Application of TaFtsZ2-1 or its biomaterials in improving stress resistance or photosynthesis of wheat

Abstract

The application provides a wheat plant TaFtsZ2-1 Or its biomaterials in improving the stress resistance or photosynthesis of wheat, and belongs to the technical field of genetic engineering and wheat breeding. Research finds that overexpression TaFtsZ2-1 Significantly increases the number of chloroplasts, optimizes the stacking structure of thylakoids, improves the net photosynthetic rate and photosynthetic efficiency, and changes the photosynthetic limitation type from RuBP regeneration rate (J max) to carbon fixation and triose phosphate utilization (TPU) cooperative limitation. TaFtsZ2-1 The overexpression of the wheat plant can effectively improve the tolerance of the wheat plant to drought and cadmium stress in the seedling stage by optimizing root growth and construction, maintaining the stability of the photosynthetic system, activating the antioxidant system of the plant and enhancing the osmotic regulation capacity.

Description

Technical Field

[0001] This invention belongs to the fields of genetic engineering and wheat breeding technology, and particularly relates to a... TaFtsZ2-1 Or the application of its biological materials in improving wheat stress resistance or photosynthesis. Background Technology

[0002] wheat( Triticum aestivum Wheat (L.) is one of the most important food crops worldwide, and its yield and quality directly affect global food security and human nutrition. Photosynthesis, as the core physiological process of energy conversion and carbon fixation in wheat, distributes its assimilated products throughout the plant via the sugar transport system, coordinating the dynamic balance between vegetative and reproductive growth and providing the material basis for grain filling and final yield formation. Studies have reported that enhancing wheat photosynthetic efficiency can effectively promote grain filling and increase yield; while abnormal photosynthetic metabolism can easily lead to insufficient carbon source supply, ultimately resulting in reduced yield. Therefore, in-depth analysis of the molecular regulatory mechanisms of photosynthetic carbon fixation and the discovery of key functional genes have significant theoretical value and application prospects for promoting high-yield molecular breeding of wheat.

[0003] Chloroplasts are the core organelles for light reactions and carbon fixation in plants, and mainly rely on... FtsZ The protein family proliferates. (Plant) FtsZ The protein family is mainly divided into FtsZ1 and FtsZ2Two subfamilies of chloroplasts, acting as self-assembling GTPases, participate in the regulation of chloroplast binary fission. They provide structural support for division by polymerizing to form Z-rings and provide contractile force during conformational changes, thereby driving chloroplast binary fission. Studies have shown that chloroplast development, quantity, and ultrastructure directly affect photosynthetic efficiency, thus regulating crop growth and yield. Overexpression or loss of function of AtFtsZ2 in Arabidopsis thaliana leads to chloroplast division inhibition, chloroplast gigantism, and decreased photosynthetic efficiency (Stokes, KD, McAndrew, RS, Figueroa, R., Vitha, S., Osteryoung, KW, 2000. Chloroplast division and morphology are differentially affected by overexpression of FtsZ1 and FtsZ2 genes in Arabidopsis 1[w] 1~10. Vitha, S., McAndrew, RS, Osteryoung, KW, and FtsZring formation at the chloroplast division site in PlantsJ 1~9.); overexpression of NtFtsZ1-2 in tobacco results in only 1~3 giant chloroplasts formed in mesophyll cells (Jeong, WJ, Park, Y.-I., Suh, K., Raven, JA, Yoo, OJ, Liu, JR, 2002. A large population of Small chloroplasts in tobacco leaf cells allow for more effective chloroplast movement than a few enlarged chloroplasts. (Plant Physiol. 129, 112-121. https: / / doi.org / 10.1104 / pp.000588). Current research has preliminarily clarified... FtsZ2-1 The gene is involved in the regulation of chloroplast division and growth and development in plants, but the molecular mechanism by which this gene regulates photosynthetic physiology and affects carbon assimilation efficiency in wheat is still a blank.

[0004] Drought has become a major factor limiting global food production. Drought stress damages chloroplast structure, leading to decreased photosynthetic efficiency and affecting wheat growth. Globally, a high percentage of wheat-producing areas are affected by drought, resulting in reduced wheat yields. Furthermore, wheat production is also severely threatened by cadmium (Cd).2+ Highly toxic and easily migrated, Cd preferentially accumulates in the root tip meristem after entering the plant, inhibiting root development by interfering with cell division and elongation. Damage to root function triggers a chain reaction in the aboveground parts, leading to the accumulation of Cd. 2+ Replacement of Mg in chlorophyll centers 2+ Drought reduces chlorophyll content and acts on photosystem II (PSII), disrupting the stability of the manganese cluster in the water-splitting complex, leading to decreased light-harvesting efficiency. The synergistic decline in root and shoot function ultimately results in reduced wheat yield, accompanied by Cd accumulation in the grains. In summary, both drought and Cd stress cause root damage, disrupt the photosynthetic system, induce oxidative stress and membrane lipid peroxidation, ultimately inhibiting wheat growth. However, TaFtsZ2-1 Whether wheat participates in stress adaptation by regulating plasmid division is currently poorly understood and has few reported research findings.

[0005] Therefore, utilizing TaFtsZ2-1 This study investigated the effects of gene knockout and overexpression on chloroplast morphology and photosynthetic parameters in wheat materials, analyzed the carbon metabolism regulatory network, and identified key functional genes and metabolites. TaFtsZ2-1 The physiological functions and regulatory mechanisms of wheat under drought and Cd stress, two abiotic stresses, provide gene targets and theoretical support for molecular breeding of high-yield and stress-resistant wheat. This is a technical problem that urgently needs to be solved. Summary of the Invention

[0006] Therefore, the object of the present invention is to provide a TaFtsZ2-1 Or the application of its biological materials in improving wheat stress resistance or photosynthesis.

[0007] To achieve the above-mentioned objectives, the present invention provides the following technical solution: This invention provides TaFtsZ2-1 Or the use of its biological materials in at least one of the following: (1) Improve wheat's resistance to adverse conditions; (2) Breeding stress-resistant transgenic wheat; (3) Increase the number of wheat chloroplasts; (4) Improve wheat photosynthesis; (5) Promotes wheat growth traits; (6) Improve the antioxidant capacity of wheat.

[0008] Preferably, the stress resistance includes resistance to drought stress and / or resistance to heavy metal stress; the growth traits include one or more of the following: single plant dry weight, plant height, root length, and number of roots.

[0009] Preferably, the biomaterial includes materials containing TaFtsZ2-1 Expression cassettes, recombinant vectors, recombinant bacteria, plant cells, plant tissues, or plant organs.

[0010] Preferably, the resistance to heavy metal stress includes resistance to Cd stress.

[0011] This invention provides a method for improving wheat traits, including increasing the content of wheat... TaFtsZ2-1 The steps to obtain transgenic wheat are: expression level and / or activity; The property includes at least one of the following S1 to S6: (S1) Improves wheat's resistance to stress; (S2) Breeding stress-resistant transgenic wheat; (S3) Increase the number of wheat chloroplasts; (S4) Promotes wheat photosynthesis; (S5) Promotes wheat growth traits; (S6) Improves the antioxidant capacity of wheat.

[0012] Preferably, the stress resistance includes resistance to drought stress and / or resistance to heavy metal stress; the growth traits include one or more of the following: single plant dry weight, plant height, root length, and number of roots.

[0013] Preferably, the improvement in wheat TaFtsZ2-1 Methods for determining the expression level and / or activity of the above-mentioned biological material include introducing the above-mentioned biological material into wheat, thereby increasing the expression level and / or activity of the material in wheat. TaFtsZ2-1 Overexpression.

[0014] This invention provides a method for breeding transgenic wheat having at least one of the traits shown in A1 to A6, comprising the following steps: Improve wheat TaFtsZ2-1 The steps to obtain transgenic wheat are: expression level and / or activity; (A1) Improves wheat's resistance to stress; (A2) Breeding stress-resistant transgenic wheat; (A3) Increase the number of chloroplasts in wheat; (A4) Promotes wheat photosynthesis; (A5) Promotes wheat growth traits; (A6) Improves the antioxidant capacity of wheat.

[0015] Preferably, the stress resistance includes resistance to drought stress and / or resistance to heavy metal stress; the growth traits include one or more of the following: single plant dry weight, plant height, root length, and number of roots.

[0016] Compared with the prior art, the present invention has the following beneficial effects: This invention provides TaFtsZ2-1 The application of its biological materials in improving wheat stress resistance or photosynthesis. This invention has found that overexpression... TaFtsZ2-1It significantly increases the number of chloroplasts, optimizes the thylakoid stacking structure, improves net photosynthetic rate and photosynthetic efficiency, and changes the photosynthetic limitation type from being dominated by RuBP regeneration rate (Jmax) to being synergistically limited by carbon fixation and triose phosphate utilization (TPU). TaFtsZ2-1 Overexpression of this gene can effectively enhance wheat seedlings' tolerance to drought and cadmium stress by optimizing root growth and formation, maintaining photosynthetic system stability, activating the plant's antioxidant system, and enhancing osmotic regulation. This indicates that the TaFtsZ2-1 gene has a dual biological function of regulating chloroplast development and mediating abiotic stress responses, expanding the physiological mechanisms by which chloroplast development genes participate in wheat stress resistance regulation. Attached Figure Description

[0017] Figure 1 For quantitative statistical analysis of chloroplast ultrastructure, A is a transmission electron microscope image of chloroplast ultrastructure; CP (chloroplast), S (starch grain), G (grana thylakoids), PG (osmophilic granules); B is the statistical analysis of chloroplast number and area, counting the number and area of ​​chloroplasts in each intact cell; C is the morphological analysis of individual chloroplasts, including perimeter, length, and breadth, with 24 independent chloroplasts counted per strain (n=24); data are expressed as mean ± standard deviation (SD), and independent samples t-tests were performed. express p <0.05, express p <0.01, express p <0.001.

[0018] Figure 2 for TaFtsZ2-1 Effects on the wheat chloroplast photosystem: A represents the total chlorophyll content, chlorophyll a content, and chlorophyll b content of different groups; B represents the electron transport rate (ETR) of different groups; C represents the Y(II), Y(NPQ), and Y(NO) of different groups; D represents the ETR-light response curves of different groups; E represents the Y(NO) light response curves of different groups; F represents the Y(NPQ) light response curves of different groups; the horizontal axis represents photosynthetically active radiation (PAR); data in the figure are expressed as mean ± standard error; significant differences were determined by independent samples t-test. express p <0.05, express p <0.01, express p <0.001.

[0019] Figure 3 for TaFtsZ2-1 The effect of chloroplast structure alterations on photosynthetic carbon fixation in wheat: A shows the light response curves for different groups; B shows the CO2 response curves for different groups; C shows the net photosynthetic rate (An) for different groups; D shows the percentages of Vcmax, Jmax, and TPU limitation for different groups; E shows the dark respiration rate (Rd); and F shows the maximum net photosynthetic rate (Anmax). Data in the figures are expressed as mean ± standard error; significant differences were determined using an independent samples t-test. This indicates that p < 0.05. This indicates that p < 0.01. This indicates that p < 0.001.

[0020] Figure 4 For transcriptome analysis, A shows principal component analysis (PCA) of transcripts from each material, where the circular icon represents the KO#2 group, the triangular icon represents the OE#1 group, and the square icon represents the WT group; B shows the Venn diagram of DEGs, with overlapping areas representing DEGs shared by both groups and non-overlapping areas representing DEGs specific to each strain; C shows the KEGG enrichment diagram of OE#1 and WT; and D shows the KEGG enrichment bubble diagram of KO#2 and WT.

[0021] Figure 5 For editing TaFtsZ2-1 Metabolomic analysis of wheat genes: A is principal component analysis (PCA); B is a pie chart showing the distribution of detected metabolite categories; C is an upset plot of differentially metabolites among different comparison groups; D is KEGG pathway enrichment analysis of differentially metabolites between OE#1 and wild-type (WT); E is KEGG pathway enrichment analysis of differentially metabolites between KO#2 and wild-type (WT).

[0022] Figure 6 For the enrichment analysis of KEGG pathways related to stress resistance in OE2 and KO2 lines and WT, in the bubble plots of KEGG enrichment analysis of differentially expressed genes in OE2 vs WT and KO2 vs WT, the X-axis represents the enrichment ratio, and the Y-axis represents the significantly enriched (Q<0.05) stress resistance-related KEGG pathways. The size of the bubble represents the number of differentially expressed genes enriched in that pathway, and the bubble color represents -log 10 (Qvalue), the number in the bubble is the number of enriched genes.

[0023] Figure 7The dynamic changes of physiological indicators of each line under PEG stress were shown in the figure. A to L represent the dynamic changes of single plant dry weight (A), root length (B), root number (C), root SOD activity (D), root POD activity (E), root MDA activity (F), plant height (G), chlorophyll content (H), leaf soluble sugar content (I), leaf SOD activity (J), leaf POD activity (K), and leaf MDA activity (L) at 0 d, 3 d, 7 d, and R3d (3 d after rehydration) under PEG stress, respectively. Among them, there were significant differences between groups represented by different letters, p<0.05.

[0024] Figure 8 The dynamic changes of physiological indicators of each line under Cd stress were shown in the figure. A to L represent the dynamic changes of single plant dry weight (A), root length (B), root number (C), root SOD activity (D), root POD activity (E), root MDA activity (F), plant height (G), chlorophyll content (H), leaf soluble sugar content (I), leaf SOD activity (J), leaf POD activity (K), and leaf MDA activity (L) at 0 d, 3 d, and 7 d, respectively. Among them, there were significant differences between groups represented by different letters, p < 0.05. Detailed Implementation

[0025] This invention provides TaFtsZ2-1 Or the use of its biological materials in at least one of the following: (1) Improve wheat's resistance to adverse conditions; (2) Breeding stress-resistant transgenic wheat; (3) Increase the number of wheat chloroplasts; (4) Promotes wheat photosynthesis; (5) Promotes wheat growth traits; (6) Improve the antioxidant capacity of wheat.

[0026] In this invention, the stress resistance includes resistance to drought stress and / or resistance to heavy metal stress. As a preferred embodiment, the heavy metal stress is cadmium stress. The drought stress is PEG-induced drought, and the PEG is PEG-6000. The growth traits include one or more of the following: single-plant dry weight, plant height, root length, and number of roots.

[0027] In this invention, the TaFtsZ2-1 Including wheat ( Triticum aestivumHomologous copies on chromosomes 2A, 2B, and 2D (L.) have IWGSC RefSeq v1.0 annotation numbers TraesCS2A02G294500, TraesCS2B02G310900, and TraesCS2D02G292200, respectively, with corresponding genomic coordinates of Chinese_Spring1.0_chr2A:507068707-507073983, Chinese_Spring1.0_chr2B:444541562-444546858, and Chinese_Spring1.0_chr2D:374104393-374109627.

[0028] In this invention, the biomaterial includes materials containing... TaFtsZ2-1 The recombinant bacteria may contain an expression cassette, a recombinant vector, a recombinant bacterium, plant cells, plant tissues, or plant organs. The backbone vector of the recombinant vector may be a plant binary expression vector known in the art, such as, but not limited to, pCambia3301, pCambia1300, or pBin19. The host bacteria of the recombinant bacteria may include *Escherichia coli* or *Agrobacterium*.

[0029] This invention provides a method for improving wheat traits, including increasing the content of wheat... TaFtsZ2-1 The steps to obtain transgenic wheat are: expression level and / or activity; The property includes at least one of the following S1 to S6: (S1) Improves wheat's resistance to stress; (S2) Breeding stress-resistant transgenic wheat; (S3) Increase the number of wheat chloroplasts; (S4) Promotes wheat photosynthesis; (S5) Promotes wheat growth traits; (S6) Improves the antioxidant capacity of wheat.

[0030] In this invention, the stress resistance includes resistance to drought stress and / or resistance to heavy metal stress. As a preferred embodiment, the resistance to heavy metal stress is resistance to cadmium stress. The drought stress is PEG-induced drought, and the PEG is PEG-6000. The growth traits include one or more of the following: single-plant dry weight, plant height, root length, and root number, overexpressed under normal conditions, drought stress, and cadmium stress. TaFtsZ2-1 Wheat plants showed increased dry weight per plant, plant height, root length, and root number. As a preferred method, the improvement in wheat... TaFtsZ2-1 Methods for determining the expression level and / or activity of the above-mentioned biological material include introducing the above-mentioned biological material into wheat, thereby increasing the expression level and / or activity of the material in wheat. TaFtsZ2-1 Overexpression.

[0031] This invention provides a method for breeding transgenic wheat having at least one of the traits shown in A1 to A6, comprising the following steps: Improve wheat TaFtsZ2-1 The steps to obtain transgenic wheat are: expression level and / or activity; (A1) Improves wheat's resistance to stress; (A2) Breeding stress-resistant transgenic wheat; (A3) Increase the number of chloroplasts in wheat; (A4) Promotes wheat photosynthesis; (A5) Promotes wheat growth traits; (A6) Improves the antioxidant capacity of wheat.

[0032] As a preferred method, the improvement of wheat TaFtsZ2-1 Methods for determining the expression level and / or activity of the above-mentioned biological material include introducing the above-mentioned biological material into wheat, thereby increasing the expression level and / or activity of the material in wheat. TaFtsZ2-1 Overexpression. The stress resistance includes resistance to drought stress and / or heavy metal stress. As a preferred embodiment, the heavy metal stress is cadmium stress. The drought stress is PEG-induced drought, and the PEG is PEG-6000. The growth traits include one or more of single-plant dry weight, plant height, root length, and root number, under normal conditions, drought stress, and cadmium stress, overexpression. TaFtsZ2-1 Wheat plants showed improvements in single-plant dry weight, plant height, root length, and root number.

[0033] In this invention, unless otherwise specified, all raw material components are commercially available products well known to those skilled in the art.

[0034] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0035] Example 1 TaFtsZ2-1 Molecular mechanisms regulating wheat chloroplast division and photosynthetic carbon assimilation efficiency 1. Materials and Methods 1.1 Materials and Growth Conditions The experimental materials were wheat Fielder wild type (WT) and two TaFtsZ2-1Overexpression lines (OE#1, OE#2) and two CRISPR / Cas9 knockout lines (KO#1, KO#2) were used; the molecular and physiological analyses in this embodiment were performed using OE#1 and KO#2 as representative lines. OE#1, OE#2, KO#1, and KO#2 are respectively OE1, OE2, KO1, and KO2 in the literature “Li, Chao, Fu, K., Wei, J., Li, G., Li, Cheng, Li, Chunyan, 2025. Impact of TaFtsZ2 overexpression and mutation on wheat starch granule characteristics and quality. CarbohydratePolymers 353, 123267. https: / / doi.org / 10.1016 / j.carbpol.2025.123267”. TaFtsZ2-1 For the document TaFtsZ2.

[0036] according to TaFtsZ2-1 gRNA target sites were designed using homologous sequences from chromosomes A, B, and D, and CRISPR / Cas9 knockout was employed. The gRNA sequence is 5'-GCTGGAATTGCCAAGTCCATGGG-3' (SEQ ID NO.1). KO#2 underwent single-base insertions at target sites 2A, 2B, and 2D, resulting in a cDNA shift mutation at position 664 and premature termination at position 714. TaFtsZ2-1 Protein function loss; construction of pCambia3301- TaFtsZ2 Overexpression vector, T2~T3 generations of OE#1 TaFtsZ2-1 The relative expression levels were more than twice that of WT.

[0037] Select plump, uniformly sized seeds, rinse three times with distilled water, blot dry with filter paper, and evenly place them in a germination box lined with moist filter paper for germination. Once the seeds have germinated and grown uniformly, transplant them into pots for further cultivation, with 6 pots for each strain and 3 seedlings per pot. All seedlings are cultivated in an artificial climate incubator under the following conditions: temperature 24.5–25.5℃, relative humidity 55%–65%, photoperiod 16h light / 8h dark, and light intensity 200 μmol·m⁻¹. -2 ·s -1 .

[0038] 1.2 Chlorophyll content determination The chlorophyll content was determined by selecting the fifth leaf at the seedling stage and using the Arnon method. Three biological replicates were set for each line.

[0039] 1.3 Observation of Chloroplast Ultrastructure The preparation of chloroplast ultrastructure samples followed the standard transmission electron microscopy (TEM) procedure. Under 8 h of illumination, the fifth leaf from the WT, OE#1, and KO#2 lines was selected. The specific procedures were as follows: Samples were fixed with 2.5% glutaraldehyde at 4°C in the dark, then rinsed three times with 0.1 M phosphate buffer (pH 7.2), 15 min each time; followed by a second fixation with 1% osmium tetroxide at room temperature for 2 h, and then rinsed with phosphate buffer to completely remove residual osmium tetroxide. Samples were dehydrated with a gradient of acetone (30%, 50%, 70%, 80%, 90%, 100%, with 100% repeated twice, 15 min each step), then subjected to a gradient permeation of acetone with Super embedding medium (3:1, 1:1, 1:3, pure embedding medium, 12 h each step), and finally embedded in pure embedding medium. Pre-curing was performed at 45°C for 12 h, followed by final curing at 60°C for 24 h. After solidification, the samples were ultrathinly sectioned and observed under a transmission electron microscope to acquire ultrastructural images. Quantitative analysis was performed using ImageJ. The total area of ​​single-cell chloroplasts was estimated as the product of the mean number of chloroplasts per cell (n = 7) and the mean area of ​​a single chloroplast (n = 24). Because the two measurements came from different samples, a nonparametric bootstrap method (10,000 resamplings with replacement) was used to estimate the 95% confidence interval of the total area and its inter-line ratio. A confidence interval for the ratio not containing 1 was considered statistically significant.

[0040] 1.4 Gas exchange parameters and chlorophyll fluorescence parameters Leaf gas exchange parameters were measured using a LI-6800 portable photosynthesis system (LI-COR, Inc., Lincoln, NE, USA). The fifth fully expanded leaf at the seedling stage was selected. Measurement parameters included steady-state photosynthetic parameters and light and CO2 response curves. Measurement conditions were set as follows: for steady-state parameter measurements, photosynthetically active radiation (PAR) was set to 1500 μmol photons / m². -2 s -1 The CO2 concentration in the leaf chamber was 400 μmol. -1 The leaf chamber temperature was 25℃, and the relative humidity was maintained at 60%~70%. The PAR gradients measured by the light response curves were 1200, 1000, 800, 600, 400, 200, 100, 50, and 0 μmol photons m -2 s -1 The intercellular CO2 concentration gradients measured by the CO2 response curves were 50, 100, 200, 300, 400, 500, 600, 800, 1000, and 1200 μmol / mol. -1 .

[0041] Chlorophyll fluorescence parameters were measured using a PAM-2500 portable modulated chlorophyll fluorometer (Heinz Walz GmbH, Germany). Leaves were placed in a dark adaptation clamp for 30 min to allow for sufficient dark adaptation. After this period, chlorophyll fluorescence parameters were measured at a PAR of 1165 μmol photons / m². -2 s -1 The parameters measured under the given conditions included electron transport rate (ETR), actual photochemical efficiency (Y(II)), photochemical quenching coefficient (qL), non-photochemical quenching quantum yield (Y(NPQ)), and unregulated energy dissipation quantum yield (Y(NO)). The PAR gradients measured by the dynamic photoresponse curves were 0, 50, 100, 200, 400, 600, 800, 1000, 1200, 1400, and 1600 μmol photons m. -2 s -1 The study focused on analyzing the response relationships of ETR, Y(II), qL, and Y(NPQ) to changes in PAR. Each strain was replicated three times.

[0042] 1.5 Transcriptome Sequencing WT, OE#1, and KO#2 lines were selected as materials, with three biological replicates for each line. Transcriptome sequencing was performed by BGI Genomics Co., Ltd. using the DNBSEQ platform: total RNA was extracted and a library was constructed. PCR products were denatured to form single-stranded DNA and then circularized into single-stranded circular DNA. DNA nanoballs (DNBs) were then generated through rolling circle replication for sequencing. Raw sequencing data were filtered using SOAPnuke to obtain clean reads conforming to the Sanger FASTQ format. HISAT was used to align clean reads to the wheat reference genome, and quality control was performed based on alignment rate and read distribution. Qualified data were further used for gene quantification, differential expression analysis, GO / KEGG functional enrichment, cluster analysis, and protein-protein interaction network construction. SNPs, InDel events, and differential splicing events were also detected simultaneously to analyze... TaFtsZ2-1 Molecular mechanisms regulating wheat physiological metabolism.

[0043] 1.6 Non-targeted metabolomics sequencing and analysis WT, OE#1, and KO#2 strains were selected as materials for non-targeted metabolomics analysis, with six biological replicates for each strain. Samples were analyzed by BGI Genomics Co., Ltd. in Shenzhen using ultra-high performance liquid chromatography-quadrupole electrostatic field orbital trap high-resolution mass spectrometry (UPLC-Q Exactive MS). Raw mass spectrometry data were imported into CompoundDiscoverer 3.3 software and analyzed using databases such as BMDB, mzCloud, and ChemSpider for metabolite identification and data analysis.

[0044] 1.7 Data Analysis and Plotting Data analysis and plotting were performed using Origin 2026b, R software (v4.3.0), and ImageJ. The significance of differences between groups was analyzed using software, employing an independent samples t-test. The significance level was determined as follows: p <0.05, p <0.01, p <0.001; ns indicates no significant difference. Bar chart data are presented as mean ± standard deviation (mean ± SD); box plot data are presented as median (interquartile range) and individual data points. The number of replicates for each experimental biology experiment was set according to the specific methodology.

[0045] 2. Results 2.1 TaFtsZ2-1 Expression dosage determines chloroplast quantity and morphogenesis Transmission electron microscopy observations showed that TaFtsZ2-1 Expression levels significantly alter the number, size, and ultrastructure of chloroplasts in mesophyll cells. Figure 1 (Middle A). The average number of chloroplasts per cell in WT was 8.14 ± 1.57 (n=7), while in the OE#1 line it increased to 12.71 ± 2.43 (an increase of 56.14% compared to WT). P <0.01), the number of KO#2 strains decreased to 3.86±0.69 (a decrease of 52.58% compared to WT), P <0.001)( Figure 1(Medium B). Morphological measurements (n=24) showed that, compared with WT, the area, perimeter, and length of individual chloroplasts in OE#1 decreased by 17.49%, 27.54%, and 37.02%, respectively, while the width increased by 28.82%, and the aspect ratio decreased from 2.68 in WT to 1.31. The chloroplasts as a whole were "small and numerous" elliptical shapes. In KO#2, the area, perimeter, length, and width of individual chloroplasts increased by 95.18%, 51.78%, 66.07%, and 21.48%, respectively, and the aspect ratio increased to 3.66. Moreover, the inter-individual variation increased significantly (area CV≈34%), exhibiting "few and giant" flattened ellipsoidal shapes. Figure 1 (C). Calculation of the total area of ​​single-cell chloroplasts shows that the WT is 91.3 μm. 2 (95% CI 78.9~104.0), OE#1 reaches 117.5 μm 2 (95% CI 101.4~136.0), an increase of 29.4% compared to WT; KO#2 was 84.3 μm. 2 (95% CI 69.9~100.4), a 7.2% reduction compared to WT, its giant size is almost completely compensated for in terms of area. In terms of ultrastructure, the thylakoid grana of OE#1 are densely stacked and regularly arranged, and a large number of electron-transparent regions are visible in the chloroplasts; while the giant KO#2 has disordered arrangement of chloroplast thylakoids.

[0046] 2.2 Chloroplast structural remodeling enhances light energy utilization efficiency and photoprotection capability TaFtsZ2-1 Overexpression significantly increased leaf chlorophyll content, while knockout significantly decreased it. Figure 2 (A). Chlorophyll fluorescence analysis showed that, compared with WT, the ETR of the OE#1 line increased by 12.15%, while the ETR of the KO#2 line decreased by 11%. The ETR-light response curves showed a "rapid rise-saturation" pattern, with the curves shifting upward in overexpression lines and downward in knockout lines, consistent with the steady-state measurement results. Figure 2 (B, D)

[0047] The results of the light energy distribution analysis show that ( Figure 2 (C, E, F). Figure 2 The data from C shows that WT's Y(II) is 6.17%, Y(NO) is 54.37%, and Y(NPQ) is 39.47%; OE#1's Y(II) is 7.37%, Y(NO) is 43.00%, and Y(NPQ) is 49.67%; and KO#2's Y(II) is 5.90%, Y(NO) is 43.43%, and Y(NPQ) is 50.63%. Figure 2 The data from the middle E shows that the range is 0~1600 μmol photons·m -2 ·s -1Within the PAR range, the Y(NO) of WT, OE#1, and KO#2 all showed a rapid increase followed by a saturation trend as PAR increased, with WT's Y(NO) being higher than OE#1 and KO#2's Y(NO) being higher than OE#1. Figure 2 The results showed that, within the same PAR range, the Y(NPQ) of WT, OE#1, and KO#2 all increased with increasing PAR, with OE#1 showing a higher Y(NPQ) than KO#2, and KO#2 showing a higher Y(NPQ) than WT. Compared to WT, the Y(NPQ) of the OE#1 strain increased from 39.47% to 49.67%, an increase of 25.84%; Y(NO) decreased from 54.37% to 43.00%, a decrease of 20.91%. Compared to WT, the Y(NPQ) of the KO#2 strain increased from 39.47% to 50.63%, an increase of 28.28%; Y(II) decreased from 6.17% to 5.90%. This indicates that the overexpression strains, while maintaining photochemical quenching, channeled more excess excitation energy into the regulatory heat dissipation pathway, reducing non-regulatory energy loss, representing active photoprotection optimization. In contrast, the KO#2 strain showed an increase in Y(NPQ) but a significant decrease in Y(II). The increased heat dissipation was a passive stress response under the background of impaired photochemical efficiency, which is fundamentally different from the active photoprotection mechanism of the overexpression strain.

[0048] 2.3 Improved carbon assimilation efficiency is accompanied by a shift in photosynthetic limitation from being dominated by Jmax to becoming more balanced. The light response curves and CO2 response curves show that all strains exhibit a "rapid rise - tending to stabilize" pattern, with the OE#1 curve shifting upwards overall and the KO#2 curve shifting downwards overall. Figure 3 (A, B). The light compensation point of the overexpression lines was higher than that of WT (21.5 μmol·m). -2 ·s -1 The dark respiration rate of OE#1 decreased significantly, and the ratio of dark respiration rate to maximum net photosynthetic rate (Rd / Anmax) decreased significantly, while low-light utilization efficiency and carbon assimilation efficiency increased simultaneously. Figure 3 E, F). Steady-state gas exchange parameters indicate ( Figure 3 Compared with WT, OE#1 showed a 9.62% increase in net photosynthetic rate (An) and a 33% increase in maximum net photosynthetic rate (Anmax), respectively.

[0049] The constraint type analysis of the A-Ci curve fitting further distinguished between the two types of strains ( Figure 3 In the D-group, photosynthetic limitation in WT was mainly due to Jmax (RuBP regeneration rate), accounting for 48%, while Vcmax and TPU limitation accounted for 31% and 21%, respectively. In the overexpression lines, photosynthetic limitation tended to be more balanced: in OE#1, the Jmax limitation rate decreased to 41%, while the Vcmax and TPU limitation rates increased to 35% and 24%, respectively. This indicates that overexpression... TaFtsZ2-1 The electron transport limitation was alleviated, and the proportion of photosynthetic product utilization in the limitation structure increased, with carbon assimilation processes becoming more synergistic. KO#2 showed the opposite trend, with Jmax limitation accounting for 63%, Vcmax limitation for 28%, and TPU limitation for 9%, indicating that the inhibited regeneration rate of RuBP was the dominant factor in the decline of photosynthesis in the knockout strain.

[0050] 2.4 Combined analysis of transcriptomics and metabolomics TaFtsZ2-1 Mediated metabolic reprogramming Transcriptome sequencing yielded 10–12 Gb of high-quality data per sample. Principal component analysis (PCA) and cluster analysis showed that the gene expression patterns of the three lines, WT, OE#1, and KO#2, were clearly separated. Figure 4 (A). Differentially expressed genes (DEGs) were screened using |log2FC|>1 and P<0.05 as the threshold. A total of 4772 DEGs were identified in the OE#1 vs WT comparison, and 1918 DEGs were identified in the KO#2 vs WT comparison. Figure 4 B). KEGG analysis showed ( Figure 4 In the C and D groups, DEGs of OE#1 were significantly enriched in the glyoxylic acid and dicarboxylic acid metabolic pathways, and were also involved in the alanine / aspartic acid / glutamate metabolic pathway, which could provide nitrogen assimilation products and carbon skeletons for chloroplast proliferation. DEGs of KO#2 were specifically enriched in plant-pathogen interaction, plant hormone signal transduction and MAPK signaling pathways, which is consistent with the oxidative stress and defense response associated with the formation of giant chloroplasts.

[0051] A total of 46,305 raw signal peaks were detected in the untargeted metabolomics, and 1,171 metabolites were annotated; PCA showed that the metabolomics profiles of the WT, OE#1, and KO#2 strains were clearly separated. Figure 5 (A). Metabolite composition analysis showed that lipids and lipid molecules accounted for the highest proportion (31.1%), followed by organic acids and their derivatives (27.4%). Figure 5 (B) Differential metabolites (DAMs) were screened using a variable importance projection (VIP) > 1 and P < 0.05 as the threshold. 183 DAMs were found between OE#1 and WT, and 240 DAMs were found between KO#2 and WT. Figure 5 KEGG enrichment analysis showed that the DAMs of OE#1 were mainly enriched in amino acid metabolism, plant secondary metabolite synthesis, and energy and cofactor metabolism pathways. Figure 5 In the D and E pathways, the enrichment of aminoacyl-tRNA biosynthesis pathway is consistent with the protein synthesis requirements of chloroplast proliferation. DAMs of KO#2 are enriched in multiple amino acid metabolic pathways, citric acid cycle, pentose phosphate pathway and α-linolenic acid metabolism, mainly oligopeptides such as ILK and amino acid derivatives, reflecting the amino acid metabolic rearrangement and defense response activation under carbon skeleton deficiency in knockout strains.

[0052] Example 2 TaFtsZ2-1 Physiological mechanisms regulating drought and cadmium stress tolerance in wheat seedlings 2.1 Materials and Methods 2.1.1 Materials and Experimental Design The material is wheat Fielder wild type (WT). TaFtsZ2-1 Overexpression lines OE2 and CRISPR / Cas9 knockout line KO2 (provided by the Winter Wheat Molecular Genetics and Breeding Laboratory, Shihezi University). KO2 in... TaFtsZ2-1 Homologous copies 2A, 2B, and 2D all carry single-base insertion mutations, resulting in loss of protein function; OE2 is derived from pCambia3301- TaFtsZ2 Obtained by vector transformation, stable overexpression in T2-T3 generations.

[0053] The KO2 and OE2 mentioned are respectively from the literature: Li, Chao, Fu, K., Wei, J., Li, G., Li, Cheng, Li, Chunyan, 2025. Impact of TaFtsZ2 overexpression and mutation on wheatstarch granule characteristics and quality. Carbohydrate Polymers 353,123267. https: / / doi.org / 10.1016 / j.carbpol.2025.123267. TaFtsZ2 That is TaFtsZ2-1 .

[0054] After the seeds were surface sterilized with 0.1% mercuric chloride, they were rinsed three times with sterile distilled water and placed in a petri dish lined with double-layered sterile filter paper. After germination, seedlings with uniform growth were selected and transplanted into a black plastic box containing Hoagland nutrient solution for hydroponics. The roots were fixed with sterile absorbent cotton and the nutrient solution was changed every 2 days. The seedlings were cultured to the three-leaf stage and then subjected to the following stress treatments: (1) PEG-6000 simulated drought stress, with 0% (control, normal 1 / 2 Hoagland nutrient solution), 15% (w / v) PEG-6000 solution, and 20% (w / v) PEG-6000 solution for 7 days. The preparation method of PEG-6000 solution is as follows: weigh PEG-6000 powder, dissolve it in 1 / 2 Hoagland nutrient solution, and make up to the required volume to obtain the corresponding concentration of PEG-6000 solution. (2) CdCl2 stress: CdCl2 solutions with a final concentration of 0 μmol / L (control, 1 / 2 Hoagland nutrient solution), 20 μmol / L, and 30 μmol / L were used to stress patients for 7 days. The CdCl2 solution was prepared by weighing CdCl2, dissolving it in distilled water and making up to the required volume to prepare the working solution with the corresponding final concentration. Each treatment was repeated 3 times.

[0055] 2.1.2 Transcriptome Sequencing Wheat leaves from WT, OE2, and KO2 seedlings cultured to the four-leaf stage (three biological replicates per line) were selected for DNBSEQ transcriptome sequencing (BGI Genomics). DNB libraries were constructed by circularization and rolling circular replication of PCR products and then sequenced. Raw data were filtered using SOAPnuke and aligned to the wheat reference genome quality control using HISAT2. Differential expression, GO / Pathway enrichment, SNP / InDel detection, and splicing analysis were then performed. TaFtsZ2-1 Molecular mechanisms regulating physiological metabolism.

[0056] 2.1.3 Measurement of growth indicators Plants were harvested at 0, 3, and 7 days after stress treatment and 3 days after rehydration (drought treatment only). The above-ground parts were separated from the roots and laid flat. Plant height (from root base to seedling leaf tip) and root length (from root tip to stem base) were measured using vernier calipers (0.1 mm accuracy), and the number of visible roots was counted. After measurement, the samples were blanched at 105℃ for 10 min, dried at 70℃ to constant weight (the difference between two weighings was <0.001 g), and the dry weight (g) of a single plant was measured using a 0.001 g electronic balance.

[0057] 2.1.4 Chlorophyll content determination Samples were taken at 0, 3, and 7 days after stress treatment and 3 days after rehydration (drought treatment only). 0.2 g of leaves were weighed and extracted with 95% ethanol solution in the dark until colorless, then diluted to 25 mL. Using 95% ethanol solution as a blank, absorbance was measured at wavelengths of 665, 649, and 470 nm. The chlorophyll concentration (mg / L FW) in the extract was calculated using the Lichtenthaler formula. Ca = 13.95A665 - 6.88A 649 ; Cb = 24.96A649 - 7.32A 665 ; CT = Ca + Cb.

[0058] In the formula: Ca is the concentration of chlorophyll a; Cb is the concentration of chlorophyll b; CT is the total chlorophyll concentration.

[0059] 2.1.5 Assay of Superoxide Dismutase (SOD) and Peroxidase (POD) Activities Samples were taken at 0, 3, and 7 days after stress treatment and 3 days after rehydration (drought treatment only). 0.1 g of fresh wheat leaves were weighed and added to pre-cooled 0.05 mol / L phosphate buffer (pH 7.0). The mixture was homogenized in an ice bath, and the volume was adjusted to 1 mL. The mixture was centrifuged at 10,000 rpm for 15 min at 4°C. The supernatant was stored at 4°C for later use. SOD activity was determined using the nitroblue tetrazolium (NBT) reduction method, and POD activity was determined using the guaiacol method.

[0060] 2.1.6 Determination of Malondialdehyde (MDA) Content Samples were taken at 0, 3, and 7 days after stress treatment and 3 days after rehydration (drought treatment only). 0.5 g of fresh wheat leaves were weighed, and the MDA content was determined using the thiobarbituric acid (TBA) method. The samples were ground with 10% trichloroacetic acid (TCA), centrifuged at 10,000 r / min for 15 min at 4℃, and the supernatant was mixed with 0.6% TBA (prepared with 10% TCA). The mixture was then boiled in a water bath for 15 min, rapidly cooled, and the absorbance was measured at 600, 532, and 450 nm.

[0061] In the formula: Vt is the total volume of the extract (mL); Vs is the volume of the extract used for the determination (mL); n is the dilution factor; FW is the fresh weight of the sample (g).

[0062] 2.1.7 Determination of soluble sugar content in leaves Samples were taken at 0, 3, and 7 days after stress treatment and 3 days after rehydration (drought treatment only). 0.1 g of fresh wheat leaves were weighed, and the soluble sugar content of the leaves was determined by the anthrone colorimetric method: 80% ethanol solution was extracted by water bath at 80℃ for 30 min, diluted, and anthrone-sulfuric acid reagent was added. After mixing, the mixture was placed in a water bath at 100℃ for 10 min, cooled to room temperature, and the absorbance at 620 nm was measured.

[0063] In the formula: C is the glucose content (µg) obtained from the standard curve; Vt is the total volume of the extract (mL); Vs is the sampling volume during the determination (mL); n is the dilution factor; W is the dry mass of the sample (mg).

[0064] 2.1.8 Data Processing Data were prepared using Excel 2019, and statistical analysis was performed using SPSS 26.0 (IBM Corporation, Armonk, NY, USA) and R 4.3.1. The main effects and interactions of genotype, stress treatment, and stress duration were analyzed using three-way ANOVA, and multiple comparisons were performed using Duncan's method (p < 0.05). Figures and tables were plotted using R 4.3.1 (ggplot2 package).

[0065] 2.2 Results 2.2.1 Transcriptome Revealed TaFtsZ2-1 Participating in counter-cyclical regulation To investigate TaFtsZ2-1 The effects of KEGG on wheat chloroplast development and photosynthetic function were investigated by transcriptome sequencing of OE2, KO2, and WT under normal growth conditions. KEGG pathway enrichment analysis revealed significant enrichment in chloroplast-related pathways (such as photosynthesis, nitrogen metabolism, and carbon fixation), and differentially expressed genes were also significantly enriched in multiple stress response-related pathways (see...). Figure 6 The KEGG assays of OE2 and WT significantly enriched pathways including carotenoid biosynthesis, propionic acid metabolism, plant hormone signal transduction, and α-linolenic acid metabolism; while those of KO2 and WT enriched pathways including plant-pathogen interaction, MAPK signaling, and plant hormone signal transduction. Notably, plant hormone signal transduction, MAPK signaling, plant-pathogen interaction, and phenylalanine metabolism were all significantly regulated in both OE2 and KO2, suggesting... TaFtsZ2-1 Under normal conditions, it participates in the regulation of multiple defense-related pathways, and overexpression and knockout exhibit different regulatory patterns.

[0066] Figure 6 The results show that TaFtsZ2-1Under normal growth conditions, it can affect the expression of pathways related to antioxidant defense, osmotic regulation (propionic acid metabolism), and hormone signaling regulation (plant hormone signal transduction), suggesting that this gene may have potential stress response regulatory functions.

[0067] To verify this hypothesis, this invention subjected three strains, OE2, KO2 and WT, to PEG-simulated drought and Cd stress treatments, and explored their differences in stress resistance response by measuring relevant physiological indicators.

[0068] 2.2.2 TaFtsZ2-1 Enhancing wheat drought stress tolerance Under normal conditions, the overall growth indicators of WT, OE2, and KO2 show the order of OE2 > WT > KO2. Figure 7 (A~C, G). Under PEG-6000 stress, at 15% PEG, all indicators of OE2 plants continued to increase. After 7 days of stress, the dry weight per plant, number of roots, root length, and plant height increased significantly by 20.0%, 13.4%, 20.6%, and 8.2% respectively compared with WT (p<0.05). After 3 days of rehydration, the dry weight increased significantly by 16.7% compared with WT (p<0.05). At 20% PEG, the differences between genotypes were more significant. After 7 days of stress, the dry weight per plant, number of roots, root length, and plant height of OE2 plants increased significantly by 29.7%, 23.1%, 11.9%, and 9.7% respectively compared with WT (p<0.05). After 3 days of rehydration, the dry weight further increased by 30.0% compared with WT. Conversely, the degree of growth inhibition in KO2 increased with increasing PEG concentration: after 3 days of 15% PEG stress, the dry weight and root number of KO2 plants were significantly reduced by 10.3% and 15.2% compared to WT, respectively (p<0.05). After 7 days, the root number was still significantly reduced by 20.0% compared to WT (p<0.05). After 3 days of rehydration, the dry weight, root length, plant height, and root number of KO2 plants were reduced by 10.0%, 7.9%, 7.0%, and 30.0% compared to WT, respectively (p<0.05). After 7 days of 20% PEG stress, the dry weight and root number of KO2 plants were significantly reduced by 10.9% and 15.2% compared to WT, respectively (p<0.05). After rehydration, the root number was still 20.0% lower than WT.

[0069] Under PEG stress, the photosynthetic and osmotic regulation abilities of different genotypes showed significant differentiation. Regarding chlorophyll content, at 15% PEG, WT initially decreased and then increased; OE2 showed a smaller decrease and recovered rapidly after rehydration; while KO2 showed a continuous decrease and did not recover after rehydration. Specifically, OE2 was significantly higher than WT at 3 days, 7 days of stress, and 3 days after rehydration, while KO2 was significantly lower than WT at all sampling points after stress. Figure 7(H in Figure 7). At 20% PEG, both WT and KO2 continuously decreased, while OE2 showed a smaller decrease, but increased significantly by 55.8% after rehydration compared to WT (p<0.05) (H in Figure 7). Leaf soluble sugar content exhibited stress-induced accumulation characteristics. OE2 showed an "increase-decrease-increase" trend, increasing significantly by 28.4% and 50.9% compared to WT after 3 and 7 days of 15% PEG stress, respectively, and by 33.5% after rehydration (p<0.05). After 7 days of 20% PEG stress, it increased significantly by 62.9% compared to WT (p<0.05). KO2 showed an "increase-decrease" trend and remained consistently lower than WT. After 7 days of 20% PEG stress, it decreased by 4.9% compared to WT, and by 11.8% after rehydration compared to WT (p<0.05). Figure 7 (I). Meanwhile, antioxidant enzyme activities showed a trend of first increasing and then decreasing with increasing PEG concentration, with significant differences among genotypes. After 7 days of 15% PEG stress, the POD and SOD activities in OE2 leaves increased significantly by 15.7% and 36.2% respectively compared to WT, and the POD and SOD activities in roots increased significantly by 24.9% and 27.5% respectively compared to WT (p<0.05); after 7 days of 20% PEG stress, the POD and SOD activities in OE2 leaves increased significantly by 18.3% and 32.8% respectively compared to WT, and the POD and SOD activities in roots increased significantly by 34.0% and 30.1% respectively compared to WT (p<0.05). Figure 7 The content of MDA was significantly lower than that of WT, and the antioxidant compensation capacity of OE2 was significantly enhanced. Figure 7 (F and L). In contrast, the antioxidant enzyme activities of KO2 were all lower than those of WT: After 7 days of 15% PEG stress, the POD and SOD activities in KO2 leaves decreased by 8.8% and 5.5% respectively compared with WT, and the POD and SOD activities in roots decreased by 8.6% and 7.8% respectively compared with WT (p<0.05); After 7 days of 20% PEG stress, the POD and SOD in leaves decreased significantly by 7.3% and 9.7% respectively compared with WT, and the POD and SOD in roots decreased significantly by 25.9% and 9.1% respectively compared with WT (p<0.05). Figure 7 The MDA content increased significantly. After 3 days of 15% PEG stress, the MDA content in leaves and roots increased significantly by 40.7% and 53.9% compared to the WT, respectively, and after 7 days, the increases were 30.6% and 53.3%, respectively. After 7 days of 20% PEG stress, the MDA content in leaves and roots increased by 33.1% and 33.7% compared to the WT, respectively. Figure 7 In the F and L cells, KO2 cells suffer severe cell membrane damage and have a weak antioxidant defense system.

[0070] In conclusion, TaFtsZ2-1Overexpression of this gene can reduce membrane lipid peroxidation damage and improve wheat drought resistance and rehydration recovery by promoting root development, maintaining chlorophyll stability, enhancing osmotic regulation, and activating the antioxidant system. Knockout of this gene, on the other hand, leads to weakened antioxidant system function, aggravated membrane damage, and increased sensitivity to drought stress.

[0071] 2.2.3 TaFtsZ2-1 Regulating wheat tolerance to cadmium stress To investigate wheat TaFtsZ2-1 Given its broad-spectrum resistance to stress, this study further investigated the function of this gene in the Cd stress response. Under normal conditions, the number of roots in OE2 was significantly increased by 18.8% compared to WT, while the dry weight of KO2 was significantly decreased by 25.0% and the root length was significantly decreased by 3.9% compared to WT. Figure 8 (A~C, G). Under Cd stress, the growth advantage of OE2 further increased with increasing concentration in terms of plant height and root length. Plant height, single plant dry weight, and root length were all significantly higher than WT: after 7 days of 20 µmol / L Cd stress, they increased significantly by 8.4%, 87.5%, and 11.3% respectively compared to WT; after 7 days of 30 µmol / L Cd stress, the increase in plant height and root length was more significant, increasing by 9.0% and 15.9% respectively, but the increase in dry weight decreased to 28.6% (…). Figure 8 (A~C, G). Conversely, the number of KO2 roots was significantly lower under Cd stress than under WT, decreasing by 11.8% and 6.0% after 7 days of stress at 20 µmol / L and 30 µmol / L, respectively. Figure 8 (A). Furthermore, the trends in chlorophyll content changes among different lines under Cd stress varied. After 7 days of stress, all OE2 concentrations were significantly higher than WT (96.3% and 22.7% higher at 20 µmol / L and 30 µmol / L, respectively), while KO2 was significantly lower than WT (14.9% and 23.3% lower, respectively). Figure 8 (H). Soluble sugars showed a continuous upward trend. After 7 days of stress at 20 µmol / L and 30 µmol / L, OE2 increased significantly by 9.3% and 8.1% compared to WT, respectively, while KO2 decreased significantly by 11.3% and 9.2% compared to WT, respectively. Figure 8 (I). Regarding antioxidant defense, POD and SOD activities showed a trend of first increasing and then decreasing. After 7 days of stress at 20 µmol / L and 30 µmol / L, the SOD and POD activities in OE2 leaves were significantly higher than those in WT (leaf SOD was 23.2% and 25.6% higher, respectively; leaf POD was 22.8% and 51.5% higher, respectively), while the increase in root SOD and POD activities was smaller. In KO2, root POD was significantly lower than in WT, decreasing by 17.4% at 20 µmol / L and by 8.9% at 30 µmol / L (p<0.05). Figure 8MDA content increased continuously with stress, with OE2 increasing at a significantly lower rate than WT, while KO2 increased at a significantly higher rate than WT. Under stress levels of 20 µmol / L and 30 µmol / L, root MDA was consistently significantly higher than WT (34.6% and 55.8% higher, respectively). Leaf MDA at 30 µmol / L had significantly higher KO2 than WT, while at 20 µmol / L, KO2 was lower than WT. Figure 8 The results (F, L) indicate that the KO2 membrane lipids suffer severe peroxidation damage and have significantly insufficient antioxidant compensation capacity.

[0072] In conclusion, TaFtsZ2-1 Overexpression significantly improves wheat's tolerance to Cd stress by enhancing root development, maintaining photosynthetic and osmotic regulation, and increasing antioxidant enzyme activity, thereby alleviating Cd-induced oxidative stress. Gene knockout, on the other hand, leads to weakened antioxidant system function, increased membrane damage, and greater sensitivity to Cd stress.

[0073] This invention shows that, TaFtsZ2-1 This gene can positively regulate wheat's tolerance to drought and cadmium stress, exhibiting significant stress-specific and organ-differentiated regulatory characteristics. Under drought stress, this gene effectively enhances wheat's drought resistance and post-stress rehydration recovery capacity by promoting root formation, strengthening osmotic regulation and antioxidant capacity, and alleviating membrane lipid peroxidation damage. Under cadmium stress, TaFtsZ2-1 It mainly improves the plant's tolerance to cadmium stress by maintaining the homeostasis and antioxidant balance of the aboveground photosynthetic system. Root number, soluble sugar content, and plant height are highly sensitive to differences in stress resistance among different genotypes and can be used as indicators for differentiation. TaFtsZ2-1 The core evaluation indicators that mediate wheat stress resistance differentiation.

[0074] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. TaFtsZ2-1 The use of its or its biomaterials in at least one of the following, characterized in that, (1) Improve wheat's resistance to adverse conditions; (2) Breeding stress-resistant transgenic wheat; (3) Increase the number of wheat chloroplasts; (4) Improve wheat photosynthesis; (5) Promotes wheat growth traits; (6) Improve the antioxidant capacity of wheat.

2. The application according to claim 1, characterized in that, The stress resistance includes resistance to drought stress and / or resistance to heavy metal stress.

3. The application according to claim 1, characterized in that, The growth traits include one or more of the following: dry weight per plant, plant height, root length, and number of roots.

4. The application according to claim 1, characterized in that, The biomaterial includes those containing TaFtsZ2-1 Expression cassettes, recombinant vectors, recombinant bacteria, plant cells, plant tissues, or plant organs.

5. The application according to claim 2, characterized in that, The resistance to heavy metal stress includes resistance to Cd stress.

6. A method for improving wheat traits, characterized in that, Including improving wheat TaFtsZ2-1 The steps to obtain transgenic wheat are: expression level and / or activity; The property includes at least one of the following S1 to S6: (S1) Improves wheat's resistance to stress; (S2) Breeding stress-resistant transgenic wheat; (S3) Increase the number of wheat chloroplasts; (S4) Promotes wheat photosynthesis; (S5) Promotes wheat growth traits; (S6) Improves the antioxidant capacity of wheat.

7. The method according to claim 6, characterized in that, The stress resistance includes resistance to drought stress and / or resistance to heavy metal stress; the growth traits include one or more of the following: single plant dry weight, plant height, root length, and number of roots.

8. The method according to claim 6, characterized in that, The improvement of wheat TaFtsZ2-1 Methods for determining the expression level and / or activity of [the substance] include introducing the biological material of claim 4 into wheat, thereby increasing the expression level and / or activity of [the substance] in wheat. TaFtsZ2-1 Overexpression.

9. A method for breeding transgenic wheat having at least one of the traits shown in A1 to A6, characterized in that, Includes the following steps: Improve wheat TaFtsZ2-1 The steps to obtain transgenic wheat are: expression level and / or activity; (A1) Improves wheat's resistance to stress; (A2) Breeding stress-resistant transgenic wheat; (A3) Increase the number of chloroplasts in wheat; (A4) Promotes wheat photosynthesis; (A5) Promotes wheat growth traits; (A6) Improves the antioxidant capacity of wheat.

10. The method according to claim 9, characterized in that, The stress resistance includes resistance to drought stress and / or resistance to heavy metal stress; the growth traits include one or more of the following: single plant dry weight, plant height, root length, and number of roots.

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