Membrane H of Phyllostachys edulis + - ATPase, its gene PePMHA9 and use thereof

Abstract

The application discloses a bamboo plasma membrane H + ATPase, its gene PePMHA9 and application, belongs to the technical field of genetic engineering, and particularly relates to a bamboo plasma membrane H + ATPase, a bamboo plasma membrane H + The amino acid sequence of the bamboo plasma membrane H ATPase is shown as SEQ ID NO. 1. A coding gene PePMHA9 of the bamboo plasma membrane H ATPase, and the nucleotide sequence of the coding gene PePMHA9 is shown as SEQ ID NO. 2. The bamboo plasma membrane H ATPase or the coding gene PePMHA9 is applied to promoting the growth of rice in a high light environment. The bamboo plasma membrane H + ATPase and its coding gene PePMHA9 are successfully separated and identified from bamboo, and are transferred into wild type rice through a transgenic technology to perform function verification, and the obtained transgenic rice has increased leaf stomatal aperture, improved photosynthetic capacity and increased biomass in the high light environment compared with the wild type rice.

Description

Technical Field

[0001] This invention belongs to the field of genetic engineering technology, and particularly relates to the H-type membrane of moso bamboo. + -ATPase, its gene PePMHA9 and its applications. Background Technology

[0002] Greenhouse gas emissions cause the greenhouse effect, leading to global warming and posing a serious threat to human survival and sustainable development. Of all greenhouse effects, CO2 is the main contributor, and its emissions have been increasing continuously in recent years. Seeking strategies to mitigate and adapt to climate change is urgent, and enhancing natural carbon sinks is a key measure for mitigating climate change. Plant photosynthesis is the largest carbon sink process in the Earth's ecosystem, and is therefore considered an effective strategy for mitigating global warming. As a fast-growing forest plant, bamboo is widely distributed and has a strong photosynthetic carbon fixation capacity, with its average carbon storage far exceeding the average of other forest types in my country. Therefore, elucidating the regulatory mechanism of bamboo photosynthesis is of great significance for revealing its carbon sink function. Leaves are the main organs of plant photosynthesis, and stomata on the leaf surface control CO2 entry into cells; therefore, the dynamic regulation of stomatal opening directly affects the efficiency of bamboo leaf photosynthesis. (Plasma membrane H...) + -ATPase mediates light-induced stomatal aperture, but the regulatory mechanism of stomatal aperture in bamboo is still unclear, especially regarding the H2O2 of the bamboo plasma membrane. + Research on the regulation of stomatal opening by ATPases and its influence on photosynthetic capacity is still lacking. Therefore, it is necessary to use modern biotechnology to analyze the H+ in the plasma membrane of bamboo plants, including bamboo. + The function of ATPase genes is of great practical significance for molecular breeding to improve photosynthetic efficiency.

[0003] bamboo membrane H + -ATPase is a key functional protein located on the plasma membrane of plant cells, belonging to the P-type ATPase superfamily. It obtains energy by hydrolyzing ATP and converting protons (H+) in the cytoplasm. + The protons pump out of the cell against their concentration gradient, thereby establishing and maintaining a proton electrochemical gradient across the plasma membrane. This gradient provides the driving force for various secondary cotransporters, influencing nutrient uptake, regulating intracellular pH homeostasis, and laying the foundation for physiological processes such as stomatal opening promoting photosynthesis and plant growth. Studies have shown that plasma membrane H... +ATPase-mediated light-induced stomatal opening was significantly enhanced by overexpression of AHA2 in guard cells of the model plant Arabidopsis thaliana, leading to improved stomatal opening, photosynthesis, and growth. Furthermore, in the mutant aha1-9, red light-dependent stomatal opening in guard cells was delayed throughout the entire leaf, demonstrating the important role of AHA1 in stomatal opening in Arabidopsis thaliana. In rice, overexpression of OsA1 enhanced light-induced stomatal opening, increased leaf photosynthetic efficiency, and consequently promoted grain yield. Additionally, the addition of an electron transport inhibitor inhibited red light-induced plasma membrane H2O. + -ATPase phosphorylation and red light-induced stomatal opening in leaves. Besides light, plasma membrane H... + -ATPase genes also respond to signals and environmental factors such as low pH, salt stress, iron deficiency, and phosphorus deficiency. Currently, plasma membrane H + The function of ATPase in bamboo has not been confirmed, and research on it remains incomplete. Therefore, analyzing the H2O content of the bamboo membrane is crucial. + The molecular mechanism of ATPase in photosynthesis is of great value for discovering and utilizing key genes and using modern biotechnology to improve the photosynthesis of plants, including bamboo. Summary of the Invention

[0004] To solve the above-mentioned technical problems, this invention proposes a bamboo membrane H... + -ATPase, its gene PePMHA9, and its applications: This invention relates to the overexpression of plasma membrane H in rice. + -ATPase or its encoding gene PePMHA9 can effectively increase the stomatal pore size of leaves under high light conditions, improve photosynthetic capacity, and promote rice growth, providing a new theoretical basis and gene resource for the genetic engineering improvement of bamboo photosynthesis.

[0005] To achieve the above objectives, the present invention provides a bamboo-based membrane H + -ATPase, the bamboo membrane H + The amino acid sequence of ATPase is shown in SEQ ID NO.1.

[0006] The present invention also provides the aforementioned bamboo membrane H + The encoding gene PePMHA9 for ATPase is shown in SEQ ID NO.2.

[0007] The present invention also provides a biological material comprising the encoding gene PePMHA9, wherein the biological material is an expression cassette, an expression vector, or an engineered bacterium.

[0008] The present invention also provides the aforementioned bamboo membrane H +- The application of ATPase, the encoding gene PePMHA9, or the biomaterial in promoting rice growth in high light environments.

[0009] Preferably, by overexpressing H in bamboo membrane + - The gene PePMHA9, which encodes the ATPase, thereby increasing the H of the bamboo membrane. + - The expression level of ATPase increases the fresh weight and dry weight of rice in high light environment, thereby promoting the growth of rice in high light environment.

[0010] The present invention also provides the aforementioned bamboo membrane H + The application of ATPase, the encoding gene PePMHA9, or the biomaterial in increasing stomatal pore size in rice leaves under high light conditions, through overexpression of bamboo membrane H... + - The gene PePMHA9, which encodes the ATPase, thereby increasing the H of the bamboo membrane. + -ATPase expression increases the stomatal diameter of rice leaves in high light environments.

[0011] The present invention also provides the aforementioned bamboo membrane H + Application of ATPase, the encoding gene PePMHA9, or the biomaterial in improving the photosynthetic capacity of rice in high-light environments.

[0012] Preferably, by overexpressing H in bamboo membrane + - The gene PePMHA9, which encodes the ATPase, thereby increasing the H of the bamboo membrane. + - ATPase expression levels increase the net photosynthetic rate of rice leaves in high light environments and improve stomatal conductance in rice leaves in high light environments, thereby enhancing the photosynthetic capacity of rice in high light environments.

[0013] The present invention also provides a method for promoting rice growth in high-light environments, comprising: overexpressing the H+ of the bamboo membrane in rice plants. + - The gene encoding ATPase, PePMHA9, increases the H2O level in bamboo membranes. + - ATPase expression level promotes rice growth in high light environment.

[0014] The present invention also provides a method for improving the photosynthetic capacity of rice in high-light environments, comprising: overexpressing the H+ of the bamboo membrane in rice plants. + - The gene encoding ATPase, PePMHA9, increases the H2O level in bamboo membranes. + - ATPase expression level improves the photosynthetic capacity of rice in high light environment.

[0015] Compared with the prior art, the present invention has the following advantages and technical effects:

[0016] 1. This invention successfully isolated and identified H from the bamboo cytoplasm membrane. + -ATPase and its encoding gene PePMHA9 were transferred into wild-type rice using transgenic technology for functional verification. The transgenic rice obtained showed increased stomatal pore size, improved photosynthetic capacity, and increased biomass in leaves under high light conditions compared to wild-type rice.

[0017] 2. This invention overexpresses bamboo membrane H in rice, a monocotyledonous model plant. + -ATPase or its encoding gene PePMHA9 can effectively increase the stomatal pore size of leaves under high light conditions, improve photosynthetic capacity, and promote rice growth, providing a theoretical basis and technical support for the subsequent use of genetic engineering technology to improve the photosynthetic capacity of moso bamboo. This invention introduces the PePMHA9 gene into wild-type rice. The resulting transgenic rice exhibits larger stomatal pore sizes under high light conditions compared to the wild type. Under high light conditions, the stomatal pore sizes of the two transgenic lines are increased by 21.5% and 31.8%, respectively, compared to the wild type. The photosynthetic capacity of the two transgenic lines is also increased under high light conditions compared to the wild type, with net photosynthetic rates increasing by 61.7% and 43.3%, respectively, and stomatal conductance increasing by 83.0% and 72.3%, respectively. Furthermore, the growth of the two transgenic lines is promoted under high light conditions compared to the wild type, with aboveground fresh weight increasing by 12.8% and 11.8%, respectively, and aboveground dry weight increasing by 22.0% and 21.8%, respectively. This demonstrates that the PePMHA9 gene of this invention has a significant effect on increasing leaf stomatal pore size, improving photosynthetic capacity, and promoting plant growth under high light conditions. Attached Figure Description

[0018] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0019] Figure 1 This is an agarose gel electrophoresis image of the PePMHA9 gene amplified according to the present invention. In the figure, 1-6 represent amplification products, and M represents the DNA molecular weight marker DL5000.

[0020] Figure 2 This is an electrophoresis diagram for verifying the double digestion of the recombinant plasmid expressing the PePMHA9 gene of this invention. Lanes 1-3 are the double digestion products of the three recombinant plasmids, and M is the DNA molecular weight marker DL15000.

[0021] Figure 3This is an agarose gel electrophoresis image of PCR amplification of the recombinant plasmid expressing the PePMHA9 gene of this invention. In the image, 1-3 represent the PCR amplification products of the three recombinant plasmids, and M represents the DNA molecular weight marker DL5000.

[0022] Figure 4 The figure shows the RT-PCR detection results of the PePMHA9 gene in the rice plants of this invention. In the figure, 1 and 2 represent different transgenic rice plants, 3 represents the pCAMBIA1300-Ubi-GFP-FLAG-PePMHA9 plasmid, 4 represents the wild-type rice plant WT, and M represents the DNA molecular weight marker DL5000.

[0023] Figure 5 This figure shows the verification results of the relative expression level of the PePMHA9 gene in rice of this invention. WT represents wild-type rice, and PePMHA9_OE1 and PePMHA9_OE2 represent transgenic rice lines. "" indicates that the difference is significant at the P ≤ 0.01 level;

[0024] Figure 6 The images show the stomatal phenotypes of rice under different light conditions according to the present invention. In the images, A represents wild-type rice in high light conditions, B represents the transgenic rice line PePMHA9_OE1 in high light conditions, C represents the transgenic rice line PePMHA9_OE2 in high light conditions, D represents wild-type rice in low light conditions, E represents the transgenic rice line PePMHA9_OE1 in low light conditions, and F represents the transgenic rice line PePMHA9_OE2 in low light conditions. The scale bar is 10 μm.

[0025] Figure 7 This is a violin plot showing the stomatal pore size of rice under different light conditions according to the present invention. In the figure, WT represents wild-type rice, and PePMHA9_OE1 and PePMHA9_OE2 represent transgenic rice lines.

[0026] Figure 8 This is a bar chart showing the net photosynthetic rate of rice under different light conditions according to the present invention. In the figure, WT represents wild-type rice, and PePMHA9_OE1 and PePMHA9_OE2 represent transgenic rice lines. "" indicates that the difference is significant at the 0.01 < P ≤ 0.05 level. "" indicates that the difference is significant at the P ≤ 0.01 level, and "ns" indicates that the difference is not significant;

[0027] Figure 9 This is a bar chart showing the stomatal conductance analysis of rice under different light conditions according to the present invention. In the figure, WT represents wild-type rice, and PePMHA9_OE1 and PePMHA9_OE2 represent transgenic rice lines. "" indicates that the difference is significant at the P ≤ 0.01 level;

[0028] Figure 10 This is a bar chart showing the fresh weight analysis of the aboveground parts of rice under different light conditions according to the present invention. In the figure, WT represents wild-type rice, and PePMHA9_OE1 and PePMHA9_OE2 represent transgenic rice lines. "" indicates that the difference is significant at the 0.01 < P ≤ 0.05 level, and "ns" indicates that the difference is not significant;

[0029] Figure 11 This is a bar chart showing the aboveground dry weight analysis of rice under different light conditions according to the present invention. In the figure, WT represents wild-type rice, and PePMHA9_OE1 and PePMHA9_OE2 represent transgenic rice lines. "" indicates that the difference is significant at the 0.01 < P ≤ 0.05 level. Detailed Implementation

[0030] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0031] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Every smaller range between any stated value or intermediate value within a stated range, and any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0032] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0033] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0034] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0035] The materials used in this invention were sourced from: bamboo leaves from the Anhui Taiping Experimental Base of the International Bamboo and Rattan Centre; pGEM-TEasy vector from Promega (A1360); Escherichia coli DH5α competent cells from Beijing Zhuangmeng International Biotechnology Co., Ltd. (ZK206); pCAMBIA1300-Ubi-GFP-FLAG vector from Miaoling Plasmid Platform (P29697); and kanamycin (50 μg·mL⁻¹). -1 The resistance plates were prepared by adding the corresponding concentration of kanamycin to LB medium. The LB medium was purchased from Beijing Coollab Technology Co., Ltd. (PM0020) and the kanamycin was purchased from Beijing Coollab Technology Co., Ltd. (CK6731).

[0036] Example 1

[0037] I. Obtaining H from bamboo membrane + - The gene encoding ATPase is PePMHA9.

[0038] Phyllostachys edulis genome data were obtained through an online platform (http: / / gigadb.org / dataset / view / id / 100498), and primers were designed based on the open reading frame sequence of PePMHA9.

[0039] The primer sequences are as follows:

[0040] Upstream primer: 5′-ATGGACGATAAGCCTGGGAACC-3′ (SEQ ID NO.3), downstream primer: 5′-TCACACGGTGTACGACTGCT-3′ (SEQ ID NO.4).

[0041] RNA was extracted from bamboo leaves and reverse transcribed into cDNA as a template for amplification. The reaction system (20 μL) consisted of: 10.0 μL of 2× Taq Master Mix (Dye Plus), 1.0 μL each of upstream and downstream primers, 1.5 μL of template, and 6.5 μL of ddH2O. The amplification program was: 95℃, 5 min; 98℃, 15 s; 62℃, 30 s; 72℃, 1 min 14 s, for 35 cycles. The PCR products were detected by agarose gel electrophoresis.

[0042] Test results as follows Figure 1As shown, the target band was excised and purified. The recovered DNA fragment was ligated into the pGEM-TEasy vector and transformed into E. coli DH5α competent cells. Plasmids were extracted from positive clones and sequenced to find that the inserted gene fragment (recovered DNA fragment) was 2877 bp, as shown in the nucleotide sequence of SEQ ID NO.2.

[0043] bamboo membrane H +

[0044] bamboo membrane H +The amino acid sequence of the -ATPase is shown in SEQ ID NO.1, SEQ ID NO.1: MDDKPGNLDAVLNESVDLENIPLEEVFENLRCNREGLSSSQAEQRLAIFGPNKLEEKKESKFLKFLGFMWNPLSWVMEAAAIMAIALANGGGKPPDWQDFVGIITLLIINSTISFIEENNAGNAAAALMARLAPKAKVLRNGRWSEEESAILVPGDIISVKLGDIIPADARLLEGDPLKIDQSALTGESLPVTKGPADGVYSGSTCKQGEIEAVVIATGVHTFFGKAAHLVDSTNQVGHFQKVLTAIGNFCICSIFLGMVVEIIVMYAIQHRAYRPGIDNLLVLLIGGIPIAMPTVLSVTMAIGSHRLAQQGAITKRMTAIEEMAGMDVLCSDKTGTLTLNKLTVDKNLIEVFQRGIDQDTVILMAARASRTENQDAIDATIVGMLADPKEARADIQELHFLPFNPTDKRTALTYLDDEGRMHRVSKGAPEQILHLAHNKSEIERRVRAVIDKFAERGLRSLGVAYQEVPEGRKESPGGPWQFIGLLPLFDPPRHDSAETIRRALNLGVNVKMITGDQLAIGKETARRLGMGTNMYPSSALLGQDKDESIVALPVDELIEKADGFAGVFPEHKYEIVKRLQARKHICGMTGDGVNDAPALKKADIGIAVADSTDAARSASDIVLTEPGLSVIISAVLTSRAIFQRMKNYTIYAVSITIRIVLGFMLLALIWKFDFPPFMVLIIAILNDGTIMTISKDRVKPSPQPDSWKLSEIFATGVVLGGYLAMMTVIFFWAAYKTNFFPRVFHVESLEKTAQDDFQKLASAVYLQVSTVSQALIFVTRSRSWSFVERPGFLLVFAFLVAQLIATLIAVYADWGFAAIKGIGWGWAGVIWLYNIVFYLPLDVIKFLIRYALSGRAWDLVLDQRIAFTRKKDFGREERELKWATAQRTLHGLQPPEAAPTDRTNRANFNELNQLAEDARRRAEMARLRELTTLKGRMESVVRQKGLDIDTIQQSYTV。

[0045] II. Constructing a plant expression vector carrying the coding gene PePMHA9.

[0046] Using moso bamboo cDNA as a template, primers were designed based on the nucleotide sequence shown in SEQ ID NO.2, and BamHⅠ and PmlⅠ restriction sites were introduced at both ends of the primers, respectively.

[0047] The primer sequences are as follows:

[0048] Upstream primer: 5′-TGTTACTTCTGCAGggatccATGGACGATAAGCCTGGGAACC-3′ (SEQ ID NO. 5) (lowercase letters at the BamHI site);

[0049] Downstream primer 5′-CTCACCATAGGCCTcacgtgCACGGTGTACGACTGCTGAAT-3′ (SEQ ID NO.6) (lowercase letters at PmlⅠ site).

[0050] The PCR amplification reaction system (20 μL) was as follows: 10.0 μL of 2×Taq Master Mix (Dye Plus), 1.0 μL each of upstream and downstream primers, 1.5 μL of template, and 6.5 μL of ddH2O. The amplification program was as follows: 95℃, 5 min; 98℃, 15 s; 55~65℃, 30 s; 72℃, 1 min 14 s, 35 cycles. The PCR amplification products were detected by agarose gel electrophoresis. The target band was excised and purified. The recovered DNA fragment was ligated into the pCAMBIA1300-Ubi-GFP-FLAG vector using a one-step cloning method under 50℃ water bath conditions for 10 min, and then transformed into *E. coli* DH5α competent cells. Kanamycin (50 μg / mL) was picked. -1 Single clones grown on resistant plates were used to extract plasmids and identify them using restriction enzyme patterns (e.g., ...). Figure 2 (As shown in the figure) and after sequencing verification, the obtained recombinant expression vector was named pCAMBIA1300-Ubi-GFP-FLAG-PePMHA9.

[0051] Figure 3 This is an agarose gel electrophoresis image of the PCR amplification of the recombinant plasmid of the PePMHA9 expression vector in this invention, where lanes 1-3 are the PCR amplification products of the three recombinant plasmids, and M is the DNA molecular weight marker DL5000.

[0052] III. Transformation of rice with the PePMHA9 gene and qPCR detection.

[0053] Since rice and bamboo are both monocotyledonous grasses and are closely related, the recombinant expression vector pCAMBIA1300-Ubi-GFP-FLAG-PePMHA9 was used to transform Agrobacterium tumefaciens into rice callus tissue. Through screening, two transgenic rice lines, PePMHA9_OE1 and PePMHA9_OE2, were obtained, and gene expression was detected. Total RNA was extracted from the transgenic rice lines PePMHA9_OE1 and PePMHA9_OE2 and wild-type rice plants (WT), and reverse transcribed into cDNA. Using this cDNA as a template, PCR detection was performed using primers with nucleotide sequences shown in SEQ ID NO.3 and SEQ ID NO.4.

[0054] like Figure 4 As shown, the results indicate that the target gene was detected in both transgenic rice lines PePMHA9_OE1 and PePMHA9_OE2, while no expression was detected in wild-type rice plant WT, proving that the gene PePMHA9 was expressed in the transgenic rice lines.

[0055] IV. Detection of PePMHA9 gene expression in transgenic rice.

[0056] The expression level of the PePMHA9 gene was determined by qPCR, with OsActin as an internal reference gene.

[0057] Depend on Figure 5 It can be seen that the expression level of the gene PePMHA9 in the two transgenic rice lines PePMHA9_OE1 and PePMHA9_OE2 is significantly higher than that in the wild-type rice plant WT.

[0058] V. Analysis of stomatal phenotype in transgenic rice.

[0059] Wild-type rice plants (WT) and two transgenic rice lines, PePMHA9_OE1 and PePMHA9_OE2, were subjected to different light treatments after 3 weeks (high light: 1000 μmol / m²). 2 / s, low light: 100μmol / m 2 / s). Two weeks later, the stomatal morphology of leaves of wild-type rice plant WT and two transgenic rice lines PePMHA9_OE1 and PePMHA9_OE2 was observed and photographed using field emission scanning electron microscopy.

[0060] The results are as follows Figure 6 China A Figure 6 B, Figure 6 C, Figure 6 D, Figure 6 China E and Figure 6As shown in Figure F, under high light conditions, the pore size of PePMHA9_OE1 and PePMHA9_OE2 is significantly larger than that of WT.

[0061] The pore size was measured using ImageJ, and the results are as follows: Figure 7 As shown, under high light conditions, the pore size of PePMHA9_OE1 and PePMHA9_OE2 is 21.5% and 31.8% larger than that of WT, respectively.

[0062] In conclusion, overexpression of the PePMHA9 gene can increase the stomatal pore size in rice.

[0063] VI. Comparative analysis of gas exchange parameters in transgenic rice.

[0064] Gas exchange parameters of wild-type rice plants (WT) and two transgenic rice lines PePMHA9_OE1 and PePMHA9_OE2 under different light treatments were measured using a portable photosynthesis system.

[0065] The results are as follows Figure 8 and Figure 9 As shown, under high light conditions, the net photosynthetic rates of PePMHA9_OE1 and PePMHA9_OE2 increased by 61.7% and 43.3% respectively compared to WT, and the stomatal conductance of PePMHA9_OE1 and PePMHA9_OE2 increased by 83.0% and 72.3% respectively compared to WT. In conclusion, overexpression of the PePMHA9 gene can improve the photosynthetic capacity of rice.

[0066] VII. Comparative analysis of biomass of transgenic rice.

[0067] The biomass of wild-type rice plants (WT) and two transgenic rice lines, PePMHA9_OE1 and PePMHA9_OE2, under different light treatments was measured.

[0068] The results are as follows Figure 10 and Figure 11 As shown, under high light conditions, the aboveground fresh weight of PePMHA9_OE1 and PePMHA9_OE2 increased by 12.8% and 11.8% respectively compared to WT, and the aboveground dry weight of PePMHA9_OE1 and PePMHA9_OE2 increased by 22.0% and 21.8% respectively compared to WT. In conclusion, overexpression of the PePMHA9 gene can promote rice growth.

[0069] In summary, this invention provides a bamboo-based membrane H + -ATPase and its encoding gene PePMHA9, overexpression of plasma membrane H in rice +The ATPase-encoding gene PePMHA9 can effectively increase the stomata of rice leaves under high light conditions, improve photosynthetic capacity, and promote rice growth, providing a new gene resource for plant material property improvement genetic engineering.

[0070] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. A bamboo membrane H + -ATPase, bamboo membrane H + The application of the PePMHA9 gene encoding ATPase or biomaterials containing the PePMHA9 gene in promoting rice growth in high-light environments, characterized by: The bamboo membrane H + The amino acid sequence of the ATPase is shown in SEQ ID NO.1, and the bamboo membrane H + The nucleotide sequence of the gene encoding ATPase PePMHA9 is shown in SEQ ID NO.2, and the biological material including the gene encoding PePMHA9 is an expression cassette, expression vector, or engineered bacteria.

2. The application according to claim 1, characterized in that, By overexpressing H in bamboo membrane + - The gene PePMHA9, which encodes the ATPase, thereby increasing the H of the bamboo membrane. + - The expression level of ATPase increases the fresh weight and dry weight of rice in high light environment, thereby promoting the growth of rice in high light environment.

3. The bamboo membrane H as described in claim 1 + -ATPase, the bamboo membrane H as described in claim 1 + The application of the PePMHA9 gene encoding ATPase or the biomaterial containing the PePMHA9 gene as described in claim 1 in increasing the stomatal pore size of rice leaves under high light conditions, characterized in that... By overexpressing H in bamboo membrane + - The gene PePMHA9, which encodes the ATPase, thereby increasing the H of the bamboo membrane. + -ATPase expression increases the stomatal diameter of rice leaves in high light environments.

4. The bamboo membrane H as described in claim 1 + -ATPase, the bamboo membrane H as described in claim 1 + The application of the PePMHA9 gene encoding ATPase or the biomaterials comprising the PePMHA9 gene as described in claim 1 in improving the photosynthetic capacity of rice in high-light environments.

5. The application according to claim 4, characterized in that, By overexpressing H in bamboo membrane + - The gene PePMHA9, which encodes the ATPase, thereby increasing the H of the bamboo membrane. + - ATPase expression levels increase the net photosynthetic rate of rice leaves in high light environments and improve stomatal conductance in rice leaves in high light environments, thereby enhancing the photosynthetic capacity of rice in high light environments.

6. A method for promoting rice growth in high-light environments, characterized in that, include: Overexpression of the bamboo membrane H described in claim 1 in rice plants + - The gene encoding ATPase, PePMHA9, increases the H2O level in bamboo membranes. + - ATPase expression level promotes rice growth in high light environment.

7. A method for improving the photosynthetic capacity of rice in high-light environments, characterized in that, include: Overexpression of the bamboo membrane H described in claim 1 in rice plants + - The gene encoding ATPase, PePMHA9, increases the H2O level in bamboo membranes. + - ATPase expression level improves the photosynthetic capacity of rice in high light environment.

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