Application of mTERF1 protein related to thermal adaptation evolution and its encoding gene

Abstract

The present invention belongs to the field of genetic engineering technology, and specifically relates to the application of mTERF1, a protein related to thermal adaptation evolution, and its encoding gene. The present invention finds through evolutionary genomics and genetic analysis that the mTEF1 gene is obviously selected by the environmental temperature in the process of thermal adaptation of plants, and tropical and subtropical plants have longer CpG islands and higher CG content. Further functional studies have shown that the mTEF1 protein is temperature responsive, and its protein participates in the maintenance and differentiation of the stability of vascular tissue stem cells, and the length of its exon CpG island determines the optimum temperature for species flowering. Therefore, through the development and utilization of the mTEF1 gene of heat-adapted species, it will not only help to increase crop yield and efficiency in high temperature environments, but will also be beneficial to the migration protection of rare species and increase biological diversity; it will even extend the growth cycle of herbaceous plants such as Arabidopsis, and promote carbon storage while increasing biomass.

Description

Technical Field

[0001] The invention belongs to the technical field of genetic engineering, and in particular relates to the application of a protein mTERF1 related to thermal adaptation evolution and a coding gene thereof. Background Art

[0002] Since the Industrial Revolution, the global carbon dioxide has increased year by year. The continuous rise of carbon dioxide in the atmosphere has caused global warming, leading to a series of disasters such as extreme temperatures, high temperatures, and droughts that are not conducive to the survival of humans and plants. Plants are the most important and basic producers in the earth's ecosystem and play an important role in the ecosystem, material cycle, and energy flow. Plants fix carbon dioxide through photosynthesis, which can not only provide nutrients for their own growth, but also provide food and energy for humans and animals. More importantly, they can preserve carbon dioxide in the form of organic matter to achieve plant "carbon sinks". According to statistics, the total global biological carbon sink is about 550 billion tons, of which plant carbon sinks are about 450 billion tons, accounting for 81%. At present, there are many ways to increase plant carbon sinks. Among them, the most commonly used is to transform plants through biotechnology innovation or functional gene mining to create perennial plants with high biomass, efficient carbon fixation, and high adaptability, which will help solve the dual carbon problem with plant science and provide a sustainable solution for carbon neutrality.

[0003] With global warming, extreme high temperature weather is becoming more and more frequent, posing an increasingly serious threat to global crops. Due to plant sessile growth, high temperatures can have a negative impact on plant growth and metabolism, such as early flowering; lack of energy to produce more seeds, resulting in low crop yields and even difficulty in survival. It is estimated that by 2050, global warming may lead to a serious reduction in the yield of crops such as wheat, rice, corn and soybeans (PMID: 33824329). Therefore, it is necessary to create plant or crop germplasm resources that can withstand higher temperatures, longer flowering times and longer growth periods to cope with severe climate change. At present, the functions and mechanisms of genes related to heat stress perception, heat-induced signals, heat stress response and heat memory in plants have been studied in depth (PMID: 37740489). These studies involve the activation of transcription factors and heat shock proteins in the early stage of stress response, the clearance of denatured toxic proteins caused by high temperature (PMID: 25985140), and epigenetic regulation and memory of recurrent heat stress conditions (PMID: 33278042). However, most of the above studies focus on the same species, such as Arabidopsis, rice or corn, and the depth and breadth of the application of the identified functional genes still need to be effectively applied in practice. With the continuous progress of evolution, many heat-resistant plants have developed a series of coping strategies that enable them to quickly sense changes in environmental temperature and make adjustments, promoting their survival and reproduction under adverse conditions (PMID: 33890138). For example, tropical woody plants such as rubber, durian, breadfruit, cocoa, coconut, coffee, cocoa, oil palm, cashew, fiber plants such as agave, sisal, sisal, abaca, and spice plants such as pepper, lemongrass, basil, and even tropical and subtropical crops such as rice, corn, peanuts, etc., all have the characteristics of high heat requirements, that is, the optimal growth temperature is high. Most of these plants or crops originated from or have been planted in tropical and subtropical regions for a long time. Although they have a certain degree of plasticity in their heat requirements and can be expanded to higher latitude climates, they have certain limits and generally show poor growth and reduced yields, indicating that their adaptation to heat is unique. It can be seen that exploring this unique heat adaptation mechanism of plants can provide good strategies for the breeding of more plants and crops, and is expected to provide an effective way to improve the heat adaptability of heat-sensitive plants. Therefore, it is necessary to explore and analyze proteins related to the evolution of heat adaptation and their encoding genes. Summary of the invention

[0004] In order to overcome the deficiencies of the above-mentioned prior art, the present invention proposes a protein mTERF1 related to the evolution of plant thermal adaptation and its encoding gene, which can be used to transform the thermal adaptability of heat-sensitive plants or cultivate high-temperature resistant plants, and has broad application prospects.

[0005] In order to achieve the above object, the technical solution adopted by the present invention is:

[0006] The first aspect of the present invention provides an mTERF1 gene related to the evolution of plant thermal adaptation, wherein the CpG island length of the mTERF1 gene exon is positively correlated with the thermal adaptability of the plant, the mTERF1 gene has a long CpG island and a high CG content, and the CpG island length of the mTERF1 gene is ≥100bp and the GC content is ≥50%.

[0007] Preferably, the mTERF1 gene encoding protein is also included. Other biological materials related to the mTERF1 gene are also included, and the other related biological materials include a recombinant vector, an expression cassette, a transgenic cell line or a recombinant bacterium containing mTERF1.

[0008] It should be understood that, taking into account the degeneracy of codons, modifications to the nucleotide sequence of the above-mentioned coding gene (such as sequence modification, replacement, and promoter replacement, etc.) without changing the amino acid sequence also fall within the scope of protection of the present invention.

[0009] Preferably, the mTERF1 gene is peanut mTER1, and the peanut mTER1 has the nucleotide sequence shown in SEQ ID No. 1 (AhmTERF1 CDS).

[0010] Preferably, the mTERF1 gene has a protein sequence as shown in SEQ ID No. 2 (AhmTERF1 protein).

[0011] Preferably, the mTERF1 gene is maize mTER1, and the maize mTER1 has the nucleotide sequence shown in SEQ ID No. 3 (ZnmTERF1 CDS).

[0012] Preferably, the corn mTER1 has a protein sequence shown in SEQ ID No. 4 (ZnmTERF1 protein).

[0013] The second aspect of the present invention further provides the use of the mTERF1 gene related to the evolution of plant thermal adaptation as described in the first aspect in transforming the thermal adaptability of heat-sensitive plants or cultivating high-temperature resistant plants.

[0014] The present invention provides a protein mTERF1 related to the evolution of plant heat adaptation, and the mTERF1 gene provided by the present invention belongs to the mitochondrial transcription termination factor gene family. Through the evolutionary analysis of mTERF1 among species, it was found that the functional specialization of mTERF1 appeared in angiosperms, and the length of the CpG island of its gene exon was positively correlated with the heat adaptation of the species; among them, the mTERF1 genes of peanut and corn both had longer CpG islands. The genes mTERF1 of peanut and corn were further transformed into Arabidopsis, and it was found that they could improve the heat resistance and biomass of Arabidopsis, and could still produce seeds normally and stably inherit offspring within a certain high temperature range (27-40°C). In addition, the AhmTERF1 of peanut was overexpressed in ferns, and it was found that it not only improved the heat resistance of ferns, but also increased the thickness of fern spore leaves, forming more veins and bifurcations. The above results indicate that mTERF1 with a long CpG island has the function of promoting the expression of genes related to vascular development and thermal adaptation, thereby increasing the thermal stability of plant vascular stem cells and can stably inherit this characteristic to offspring, which has great application value for species transformation and promoting plant carbon sequestration.

[0015] The third aspect of the present invention also provides a method for improving the thermal adaptability of heat-sensitive plants, namely, using the primer pair Fern-EcoR1-R / Fern-Sal1-F shown in SEQ ID No.5 and SEQ ID No.6 or the primer pair zea-EcoR1-R / zea-Sal1-F shown in SEQ ID No.7 and SEQ ID No.8 to overexpress peanut mTERF1 or corn mTERF1 in sensitive plants, thereby improving the thermal adaptability of heat-sensitive plants.

[0016] Preferably, the above method for improving the thermal adaptability of heat-sensitive plants is specifically as follows: first, the mTERF1 of peanut or corn is cloned respectively by primer pair Fern-EcoR1-R / Fern-Sal1-F or zea-EcoR1-R / zea-Sal1-F, and then connected to the pGreen-C18 vector by homologous recombinase, and the obtained pGreen-AhmTERF1-C18 and pGreen-ZnmTERF1-C18 plasmids are transformed into plants by gene gun or Agrobacterium, and finally transgenic plants can be obtained by resistance screening.

[0017] Preferably, the heat-sensitive plants include (but are not limited to) ferns and Arabidopsis thaliana.

[0018] Compared with the prior art, the present invention has the following beneficial effects:

[0019] The present invention finds through evolutionary genomics and genetic analysis that the functional specificity of mTEF1 protein first appeared in angiosperms and belongs to the mitochondrial transcription termination factor (mTERF) family. The exons of mTEF1 of different species have CpG islands of different lengths, but heat-sensitive plants do not have CpG islands, while tropical and subtropical plants have longer CpG islands and higher CG content (length ≥100bp, GC content ≥50%), and the copy number of mTEF1 is related to the degree of vascular development of plants, indicating that the gene is obviously selected by the environmental temperature in the process of heat adaptation of plants. Further functional studies have shown that mTEF1 protein has temperature response, and its protein participates in the maintenance and differentiation of the stability of vascular tissue stem cells, and the length of its exon CpG island determines the optimal temperature for species flowering. Since the success or failure of flowering will determine whether it can be normal to set seeds and reproduce offspring, that is, before the formation of seeds, mTEF1 has written this molecular "memory" into the embryo and passed it on to offspring, which is preserved and inherited from generation to generation, laying a molecular foundation for the formation and development of this species. Therefore, the development and utilization of the mTEF1 gene in heat-adapted species, such as the transformation of heat-sensitive species, will not only help increase crop yields and efficiency in high temperature environments, but will also be beneficial for the migration and protection of rare species and increase biological diversity; it will even extend the growth cycle of herbaceous plants such as Arabidopsis, thereby increasing biomass and promoting carbon storage. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 The relationship between mTERF1 and thermal adaptation evolution in different species.

[0021] Figure 1 In the figure, a is the CG content (%) of the corresponding mTERF1 gene exon in each species. The higher the CG content, the longer the corresponding CpG island. bd are the 2 Analysis of the correlation between the length of the CpG island in the exon of the mTERF1 gene and the species evolution time (b), genome size (c) and genome GC content (d). 2 The larger the value, the stronger the correlation, and vice versa. e is the relationship between the length of the CpG island of the mTERF1 gene exon and the most suitable flowering temperature of flowering plants. f is the evolutionary tree analysis based on the mTERF1 protein, which shows that the mTERF1 protein with CpG island and specialized functions is mainly concentrated in flowering plants (monocotyledons and dicotyledons).

[0022] Figure 2 Schematic diagram of the construction of peanut AhmTERF1 vector.

[0023] Figure 3 Schematic diagram of the construction of the ZnmTERF1 vector for maize.

[0024] Figure 4CpG island prediction results of AhmTERF1 (A) and ZnmTERF1 (B).

[0025] Figure 5 Overexpression of AhmTERF1 and ZnMTERF1 increases the biomass of Arabidopsis thaliana.

[0026] Figure 5 In the figure, a is the root system of wild-type Arabidopsis at 25 days of growth and the root system of AhmTERF1-overexpressing Arabidopsis at 50 days of growth; b is the root system of wild-type Arabidopsis at 25 days and 50 days of growth and the root system of ZnmTERF1-overexpressing Arabidopsis at 50 days of growth; c is the dry seeds of wild-type, AhmTERF1 and ZnmTERF1-overexpressing Arabidopsis (scale bar is 1 mm).

[0027] Figure 6 To overexpress ZnMTERF1 to increase the suitable flowering temperature in Arabidopsis.

[0028] Figure 6 In the figure, a shows ZnMTERF1-overexpressing Arabidopsis grown at 22±2℃ for 50 days, b shows ZnMTERF1-overexpressing Arabidopsis transferred to 33℃±2℃ for three days to induce flowering and normal seed setting, c shows ZnMTERF1-overexpressing Arabidopsis plants grown at 22±2℃ for 50 days and then transferred to 33±2℃ for two weeks to start bolting, flowering and normal seed setting, d shows bolting wild-type Arabidopsis transferred to 33±2℃ and then bolted and flowered quickly, but the leaves turned yellow and the pods were mostly aborted, while ZnMTERF1-overexpressing Arabidopsis plants could set seeds normally).

[0029] Figure 7 The suitable flowering temperature of Arabidopsis thaliana was also increased to overexpress AhMTERF1 (the figure shows the growth and flowering process of AhMTERF1-overexpressing Arabidopsis plants at different times after being transposed from 22°C to 27°C).

[0030] Figure 8 AhmTERF1 significantly improves the heat tolerance and strengthens the vascular system of ferns.

[0031] Figure 8 In the figure, the upper part of a is the leaf development process of the wild type (WT), and the lower part is the leaf development process of the AhMTERF1 overexpressing fern; bc is the enlarged image of the AhMTERF1 overexpressing fern leaf in the corresponding figure a. de is the phenotype of the wild type and AhMTERF1 overexpressing fern after being cultured at 22±2℃ for one month and then at 33±2℃ for one month. fg is the phenotype of the wild type and AhMTERF1 overexpressing fern gametophyte cultured at 22±2℃ (two months) and 33±2℃ (four months). DETAILED DESCRIPTION

[0032] The specific embodiments of the present invention are further described below. It should be noted that the description of these embodiments is used to help understand the present invention, but does not constitute a limitation of the present invention. In addition, the technical features involved in each embodiment of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0033] The experimental methods in the following examples are conventional methods unless otherwise specified, and the experimental materials used in the following examples are commercially available unless otherwise specified.

[0034] Example: Mining and analysis of proteins related to thermal adaptation evolution and their encoding genes

[0035] 1. Evolutionary analysis of mTERF genes

[0036] The length and number of CpG islands in the mTEF1 gene of 94 species of boreal, temperate, subtropical and tropical plants were analyzed. The results showed that the exons of the mTEF1 gene in subtropical and tropical plants had longer CpG islands and higher CG content (length ≥100bp, GC content ≥50%; see Table 1).

[0037] Table 1 Analysis of the length and number of CpG islands in the mTEF1 gene of 194 species of frigid, temperate, subtropical and tropical plants

[0038]

[0039]

[0040]

[0041]

[0042] 2. The length of the CpG island in the mTEF1 gene exon is positively correlated with species thermal adaptation

[0043] Further correlation analysis was performed on the length of the exon CpG island of the mTEF1 gene in different flowering plants, the optimal growth temperature, and the evolutionary time. The results showed that the length of the exon CpG island of the mTEF1 gene in flowering plants was positively correlated with the optimal flowering temperature (R 2 =0.8901, the larger the R value, the stronger the correlation), and the evolutionary time of species (R 2 =0.0264), genome size (R 2 =0.01), genomic GC content (R 2 =0.0366) are all lowly correlated ( Figure 1 ).

[0044] 3. Construction of CpG island-rich mTEF1 overexpression vector (taking peanut and corn as examples)

[0045] The peanut AhmTERF1 (AhmTERF66) was cloned from peanut (Yueyou No. 7, bred by Guangdong Academy of Agricultural Sciences) and the maize ZnmTERF1 (Zm00001d029242) was cloned from maize (Jingjingnuo, Guangzhou Lvba Company) using primer pairs Fern-EcoR1-R / Fern-Sal1-F and zea-EcoR1-R / zea-Sal1-F, respectively. IIOne Step Cloning Kit, Vazyme) was connected to the pGreen-C18 vector (provided by Professor Liu Xu of South China Botanical Garden, Chinese Academy of Sciences, and the relevant information of the vector can be found in the document "PMID: 36672950"); the obtained pGreen-AhmTERF1-C18 and pGreen-ZnmTERF1-C18 plasmids ( Figure 2 , 3 ) Transform Pteris crenata by gene gun, or soak Arabidopsis thaliana buds with Agrobacterium, and obtain transgenic plants by resistance screening.

[0046] Among them, Fern-EcoR1-R: TCCCCCGGGCTGCAGGAATTCAACACTAACCTGAGCCTTTGC (SEQ IDNo. ​​5);

[0047] Fern-Sal1-F: AGCTACGCGTCTCGAGGTCGACATGGCAGCCGTGGTG (SEQ ID No. 6);

[0048] zea-EcoR1-R:TCCCCCGGGCTGCAGGAATTCCACCGTGGTCTCCTTCG (SEQ IDNo.7);

[0049] zea-Sal1-F: GCTACGCGTCTCGAGGTCGACATGCGGTGGTTCGACC (SEQ ID No. 8); AhmTERF1 CDS (SEQ ID No. 1):

[0050] ATGCTTCTCTCTTTACACCTCCCTTCACCATCCCCTTCTTCTTACCACAC TCCTTCTAGAACCTTCCA CAACAAACCCCACTACATCAAGTTCCGTACATCTTACCGCCAAAATCTCCGTTACCTCCAACACCTAACCGTTATC CCGCCCGACTCCCTCCCTGACTCCGACACTCTCCACCGCATATTCACCATCGTTAACCTCCTCAAATCCCACTCCC TCTCCGACTCCGACATCCCCCGCATCGCCGCTGCTGCACCCTCCCTCTTCTCCCCCTCCTCCGCTGCCTTCTCCCC CGATAACATATCCGCCGTGTTCCACTTCCTCGCCACCGACGTCGCCGCCTCCTCCGCCGAGTCCCGCGGCCTCGTC CTGCGCTGCCCTCACCTACTCCTCTCCCCGCACCCCGACCTCTGCCTCCGCCCAACGCTCTACTTCCTCCGCGACG AGATCGGGCTCTCCGGTCTGAACCGGCCGACGAACAGGAACGCGCATCTGCTGAACACGCGCGTGGAGGATCTGCG CGTGAGGGTGGAGTTTTTGACGGAGGAGGTTGGGTTTGAGTATGAGGAGGCGGTTAGGGCGTTGGTACGGGTACCGGCTATATTCGGTTACGGTTTGGAAAGT AATTTGAGGCCGAAATTTGAGTATTTGGTTGGTGAGATGGGTAGGGATGTTGAAGAACTGAAAAAGTTTCCACACTACTTTGGGTTTAGCTTGCGTAACAAAATTGTGCCAAGGCATTTACATTTGAAGCAAATGGGTGTTGTTGATGTTCCTTTGAATAAGATGCTTATGTGGCGTGATGAAAGATTCTATGCAAAATGGAAGTGA (The underlined part is the CpG island, as shown in Figure 4 Figure A, located in the 51 - 627 section, with a total of 577 bp and a GC content of 54.14).

[0051] AhmTERF1 protein (SEQ ID No.2):

[0052] MLLSLHLPSPSPSSYHTPSRTFHNKPHYIKFRTSYRQNLRYLQHLTVIPPDSLPDSDTLHRIFTIVNLLKSHSLSDSDIPRIAAAAPSLFSPSSAAFSPDNISAVFHFLATDVAASSAESRGLVLRCPHLLLSPHPDLCLRPTLYFLRDEIGLSGLNRPTNRNAHLLNTRVEDLRVRVEFLTEEVGFEYEEAVRALVRVPAIFGYGLESNLRPKFEYLVGEMGRDVEELKKFPHYFGFSLRNKIVPRHLHLKQMGVVDVPLNKMLMWRDERFYAKWK.

[0053] ZnmTERF1 CDS (SEQ ID No.3):

[0054] ATGCGGTGGTTCGACCAGATCCATCTCACACCCTTATCCTCTTTACGCC TCACCACTACAGAGAAAAA GAGAAACCTGCGAGCGACGGGGACTATGGCCGCCACCCGCACCGCCCTGGCCCTGCTAGGGCACGAGGGCATGGAA TCTGGGCCCAGGCCGGCGGCGCCGCGCCGCAGCAGGCTACGCGTCGTCGCCGTCGCGCTGCGGACCAGGCCCACCA GCCTCGCGGCCCCGGGTCTCCCGCCCGCGGCGCCGGAGCCCGTGCTGCCGTCGCCGCCCGTGGCCGCGGGCGCCGC GGCGGTGCTGCTCGAGGCGGGCGTGCCGCCTGCCGACCTCCGGCGCGCGGCGGGGATGTGCCCGGAGCTGCTGTCC GTGTCCGCGGAAGCCGTCGAGGCCGCGCTCCGGTTCCTCACGGAGGAGGCGGGCGTCGCCGAGGCCGACCTCCCGC GCGTGCTCCGGCGGCGCCCGCGCCTGCTCGTGTCTCCCGTGGCGGCGCGGCTGCGGCCCACGCTCTACTTCCTGCG CGCGCTCGGCGTGCCCGACCTGCACCGCCGCGCCGACCTGCTGTCCTTCTCCGTGGAGGGCAAGCTGCTGCCGCGG CTCGAGTTCCTCGAGTCGCTGGGCCTGCCGGCGCGCGCCGCGCGCTCCATGGCGCGCCGCTTCCCCGCGCTCTTCG GCTACGGCGTGGAGGGGAACATGCGGCCCAAGGCGGACTACCTCCTGGGCGCCATGGCCCGCCGCGCCGACGAGCT GTACGACTTCCCCGAGTACTTCTCCTACGCGCTGGCCACGCGCATCGTGCCGCGTTACGAGGCCTGCGCCGCGCGC GGGGTCAGCAGGCTGCCGCTTCCCGCCATGCTCCGCCCCGGGGACGCCAAGTTCCGCTCCACTCTCACCAGCTGCGTCGGGTCCATGCTGCCCCGGAGGCGGTCGGCGCTGTGGCACGCCACGTGGGTGGATGACGACGCGGCGGCGGCGGCGGCGGCGAAGGAGACCACGGTGTGA (the underlined part is the CpG island, such as Figure 4 B, located in the 50-950 segment, a total of 901 bp, with a GC content of 74.83).

[0055] ZnmTERF1 protein(SEQ ID No.4):

[0056] MRWFDQIHLTPLSSLRLTTTEKKRNLRATGTMAATRTALALLGHEGMESGPRPAAPRRSRLRVVAVALRTRPTSLAAPGLPPAAPEPVLPSPPVAAGAAAVLLEAGVPPADLRRAAGMCPELLSVSAEAVEAALRFLTEEAGVAEADLPRVLRRRPRLLVSPVAARL RPTLYFLRALGVPDLHRRADLLSFSVEGKLLPRLEFLESLGLPARAARSMARRFPALFGYGVEGNMRPKADYLLGAMARRADELYDFPEYFSYALATRIVPRYEACAARGVSRLPLPAMLRPGDAKFRSTTLTSCVGSMLPRRRSALWHATWVDDDAAAAAAAKETTV.

[0057] 4. Overexpression of AhmTERF1 and ZnMTERF1 in Arabidopsis thaliana significantly increased biomass and growth period

[0058] AhmTERF1 and ZnMTERF1 were overexpressed in Arabidopsis thaliana and then planted in an artificial climate chamber (22±2℃, 16h / 8h light / dark). At temperatures below the most suitable flowering time (27℃), they could maintain multi-year growth without flowering, but their biomass (roots, stems, leaves and seeds) increased significantly ( Figures 5 - 7 ).

[0059] 5. Overexpression of AhmTERF1 and ZnMTERF1 confers a higher optimum flowering temperature

[0060] The most suitable growth and flowering temperature for Arabidopsis is 22±2℃. Too high temperature will lead to early flowering and reduced fertility. The most suitable growth and flowering temperature for peanut and corn are ≥27±2℃. ZnMTERF1 overexpressing Arabidopsis grown at 22±2℃ for 50 days ( Figure 6a) Place it at 33℃±2℃ for three days to induce flowering and normal seed setting ( Figure 6 b), but when wild-type Arabidopsis thaliana was placed at this temperature during the bolting period, it showed obvious sterility. Arabidopsis thaliana overexpressing AhmTERF1 did not flower after growing at 22±2℃ for 105 days, and showed growth inhibition. However, when the plant was placed at 27±2℃ for a week, it could be induced to flower and set seeds normally, and the growth was good. The above results show that overexpression of AhmTERF1 and ZnMTERF1 both endow Arabidopsis thaliana with a higher suitable temperature for flowering, that is, higher thermal adaptability, and this characteristic can be inherited by offspring, and the trait is stably inherited.

[0061] At the same time, ZnMTERF1-overexpressing Arabidopsis plants were grown at 22±2°C for 50 days and then transferred to 33±2°C for two weeks ( Figure 6 c), it began to bolt, flower and set seeds normally. However, when the wild-type Arabidopsis thaliana that was about to bolt was transferred to 33±2℃ for cultivation, it bolted and flowered quickly, but the leaves turned yellow and most of the pods were aborted ( Figure 6 d (indicated by white arrow).

[0062] In addition, AhMTERF1-overexpressing Arabidopsis plants planted at 22±2°C did not flower after 105 days, and this state could be maintained for two to three years. However, after being transferred to 27±2°C for two weeks, they began to bolt, flower, and set seeds normally ( Figure 7 ).

[0063] 6. AhmTERF1 significantly improves heat tolerance and strengthens the vascular system of ferns

[0064] Ferns grow best in warm environments, with an optimum temperature range of 15-25°C. Wild-type and AhMTERF1-overexpressing ferns were planted in MS medium and cultured at 22±2°C. Compared with the wild-type (WT), AhMTERF1-overexpressing ferns had thicker leaves, thicker veins, and two or three forks ( Figure 8 ad), but their gametophytes showed no significant differences ( Figure 8 f). At the same time, AhMTERF1-overexpressing ferns and wild-type ferns were cultured at 33±2℃. It can be seen that the growth of the wild-type ferns was inhibited, and the gametophytes and leaves withered, while the growth of the AhMTERF1-overexpressing ferns was good ( Figure 8 e&g).

[0065] In summary, mTERF1, represented by peanut AhmTERF1 and corn ZnmTERF1, is closely related to the evolution of thermal adaptation of plants, and is expected to be used to transform the thermal adaptability of heat-sensitive plants or cultivate high-temperature resistant plants, which will help increase crop yields and efficiency in high-temperature environments, and promote plant carbon sequestration, which has great application value.

[0066] The embodiments of the present invention are described in detail above, but the present invention is not limited to the described embodiments. For those skilled in the art, various changes, modifications, substitutions and variations of these embodiments are made without departing from the principles and spirit of the present invention, and still fall within the protection scope of the present invention.

Claims

1. Application of the mTERF1 gene related to plant thermal adaptation evolution in modifying thermal adaptability of heat-sensitive plants or cultivating high temperature resistant plants, characterized in that: The mTERF1 gene is peanut mTER1, the nucleotide sequence of the peanut mTER1 is shown in SEQ ID No. 1, and the heat-sensitive plant is a fern or Arabidopsis thaliana.

2. The use of the mTERF1 gene related to the evolution of plant thermal adaptation in modifying the thermal adaptability of heat-sensitive plants or cultivating high-temperature resistant plants, characterized in that: The mTERF1 gene is maize mTER1, the nucleotide sequence of the maize mTER1 is shown in SEQ ID No. 3, and the heat-sensitive plant is Arabidopsis thaliana.

3. The use according to claim 1, characterized in that: The protein sequence of peanut mTERF1 is shown as SEQ ID No.

2.

4. The use according to claim 2, characterized in that: The protein sequence of the corn mTER1 is shown in SEQ ID No.

4.

5. The use according to claim 1, characterized in that: By overexpressing peanut mTERF1 in sensitive plants using the primer pair Fern-EcoR1-R / Fern-Sal1-F shown in SEQ ID No.5 and SEQ ID No.6, the thermal adaptability of heat-sensitive plants can be improved.

6. The use according to claim 2, characterized in that: By overexpressing the maize mTERF1 in sensitive plants using the primer pair zea-EcoR1-R / zea-Sal1-F shown in SEQ ID No.7 and SEQ ID No.8, the thermal adaptability of heat-sensitive plants can be improved.

7. The use according to claim 5, characterized in that: Peanut mTERF1 was first cloned using the primer pair Fern-EcoR1-R / Fern-Sal1-F, and then connected to the pGreen-C18 vector using the homologous recombinase. The obtained pGreen-AhmTERF1-C18 plasmid was transformed into plants using a gene gun or Agrobacterium, and finally transgenic plants were obtained through resistance screening.

8. The use according to claim 6, characterized in that: First, corn mTERF1 was cloned using the primer pair zea-EcoR1-R / zea-Sal1-F, and then connected to the pGreen-C18 vector using the homologous recombinase. The obtained pGreen-ZnmTERF1-C18 plasmid was transformed into plants using a gene gun or Agrobacterium, and finally transgenic plants were obtained through resistance screening.

Images