A small peptide hormone for improving rice's tolerance to high-temperature stress and / or rice yield under high-temperature stress and its application.

Abstract

This invention discloses a small peptide hormone that enhances the rice's tolerance to high-temperature stress and / or its yield under high-temperature stress. The amino acid sequence of the small peptide hormone is shown in Seq ID No:1, and it is an artificially synthesized exogenous sprayable small peptide hormone. This invention also discloses a composition containing the above-mentioned small peptide hormone and its preparation method. Furthermore, this invention discloses the method and application of using the above-mentioned small peptide hormone or small peptide hormone composition. It is applied in solution form to the panicle, leaves, and stems of rice plants during the flowering period and before high-temperature stress, and can be used to improve the rice's tolerance to high-temperature stress and / or its yield (including grain filling rate and thousand-grain weight) under high-temperature stress. This invention can significantly improve the rice's resistance to high-temperature disasters, especially its resistance during the flowering period. It can increase the grain filling rate and thousand-grain weight when high-temperature disasters occur, reducing yield losses caused by high temperatures, and is of great significance for mitigating high-temperature disasters affecting rice in summer in my country.

Description

Technical Field

[0001] This invention relates to the field of small peptide hormones, and in particular to a small peptide hormone that improves the rice's tolerance to high temperature stress and / or the rice yield under high temperature stress, and its application. Background Technology

[0002] In recent decades, my country has experienced frequent high temperatures in summer, which has seriously affected the country's summer rice production. In particular, prolonged high temperatures during the rice flowering period can lead to a decrease in the rice grain filling rate, reduced yield, and production losses.

[0003] Therefore, there is an urgent need in production for products and methods that can improve the rice's tolerance to high-temperature stress, especially its tolerance during the flowering period, and to resist high-temperature disasters during the rice flowering stage. Currently, there are no reports on using small peptide hormones to improve the rice's tolerance to high-temperature stress or its yield under high-temperature stress. Summary of the Invention

[0004] The technical problem to be solved by the present invention is to provide a small peptide hormone and its application to improve the rice's resistance to high temperature stress and / or rice yield under high temperature stress, so as to greatly improve the rice's resistance to high temperature disasters, especially the resistance to high temperature disasters during the flowering period, increase the rice seed setting rate and thousand-grain weight when high temperature disasters occur, and reduce the yield loss caused by high temperature.

[0005] To solve the above-mentioned technical problems, the present invention adopts the following technical solution:

[0006] On the one hand, the present invention provides a small peptide hormone that improves the rice's resistance to high temperature stress and / or the rice yield under high temperature stress, the amino acid sequence of which is shown in Seq ID No:1.

[0007] As a further improvement of the present invention, the small peptide hormone is an artificially synthesized exogenous sprayed small peptide hormone.

[0008] In a second aspect, the present invention provides a small peptide hormone composition for improving the rice's resistance to high temperature stress and / or the rice yield under high temperature stress, comprising the above-mentioned small peptide hormone and a solvent for dissolving the small peptide hormone.

[0009] Furthermore, the solvent for dissolving the small peptide hormone is acetonitrile.

[0010] Furthermore, the small peptide hormone is dissolved in acetonitrile solvent to prepare a 10 μM solution.

[0011] Furthermore, the acetonitrile solvent is a 25% acetonitrile solvent, which is a solvent prepared by mixing water and acetonitrile in a volume ratio of 3:1.

[0012] Thirdly, the present invention provides a method for preparing the above-mentioned small peptide hormone composition for improving rice's resistance to high-temperature stress and / or rice yield under high-temperature stress, comprising the following steps:

[0013] (1) The amino acid sequence shown in Seq ID No:1 was artificially synthesized;

[0014] (2) Dissolve the artificially synthesized amino acid sequence powder in a solvent to prepare a spray solution.

[0015] Fourthly, the present invention provides a method for using the above-mentioned small peptide hormone or small peptide hormone composition, wherein the method is: spraying the rice plant panicle, leaves and stem surface in solution form during the rice flowering period and before high temperature stress.

[0016] Fifthly, the present invention provides an application of the above-mentioned small peptide hormone or small peptide hormone composition, the application being to improve the rice's resistance to high temperature stress and / or the rice yield under high temperature stress.

[0017] Furthermore, the application is to improve the rice's tolerance to high-temperature stress and / or rice yield under high-temperature stress by external spraying in solution form; improving the rice yield under high-temperature stress includes increasing the rice seed setting rate and thousand-grain weight under high-temperature stress.

[0018] This invention has discovered a small peptide hormone that can greatly improve the resistance of rice to high-temperature disasters, especially during the flowering period. It can be used to reduce the damage caused by high temperatures in summer in my country, increase the rice seed setting rate and thousand-grain weight when high-temperature disasters occur, and reduce yield losses caused by high temperatures, which is of great significance to my country's food security. Attached Figure Description

[0019] The above is merely an overview of the technical solution of the present invention. In order to better understand the technical means of the present invention, the present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments.

[0020] Figure 1 Figure 1 shows the changes in seed setting rate of Huazhan and Huazhan GS2 under high temperature treatment; (a) Comparison of Huazhan plants at maturity (top) and after high temperature treatment (bottom); (b) Comparison of Huazhan GS2 plants at maturity (top) and after high temperature treatment (bottom); (c) Photographs of the ears of Huazhan (left) and Huazhan GS2 (right) under normal temperature treatment at maturity; (d) Photographs of the ears of Huazhan (left) and Huazhan GS2 (right) under high temperature treatment at maturity; (e) Comparison of seed setting rate of Huazhan and Huazhan GS2 at maturity under normal temperature (control) and high temperature treatment.

[0021] Figure 2This is a heatmap showing the expression levels of 454 genes detected by transcriptome analysis. On the left, Log2FC_HZHS2_vs_HZCK2 represents the Log2 Fold change value between the Huazhan high-temperature treatment and the Huazhan control, obtained by dividing the expression level of the Huazhan high-temperature treatment by the expression level of the Huazhan control, followed by Log2 transformation. On the right, Log2FC_GS2HS_vs_GS2CK represents the Log2 Fold change value between the Huazhan GS2 high-temperature treatment and the control, obtained by dividing the expression level of the Huazhan GS2 high-temperature treatment by the expression level of the Huazhan GS2 control, followed by Log2 transformation.

[0022] Figure 3 A heatmap showing the expression levels of 257 proteins that were upregulated after high-temperature treatment of Huazhan and downregulated after high-temperature treatment of Huazhan GS2, as detected by proteomic analysis.

[0023] Figure 4 Comparison of seed setting rate of Huazhan and Huazhan GS2 after spraying with small peptide hormone at high temperature; (a) Comparison of ears at maturity of Huazhan at normal temperature (control), blank group without amino acid sequence spraying at high temperature, and spraying group with amino acid sequence spraying at high temperature; (b) Comparison of ears at maturity of Huazhan GS2 at normal temperature (control), blank group without amino acid sequence spraying at high temperature, and spraying group with amino acid sequence spraying at high temperature. Detailed Implementation

[0024] First, it should be noted that this invention was discovered accidentally during the exploration of the heat resistance mechanism of Huazhan and Huazhan GS2. The rice restorer line Huazhan was obtained from the National Crop Germplasm Resource Rice Mid-term Bank of the China National Rice Research Institute. The near-isogenic line "Huazhan GS2" was obtained by using Kuangsi Huaju as the donor parent and Huazhan as the recurrent parent, through hybridization and four backcrosses, and multiple consecutive self-pollination homozygotes from a large-grained single plant selected from the BC4F1 generation. "Huazhan GS2" was provided by the National Rice Improvement Center of the China National Rice Research Institute.

[0025] During the flowering period, Huazhan and Huazhan GS2 rice varieties were placed in an artificial climate chamber for high-temperature treatment. The high-temperature treatment method was as follows: 7:01 AM to 9:34 AM; 9:01 AM to 2:38 PM; 2:01 PM to 5:36 PM; 5:01 PM to 9:33 PM; 9:01 PM to 7:27 AM the next day. After 7 days of high-temperature treatment, Huazhan and Huazhan GS2 rice varieties were transferred to normal rice growth temperature conditions, with the following normal temperature adjustments: 7:01 AM to 9:26 AM; 9:01 AM to 2:30 PM; 2:01 PM to 5:28 PM; 5:01 PM to 9:25 PM; 9:01 PM to 7:23 AM the next day. Huazhan and Huazhan GS2 rice varieties were placed under these normal temperature conditions until the rice matured and was harvested. After maturation, the fruit setting rate of Huazhan and Huazhan GS2 was examined. It was found that the fruit setting rate of Huazhan after high-temperature treatment was 38.3%, while that of Huazhan GS2 was 20.7%. Huazhan is more heat-resistant than Huazhan GS2. Figure 1 To explore the mechanism by which Huazhan GS2 rice is more heat-tolerant, transcriptome RNA sequencing and proteomics analysis were performed simultaneously on four samples: Huazhan rice and Huazhan GS2 rice after high-temperature treatment, and Huazhan rice and Huazhan GS2 rice without high-temperature treatment (control). The hypothesis was that the greater heat tolerance of Huazhan rice and Huazhan GS2 rice is due to the existence of genes positively regulating heat tolerance, i.e., the expression level of Huazhan rice was upregulated after high-temperature treatment, while the expression level of Huazhan GS2 rice was downregulated. Based on this hypothesis, genes whose expression level was upregulated in Huazhan rice and downregulated in Huazhan GS2 rice after high-temperature treatment were screened from the transcriptome RNA sequencing results. A total of 454 related genes were identified. The gene numbers and Log2 Fold change values ​​of these 454 genes are shown in Table 1. These 454 genes are likely related to heat tolerance in rice. A heatmap of the Log2 Fold change values ​​of these 454 genes is shown in [Table 1]. Figure 2 .

[0026] In addition, proteomics analysis was used to screen for proteins whose expression was upregulated after high-temperature treatment of Huazhan rice and proteins whose expression was downregulated after high-temperature treatment of Huazhan GS2 rice. A total of 257 proteins were screened. The gene codes and Log2 Fold change values ​​of the encoding genes of these 257 proteins are shown in Table 2. Therefore, the 257 protein encoding genes listed in Table 2 are likely related to the heat tolerance of rice. The expression heatmaps of these 257 proteins are shown below. Figure 3 .

[0027] Through combined transcriptomics and proteomics analysis—comparing the 454 possible heat-resistance-related genes detected by transcriptomics RNA sequencing listed in Table 1 and the 257 possible heat-resistance protein-coding genes detected by proteomics analysis listed in Table 2—six genes were found to be co-detected by both transcriptomics RNA sequencing and proteomics analysis. These six common genes are listed in Table 3, and they were used as candidate heat-resistance genes for further analysis. Further analysis of the conserved domains of the proteins encoded by these six candidate genes revealed that the amino acid sequences of these conserved domains are shown in Table 4. Amino acid sequence fragments of the conserved domains of the proteins encoded by the six genes listed in Table 4 were synthesized and applied externally to rice plant leaves for high-temperature experiments. The results showed that the amino acid sequence fragment of the BGIOSGA018627 gene can improve the heat resistance of rice (specific experimental results are shown in Tables 5 and 6). This is the reason why the BGIOSGA018627 gene was selected as a small peptide hormone in this invention.

[0028] Table 1. Log2FC values ​​of expression levels of 454 genes detected by transcriptome sequencing (It should be noted that this table only lists a portion of the 454 genes as an example).

[0029] BGIOSGA024495 1.0120 -6.8728 BGIOSGA007554 0.8304 -4.6702 BGIOSGA038856 0.7370 -4.2479 BGIOSGA039926 0.7370 -4.2479 BGIOSGA038975 0.5850 -3.9070 BGIOSGA023050 1.6471 -3.7005 BGIOSGA010977 2.8651 -3.5443 BGIOSGA013682 3.3552 -3.0780 BGIOSGA031864 0.9889 -2.9100 BGIOSGA001493 1.5185 -0.9286 BGIOSGA015866 0.9419 -0.9275 BGIOSGA026208 1.1627 -0.9209 BGIOSGA021881 0.9664 -0.9094 BGIOSGA004837 0.7843 -0.9069 BGIOSGA027545 0.6401 -0.9030 BGIOSGA027908 1.1903 -0.9012 BGIOSGA013156 0.6897 -0.9005 BGIOSGA036918 0.5854 -0.8953 BGIOSGA001669 1.2224 -0.8931 BGIOSGA026881 2.1481 -0.8911 BGIOSGA004587 1.5763 -0.8845 BGIOSGA028786 1.0302 -0.8832 BGIOSGA038665 0.7776 -0.8745 BGIOSGA022102 1.3219 -0.8745 BGIOSGA026503 1.3219 -0.8745 BGIOSGA021071 0.5380 -0.8731 BGIOSGA009512 0.9717 -0.8655 BGIOSGA011166 0.7226 -0.8631 BGIOSGA005859 1.0589 -0.8625 BGIOSGA030260 1.9696 -0.8625 BGIOSGA021710 1.5729 -0.8601 BGIOSGA012554 1.0000 -0.8580 BGIOSGA025940 2.0704 -0.8480 BGIOSGA027487 0.9511 -0.8480 BGIOSGA001683 0.7885 -0.8480 BGIOSGA006410 0.9270 -0.8420 BGIOSGA015024 1.3347 -0.8415 BGIOSGA025447 0.9783 -0.8349 BGIOSGA015888 1.6674 -0.8329 BGIOSGA035438 0.7539 -0.8295 ENSRNA049493369 0.5338 -0.8282 BGIOSGA033602 1.2828 -0.5603 ENSRNA049493884 0.6761 -0.5557 BGIOSGA020027 0.5454 -0.5536 BGIOSGA040654 0.9492 -0.5527 BGIOSGA038955 1.3026 -0.5475 ENSRNA049493745 1.1435 -0.5424 BGIOSGA010696 1.0587 -0.5391 BGIOSGA004350 0.6088 -0.5389 BGIOSGA030908 0.9868 -0.5348 BGIOSGA004220 1.2224 -0.5343 BGIOSGA013896 1.2591 -0.5334 BGIOSGA026043 1.1170 -0.5332 BGIOSGA004967 0.5577 -0.5317 BGIOSGA032911 0.6038 -0.5313 BGIOSGA031601 0.6289 -0.5289 [[ID=]]BGIOSGA023635 0.7859 -0.5272 BGIOSGA024506 1.0001 -0.5146 BGIOSGA026005 0.6057 -0.5131 BGIOSGA020294 0.5487 -0.5129 BGIOSGA031966 0.9034 -0.5120 BGIOSGA026648 0.7966 -0.5090 BGIOSGA0050 1.2856 -0.5090 BGIOSGA008892 0.7755 -0.5068 BGIOSGA024757 0.7878 -0.5047 BGIOSGA028367 0.9184 -0.5041 BGIOSGA028152 0.5640 -0.5033 BGIOSGA037454 0.6002 -0.5033 BGIOSGA016925 0.7193 -0.5031 BGIOSGA004075 0.6630 -0.5025 BGIOSGA014846 0.6642 -0.5006 BGIOSGA029682 1.8433 -0.4975 BGIOSGA004059 0.9635 -0.4926 BGIOSGA024963 0.5057 -0.3632 BGIOSGA018627 0.5731 -0.3617 BGIOSGA012816 0.6691 -0.3614 BGIOSGA008984 0.5205 -0.3550 BGIOSGA013326 1.4753 -0.3548 BGIOSGA011711 0.5095 -0.3541 BGIOSGA004014 0.5829 -0.3540 BGIOSGA005771 0.9392 -0.3538 BGIOSGA032418 2.9329 -0.3536 BGIOSGA023536 0.6582 -0.3515 BGIOSGA016351 0.6633 -0.3492 BGIOSGA016285 0.5938 -0.3484 BGIOSGA002880 0.5361 -0.3479 BGIOSGA007045 0.6766 -0.3466 BGIOSGA009279 1.1134 -0.3449 BGIOSGA018226 0.6650 -0.3435 BGIOSGA035758 0.6951 -0.3410 BGIOSGA035952 0.7254 -0.3395 BGIOSGA028223 1.6881 -0.3395 BGIOSGA016999 0.8163 -0.3379 BGIOSGA037914 0.7437 -0.3370 BGIOSGA037628 1.1196 -0.3367 BGIOSGA024365 1.7843 -0.3344 BGIOSGA015714 0.6781 -0.3344 BGIOSGA029510 1.0721 -0.3326 BGIOSGA025898 2.0641 -0.3316 BGIOSGA037439 0.7413 -0.3288 BGIOSGA035607 1.2945 -0.3267 BGIOSGA022313 1.1118 -0.3259 BGIOSGA009251 0.5301 -0.3255 BGIOSGA031032 0.9103 -0.3248 BGIOSGA004596 0.7196 -0.3234 BGIOSGA013880 1.0506 -0.3219 BGIOSGA021042 1.5106 -0.3219 BGIOSGA013272 0.8442 -0.3219 BGIOSGA028957 0.5815 -0.3203 BGIOSGA019001 0.6613 -0.3166 BGIOSGA031016 0.6113 -0.3162 BGIOSGA022577 1.2424 -0.3160 BGIOSGA013984 0.8797 -0.3148 BGIOSGA037878 0.5075 -0.3138 BGIOSGA023129 0.6826 -0.3120 BGIOSGA015674 0.5172 -0.3114 BGIOSGA009327 0.9733 -0.3075 BGIOSGA018383 0.9171 -0.3073 BGIOSGA025947 0.5087 -0.3035 BGIOSGA017894 0.5957 -0.3029 BGIOSGA027977 2.6349 -0.2990 BGIOSGA019454 0.7954 -0.2982 BGIOSGA035432 1.0780 -0.2964 BGIOSGA021222 0.8162 -0.2958 BGIOSGA006084 0.5382 -0.2930 BGIOSGA015151 0.6443 -0.2921 BGIOSGA035946 0.8707 -0.2918 BGIOSGA020953 0.6681 -0.2914 BGIOSGA008153 0.7255 -0.2904 BGIOSGA021822 1.9143 -0.2895 BGIOSGA040312 2.1699 -0.2895 BGIOSGA009425 0.6687 -0.2869 BGIOSGA030561 0.7256 -0.2843 BGIOSGA022593 0.9602 -0.2829 BGIOSGA013751 0.6172 -0.2809 BGIOSGA011997 0.5034 -0.2775 BGIOSGA009908 0.5716 -0.2758

[0030] In Table 1 above, the Log2FC_HZHS2_vs_HZCK2 value is greater than 0.5, and the Log2FC_GS2HS2_vs_GS2CK2 value is negative. Log2FC_HZHS2_vs_HZCK2 refers to the Log2 Fold change value between the high-temperature treatment and control of Huazhan (a specific gene), obtained by dividing the gene expression level after high-temperature treatment of Huazhan by the gene expression level of the Huazhan control, and then performing a Log2 transformation. A positive Log2FC_HZHS2_vs_HZCK2 value indicates an increase in gene expression after high-temperature treatment. This table only screens genes with a Log2FC_HZHS2_vs_HZCK2 value greater than 0.5, aiming to detect genes whose expression levels are upregulated after high-temperature treatment in Huazhan. Log2FC_GS2HS_vs_GS2CK refers to the Log2 Fold change value between the high-temperature treatment and control of Huazhan (a specific gene). The change value is obtained by dividing the gene expression level after high-temperature treatment with Huazhan GS2 by the gene expression level of the Huazhan GS2 control, and then performing a Log2 transformation. A negative Log2FC_GS2HS_vs_GS2CK value indicates that the gene expression level decreased after high-temperature treatment. This table only screens genes with negative Log2FC_GS2HS_vs_GS2CK values, aiming to detect genes whose Huazhan GS2 expression level was downregulated after high-temperature treatment.

[0031] Table 2. Log2FC values ​​of expression levels of 257 proteins detected by proteomics (It should be noted that this table only lists a portion of the 257 proteins as an example).

[0032] BGIOSGA033362 0.9421 -2.0592 BGIOSGA014499 0.2596 -1.6173 BGIOSGA000030 0.7893 -1.4267 BGIOSGA026981 0.2333 -1.2379 BGIOSGA014739 0.3549 -1.0167 BGIOSGA002246 0.4537 -0.9786 BGIOSGA006294 0.4201 -0.9061 BGIOSGA028896 0.7520 -0.9013 BGIOSGA007794 0.5065 -0.8823 BGIOSGA005843 0.4934 -0.8527 BGIOSGA008199 0.3703 -0.8363 BGIOSGA017328 0.2710 -0.8191 BGIOSGA028312 0.3837 -0.6420 BGIOSGA034953 0.3762 -0.5835 BGIOSGA013765 0.2534 -0.5699 BGIOSGA023570 0.3716 -0.5608 BGIOSGA007125 0.6716 -0.5440 BGIOSGA013586 0.3241 -0.5324 BGIOSGA015153 0.3256 -0.5282 BGIOSGA003143 0.6876 -0.4924 BGIOSGA017671 0.2765 -0.4892 BGIOSGA001442 0.5170 -0.4703 BGIOSGA025258 0.2038 -0.4618 BGIOSGA008354 0.3107 -0.4597 BGIOSGA007377 0.2667 -0.4437 BGIOSGA017665 0.3546 -0.4364 BGIOSGA003089 0.3070 -0.4282 BGIOSGA018248 0.4753 -0.4173 BGIOSGA011874 0.2240 -0.4129 BGIOSGA013984 0.5010 -0.4037 BGIOSGA034553 0.3665 -0.4001 BGIOSGA024393 0.3531 -0.3916 BGIOSGA026376 0.4503 -0.3914 BGIOSGA037861 0.2205 -0.3825 BGIOSGA012069 0.2013 -0.3761 BGIOSGA031298 0.2620 -0.3736 BGIOSGA003033 0.2266 -0.3735 BGIOSGA018322 0.2139 -0.3673 BGIOSGA034735 0.3126 -0.3608 BGIOSGA018906 0.3595 -0.3605 BGIOSGA021986 0.2044 -0.3573 BGIOSGA010714 0.3656 -0.3444 BGIOSGA005713 0.7074 -0.3382 BGIOSGA003920 0.3163 -0.3339 BGIOSGA010966 0.4458 -0.3160 BGIOSGA008403 0.2083 -0.3126 BGIOSGA025371 0.4887 -0.3065 BGIOSGA018537 0.7416 -0.3041 BGIOSGA027974 0.2159 -0.3020 BGIOSGA001422 0.3715 -0.2805 BGIOSGA014843 0.2078 -0.2799 BGIOSGA009714 0.2105 -0.2782 BGIOSGA003388 1.6209 -0.2731 BGIOSGA016344 0.8181 -0.1425 BGIOSGA006015 0.2945 -0.1417 BGIOSGA000298 0.2317 -0.1410 BGIOSGA017393 0.4021 -0.1390 BGIOSGA009223 0.2641 -0.1383 BGIOSGA005337 0.2639 -0.1357 BGIOSGA005429 0.2227 -0.1347 BGIOSGA037619 0.2891 -0.1311 BGIOSGA033656 0.2594 -0.1299 BGIOSGA003165 0.2798 -0.1254 BGIOSGA036566 0.5260 -0.1240 BGIOSGA034506 0.2045 -0.1225 BGIOSGA032154 0.2399 -0.1208 BGIOSGA008103 0.7705 -0.1205 BGIOSGA034973 0.3251 -0.1194 BGIOSGA028103 0.2711 -0.1189 BGIOSGA020252 0.4251 -0.1175 BGIOSGA014061 0.2165 -0.1155 BGIOSGA010029 0.2039 -0.1137 BGIOSGA004612 0.3332 -0.1123 BGIOSGA001119 0.3405 -0.1113 BGIOSGA029702 0.2296 -0.1103 BGIOSGA001493 0.2482 -0.1099 BGIOSGA011003 0.2196 -0.1091 BGIOSGA021800 0.2657 -0.1064 BGIOSGA035694 0.2272 -0.1028 BGIOSGA012909 0.3124 -0.1026 BGIOSGA016159 0.2459 -0.1025 BGIOSGA033602 0.2074 -0.1023 BGIOSGA019738 0.3490 -0.1007 BGIOSGA008825 0.2974 -0.1000 BGIOSGA002757 0.2971 -0.0998 BGIOSGA034009 0.2481 -0.0993 BGIOSGA000683 0.2150 -0.0985 BGIOSGA003516 0.3627 -0.0969 BGIOSGA026971 0.3396 -0.0952 BGIOSGA017314 0.3013 -0.0949 BGIOSGA019115 0.2197 -0.0948 BGIOSGA009635 0.4201 -0.0927 BGIOSGA015767 0.3429 -0.0924 BGIOSGA036042 0.2770 -0.0919 BGIOSGA008611 0.3274 -0.0905 BGIOSGA039972 0.3600 -0.0903 BGIOSGA009876 0.2577 -0.0903 BGIOSGA008933 0.3107 -0.0902 BGIOSGA026489 0.5549 -0.0902 BGIOSGA012464 0.3099 -0.0898 BGIOSGA025982 0.2019 -0.0880 BGIOSGA003260 0.2136 -0.0872 BGIOSGA020199 0.2510 -0.0872 BGIOSGA009440 0.3055 -0.0861 BGIOSGA026262 0.2002 -0.0849 BGIOSGA013561 0.2015 -0.0841 BGIOSGA027802 0.4019 -0.0828 BGIOSGA020294 0.3993 -0.0824 BGIOSGA035157 0.2305 -0.0824 BGIOSGA017624 0.2968 -0.0805 BGIOSGA009404 0.2157 -0.0794 BGIOSGA000588 0.3342 -0.0793 BGIOSGA008798 0.2818 -0.0789 BGIOSGA025435 0.3236 -0.0784 BGIOSGA013052 0.4215 -0.0782 BGIOSGA022532 0.2028 -0.0775 BGIOSGA023250 0.2219 -0.0751 BGIOSGA033494 0.2244 -0.0097 BGIOSGA017923 0.2631 -0.0082 BGIOSGA010628 0.2250 -0.0081 BGIOSGA031512 0.2600 -0.0071 BGIOSGA035064 0.2626 -0.0054 BGIOSGA008648 0.2027 -0.0054 BGIOSGA018627 0.3269 -0.0054 BGIOSGA025715 0.3748 -0.0044 BGIOSGA025155 0.2103 -0.0022 BGIOSGA021071 0.2868 -0.0016 BGIOSGA011269 0.2185 -0.0012 BGIOSGA037317 0.2892 -0.0003

[0033] In Table 2 above, the Log2FC_HZHS2_vs_HZCK2 value is greater than 0.2, and the Log2FC_GS2HS2_vs_GS2CK2 value is negative. Log2FC_HZHS2_vs_HZCK2 refers to the Log2 Fold change value between the high-temperature treatment and control of Huazhan protein. It is obtained by dividing the protein expression level after high-temperature treatment of Huazhan protein by the protein expression level of the Huazhan control protein, and then performing a Log2 transformation. A positive Log2FC_HZHS2_vs_HZCK2 value indicates an increase in the expression level of the protein after high-temperature treatment. This table only screens protein-coding genes with a Log2FC_HZHS2_vs_HZCK2 value greater than 0.2, aiming to detect proteins whose expression levels are upregulated after high-temperature treatment in Huazhan protein. Log2FC_GS2HS_vs_GS2CK refers to the Log2 Fold change value between the high-temperature treatment and control of Huazhan GS2 protein. The change value is obtained by dividing the protein expression level of Huazhan GS2 after high-temperature treatment by the expression level of the Huazhan GS2 control protein, and then performing a Log2 transformation. A negative Log2FC_GS2HS_vs_GS2CK value indicates that the expression level of the gene decreased after high-temperature treatment. This table only screens protein-coding genes with negative Log2FC_GS2HS_vs_GS2CK values, aiming to detect proteins whose expression level of Huazhan GS2 is downregulated after high-temperature treatment.

[0034] Table 3. Six genes detected simultaneously by combined transcriptomics and proteomics analysis, i.e., genes appearing in both Table 1 and Table 2.

[0035] BGIOSGA013984 0.5010 -0.4037 0.8797 -0.3148 BGIOSGA001493 0.2482 -0.1099 1.5185 -0.9286 BGIOSGA033602 0.2074 -0.1023 1.2828 -0.5603 BGIOSGA020294 0.3993 -0.0824 0.5487 -0.5129 BGIOSGA018627 0.3269 -0.0054 0.5731 -0.3617 BGIOSGA021071 0.2868 -0.0016 0.5380 -0.8731

[0036] Table 4. Amino acid sequences of conserved domains of proteins encoded by six candidate genes obtained from combined transcriptomic and proteomic analysis.

[0037] BGIOSGA013984 RLIMQGRNVKLNEKVKEHIEEKAGRAVAKHSQLVKEVDVRLSARGGELSRGPKICRCEITLFTKRHGVIRAEEDAESTYASIDLASSIIKRKLRKIKEK BGIOSGA001493 LQDEYRVRFHYGHPDIFDRLFHITRGGISKASKTINLSEDIFSGFNSTMREGNVTHHEYMQVGKGRDVGMNQISSFEAKVANGNGEQTLSRDIYRLGRRFDFYRMLSFYFTTVGFYFSSMV BGIOSGA033602 ALQAWRRAFYSDPKGYTNNWTGNDVCSYNGVIC BGIOSGA020294 ECAWTCDHVAAGNKKMCNTLRKLPGVSSPKELLTAAVKLSMRKAKAARARFEAAARAAEKGTPMESILDTCKEGYDSTVSALQEVQRCIDANDSKASLITKMSAATTFTGDCGNAYEERELEPSLALKATKNNVNRVVTGALAIAAKLK BGIOSGA018627 RRALAARKPTNRYVSYSALDANKVPCNKRGQTYYQNCASQQAANPYRRGCSAITRCSR BGIOSGA021071 KTCNPLYIVTPTDVKHIQVAVSCGRRHNVRIRVRSGGHDYEGLSYRSEIPEPFAIVDLVNMRNVTVDGKARTAWVESGAQIGELYYGISKASPTLAFPAGVCPTIGVGGHFSGGGFGMLLRKFGLASDNVLDVKVVDANGKVQDRKSMGEDYLWAVRGGGGSSFGIVVSWKLRLLPV

[0038] This invention screens tens of thousands of rice genes through transcriptome RNA sequencing and proteomics analysis. Six candidate genes were selected through combined transcriptome and proteome analysis. Amino acid sequence fragments of the conserved structural domains of the proteins encoded by the six candidate genes were synthesized. After spraying different amino acid sequence fragments, high-temperature identification and comparison experiments were conducted. Finally, an amino acid sequence fragment (small peptide hormone) that improves rice's tolerance to high-temperature stress and rice yield under high-temperature stress was obtained. The process of obtaining the source gene BGIOSGA018627 of this small peptide hormone is described above. This result was an unexpected discovery during the research process, and the specific process of discovery has been described above.

[0039] The amino acid sequence of the small peptide hormone in this invention comes from the rice BGIOSGA018627 gene. It is a 58-amino acid sequence extracted from the full-length 137-amino acid coding sequence of the BGIOSGA018627 gene. The small peptide hormone is obtained by in vitro artificial synthesis.

[0040] The present invention found that the exogenous spraying of the above-mentioned amino acid sequence fragment solution can improve the rice's ability to withstand high temperature stress. Experiments were conducted to verify this, further proving the reliability of its application.

[0041] The experimental method of this invention is as follows:

[0042] Step (1): The 58-amino acid sequence RRALAARKPTNRYVSYSALDANKVPCNKRGQTYYQNCASQQAANPYRRGCSAITRCSR was synthesized by Hefei Hesheng Biotechnology Co., Ltd.

[0043] Step (2): The synthesized 58-length amino acid sequence powder is dissolved in 25% acetonitrile solvent to prepare a 10 μM solution. The 25% acetonitrile solvent is a solution of water and acetonitrile in a 3:1 volume ratio. The 58-length amino acid sequence powder (i.e., the small peptide hormone) is the active substance, and the main function of the acetonitrile solvent is to dissolve the small peptide hormone. Those skilled in the art will understand that other conventional solvents suitable for plant spraying and capable of dissolving amino acid sequence powder can also achieve the purpose of this invention.

[0044] Step (3): Select the rice restorer line "Huazhan" and its near-isogenic line "Huazhan GS2" as experimental materials. During the flowering period of Huazhan and Huazhan GS2, one hour before the onset of high-temperature stress, spray a solution containing an amino acid sequence of length 58 on the surface of the panicles, leaves, and stems of the Huazhan and Huazhan GS2 plants. The purpose is to allow the rice plants, including the panicles, leaves, and stems, one hour to fully absorb the sprayed amino acid sequence. Those skilled in the art will understand that the above-mentioned rice can be different types of rice materials such as indica rice, japonica rice, hybrid rice, conventional rice, sterile lines, and restorer lines. This embodiment only uses Huazhan and Huazhan GS2 as examples for illustration.

[0045] Step (4): Huazhan and Huazhan GS2 were placed in an artificial climate chamber for 7 consecutive days of high-temperature treatment. The high-temperature treatment method was as follows: 7:01 a.m. to 9:34 a.m.; 9:01 a.m. to 2:38 p.m.; 2:01 p.m. to 5:36 p.m.; 5:01 p.m. to 9:33 p.m.; 9:01 p.m. to 7:27 a.m. the next day; for a total of 168 hours of continuous treatment over 7 days. The purpose of this step was to use an artificial climate chamber to fully simulate the damage caused by continuous high temperatures of 38°C for 7 days in the summer in rice-growing areas of my country.

[0046] Step (5): After 7 days of high-temperature treatment, Huazhan and Huazhan GS2 rice were placed in an artificial climate chamber under normal natural temperature conditions until maturity. The purpose of this step was to simulate the situation after 7 consecutive days of high-temperature damage at 38℃ in rice-growing areas of my country in summer. Normal natural temperature conditions refer to the following temperature conditions: 7:01 AM to 9:26 AM; 9:01 AM to 2:30 PM; 2:01 PM to 5:28 PM; 5:01 PM to 9:25 PM; 9:01 PM to 7:23 AM the next day. This temperature cycle was repeated daily until the rice matured.

[0047] The above method will be described in detail below through examples:

[0048] 1. The rice restorer line Huazhan used in this embodiment was obtained from the National Crop Germplasm Resource Rice Mid-term Bank of the China National Rice Research Institute. The near-isogenic line "Huazhan GS2" was obtained by using Kuangsi Huaju as the donor parent and Huazhan as the recurrent parent. Through hybridization and four backcrosses, a large-grained single plant selected from the BC4F1 generation was subjected to multiple consecutive self-pollination homozygotes. "Huazhan GS2" was provided by the National Rice Improvement Center of the China National Rice Research Institute. Huazhan and Huazhan GS2 were cultured in pots. The method was as follows: On May 21, 2024, Huazhan and Huazhan GS2 seedlings were sown at the Experimental Farm of China National Rice Research Institute in Fuyang District, Hangzhou City, Zhejiang Province. Twenty-two days after sowing, Huazhan seedlings were transplanted into 12 plastic buckets (20cm high and 20cm in diameter), and Huazhan GS2 seedlings were transplanted into another 12 plastic buckets. Three Huazhan or Huazhan GS2 seedlings were planted in each bucket, resulting in a total of 36 Huazhan seedlings in the 12 buckets and 36 Huazhan GS2 seedlings in the other 12 buckets. Six buckets were used as treatments sprayed with a solution containing the amino acid sequence RRALAARKPTNRYVSYSALDANKVPCNKRGQTYYQNCASQQAANPYRRGCSAITRCSR, and the other six buckets served as untreated treatments, with six replicates per treatment. The potting soil is made of paddy field clay loam, with each bucket containing approximately 4 kg of dry soil. The soil contains 36.1 g / kg of organic matter, 2.70 g / kg of total nitrogen, 0.62 g / kg of total phosphorus, 20.4 g / kg of total potassium, 239 mg / kg of available nitrogen, 9.8 mg / kg of ammonium nitrogen, 24.1 mg / kg of available phosphorus, 62 mg / kg of available potassium, and has a pH of 6.5.

[0049] 2. Place the plastic buckets planted with Huazhan or Huazhan GS2 in a well-ventilated greenhouse with normal natural light and temperature conditions, and manage water and fertilizer normally until Huazhan or Huazhan GS2 heads and flowers.

[0050] 3. Starting from the emergence of the main spike of Huazhan or Huazhan GS2, the spike, leaves and stem surfaces of the plant were treated as follows: (1) Spraying group with 10 μM concentration of amino acid sequence RRALAARKPTNRYVSYSALDANKVPCNKRGQTYYQNCASQQAANPYRRGCSAITRCSR solution, with 6 replicates (i.e., spraying Huazhan or Huazhan GS2 planted in 6 plastic buckets, each plastic bucket is a replicate, and 5 ml is sprayed per replicate); (2) Blank group sprayed with distilled water, with 6 replicates (i.e., spraying Huazhan or Huazhan GS2 planted in 6 plastic buckets, each plastic bucket is a replicate, and 5 ml is sprayed per replicate). One hour after spraying the rice plants, 12 plastic buckets containing Huazhan (6 replicates in the spraying group and 6 replicates in the control group) and 12 plastic buckets containing Huazhan GS2 (6 replicates in the spraying group and 6 replicates in the control group) were placed in an artificial climate chamber for high-temperature treatment. The treatment temperatures were: 7:01 AM to 9:34 AM; 9:01 AM to 2:38 PM; 2:01 PM to 5:36 PM; 5:01 PM to 9:33 PM; and 9:01 PM to 7:27 AM the next day, for a total of 168 hours over 7 days.

[0051] 4. After 7 days of high-temperature treatment, the temperature of the artificial climate chamber was set as follows: 7:01 AM to 9:26 AM; 9:01 AM to 2:30 PM; 2:01 PM to 5:28 PM; 5:01 PM to 9:25 PM; 9:01 PM to 7:23 AM the next day. This temperature cycle was repeated daily until the rice matured.

[0052] 5. After the Huazhan or Huazhan GS2 ears matured and were harvested, all ears were collected, and the seed setting rate, thousand-grain weight, and yield were measured. Table 5 shows that the seed setting rate of the Huazhan ear sprayed with the amino acid sequence was 50.69%, higher than the 37.92% of the control group without the amino acid sequence. The thousand-grain weight of the Huazhan ear sprayed with the amino acid sequence was 17.72 grams, also higher than the 16.95 grams of the control group without the amino acid sequence. The yield per bucket of Huazhan ear sprayed with the amino acid sequence was 13.52 grams, also higher than the 10.41 grams of the control group without the amino acid sequence. Table 6 shows that the seed setting rate of the Huazhan GS2 ear sprayed with the amino acid sequence was 40.26%, higher than the 20.0% of the control group without the amino acid sequence. The thousand-grain weight of Huazhan GS2 treated with the amino acid sequence was 26.66 grams, which was higher than the 26.03 grams of the control group without the amino acid sequence. The yield per barrel of Huazhan GS2 treated with the amino acid sequence was 15.45 grams, which was also higher than the 7.76 grams of the control group without the amino acid sequence. Photos of the ears of Huazhan and Huazhan GS2 after high-temperature treatment are shown below. Figure 4 .

[0053] Table 5. Comparison of seed setting rate, thousand-grain weight and yield of Huazhan under two different spraying treatments during the flowering period, after 7 days of high temperature treatment and until maturity under normal temperature (6 replicates per treatment).

[0054] Amino acid sequence for spraying (spraying group) 50.69±7.43 17.72±1.25 13.52±0.83 Amino acid sequence not sprayed (blank group) 37.92±6.52 16.95±0.95 10.41±0.73

[0055] Table 6 Comparison of seed setting rate, thousand-grain weight and yield of Huazhan GS2 under two different spraying treatments during the flowering period, after 7 days of high temperature treatment and until maturity under normal temperature (6 replicates per treatment).

[0056] Amino acid sequence for spraying (spraying group) 40.26±10.56 26.66±1.56 15.45±1.77 Amino acid sequence not sprayed (blank group) 20.0±5.27 26.03±1.44 7.76±0.82

[0057] In summary, this embodiment simulates the sustained high-temperature disaster of 38℃ encountered in rice-growing areas of my country during the summer by subjecting two rice materials, Huazhan and Huazhan GS2, to continuous high-temperature treatment for 7 days in an artificial climate chamber. The experimental results have important practical significance and application value. Spraying a solution containing the amino acid sequence RRALAARKPTNRYVSYSALDANKVPCNKRGQTYYQNCASQQAANPYRRGCSAITRCSR can greatly improve the seed setting rate, thousand-grain weight, and yield of rice plants, which is of great significance for mitigating the damage caused by high-temperature disasters in rice in summer in my country.

[0058] The above description is merely an embodiment of the application of the present invention in the rice materials Huazhan and Huazhan GS2, and is not intended to limit the present invention in any way. Any simple modifications, equivalent changes or alterations made by those skilled in the art to other rice materials based on the above-disclosed technical content shall fall within the protection scope of the present invention.

Claims

1. A method for using a small peptide hormone to improve the rice's tolerance to high-temperature stress and / or its yield under high-temperature stress, characterized in that, The amino acid sequence of the small peptide hormone is shown in Seq ID No:1; it is applied in solution form to the surface of the panicle, leaves and stems of rice plants during the flowering period and before high temperature stress.

2. A method of using a small peptide hormone composition to improve rice's tolerance to high-temperature stress and / or rice yield under high-temperature stress, characterized in that, The small peptide hormone composition comprises a small peptide hormone and a solvent for dissolving the small peptide hormone; the amino acid sequence of the small peptide hormone is shown in Seq ID No:1; Apply the solution to the surface of the panicle, leaves and stems of rice plants during the flowering period and before high temperature stress.

3. The method of using the small peptide hormone composition for improving rice's tolerance to high-temperature stress and / or rice yield under high-temperature stress according to claim 2, characterized in that, The solvent used to dissolve the small peptide hormone is acetonitrile.

4. The method of using the small peptide hormone composition for improving rice's tolerance to high-temperature stress and / or rice yield under high-temperature stress according to claim 3, characterized in that, The small peptide hormone was dissolved in acetonitrile solvent to prepare a 10 μM solution.

5. The method of using the small peptide hormone composition for improving rice's tolerance to high-temperature stress and / or rice yield under high-temperature stress according to claim 3, characterized in that, The acetonitrile solvent is a 25% acetonitrile solvent, which is a solvent prepared by mixing water and acetonitrile in a volume ratio of 3:

1.

6. The application of a small peptide hormone, characterized in that, The amino acid sequence of the small peptide hormone is shown in Seq ID No:1; the application is to improve the rice's resistance to high temperature stress and / or the rice yield under high temperature stress.

7. The application of the small peptide hormone according to claim 6, characterized in that, The application is to improve the rice's tolerance to high temperature stress and / or rice yield under high temperature stress by applying an external spray in solution form; Improving rice yield under high temperature stress includes increasing the seed setting rate and thousand-grain weight of rice under high temperature stress.

8. The application of a small peptide hormone composition, characterized in that, The application is to improve the rice's tolerance to high-temperature stress and / or the rice yield under high-temperature stress; The small peptide hormone composition comprises a small peptide hormone and a solvent for dissolving the small peptide hormone; the amino acid sequence of the small peptide hormone is shown in Seq ID No:

1.

9. The application of the small peptide hormone composition according to claim 8, characterized in that, The solvent used to dissolve the small peptide hormone is acetonitrile.

10. The application of the small peptide hormone composition according to claim 8, characterized in that, The small peptide hormone was dissolved in acetonitrile solvent to prepare a 10 μM solution.

11. The application of the small peptide hormone composition according to claim 10, characterized in that, The acetonitrile solvent is a 25% acetonitrile solvent, which is a solvent prepared by mixing water and acetonitrile in a volume ratio of 3:

1.

12. The application according to any one of claims 8-11, characterized in that, The application is to improve the rice's tolerance to high temperature stress and / or rice yield under high temperature stress by applying an external spray in solution form; Improving rice yield under high temperature stress includes increasing the seed setting rate and thousand-grain weight of rice under high temperature stress.

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