe Lys Asp Asn
210 215 220
Pro Gly Val Val Asp Ser Gly Asn Gly Leu Lys Tyr Ser Ser Leu Leu
225 230 235 240
Glu Ala Gln Ile Asp Ala Val Phe Ala Ala Met Ser Ala Ile Lys Tyr
245 250 255
Asp Asp Val Lys Met Val Val Thr Glu Thr Gly Trp Pro Ser Met Gly
260 265 270
Asp Glu Asp Glu Ile Gly Ala Ser Glu Ser Asn Ala Ala Ser Tyr Asn
275 280 285
Gly Asn Leu Val Arg Lys Val Leu Thr Gly Asn Gly Thr Pro Leu Arg
290 295 300
Pro Gln Asp Pro Leu Asn Val Tyr Leu Phe Ala Leu Phe Asn Glu Asn
305 310 315 320
Lys Lys Pro Gly Pro Thr Ser Glu Arg Asn Tyr Gly Leu Phe Tyr Pro
325 330 335
Asn Glu Gln Lys Val Tyr Asn Ile Pro Leu Thr Lys Glu Glu Ala Lys
340 345 350
Thr Gly Gln Ser Thr Pro Val Asn Ser Asn Thr Ser Gln Val Pro Val
355 360 365
Ala Gly Glu Val Ser Lys Ala Lys Val Gly Gln Thr Trp Cys Val Ala
370 375 380
Asn Gly Lys Ala Asp Glu Lys Lys Leu Gln Ala Ala Leu Asp Tyr Ala
385 390 395 400
Cys Gly Glu Gly Lys Ala Asp Cys Ser Pro Ile Gln Pro Gly Ala Thr
405 410 415
Cys Tyr Asn Pro Asn Thr Leu Glu Ala His Ala Ser Tyr Ala Phe Asn
420 425 430
Ser Tyr Tyr Gln Lys Asn Thr Arg Val Thr Gly Thr Cys Glu Phe Gly
435 440 445
Gly Ala Ala Tyr Val Val Ser Gln Arg Pro Thr Tyr Gly Ser Cys Glu
450 455 460
Phe Pro Thr Gly His
465
<210> 4
<211> 469
<212> PRT
<213> Cotton (Gossypium spp)
<400> 4
Met Ala Ser Gln Ala Phe Tyr Leu Ile Val Ser Ile Ile Val Leu Leu
1 5 10 15
Ser Ala Ile Val Val Ser Gly Ser Gly Ser Val Gly Ile Asn Tyr Gly
20 25 30
Arg Val Ala Asn Asn Leu Pro Ser Pro Glu Lys Val Val Glu Leu Leu
35 40 45
Lys Ser Gln Gly Ile Asn Lys Val Lys Leu Tyr Asp Thr Asp Ala Thr
50 55 60
Val Leu Thr Ala Leu Ala Asp Ser Gly Ile Thr Val Val Val Ala Leu
65 70 75 80
Pro Asn Glu Leu Leu Ser Ser Ile Ala Ala Asp Gln Ser Phe Ala Asp
85 90 95
Asn Trp Val Glu Ala Asn Ile Thr Lys Phe Tyr Pro Lys Thr Lys Ile
100 105 110
Glu Ala Ile Ala Val Gly Asn Glu Val Phe Val Asp Pro Ala Asn Thr
115 120 125
Thr Lys Tyr Leu Val Pro Ala Met Lys Asn Ile His Ala Ser Leu Val
130 135 140
Lys Ser Lys Leu Asp Ser Ala Ile Lys Ile Ser Ser Pro Ile Ala Phe
145 150 155 160
Ser Ala Leu Lys Thr Ser Tyr Pro Ser Ser Ala Gly Ser Phe Lys Pro
165 170 175
Glu Leu Ile Glu Pro Val Ile Lys Pro Met Leu Asp Phe Leu Lys Gln
180 185 190
Thr Gly Ser Tyr Leu Met Val Asn Ala Tyr Pro Phe Phe Ala Tyr Thr
195 200 205
Ala Asn Ser Asp Gln Ile Ser Leu Asp Tyr Ala Leu Phe Lys Glu Asn
210 215 220
Pro Gly Val Val Asp Ser Gly Asn Gly Leu Lys Tyr Ser Ser Leu Phe
225 230 235 240
Glu Ala Gln Ile Asp Ala Val Phe Ala Ala Met Ser Ala Ile Gln Tyr
245 250 255
Asp Asp Val Lys Met Val Val Thr Glu Thr Gly Trp Pro Ser Met Gly
260 265 270
Asp Gln Asp Glu Lys Gly Ala Ser Glu Ser Asn Ala Ala Ser Tyr Asn
275 280 285
Gly Asn Leu Val Arg Lys Val Leu Thr Gly Asn Gly Thr Pro Leu Arg
290 295 300
Pro Gln Asp Pro Leu Asn Val Tyr Leu Phe Ala Leu Phe Asn Glu Asn
305 310 315 320
Gln Lys Pro Gly Pro Thr Ser Glu Arg Asn Tyr Gly Leu Phe Tyr Pro
325 330 335
Asn Glu Gln Lys Val Tyr Thr Ile Pro Leu Thr Lys Glu Glu Ala Lys
340 345 350
Thr Gly Glu Ser Thr Gln Val Asn Thr Asn Thr Ser Leu Ala Pro Val
355 360 365
Ala Gly Glu Val Ser Lys Ala Lys Val Gly Gln Thr Trp Cys Val Ala
370 375 380
Asn Glu Lys Ala Asp Glu Lys Lys Leu Gln Ala Ala Leu Asp Tyr Ala
385 390 395 400
Cys Gly Glu Gly Glu Ala Asp Cys Ser Pro Ile Gln Pro Gly Ala Thr
405 410 415
Cys Tyr Asn Pro Asn Thr Leu Glu Ala His Ala Ser Tyr Ala Phe Asn
420 425 430
Ser Tyr Tyr Gln Lys Asn Thr Arg Ala Thr Gly Thr Cys Glu Phe Gly
435 440 445
Gly Ala Ala Tyr Val Val Thr Gln Arg Pro Thr Tyr Gly Asn Cys Glu
450 455 460
Phe Pro Thr Gly His
465

Claims (4)

1. CottonGhGLU19Use of genes for increasing cotton yield or for breeding new varieties of cotton with increased yield by inhibiting cotton growthGhGLU19The expression quantity of the gene improves the cotton yield or cultivates a new cotton seed with improved yield;
the cottonGhGLU19The gene is any one of (1) to (2):
(1) The gene is in a Germinatum TM-1A subgroup, and the nucleotide sequence of the full-length cDNA ORF is shown as SEQ ID NO. 1;
(2) The gene is in a Gekko cotton TM-1D subgroup, and the nucleotide sequence of the full-length cDNA ORF of the gene is shown as SEQ ID NO. 2.
2. The use of claim 1, wherein said cotton is inhibitedGhGLU19The expression level of the gene is determined by using a plant expression vector pBI121GhGLU19Integrating gene sequence on the carrier in antisense direction, constructing the antisense expression carrier of said gene, transforming the antisense expression carrier into cotton by agrobacterium mediation method to obtainGhGLU19Transgenic cotton material with significantly reduced gene expression.
3. Use according to claim 1, wherein the improvement in cotton yield is an increase in cotton yield trait as defined by: (1) increasing the ring weight; (2) increasing the number of seeds per bell; (3) increasing the effective bell number of the single plant; (4) increasing the clothes score; and (5) enhancing the seed vigor.
4. A method for increasing cotton yield by inhibiting cotton growthGhGLU19The expression quantity of the gene improves the cotton yield; the cottonGhGLU19The gene is any one of (1) to (2):
(1) The gene is in a Gekko cotton TM-1A subgroup, and the nucleotide sequence of the full-length cDNA ORF of the gene is shown as SEQ ID NO. 1;
(2) The gene is in a Germinatum TM-1D subgroup, and the nucleotide sequence of the full-length cDNA ORF is shown as SEQ ID NO. 2.
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