A growth-promoting Chlorella vulgaris E1 strain and its applications

Abstract

This invention discloses a strain of Chlorella vulgaris E1, which can increase soil organic matter, available nitrogen, available phosphorus, and available potassium, as well as total nitrogen and total phosphorus, and improve soil microbial biomass carbon, microbial biomass nitrogen, and microbial biomass phosphorus, thereby promoting the availability of soil nutrients. Simultaneously, this strain can produce metabolites that can be used as plant growth regulators, as well as metabolites that promote plant growth or enhance plant stress resistance, thereby promoting plant growth and improving plant salt tolerance. Furthermore, this strain can produce metabolites that can be used as insecticides, thus having the potential to enhance plant insect resistance.

Description

Technical Field

[0001] This invention belongs to the field of microbial technology, specifically a growth-promoting Chlorella vulgaris E1 strain and its applications. Background Technology

[0002] Soil salinity stress is one of the important abiotic factors restricting plant growth and development, and also one of the serious problems limiting agricultural development. Saline-alkali land is the main soil type of low- and medium-yield fields in my country, and generally suffers from poor soil physical and chemical properties, such as ① low organic matter content ② poor nutrient conditions ③ low number and low activity of microorganisms (respiration, soil organic nitrogen mineralization activity, nitrification activity, etc.) ④ degradation of physical properties (destruction of aggregate structure; reduced porosity; inhibited water and air transport, weakened permeability), which in turn leads to poor crop growth, seedling loss and reduced yield.

[0003] Microalgae are a type of autotrophic plant that reproduces rapidly and is highly adaptable to different environments, playing a vital role in soil and plant health. They improve soil structure, enhance soil fertility, balance the soil micro-ecosystem, promote crop growth, increase crop yield and quality, and improve crop adaptability to environmental stresses through various pathways, including releasing plant hormones, vitamins, polypeptides, organic acids, various volatiles, and osmotic regulators; regulating plant antioxidant enzyme activity; improving soil nutrient availability; and promoting soil microbial activity.

[0004] Currently, the discovered Chlorella microalgae species have wide applications in aquaculture, food, medicine, and environmental protection, but their application in agriculture is relatively lagging and scarce. Specifically, for the Chlorella vulgaris species, no research has yet indicated that it can increase soil microbial biomass carbon, microbial biomass nitrogen, and microbial biomass phosphorus, thereby promoting the availability of soil nutrients. Meanwhile, no studies have yet indicated that it can produce metabolites that can be used as plant growth regulators, including diethylene glycol, isophorone, erucamide, hexadecylamide, allantoic acid, L-serine, ethephon, glutamine-arginine, and lauramide; or produce metabolites that promote plant growth or enhance plant stress resistance, including linoleic acid, methionine, N-acetyl-L-glutamic acid, palmitic acid, L-glutamic acid, valeramide, betaine, phenylalanine-tyrosine, phenylalanine-glutamic acid, L-threonine, leucine, proline, and L-isoleucine-L-threonine; thereby promoting plant growth, enhancing plant salt tolerance, and reducing the degree of lipid peroxidation in wheat stem and leaf cell membranes under high salt stress conditions of 8.72 g / kg (NaCl / dry soil), thus alleviating the damage of salt stress to plants. Summary of the Invention

[0005] The purpose of this invention is to overcome the shortcomings of existing technologies and provide a strain of Chlorella vulgaris E1 that can increase soil organic matter, available nitrogen, available phosphorus, and available potassium, as well as total nitrogen and total phosphorus, and improve soil microbial biomass carbon, microbial biomass nitrogen, and microbial biomass phosphorus, thereby promoting the availability of soil nutrients. Simultaneously, this strain can produce metabolites that can be used as plant growth regulators, as well as metabolites that promote plant growth or enhance plant stress resistance, thereby promoting plant growth and improving plant salt tolerance. Furthermore, this strain can produce metabolites that can be used as insecticides, thus having the potential to improve plant insect resistance.

[0006] This invention is achieved through the following technical solution:

[0007] This invention discloses Chlorella vulgaris E1 isolated from saline-alkali soil in Haixing County, Cangzhou City, Hebei Province, China. This Chlorella has been deposited at the China Center for Type Culture Collection (CCTCC), located at Luojia Mountain, Bayi Road, Wuchang District, Wuhan City, Hubei Province, on December 20, 2024, with accession number CCTCCNO: M20242875.

[0008] The aforementioned Chlorella vulgaris E1 colony is dark green and protruding with smooth edges.

[0009] The growth-promoting Chlorella vulgaris E1 described herein has the tufA gene sequence shown in SEQ ID NO.1 and belongs to Chlorella vulgaris.

[0010] The growth-promoting Chlorella vulgaris E1 described herein has an 18S rDNA sequence as shown in SEQ ID NO.2 and belongs to Chlorella vulgaris.

[0011] The aforementioned Chlorella vulgaris E1 can regulate soil nutrients and promote their availability. This is mainly manifested in an increase of 2.93%–7.41% in soil organic matter, 1.34%–21.49% in available nitrogen, 24.29%–31.97% in available phosphorus, 11.22%–12.44% in available potassium, 1.50%–2.80% in total nitrogen, and 0.45%–9.43% in total phosphorus.

[0012] The aforementioned growth-promoting Chlorella vulgaris E1 can promote plant growth and improve plant salt tolerance under salt stress conditions of 8.72 g / kg (NaCl / dry soil). Taking wheat as an example, the main effects are an increase in aboveground fresh weight of 2.70%–43.24%, an increase in plant height of 5.52%–5.88%, a decrease in malondialdehyde (MDA) content in stems of 29.95%–31.13%, and a decrease in MDA content in leaves of 7.33%–13.25%.

[0013] The aforementioned growth-promoting Chlorella vulgaris E1 can produce metabolites that can be used as plant growth regulators, including diethylene glycol, isophorone, erucamide, hexadecanoamide, allantoic acid, L-serine, ethephon, glutamine-arginine, and laurylamide.

[0014] The aforementioned growth-promoting Chlorella vulgaris E1 can produce metabolites that promote plant growth or enhance plant stress resistance, including linoleic acid, methionine, N-acetyl-L-glutamic acid, palmitic acid, L-glutamic acid, valeramide, betaine, phenylalanine-tyrosine, phenylalanine-glutamic acid, L-threonine, leucine, proline, and L-isoleucine-L-threonine.

[0015] The aforementioned growth-promoting Chlorella vulgaris E1 can produce metabolites that can be used as insecticides, including capsaicin.

[0016] In another aspect of the present invention, the present invention provides a Chlorella preparation comprising the growth-promoting Chlorella vulgaris E1 described in the above technical solution.

[0017] In another aspect of the invention, a microbial fertilizer is provided, comprising at least one of the following:

[0018] The above technical solution describes the growth-promoting Chlorella vulgaris E1, its culture, or its processed products;

[0019] The Chlorella preparation described in the above technical solution.

[0020] In another aspect of the invention, a soil conditioner is provided, comprising at least one of the following:

[0021] The above technical solution describes the growth-promoting Chlorella vulgaris E1, its culture, or its processed products;

[0022] The Chlorella preparation described in the above technical solution.

[0023] In another aspect, the present invention provides a plant growth regulator comprising at least one of the following:

[0024] The above technical solution describes the growth-promoting Chlorella vulgaris E1, its culture, or its processed products;

[0025] The Chlorella preparation described in the above technical solution.

[0026] In another aspect of the invention, a biostimulant is provided, comprising at least one of the following:

[0027] The above technical solution describes the growth-promoting Chlorella vulgaris E1, its culture, or its processed products;

[0028] The Chlorella preparation described in the above technical solution.

[0029] In another aspect of the present invention, a biological pesticide is proposed, comprising a carrier and an active ingredient; the active ingredient comprises: the growth-promoting Chlorella vulgaris E1 described in the above technical solution, its culture or its processed form.

[0030] In another aspect of the present invention, the present invention proposes the use of the Chlorella vulgaris E1 growth-promoting agent, the Chlorella preparation, or the microbial fertilizer described in the above-mentioned technical solutions in at least one of the following:

[0031] It enhances crop resistance to adverse conditions, especially salt tolerance;

[0032] Promote crop growth;

[0033] Soil improvement.

[0034] In another aspect of the invention, the invention proposes a method for improving crop stress resistance, particularly salt tolerance; promoting crop growth; and improving soil. According to an embodiment of the invention, the method includes adding at least one of the following to the soil where the crop is grown: the Chlorella vulgaris E1 growth-promoting agent described in the above-mentioned technical solutions, a Chlorella preparation, or a microbial fertilizer.

[0035] In another aspect of the invention, a method for preparing a biopesticide is provided, wherein the biopesticide is prepared by using the growth-promoting Chlorella vulgaris E1, its culture or its processed products.

[0036] The advantages and beneficial effects of this invention are as follows:

[0037] The *Chlorella vulgaris* E1 described in this invention can increase soil organic matter, available nitrogen, available phosphorus, and available potassium, as well as total nitrogen and total phosphorus, and improve soil microbial biomass carbon, microbial biomass nitrogen, and microbial biomass phosphorus, thus promoting the availability of soil nutrients. Therefore, it can be used as a soil conditioner, suitable for medium- and low-yielding saline-alkali soils with various problems such as ① low organic matter content, ② poor nutrient conditions, and ③ low number and activity of microorganisms (respiration, soil organic nitrogen mineralization activity, nitrification activity, etc.), which is beneficial to improving the soil's ability to provide nutrients to plants.

[0038] The *Chlorella vulgaris* E1 described in this invention exhibits excellent properties in promoting plant growth and enhancing salt tolerance under salt stress conditions of 8.72 g / kg (NaCl / dry soil). This is primarily manifested in increased aboveground fresh weight and plant height of wheat, as well as decreased malondialdehyde (MDA) content in stems and leaves. Therefore, it is suitable for preparing microbial inoculants and biofertilizers, providing resource and technical support for the use of chemical fertilizers as a substitute or partial substitute, and for the development of crop salt-tolerant and yield-increasing technologies based on microalgae.

[0039] Furthermore, the *Chlorella vulgaris* E1 described in this invention can produce metabolites that can be used as plant growth regulators, including diethylene glycol, isophorone, erucamide, hexadecanoamide, allantoic acid, L-serine, ethephon, glutamine-arginine, and laurylamide; it can also produce metabolites that promote plant growth or enhance plant stress resistance, including linoleic acid, methionine, N-acetyl-L-glutamic acid, palmitic acid, L-glutamic acid, valeramide, betaine, phenylalanine-tyrosine, phenylalanine-glutamic acid, L-threonine, leucine, proline, and L-isoleucine-L-threonine; thereby promoting plant growth and, under salt stress conditions, enhancing plant salt tolerance, reducing the degree of lipid peroxidation in wheat stem and leaf cell membranes under salt stress conditions, and alleviating the damage caused by salt stress to plants. In addition, this algal strain can produce capsaicin, a metabolite that can be used as an insecticide, thus having the potential to enhance plant insect resistance. Therefore, it provides resource support for the preparation of plant growth regulators and biostimulants.

[0040] Preservation Information

[0041] Chlorella vulgaris E1 strain has been deposited at the China Center for Type Culture Collection (CCTCC), Wuhan University, Wuhan, China, on December 20, 2024, with accession number CCTCCNO: M 20242875. Attached Figure Description

[0042] Figure 1These are colony observation images of Chlorella vulgaris E1 from this invention, (a) front view, (b) back view;

[0043] Figure 2 This is a phylogenetic tree of the tufA gene sequence of Chlorella vulgaris E1, which is the subject of this invention;

[0044] Figure 3 This is an observational image showing how algal cells of Chlorella vulgaris E1 promote wheat growth under salt stress conditions, according to the present invention.

[0045] Figure 4 This is an observational diagram showing the effect of the supernatant of the culture of Chlorella vulgaris E1 on promoting wheat growth under salt stress.

[0046] Figure 5 This invention relates to the effects of algal cells and culture supernatant of Chlorella vulgaris E1 on wheat plant height and aboveground fresh weight under salt stress conditions;

[0047] Figure 6 This invention relates to the effects of algal cells and culture supernatants of Chlorella vulgaris E1 on the malondialdehyde (MDA) content in wheat stems and leaves under salt stress conditions.

[0048] Figure 7 This invention relates to the effects of algal cells and culture supernatant of Chlorella vulgaris E1 on soil organic matter content;

[0049] Figure 8 This invention relates to the effects of algal cells and culture supernatant of Chlorella vulgaris E1 on soil nutrient (alkaline nitrogen, available phosphorus, and available potassium) content;

[0050] Figure 9 This invention relates to the effects of algal cells and culture supernatant of Chlorella vulgaris E1 on soil nutrient (total nitrogen, total phosphorus, and total potassium) content;

[0051] Figure 10 This invention relates to the effects of algal cells and culture supernatant of Chlorella vulgaris E1 on soil microbial biomass carbon, nitrogen, and phosphorus.

[0052] For those skilled in the art, other related figures can be obtained from the above figures without any creative effort. Detailed Implementation

[0053] To enable those skilled in the art to better understand the present invention, the technical solution of the present invention will be further described below with reference to specific embodiments.

[0054] In this article, the terms “Chlorella vulgaris E1”, “Chlorella vulgaris E1”, “Chlorella vulgaris E1” and “E1” are interchangeable.

[0055] Example 1: Isolation, purification and culture of Chlorella vulgaris E1

[0056] Sample source: Saline-alkali soil from Haixing County, Cangzhou City, Hebei Province, China.

[0057] Preparation of A&A medium: Refer to the patent "A Chlamydomonas aeruginosa strain m9v and its application", China, patent number: ZL201811554750.8; authorization date: 2022.4.19.

[0058] Preparation of BG11 medium: Prepare according to the usage requirements of BG11 medium (Haibo Biotechnology, catalog number HB8793), autoclave, and then cool to room temperature.

[0059] Separation and purification steps: Refer to the patent "A Chlamydomonas aeruginosa strain m9v and its application", China, patent number: ZL201811554750.8; authorization date: 2022.4.19.

[0060] Example 2: Identification of Chlorella vulgaris E1

[0061] 1) Identification of biological characteristics

[0062] Chlorella vulgaris E1 colonies on BG11 medium appear as dark green raised bumps with smooth edges. Figure 1 ).

[0063] 2) System classification and identification

[0064] Take an appropriate amount of microalgae E1 culture medium into a sterile 2mL centrifuge tube, centrifuge repeatedly, and discard the supernatant until a certain amount of microalgae E1 cells are obtained. Transfer the microalgae E1 cells to FastDNA. ○RGenomic DNA of microalgae E1 was extracted into LysingMatrix E tubes using the SPIN kit (MP Biomedicals, USA) following the prescribed procedure. The tufA gene sequence was amplified using primers tufAF (5'-GGNGCNGCNCAAATGGAYGG-3') and tufAR (5'-CCTTCNCGAATMGCRAAWCGC-3') (Ao Yuan, 2020). The 18S gene sequence was amplified using primers 18F (5'-TGGTTGATCCTGCCAGT-3') and 18R (5'-TGATCCTTCTGCAGGTTCACC-3') (Medlin et al., 1988; Song et al., 2016). The PCR reaction system was 50 μL, containing 10 μL EasyTaq buffer, 5 μL dNTPs, 0.5 μL primer 18F, 0.5 μL primer 18R, 1.5 μL DNA template, and 2 μL EasyTaq DNA polymerase. Sterile ultrapure water was added to bring the final volume to 50 μL. The PCR reaction program for the tufA gene was: (i) 95℃ for 3 min pre-denaturation; (ii) 35 cycles of 94℃ for 30 s denaturation, 56℃ for 30 s annealing, and 72℃ for 1 min extension; (iii) final extension at 72℃ for 7 min. The PCR reaction program for the 18S gene was: (i) 94℃ for 5 min pre-denaturation; (ii) 32 cycles of 94℃ for 50 s denaturation, 55℃ for 50 s annealing, and 72℃ for 90 s extension; (iii) final extension at 72℃ for 10 min. The PCR amplification products were submitted to Sangon Biotech (Shanghai) Co., Ltd. for sequencing. The tufA gene sequence of microalga E1 obtained by sequencing is shown in SEQ ID NO.1 of the sequence listing, and the 18S gene sequence of microalga E1 obtained by sequencing is shown in SEQ ID NO.2 of the sequence listing.

[0065] The sequenced tufA gene sequence was homology-aligned using the NCBI (National Center for Biotechnology Information) database website (http: / / www.ncbi.nlm.nih.gov / ), and a phylogenetic tree was constructed. Figure 2The results showed that the tufA gene of microalga E1 shared over 99% similarity with nine tufA gene sequences from Chlorella vulgaris clone ITBBA3-12, Chlorella vulgaris strains UTEX259, NJ-7, and IRK-A 46, and Chlorella vulgaris isolates 962-2 and 962-1, and formed a stable evolutionary clade (bootstrap value 100) with the tufA gene sequence of Chlorella vulgaris strain NJ-7. Furthermore, homology comparison of the sequenced 18S gene sequences showed that the 18S gene of microalga E1 shared over 99% similarity with 57 18S gene sequences from Chlorella vulgaris strains CCAP211 / 35, KNUA027, and ACSSI 361. Therefore, the microalga E1 obtained in this invention is identified as *Chlorella vulgaris*, belonging to the genus *Chlorella* of the phylum Chlorophyta, and is named *Chlorella vulgaris* E1.

[0066] Example 3: Effects of Chlorella vulgaris E1 on wheat growth and salt tolerance under 8.72 g / kg (NaCl / dry soil) salt stress.

[0067] Wheat pot experiment: A negative control (represented by Control) and a positive control (represented by BG11) were set up (the positive control was mainly used to compare with the culture supernatant to exclude the influence of the culture medium and to demonstrate the role of E1 metabolites). The experimental groups were treated with Chlorella vulgaris E1 algal cells (represented by E1 (algal cells)) and Chlorella vulgaris E1 culture supernatant (represented by E1 (supernatant)). A total of four treatments were conducted, with each treatment repeated three times. Figure 3 and Figure 4Saline-alkali farmland soil, sieved through a 2mm sieve and thoroughly mixed, with a salt content of 0.9 g / kg (NaCl / dry soil), was used. The soil was divided into 330g portions per pot, and "Malan No. 1" wheat was planted. Each pot contained 16 seeds (recorded as day 1). After uniform emergence, thinning began, leaving 6 wheat seedlings per pot. The cultivation conditions were 25℃, 16h light / 8h dark. Initially, sterile water was used for irrigation. Once the wheat seedlings were robust, a 4g / L NaCl solution was used for salt stress treatment. Negative and positive controls were provided using sterile water and BG11 medium, respectively. Irrigation was carried out every 1–3 days until the soil NaCl content reached approximately 8.72 g / kg (NaCl / dry soil), at which point irrigation with the NaCl solution was discontinued. Meanwhile, 20 mL of Chlorella vulgaris E1 algal cells and Chlorella vulgaris E1 supernatant were poured into the soil, and sterile water and BG11 medium were poured into the soil as negative and positive control treatments, respectively, for a total of 5 pours. The experimental results showed that: (1) Under severe salt stress (8.72 g / kg NaCl / dry soil), both E1 (algal cells) and E1 (supernatant) could promote wheat growth, alleviate the degree of lipid peroxidation of stem and leaf cell membranes of wheat under salt stress, and improve the salt resistance of wheat. Figure 3 , Figure 4 and Figure 5 (2) The aboveground fresh weight of wheat treated with E1 (algal cells) increased by 2.70% compared with the Control treatment. Figure 3 (3) The aboveground fresh weight of the E1 (supernatant) treatment increased by 43.24% compared with the Control treatment and by 20.45% compared with the BG11 treatment; the plant height of the E1 (supernatant) treatment increased by 5.52% compared with the Control treatment and by 5.88% compared with the BG11 treatment. Figure 4 ).

[0068] In summary, the Chlorella vulgaris E1 algal cells and supernatant described in this invention can promote wheat growth under severe salt stress (8.72 g / kg NaCl / dry soil). These promoting effects are mainly manifested in an increase in aboveground fresh weight and / or plant height, and a decrease in malondialdehyde (MDA) content in stems and / or leaves. Figure 6 This reduces the degree of lipid peroxidation in the stem and / or leaf cell membranes of wheat under salt stress, thus alleviating the damage caused by salt stress to wheat.

[0069] Example 4: Effects of Chlorella vulgaris E1 on soil nutrients under salt stress of 8.72 g / kg (NaCl / dry soil)

[0070] Soil samples from the wheat pot experiment in Example 3 were collected, and 10 indicators, including soil organic matter, available nitrogen, available phosphorus, available potassium, total nitrogen, total phosphorus, total potassium, microbial biomass carbon, microbial biomass nitrogen, and microbial biomass phosphorus, were measured. The results showed that: (1) Under severe salt stress (8.72 g / kg NaCl / dry soil), both E1 (algal cells) and E1 (supernatant) increased soil organic matter, available nitrogen, available phosphorus, available potassium, total nitrogen, and total phosphorus, and improved soil microbial biomass carbon, microbial biomass nitrogen, and microbial biomass phosphorus, thus improving the soil's effectiveness in providing nutrients for plant growth. Figure 7 , Figure 8 , Figure 9 and Figure 10 (2) Compared with the Control treatment, the soil organic matter, available nitrogen, available phosphorus, available potassium, total nitrogen, and total phosphorus increased by 7.41%, 15.80%, 24.29%, 25.74%, 2.80%, and 9.43% respectively in the E1 (algal cell) treatment. Figure 7 , Figure 8 , Figure 9 and Figure 10 In the E1 (supernatant) treatment, soil organic matter, available nitrogen, available phosphorus, available potassium, total nitrogen, and total phosphorus increased by 2.93%, 38.81%, 31.56%, 12.44%, 1.50%, and 12.24%, respectively. Figure 7 , Figure 8 , Figure 9 and Figure 10 (3) Compared with the BG11 treatment, the soil available nitrogen, available potassium, and total potassium in the E1 (algal cell) treatment increased by 1.34%, 11.22%, and 9.32%, respectively. Figure 7 , Figure 8 , Figure 9 and Figure 10 In the E1 (supernatant) treatment, the available nitrogen, available phosphorus, available potassium, total phosphorus, and total potassium in the soil increased by 21.49%, 31.97%, 31.56%, 12.41%, 0.45%, and 8.71%, respectively. Figure 7 , Figure 8 , Figure 9 and Figure 10 ).

[0071] Example 5: Determination of E1 metabolites from Chlorella vulgaris

[0072] Single colonies of Chlorella vulgaris E1 were picked from solid BG11 medium and inoculated into 50 mL of liquid BG11 medium. The culture was incubated for 14 days at 25°C, 160 rpm, with a 12-hour light / 12-hour dark cycle. An appropriate amount of Chlorella vulgaris E1 algal culture was inoculated into 50 mL of liquid BG11 medium, with six replicates. The cultures were incubated at 25°C, 160 rpm, with a 12-hour light / 12-hour dark cycle for a certain period. The supernatant (denoted as E1) was collected by centrifugation at 8000 rpm for 5 min. Simultaneously, 50 mL of uninoculated Chlorella vulgaris E1 liquid BG11 medium was used as a control (denoted as Control), with six replicates. The supernatants and control samples were stored at -80°C and submitted to Shanghai Paisennuo Biotechnology Co., Ltd. for metabolomics analysis. The results showed that the *Chlorella vulgaris* E1 strain described in this invention can produce metabolites that can be used as plant growth regulators, including diethylene glycol, isophorone, erucamide, hexadecanoamide, allantoic acid, L-serine, ethephon, glutamine-arginine, and lauramide; it can also produce metabolites that promote plant growth or enhance plant stress resistance, including linoleic acid, methionine, N-acetyl-L-glutamic acid, palmitic acid, L-glutamic acid, valeramide, betaine, phenylalanine-tyrosine, phenylalanine-glutamic acid, L-threonine, leucine, proline, and L-isoleucine-L-threonine; thus, it promotes plant growth and, under salt stress conditions, enhances the plant's salt tolerance, reduces the degree of lipid peroxidation in the stem and leaf cell membranes of wheat under salt stress conditions, and alleviates the damage caused by salt stress to plants. Furthermore, this algal strain can produce capsaicin, a metabolite that can be used as an insecticide, thus having the potential to enhance plant insect resistance.

[0073] SEQ ID NO:1

[0074] >E1_tufA

[0075] >E1_816bp

[0076] TTGAATAAGTTCTACAATCATTTTAATACGATCTCCAGGCATTACCATTTGAGTTGCAGTG

[0077] TTATCATCCGCACGGAAAGATTCAATTTTTCCTGTAACATCTGTTGTGCGAACATAAAAC

[0078] TGTGGACGATATCCAGGGAAAAAAGGAGTATGACGCGCCACCTTCTTCTTTATTTAACACG

[0079] TAAACTTGTGCTTCAAATTTGGTGTGTGGTAAAATACTTCCTGGTTTTGCTAGAACCATA

[0080] CCACGTTCAATATCAATTTTCTGAACCCCACGTAATAAAATACCTACATTGTCTCCAGCTA

[0081] CACTTTCGTCTAACGTTTTTTGGAACATTTCTAAACCTGTAACAGTAGTTGTTTTAGTATC

[0082] ACGTAGTCCAACAAGTTCCACAGTATCGCCAATTTTTACACATCCACGTTCTACACGCCC

[0083] TGTAGCTACAGTACCACGACCAGTGATTGAAAAAACGTCTTCAACAGCCATTAAATG

[0084] GTTTCTCAGTTTCACGTTCGGGTGTTGGGATATATGAATCAACTTGATCCATCAGATTATA

[0085] AATTTTATCTACCCATTTGTTATCACCAGGTTTTGTTTCTGGGTTTTCAGTTAATGCTTCTA

[0086] AAGCTAAAAGCGCTGAACCAGAAATAATTGGAATTTCATCACCAGGGAATTCATATTTAT

[0087] CTAATGTTTCACGAATTTCTAGTTCTACAAGTTCAAGTAATTCTGCATCATCAACTTGATC

[0088] TTCTTTATTTAAAAATACCACGATATTTGGAACACCAACTTGTTTTGCTAATAAGAGATGT

[0089] TCTTTTGTTTGTGGCATAGTCCCATCCGC

[0090] SEQ ID NO:2

[0091] >E1_18S

[0092] >E1_1703 bp

[0093] CTCTTCCTCTAGGTGGGAGGGTTTAATGAACTTCTCGGCGGCCGAGAGCGGAGACCGCC

[0094] CCAGGTCGCCAATCCGAACACTTCACCAGCACACCCAATCGGTAGGAGCGACGGGCGG

[0095] TGTGTACAAAGGGCAGGGACGTAATCAACGCAAGCTGATGACTTGCGCTTACTAGGCAT

[0096] TCCTCGTTGAAGATTAATAATTGCAATAATCTATCCCCATCACGATGCAGTTTCGAAGATT

[0097] ACCCGGGCCTCTCGGCCAAGGCTAGGCTCGTTGAATGCATCAGTGTAGCGCGCGTGCGG

[0098] CCCAGAACATCTAAGGGCATCACAGACCTGTTATTGCCTCATGCTTCCATTGGCTAGTCG

[0099] CCAATAGTCCCTCTAAGAAGTCCGCCGGCTGGCGAACCAACCGTGACTATTTAGCAGGC

[0100] TGAGGTCTCGTTCGTTACCGGAATCAACCTGACAAGGCAACCCACCAACTAAGAACGG

[0101] CCATGCACCACCACCCATAGAATCAAGAAAGAGCTCTCAATCTGTCAATCCTCACTATGT

[0102] CTGGACCTGGTAAGTTTTCCCGTGTTGAGTCAAATTAAGCCGCAGGCTCCACGCCTGGT

[0103] GGTGCCCTTCCGTCAATTCCTTTAAGTTTCAGCCTTGCGACCATACTCCCCCCGGAACCC

[0104] AAAAACTTTGATTTCTCATAAGGTGCCGGCGGAGTCATCGAAGAAACATCCGCCGATCC

[0105] CTAGTCGGCATCGTTTATGGTTGAGACTAGGACGGTATCTAATCGTCTTCGAGCCCCCAA

[0106] CTTTCGTTCKKGATTAATGAAAACATCCTTGGCAAATGCTTTCGCAGTAGTTCGTCTTTC

[0107] ATAAATCCAAGAATTTCACCTCTGACAATGAAATACGAATGCCCCCGACTGTCCCTCTTA

[0108] ATCATTACTCCGGTCCTACAGACCAACAGGATAGGCCAGAGTCCTATCGTGTTATTCCAT

[0109] GCTAATGTATTCAGAGCGTAGGCCTGCTTTGAACACTCTAATTTACTCAAAGTAACAGCG

[0110] CCGACTCCGAGTCCCGGACAGTGAAGCCCAGGAGCCCGTCCCCGGCAACAAGGTGAGC

[0111] CCTGCCAGTGCACACCGAAACGGCGGACCGGCAGGTCCCACCCGAAATCCAACTACGA

[0112] GCTTTTTAACTGCAGCAACTTAAATATACGCTATTGGAGCTGGAATTACCGCGGCTGCTG

[0113] GCACCAGACTTGCCCTCCAATTGATCCTCGTTAAGGGGTTTAGATTGTACTCATTCCAATT

[0114] ACCAGACCTGAAAAGGCCCAGTATTGTTATTTATTGTCACTACCTCCCTGTGTCAGGATT

[0115] GGGTAATTTGCGCGCCTGCTGCCTTCCTTGGATGTGGTAGCCGTTTCTCAGGCTCCCTCT

[0116] CCGGAATCGAACCCTAATCCTCCGTCACCCGTTACCACCATGGTAGGCCTCTATCCTACC

[0117] ATCAAAAGTTGATAGGGCAGAAATTTGAATGAAACATCGCCGGCACAAGGCCATGCGAT

[0118] TCGTGAAGTTATCATGATTCACCGCGAGTCGGGCAGAAGCCCGGTCGGCCTTTTATCTAA

[0119] TAAATACGTCCCTTCCAGAAGTCGGGATTTACGCACGTATTAGCTCTAGATTTACTACGG

[0120] GTATCCGAGTAGTAAGTACCATCAAATAAACTATAACTGATTTAATGAGCCATTCGCAGTT

[0121] TCACAGTATAAAGCAGTTTTATACTTAGACATGCAT

[0122] The present invention has been described above by way of example. It should be noted that any simple modifications, alterations or other equivalent substitutions that can be made by those skilled in the art without creative effort without departing from the core of the present invention fall within the protection scope of the present invention.

Claims

1. A strain of Chlorella vulgaris E1, characterized in that, CCTCC NO: M 20242875 on December 20, 2024.

2. A Chlorella formulation, characterized in that, The Chlorella vulgaris E1 according to claim 1.

3. A microbial fertilizer, characterized by, comprising at least one of the following: The Chlorella vulgaris E1 according to claim 1 or its culture or the Chlorella preparation according to claim 2.

4. A soil conditioner, characterized in that, comprising at least one of the following: The Chlorella vulgaris E1 according to claim 1 or its culture or the Chlorella preparation according to claim 2.

5. A plant growth regulator for use in salt stress conditions in wheat, characterized in that, comprising at least one of the following: The Chlorella vulgaris E1 according to claim 1 or its culture or the Chlorella preparation according to claim 2.

6. The Chlorella vulgaris E1 according to claim 1, the Chlorella preparation according to claim 2 or the microbial fertilizer according to claim 3 for use in improving salt resistance of wheat, promoting growth of wheat under salt stress or improving saline-alkali soil.

7. Use according to claim 6, characterized in that, The Chlorella vulgaris E1 according to claim 1, the Chlorella preparation according to claim 2 or the microbial fertilizer according to claim 3 is added to the soil for planting wheat.

Images