CN117925639A - PATL1 gene and application thereof in regulating and controlling plant vesicle transport or plant heat resistance - Google Patents

Abstract

The invention discloses PATL1 gene and application thereof in regulating and controlling plant vesicle transport or plant heat resistance, wherein the heat stress tolerance of plants is enhanced by reducing or silencing the expression of PATL1 homologous genes in the plants; and promoting the trafficking of vesicles and/or the accumulation of heat shock proteins in plant cells by reducing the expression of the PATL1 gene in the plant. The invention has important scientific research and practical value in the aspect of culturing heat-resistant plant varieties.

Description

PATL1 gene and application thereof in regulating and controlling plant vesicle transport or plant heat resistance

Technical Field

The invention relates to the technical field of molecular biology, in particular to PATL1 gene and application thereof in regulating and controlling plant vesicle transport or plant heat resistance.

Background

With the increasing influence of the greenhouse effect, the integration of temperature signals is crucial for the survival and adaptation of plants (Kaiserli, 2021). An estimate of the specialized committee for climate change (IPCC 2019) shows that the global average air temperature is expected to rise by 0.3 ℃ every 10 years, which will result in an average temperature rise of about 1-3 ℃ (Janni et al 2020) on the earth's surface from 2025 to 2100 years. The main effect of the presence of heat stress is that the actual survival temperature of the plant is 10-15 ℃ higher than the optimal growth temperature, and the heat stress problem is further aggravated according to the current predicted temperature rising rate and duration. It is estimated that wheat yield decreases by 6% (Asseng et al, 2014;Zaveri et al, 2019) for every 1 degree celsius increase in global air temperature. In cereals, peas, lentils and chickpeas, brief heat stress (> 24 ℃) during reproduction leads to reduced flower fertility, whereas an average temperature lasting 35 ℃ may lead to overall crop failure (Janni et al, 2020). The heat stress comprises the change of growth mode, the sensibility to plant diseases and insect pests, the plant climate, the flowering and mature period shortening, the grain grouting and the aging aggravation, etc. which can lead to the yield reductionEt al, 2010; jha et al., 2014). In addition, the early flowering type in spring is also more sensitive to high temperatures (Wolkovich et al, 2012).

At the same time, many aspects of plant growth, development, morphology, biochemical and physiological processes are affected by heat stress and often are negative. For the different stages of plant growth, the bud germination stage is the most affected stage by heat stress, as high temperatures can lead to reduced germination rate, root conductivity, leaf moisture content, resulting in malformation, reduced vigor, poor growth of seedlings (PIRAMILA ET al, 2012; rai et al, 2020). Heat stress can promote early flowering of plants and disrupt seasonal growth of certain species. In plant morphology, heat stress can lead to the situation that the overground parts of plants are burnt and sunburned, premature aging and falling-off phenomena can occur at leaf parts, the leaf morphology is curled and dried, and fruits can be discolored and damaged (Hasanuzzaman et al., 2013). In addition, heat stress can lead to reduced nutrient growth, such as temperatures above 35 ℃ can adversely affect nutrient and reproductive growth of corn (HATFIELD ET al., 2011). Heat stress can consume carbohydrates accumulated in plants, and long-term heat stress can even consume carbohydrate reserves, and as a result, different effects can be generated on the size, quantity, grouting degree and quality of seeds, so that seed yield is reduced. Another physiological process in plants that is sensitive to heat is photosynthesis (Brandner and Saluci, 2002), where the matrix where carbon metabolism occurs and the thylakoids where photochemical reactions exist are the main sensitive sites, whereas high temperatures can cause damage to the plant photosynthetic process by reducing synthesis or accelerating degradation (Fahad et al., 2017). Ultimately, the yield and transport of photosynthesis assimilates is low and precursors for seed reserves, mineral and other functional ingredient biosynthesis are removed (Sehgal et al 2018), resulting in problems with seed or grain filling. The effects of heat stress on photosynthesis, essential metabolism and plant development of crops have led to reduced annual crop yields (Moore et al, 2021). There is growing evidence that plants undergo a complex series of signal transduction processes in high temperature environments that may involve changes in plant hormone signals, coordinated regulation of light signals, and effects on disease resistance and Ciradian rhythms in plants.

In summary, in the process of crop growth, abiotic stress such as heat, cold, salt, drought and the like can influence the crop growth, and the abiotic stress has a non-negligible influence on the crop yield; at the same time, temperature changes caused by global warming have also evolved into a problem that is not negligible for plant growth. Therefore, the development of a gene tool capable of regulating and controlling plants to cope with heat stress on a molecular level is of important scientific research and application value.

Disclosure of Invention

The invention aims to provide a PATL1 gene and application thereof in regulating and controlling plant vesicle transport or plant heat resistance.

In order to solve the technical problems, the technical scheme adopted by the invention is as follows.

The kit is used for regulating and controlling the resistance of plants to heat stress, and is characterized in that: the kit comprises a molecular biological element capable of regulating and controlling the expression level of a specific gene; the specific gene is a gene related to plant response to heat stress; the molecular biological element optionally comprises: the over-expression element combination of the specific gene, and/or the element combination for inhibiting or reducing the expression amount of the specific gene, and/or the element combination for silencing the expression of the specific gene.

As a preferable technical scheme of the invention, the specific gene is PATL1 gene, homologous gene thereof or equivalent gene with equivalent plant physiological function; the PATL1 gene is: at1g72150.

As a preferred embodiment of the present invention, the molecular biological element is an element combination that suppresses or reduces the expression level of the specific gene, and/or an element combination that silences the expression of the specific gene.

The invention also comprises the following technical scheme: a recombinant expression vector comprising the PATL1 gene or a homologous gene thereof.

As a preferred technical scheme of the invention, the recombinant expression vector is a silencing vector.

A method of enhancing heat stress tolerance in a plant, reducing or silencing expression of a PATL1 homologous gene in the plant to enhance heat stress tolerance in the plant.

A method of increasing crop yield by reducing or silencing expression of a PATL1 homologous gene in the crop to enhance heat stress tolerance of the corresponding crop, thereby increasing yield of the corresponding crop.

The invention also includes the use of the PATL1 gene to regulate the heat tolerance of a plant.

The invention also includes the use of the PATL1 gene to regulate the trafficking of vesicles and/or the accumulation of heat shock proteins within plant cells.

As a preferred embodiment of the present invention, the transport of vesicles and/or accumulation of heat shock proteins in plant cells is promoted by decreasing the expression of PATL1 gene in plants.

The beneficial effects of adopting above-mentioned technical scheme to produce lie in: the invention introduces PATL1 genes into plant materials to obtain a recovery line and an over-expression transgenic line, analyzes the heat stress tolerance of PATL mutants, recovery (COM 1/2) and over-expression lines (OE 1/2), and discovers that the PATL genes can negatively regulate and control the heat resistance of plants; furthermore, the present invention also investigated and confirmed the regulation of vesicle transport by PATL gene expression under heat stress.

Based on the scientific findings, the invention is created; the invention has important scientific research and practical value in the aspect of culturing heat-resistant plant varieties.

Drawings

FIG. 1 is an identification chart of Arabidopsis patl1 mutant; in the figure: (A) DNA level detection T-DNA insertion mutant patl (SALK_080204C); (B) Real-time PCR detects the transcription level, and RNA level identifies AtPATL1 expression level.

FIG. 2 is an expression diagram of heat shock at different times AtPATL 1; in the figure: * P <0.05, < P <0.01, < P <0.001, n=3.

FIG. 3 is a graph showing the effect of heat stress on the survival of restorer seedlings. In the figure: (A and B) Col, patl, COM1 and COM2 Arabidopsis seedlings were grown normally at 22℃for 10 days (Control) and at 22℃for 6 days, heat-shock treated at 45℃for 60 minutes and then recovered in a 22℃incubator for 5 days (HS), and survival rates were compared; (1, col, 2, patl1, 3, COM1, 4, COM2); (C) Survival rates of the Col, patl, COM1 and COM2 Arabidopsis seedlings were counted. Values shown are mean ± standard deviation P < 0.05.

FIG. 4 is a graph of RNA and protein level identification PATL1 overexpressing strains. In the figure: protein level detection of PATL1-cMyc expression level. (B) RNA level detection of the expression level of the PATL1 Gene. Wild type was assigned 1, patl1 was used as negative control and 18s was used as internal reference. All values represent mean ± standard deviation, P <0.01, P <0.001, n > 3.

FIG. 5 is a graph of survival rate of Arabidopsis seedlings under heat stress. In the figure: (A) Photographing and recording data of seedlings (1, WT;2, patl1;3, OE1;4, OE2) grown at normal temperature for 11 d; (B) Seedlings (1, WT;2, patl1;3, OE1;4, OE2) grown for 6d at normal temperature were subjected to 45℃for 1 hour and then were photographed and recorded after growth was resumed for 5d at 22 ℃; (C) analyzing heat stress survival data of the seedlings; * P <0.05, < P <0.01, < P <0.001.

FIG. 6 is a graph showing the negative effects of PATL1 during vesicle transport under heat stress. (A) Seedlings grown for 4 days were exposed to 45 ℃ (HS) or maintained at 22 ℃ (control) for 60 minutes, FM4-64 stained for 10 minutes, and observed waiting for 15 minutes. (B) Bar graphs represent quantification of intracellular FM4-64 signals in (a) using ImageJ software (n=60). The different letters represent significant differences in P <0.05 in each group (one-way analysis of variance using Tukey multiple comparison test).

Detailed Description

The following examples illustrate the application in detail. The raw materials and the equipment used by the application are conventional commercial products, and can be directly obtained through market purchase. In the following description of embodiments, for purposes of explanation and not limitation, specific details are set forth, such as particular system architectures, techniques, etc. in order to provide a thorough understanding of the embodiments of the application. It will be apparent, however, to one skilled in the art that the present application may be practiced in other embodiments that depart from these specific details. In other instances, detailed descriptions of well-known systems, devices, and methods are omitted so as not to obscure the description of the present application with unnecessary detail.

It should be understood that the terms "comprises" and/or "comprising," when used in this specification and the appended claims, specify the presence of stated features, integers, steps, operations, elements, and/or components, but do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, and/or groups thereof. It should also be understood that the term "and/or" as used in the present specification and the appended claims refers to any and all possible combinations of one or more of the associated listed items, and includes such combinations. As used in the present description and the appended claims, the term "if" may be interpreted as "when..once" or "in response to a determination" or "in response to detection" depending on the context. Similarly, the phrase "if a determination" or "if a [ described condition or event ] is detected" may be interpreted in the context of meaning "upon determination" or "in response to determination" or "upon detection of a [ described condition or event ]" or "in response to detection of a [ described condition or event ]".

Furthermore, the terms "first," "second," "third," and the like in the description of the present specification and in the appended claims, are used for distinguishing between descriptions and not necessarily for indicating or implying a relative importance. Reference in the specification to "one embodiment" or "some embodiments" or the like means that a particular feature, structure, or characteristic described in connection with the embodiment is included in one or more embodiments of the application. Thus, appearances of the phrases "in one embodiment," "in some embodiments," "in other embodiments," and the like in the specification are not necessarily all referring to the same embodiment, but mean "one or more but not all embodiments" unless expressly specified otherwise. The terms "comprising," "including," "having," and variations thereof mean "including but not limited to," unless expressly specified otherwise.

Example 1 planting and cultivation of Arabidopsis thaliana

The seeds are sterilized by shaking with 75% alcohol for 3 times and absolute alcohol for 1 time, and then the seeds are placed on sterile filter paper for airing, and the toothpick is inoculated. The culture medium was placed in a refrigerator at 4℃for vernalization for 2D, and then transferred to an incubator at 22℃for 16L/8D cycles.

Heat stress treatment: seedlings grown in the culture medium for 6d are subjected to heat treatment (45 ℃) for 1h and then returned to 22 ℃ for 5d recovery, and the statistical survival rate is calculated.

Transplanting: plants were grown in medium for about 10d, and when two separate round leaves were grown, they were transplanted to 1:1, in an exosome extraction experiment, rosette leaves which grow in the soil for about 6 weeks and are not bolting yet are adopted.

Example 2 obtaining transgenic plants

2.1 Extraction of DNA by CTAB method. The leaves of Arabidopsis thaliana were taken, ground at low temperature in liquid nitrogen, 400. Mu.L of CTAB buffer was added thereto, 1h at 65℃and 400. Mu.L of chloroform were further added thereto, and the supernatant was collected by centrifugation at the highest speed. 400. Mu.L of isopropanol was added and mixed by shaking at-20℃for 20min. Centrifuging at the highest speed for 10min to leave a precipitate. The precipitate was then sufficiently sprung with 800. Mu.L of 75% alcohol. Centrifuging at high speed for 5min, collecting supernatant, optionally placing in oven for 15min, adding 30uL of ultrapure water, centrifuging at room temperature for 2min, and measuring concentration.

2.2 Obtaining the desired fragment. Primers are designed according to the target fragment nucleic acid sequence, and amplification is carried out through a PCR reaction system.

TABLE 2.2PCR reaction System

And (3) performing nucleic acid gel running on the PCR product, and recovering the target fragment through gel recovery to obtain the target fragment DNA.

2.3 Enzyme digestion connection, transformation and identification. Selecting proper enzyme cutting sites, selecting corresponding endonucleases in target fragments and vectors, and performing enzyme cutting at 37 ℃ for 15min to inactivate. Adding ligase, and connecting for 2 hours at room temperature. 1. Mu.L of the well-connected product is added into DH5 alpha according to the ratio of 1:50, and after half an hour of ice bath, the mixture is subjected to heat shock at 42 ℃ for 30s, and after 2 minutes of ice bath, 500. Mu.L of LB is added without resistance. Incubation was performed for 1h at 37℃with shaking at 100rpm, spread on a resistance plate and incubated upside down at 37 ℃.

Designing a primer for colony PCR, carrying out enzyme digestion identification on plasmid, and carrying out sequencing.

2.4 Agrobacterium transfection and identification. 1 mu L of the successfully sequenced plasmid is added into GV3101, ice-bathed for 30min, quick frozen for 5min, then placed into a metal bath at 37 ℃ for 5min, and then ice-bathed for 3-5min. After adding 500. Mu.L of the non-antibiotic YEB culture medium and 3 hours at 28 ℃ and 150rpm, the mixture is evenly spread on the antibiotic YEB culture medium and is cultivated upside down at 30 ℃. And (3) identification: is consistent with the identification method of the escherichia coli.

2.5 Arabidopsis transformation and positive plant selection. And (3) performing expansion culture on the converted agrobacterium tumefaciens bacterial liquid, and collecting bacteria at 600rpm for 5min when the bacterial liquid presents orange color. The pellet was resuspended with transformation medium and then OD 600 = 0.8. The plants were transfected by the floral dip method for 5min.

And (3) recovering transgenic plant seeds from the single plant, carrying out resistance screening, culturing for about 14d under normal light after vernalization, and transplanting seedlings. And screening the resistance of the single plant after harvest, wherein the isolation ratio of the grown positive plants is 3:1, which indicates that the single sequence is inserted into the plants. And screening the resistance of the single plant after harvesting, screening all positive plants, and carrying out DNA, RNA and protein level identification.

Example 3 RNA extraction and reverse rotation, real-time fluorescence quantitative PCR

3.1RNA extraction. The seedling material is grown for 10d, and the seedling is formed by adopting leaves. RNA extraction was performed using Eastep Super kit after liquid nitrogen milling and concentration was measured.

3.2 Reverse transcription. The materials were mixed and subjected to the inversion procedure according to the following table.

TABLE 2.3 reverse transcription System

The inversion procedure was performed after mixing the materials according to the above table: pre-denaturation; the conditions were 95℃for 30s, the number of cycles, 1. Amplifying; the conditions were 95℃for 5s,60℃for 34s and the number of cycles was 39.

3.3QPCR. After the obtained cDNA was diluted to 100. Mu.L, it was mixed with the following reagents, and QPCR was performed according to the procedure.

TABLE 2.4 quantitative PCR System

The PCR procedure was set as follows: (1) 95 ℃ for 10s; (2) 95℃for 5s; at 60℃34s 40 cycles.

Example 4 plant Total protein extraction and Western-Blot

The seedling material is grown for 10d, and the seedling is formed by adopting leaves. Grinding the material in liquid nitrogen by a grinder, adding 2 XSDS buffer, boiling, centrifuging at maximum rotation speed for 5min at 100 ℃ and preserving at-80 ℃.

And loading the sample in an equal volume in the protein glue, and firstly, running the glue by 80v, and converting into 120v after the red Mark appears. NC film with the same size as the glue is taken, and film is transferred on a semi-dry film transfer instrument, wherein the size of the NC film is about 22v 45 min. After the membrane was placed in a blocking solution and shaken for 2h, the primary antibody was changed to the primary antibody and shaken for 2h, the membrane was washed 3 times with PBST solution for 10min each. Then placing the sample in a secondary antibody, shaking for 45min, and washing by using PBST (PBST) to perform chemiluminescence observation.

EXAMPLE 5 identification of Arabidopsis patl mutant

To verify if patl1 mutants responded to heat, we identified the T-DNA insertion mutant of patl1 (SALK_080204C) and that mutant patl1 was a homozygous strain by DNA level (FIG. 1A; DNA level detection T-DNA insertion mutant patl; SALK_080204C). Then we carried out the identification of the transcription level (FIG. 1B; real-time PCR detection of the transcription level, RNA level identification AtPATL1 expression level), found that the expression level of gene AtPATL1 in the three mutant plants was lower than that of the wild type, and the No. 1 plant and the No. 2 plant were identified as AtPATL1 knock-out plants. Since the expression level of gene AtPATL1 in plant No. 1 is the lowest, plant No. 1 was selected for the subsequent experiments.

EXAMPLE 6 Heat shock different Gene expression levels

To demonstrate whether heat shock treatment affects gene transcription, the following experiments were designed and performed. Seedlings which normally grow for 7 days in a 22 ℃ incubator are subjected to heat shock for 0-60 min at 45 ℃ and are obtained once every 10 min. Real-time PCR detection of AtPATL1 expression levels in wild-type plants, 18SrRNA as an internal reference.

The results are shown in FIG. 2; expression of heat shock at different times AtPATL 1P <0.05, P <0.01, P <0.001, n=3 are shown. This result indicates that AtPATL1 expression generally decreased after heat shock.

Example 7 Effect of Heat stress on survival of restorer seedlings

To genetically verify the role of PATL1 in the heat shock signaling pathway, we obtained restorer plants PATL/pPATL 1:PATL1-Myc (COM 1/2). Furthermore, we examined the survival rate of plants under heat shock and performed statistics. Seedlings of Col, patl1 mutant, COM1 and COM2 grown normally on MS medium for 6 days were heat-shock treated at 45℃for 60 minutes and then after recovery in a 22℃incubator for 5 days, survival statistics were performed.

See fig. 3; showing the effect of heat stress on the survival rate of restorer seedlings; in the figure, a and B show: col, patl, 1COM1 and 1COM2 Arabidopsis seedlings were grown normally at 22℃for 10 days (Control) and at 22℃for 6 days, heat-shock treated at 45℃for 60 minutes and then restored in a 22℃incubator for 5 days (HS), and survival rates were compared. (1, col, 2, patl1, 3,1COM1, 4,1COM2); c shows: survival rates of the Col, patl, COM1 and COM2 Arabidopsis seedlings were counted. Values shown are mean ± standard deviation P < 0.05.

It can be seen that after heat shock treatment, the survival rate of Col was 55%, the survival rate of patl mutant was 63%, and the survival rates of both restorer plants COM1 and COM2 were 53% and 55%, respectively, and recovered to Col levels. It was demonstrated that PATL1 is indeed affected by heat stress.

Example 8 purification and characterization of PATL1 overexpressing lines in wild-type background

To further verify the relationship between PATL1 and HS at the genetic level. The super-expression strain of Col/pPATL1:PATL1-Myc (OE 1/2) is obtained by genetic means in this section, and RNA and protein levels are identified. Protein extraction and Western Blotting (WB) detection were performed by milling the material grown in the same period.

The results are shown in FIG. 4. Tubulin was found to act as an internal control protein in this process, and as a result a distinct Myc band was observed in the over-expressed strain OE1/2, but not in the WT. The expression level of PATL1 in the strain was analyzed by extracting RNA and real-time fluorescent quantitative PCR (qPCR). The results show that the expression level of PATL1 in different over-expression strains OE1/2 is obviously higher than that of the wild type strain, the expression level of PATL1 in OE1 is 2.16 times that of the wild type strain, and the expression level of OE2 is 1.81 times that of the wild type strain. The experiment shows that the pure strain of the PATL1 gene overexpression in the wild type background is successfully obtained, and can be used for subsequent experiments.

Example 9 survival analysis of OE1/2 seedlings under Heat stress

In order to explore the relationship between PATL1 and HS, the over-expressed pure and strain OE1/2 obtained in the upper section was cultured for 6d on a medium for normal growth, and after heat stress treatment (45 ℃ for 60 min) was applied with WT and PATL1 as a control, and then was recovered to 22 ℃ for 5d in a normal environment for observation and statistics.

As a result, as shown in FIG. 5, the survival rates of each of the WT, patl1, OE1/2 overexpressing plants were 56%, 64%, 40% and 42%, respectively. Survival rates for both overexpressing pure and lines were significantly lower than WT levels. This result also further demonstrates that PATL1 plays a negative regulatory role in heat stress signaling.

Example 10 survival analysis of OE1/2 seedlings under Heat stress

In order to explore the accumulation condition of plant vesicle transport, a lipophilic styrene compound FM4-64 which can be dissolved in water is adopted, when the plasma membrane is complete, the FM4-64 can be combined with phospholipid molecules on the cell membrane, and can enter the inside of the cell along with the movement of the plasma membrane in the endocytosis process, then the plant vesicle transport is further carried out to a Golgi body, finally the plant vesicle transport is stopped at a vacuole, and the positioning observation of the vesicle structure can be realized through a fluorescence observation mode. In the experiment, col, patl1, COM1/2 and OE1/2 plant materials of 4d are selected for experiment, dyeing treatment is carried out, and after the plant materials are placed in a culture solution for 15min, the plant materials are respectively observed, and statistical analysis is carried out on the relative fluorescence intensity in cells of different strains.

Referring to fig. 6, the results show that red-labeled vesicle structure increases with the time of staining treatment. As a result of statistics for 15min, the fluorescent signal in patl1 mutants was 1.5 times that of Col, and the relative fluorescence intensity of OE1/2 was about 0.6. And (3) analyzing strain difference results, wherein compared with Col vesicle results, the COM1/2 conditions are relatively consistent with wild type results, the vesicle transport in patl mutants is obviously increased, and the vesicle transport results in OE1/2 are obviously reduced and almost no vesicle transport is observed. The above results demonstrate that PATL1 pair is involved in the vesicle transport process that occurs in plant cells and is capable of exerting negative regulatory effects.

In summary, in the present invention, we have found that PATL1, as a negative thermal response factor, reduces HSP transport through vesicles. We studied the effect of HS on plant survival expressing modified versions of PATL 1. PATL1 expression is inversely proportional to plant survival, indicating that PATL1 negatively regulates heat resistance. PATL1 comprises SEC14 and GOLD domains, indicating that it is involved in membrane transport. As a physical signal, temperature changes the flow of the biofilm and remodeling of the membrane micro-regions. Our results also indicate that heat promotes the efficiency of vesicle transport. Deletion of PATL1 increases vesicle transport, while overexpression of PATL1 decreases vesicle transport, indicating that PATL1 plays a negative role in vesicle transport.

In the foregoing embodiments, the descriptions of the embodiments are emphasized, and in part, not described or illustrated in any particular embodiment, reference is made to the related descriptions of other embodiments.

The above embodiments are only for illustrating the technical solution of the present invention, and not for limiting the same; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit and scope of the technical solutions of the embodiments of the present invention, and are intended to be included in the scope of the present invention.

Claims (10)

1. The kit is used for regulating and controlling the resistance of plants to heat stress, and is characterized in that: the kit comprises a molecular biological element capable of regulating and controlling the expression level of a specific gene; the specific gene is a gene related to plant response to heat stress; the molecular biological element optionally comprises: the over-expression element combination of the specific gene, and/or the element combination for inhibiting or reducing the expression amount of the specific gene, and/or the element combination for silencing the expression of the specific gene.

2. The kit of claim 1, wherein: the specific gene is PATL1 gene, homologous gene thereof or equivalent gene with equivalent plant physiological function; the gene number of the PATL1 gene is specifically as follows: at1g72150.

3. The kit of claim 1, wherein: the molecular biological element is an element combination that inhibits or reduces the expression of the specific gene, and/or an element combination that silences the expression of the specific gene.

4. A recombinant expression vector comprising the PATL1 gene or a homologous gene thereof.

5. The recombinant expression vector of claim 4, wherein: the recombinant expression vector is a silencing vector.

6. A method of enhancing heat stress tolerance in a plant, comprising: reducing or silencing expression of a PATL1 homologous gene in the plant to enhance heat stress tolerance of the plant.

7. A method for increasing crop yield, comprising: reducing or silencing expression of the PATL1 homologous gene in the crop to enhance heat stress tolerance of the corresponding crop, thereby increasing yield of the corresponding crop.

Use of the patl1 gene for modulating heat tolerance in plants.

Use of the patl1 gene for modulating vesicle transport and/or heat shock protein accumulation in a plant cell.

10. Use according to claim 9, characterized in that: the transport of vesicles and/or accumulation of heat shock proteins within plant cells is facilitated by reducing expression of the PATL1 gene in plants.