Growth-promoting chlorella strain Chlorella vulgaris E1 and application thereof

Abstract

The invention discloses a strain of chlorella vulgaris E1, which can increase soil organic matters, alkali-hydrolyzable nitrogen, available phosphorus, quick-acting potassium, total nitrogen and total phosphorus, improve soil microbial biomass carbon, microbial biomass nitrogen and microbial biomass phosphorus, and further promote the effectiveness of soil nutrients. Meanwhile, the algal strain can generate a metabolite which can be used as a plant growth regulator and a metabolite which has the effect of promoting plant growth or improving plant stress resistance, so that the plant growth is promoted, and the salt resistance of the plant is improved. In addition, the algal strain can generate metabolites which can be used as insecticides, so that the algal strain has a potential effect of improving the insect resistance of plants.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms and specifically discloses a growth-promoting chlorella Chlorella vulgaris E1 and application thereof. Background Art

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

[0003] Microalgae are a type of autotrophic plants that reproduce quickly and have strong environmental adaptability. They play an important role in soil health and plant health. They improve soil structure, soil fertility, balance soil micro-ecological systems, promote crop growth, increase crop yield and quality, and improve crop adaptability to environmental stress by releasing plant hormones, vitamins, peptides, organic acids, various volatiles and osmotic regulating substances, regulating plant antioxidant enzyme activity, improving soil nutrient availability, and promoting soil microbial activity.

[0004] The Chlorella microalgae species discovered so far are widely used in aquaculture, food, medicine and environmental protection, but their application in agriculture is relatively lagging and scarce. Among them, no research has shown that Chlorella vulgaris can increase soil microbial biomass carbon, microbial biomass nitrogen and microbial biomass phosphorus, and promote the effectiveness of soil nutrients. At the same time, no research has pointed out that it can produce metabolites that can be used as plant growth regulators, including diethylene glycol, isophorone, erucamide, hexadecanoic acid, allantoic acid, L-serine, ethephon, glutamine-arginine and lauramide; and metabolites that have the effect of promoting plant growth or improving plant stress resistance, including linoleic acid, methionine, N-acetyl-L-glutamate, palmitic acid, L-glutamic acid, valeramide, betaine, phenylalanine-tyrosine, phenylalanine-glutamate, L-threonine, leucine, proline, L-isoleucine-L-threonine; thereby, promoting plant growth, improving plant salt resistance, and under high salt stress conditions of 8.72g / kg (NaCl / dry soil), reducing the degree of membrane lipid peroxidation of wheat stems and leaves, and alleviating the damage of salt stress to plants. Summary of the invention

[0005] The purpose of the present invention is to overcome the deficiencies of the prior art and provide a strain of Chlorella vulgarisE1, which can increase soil organic matter, alkaline nitrogen, available phosphorus and available potassium, as well as total nitrogen and total phosphorus, improve soil microbial biomass carbon, microbial biomass nitrogen and microbial biomass phosphorus, and thus promote the effectiveness of soil nutrients. At the same time, the algae strain can produce metabolites that can be used as plant growth regulators, as well as metabolites that have the effect of promoting plant growth or improving plant stress resistance, thereby promoting plant growth and improving plant salt resistance. In addition, the algae strain can produce metabolites that can be used as insecticides, thereby having the potential to improve plant insect resistance.

[0006] The present invention is achieved through the following technical solutions:

[0007] The present invention discloses Chlorella vulgaris E1 isolated from saline-alkali soil in Haixing County, Cangzhou City, Hebei Province, China. The Chlorella vulgaris E1 has been preserved in China Center for Type Culture Collection (CCTCC), with a preservation address of Luojia Mountain, Bayi Road, Wuchang District, Wuhan City, Hubei Province, a preservation date of December 20, 2024, and a preservation number of CCTCCNO: M20242875.

[0008] The colony of the growth-promoting chlorella Chlorella vulgaris E1 is dark green and protruding with smooth edges.

[0009] The growth-promoting chlorella Chlorella vulgaris E1 has a tufA gene sequence as shown in SEQ ID NO.1, and belongs to Chlorella vulgaris.

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

[0011] The growth-promoting Chlorella vulgaris E1 can regulate soil nutrients and promote the effectiveness of soil nutrients, which is mainly manifested in an increase of 2.93% to 7.41% in soil organic matter, 1.34% to 21.49% in alkaline nitrogen, 24.29% to 31.97% in available phosphorus, 11.22% to 12.44% in available potassium, 1.50% to 2.80% in total nitrogen, and 0.45% to 9.43% in total phosphorus.

[0012] The growth-promoting Chlorella vulgaris E1 can promote plant growth and improve plant salt resistance under 8.72g / kg (NaCl / dry soil) salt stress conditions. Taking wheat plants as an example, the main performance is that the aboveground fresh weight increases by 2.70% to 43.24%, the plant height increases by 5.52% to 5.88%, the malondialdehyde content in the stems decreases by 29.95% to 31.13%, and the malondialdehyde content in the leaves decreases by 7.33% to 13.25%.

[0013] The growth-promoting chlorella Chlorella vulgaris E1 can produce metabolites that can be used as plant growth regulators, including diethylene glycol, isophorone, erucic acid amide, hexadecanoic acid amide, allantoic acid, L-serine, ethephon, glutamine-arginine and lauramide.

[0014] The growth-promoting chlorella Chlorella vulgaris E1 can produce metabolites that promote plant growth or improve 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 growth-promoting chlorella Chlorella vulgaris E1 can produce metabolites that can be used as pesticides, 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 present invention, the present invention provides a microbial fertilizer comprising at least one of the following:

[0018] The growth-promoting chlorella Chlorella vulgaris E1, its culture or its processed product described in the above technical solution;

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

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

[0021] The growth-promoting chlorella Chlorella vulgaris E1, its culture or its processed product described in the above technical solution;

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

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

[0024] The growth-promoting chlorella Chlorella vulgaris E1, its culture or its processed product described in the above technical solution;

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

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

[0027] The growth-promoting chlorella Chlorella vulgaris E1, its culture or its processed product described in the above technical solution;

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

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

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

[0031] Improve crop stress resistance, especially salt resistance;

[0032] Promote crop growth;

[0033] Soil improvement.

[0034] In another aspect of the present invention, the present invention provides a method for improving crop stress resistance, in particular, salt resistance; promoting crop growth; and a method for soil improvement. According to an embodiment of the present invention, the method comprises: adding at least one of the following to the soil where crops are planted: the growth-promoting Chlorella vulgaris E1, the Chlorella preparation, or the microbial fertilizer described in the above technical solution.

[0035] In another aspect of the present invention, the present invention provides a method for preparing a biopesticide, wherein the biopesticide is prepared by using the growth-promoting chlorella Chlorella vulgaris E1, a culture thereof or a processed product thereof.

[0036] The advantages and beneficial effects of the present invention are:

[0037] The Chlorella vulgaris E1 of the present invention can increase soil organic matter, alkaline nitrogen, available phosphorus and available potassium, as well as total nitrogen and total phosphorus, improve soil microbial biomass carbon, microbial biomass nitrogen and microbial biomass phosphorus, and promote the effectiveness of soil nutrients. Therefore, it can be used as a soil conditioner, suitable for medium and low-yield saline-alkali soils with multiple problems such as 1. low organic matter content 2. poor nutritional conditions 3. small number of microorganisms and low activity (respiration, soil organic nitrogen mineralization activity, nitrification activity, etc.), and is conducive to improving the ability of the soil to provide nutrients for plants.

[0038] The Chlorella vulgaris E1 of the present invention has the excellent characteristics of promoting plant growth and improving plant salt resistance under 8.72g / kg (NaCl / dry soil) salt stress conditions, which is mainly manifested in the increase of wheat aboveground fresh weight and plant height, and the reduction of malondialdehyde (MDA) content in stems and leaves. Therefore, it is suitable for preparing microbial agents and biofertilizers, and provides resources and technical support for the use of replacing or partially replacing chemical fertilizers and developing crop salt resistance and yield increase technology based on microalgae.

[0039] In addition, the chlorella vulgaris E1 of the present invention can produce metabolites that can be used as plant growth regulators, including diethylene glycol, isophorone, erucamide, hexadecanoic acid, allantoic acid, L-serine, ethephon, glutamine-arginine and lauramide; can produce metabolites that promote plant growth or improve 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, L-isoleucine-L-threonine; and then, promote plant growth, and under salt stress conditions, improve plant salt resistance, reduce the degree of peroxidation of stem and leaf cell membrane lipids of wheat under salt stress conditions, and alleviate the damage of salt stress to plants. In addition, the algae strain can produce a metabolite capsaicin that can be used as an insecticide, and then, it has the potential to improve the insect resistance of plants. Therefore, it provides resource support for the preparation of plant growth regulators and biostimulants.

[0040] Preservation Information

[0041] Chlorella vulgaris E1, this strain has been deposited, the depository unit: China Center for Type Culture Collection, the deposit address: Wuhan University, Wuhan, China, the deposit date: December 20, 2024, the deposit number: CCTCCNO: M 20242875. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] Figure 1It is a colony observation diagram of Chlorella vulgaris E1 of the present invention, (a) front side, (b) back side;

[0043] Figure 2 is a phylogenetic tree of the tufA gene sequence of Chlorella vulgaris E1 of the present invention;

[0044] Figure 3 This is an observation diagram of the algae cells of Chlorella vulgaris E1 of the present invention promoting wheat growth under salt stress conditions;

[0045] Figure 4 This is an observation diagram showing that the culture supernatant of Chlorella vulgaris E1 of the present invention promotes wheat growth under salt stress conditions;

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

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

[0048] Figure 7 The effect of the algae cells and culture supernatant of Chlorella vulgaris E1 of the present invention on the content of soil organic matter;

[0049] Figure 8 The effect of the algae cells and culture supernatant of Chlorella vulgaris E1 of the present invention on the content of soil nutrients (alkaline nitrogen, available phosphorus and available potassium);

[0050] Fig. 9 The effect of the algae cells and culture supernatant of Chlorella vulgaris E1 of the present invention on the content of soil nutrients (total nitrogen, total phosphorus and total potassium);

[0051] Fig.10 The present 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 ordinary technicians in this field, other relevant drawings can be obtained based on the above drawings without any creative work. DETAILED DESCRIPTION

[0053] In order to enable those skilled in the art to better understand the technical solution of the present invention, the technical solution of the present invention is further described below in conjunction with specific embodiments.

[0054] As used herein, the terms “growth-promoting Chlorella vulgaris E1”, “Chlorella vulgaris E1”, “Chlorella vulgaris E1” and “E1” are interchangeable.

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

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

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

[0058] Preparation of BG11 medium: Prepare according to the requirements of BG11 medium (Haibo Biotechnology, catalog number HB8793), sterilize with high-pressure steam, and cool to room temperature.

[0059] Separation and purification steps: refer to the patent "A growth-promoting Chlamydomonas 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] The colonies of Chlorella vulgaris E1 on BG11 medium are dark green and protruding, and the edges of the colonies are smooth ( Figure 1 ).

[0063] 2) Systematic classification and identification

[0064] Take an appropriate amount of microalgae E1 culture medium into a sterile 2mL centrifuge tube, centrifuge repeatedly, and remove the supernatant until a certain amount of microalgae E1 cells are obtained. Transfer microalgae E1 cells to FastDNA ○RThe microalgae E1 genomic DNA was extracted in the LysingMatrix E tube provided by the SPIN kit (MP Biomedicals, USA) according to its operation steps. 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, which contained 10 μL EasyTaq buffer, 5 μL dNTPs, 0.5 μL primer 18F, 0.5 μL primer 18R, 1.5 μL DNA template, 2 μL Easy Taq DNA polymerase, and finally sterile ultrapure water was added to 50 μL. The PCR reaction program of the tufA gene was: (i) 95℃3min pre-denaturation; (ii) 35 cycles of 94℃30s denaturation, 56℃30s annealing, 72℃1min extension; (iii) the final extension condition was 72℃7min. The PCR reaction program of the 18S gene was: (i) 94℃5min pre-denaturation; (ii) 32 cycles of 94℃50s denaturation, 55℃50s annealing, 72℃90s extension; (iii) the final extension condition was 72℃10min. The PCR amplification product was submitted to Sangon Biotech (Shanghai) Co., Ltd. for sequencing. The tufA gene sequence of microalgae E1 obtained by sequencing is as shown in the sequence list SEQ ID NO.1, and the 18S gene sequence of microalgae E1 obtained by sequencing is as shown in the sequence list SEQ ID NO.2.

[0065] The tufA gene sequences obtained by sequencing were compared for homology using the NCBI (National Center for Biotechnology Information) database website (http: / / www.ncbi.nlm.nih.gov / ), and a phylogenetic tree was constructed ( Figure 2), the results showed that the tufA gene of microalgae E1 had a similarity of more than 99% with 9 tufA gene sequences from Chlorella vulgaris clone ITBBA3-12, Chlorella vulgaris strains UTEX259, NJ-7 and IRK-A 46, and Chlorella vulgarisisolate 962-2 and 962-1, and formed a stable evolutionary branch with the tufA gene sequence of Chlorellavulgaris strain NJ-7 (bootstrap value was 100). In addition, the 18S gene sequence obtained by sequencing was compared for homology, and the results showed that the 18S gene of microalgae E1 had a similarity of more than 99% with 57 18S gene sequences from Chlorellavulgaris strains CCAP211 / 35, KNUA027 and ACSSI 361. Therefore, the microalgae E1 obtained in the present invention was determined to be vulgaris of the genus Chlorella of the phylum Chlorella, and was named Chlorella vulgaris E1.

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

[0067] Wheat pot experiment: set up negative control (represented by Control) and positive control (represented by BG11) (the role of the positive control here is mainly to control the culture supernatant, excluding the role of the culture medium, which proves that the E1 metabolite works), and the experimental group applied Chlorella vulgaris E1 algae cells (represented by E1 (algae cells)) and applied Chlorella vulgaris E1 culture supernatant (represented by E1 (supernatant)). A total of 4 treatments, each treatment was repeated 3 times ( Figure 3 and Figure 4). The saline-alkali farmland soil that had been sieved through a 2mm sieve and fully mixed was used. The salt content of the soil was 0.9g / kg (NaCl / dry soil). It was packaged according to 330g of dry soil per pot and planted with "Malan No. 1" wheat. 16 seeds were planted in each pot (recorded as the first day). Thinning began after the seedlings emerged neatly, and 6 wheat seedlings were retained in each pot. The culture conditions were 25°C, 16h light / 8h dark. Sterile water was used for irrigation in the early stage. When the wheat seedlings were thriving, 4g / LNaCl solution was used for salt stress treatment, and sterile water and BG11 culture medium were used for negative control treatment and positive control treatment, respectively. Watering was performed every 1 to 3 days. After continuous watering, the soil NaCl content reached about 8.72g / kg (NaCl / dry soil), and then the use of NaCl solution for irrigation was stopped. At the same time, 20 mL of Chlorella vulgaris E1 algae cells and Chlorella vulgaris E1 supernatant were irrigated into the soil, and sterile water and BG11 culture medium were irrigated for negative control and positive control treatments, respectively, for a total of 5 times. The experimental results showed that: (1) Under severe salt stress (8.72 g / kg NaCl / dry soil), E1 (algae cells) and E1 (supernatant) can promote wheat growth, alleviate the degree of membrane lipid peroxidation of wheat stems and leaves under salt stress, and improve wheat salt resistance ( Figure 3 , Figure 4 and Figure 5 (2) The aboveground fresh weight of wheat treated with E1 (algae cells) increased by 2.70% compared with that of the control treatment ( Figure 3 (3) The aboveground fresh weight of 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 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 algae cells and supernatant of the present invention can promote the growth of wheat under severe salt stress (8.72 g / kg NaCl / dry soil), and these promoting effects are mainly manifested in the increase of aboveground fresh weight and / or plant height of wheat plants, the decrease of malondialdehyde (MDA) content in stems and / or leaves ( Figure 6 ), reduced the degree of membrane lipid peroxidation in stems and / or leaves of wheat under salt stress conditions, and alleviated the damage of salt stress to wheat.

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

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

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

[0072] Pick a single colony of Chlorella vulgaris E1 on solid BG11 medium, inoculate it into 50mL liquid BG11 medium, and culture it for 14 days at 25℃, 160r / min, 12h light / 12h dark. Take an appropriate amount of Chlorella vulgaris E1 algae liquid and inoculate it into 50mL BG11 liquid medium, set up 6 replicates, culture it for a certain period of time at 25℃, 160r / min, 12h light / 12h dark, and collect the supernatant (represented by E1) by centrifugation at 8000rpm and 5min. At the same time, 50mL BG11 liquid medium without inoculation of Chlorella vulgaris E1 algae liquid was used as a control (represented by Control), and 6 replicates were set. The above supernatant and control samples were stored in a -80℃ refrigerator and submitted to Shanghai Paisonno Biotechnology Co., Ltd. for metabolomics detection. The results show that the Chlorella vulgaris E1 of the present invention can produce metabolites that can be used as plant growth regulators, including diethylene glycol, isophorone, erucamide, hexadecanoic acid, allantoic acid, L-serine, ethephon, glutamine-arginine and lauramide; can produce metabolites that promote plant growth or improve 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, L-isoleucine-L-threonine; further, promote plant growth, and under salt stress conditions, improve plant salt resistance, reduce the degree of peroxidation of stem and leaf cell membrane lipids of wheat under salt stress conditions, and alleviate the damage of salt stress to plants. In addition, the algae strain can produce the metabolite capsaicin that can be used as an insecticide, and further, has the potential to improve 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] AATTTTATCTACCCATTTGTTATCACCAGGTTTTGTTTCTGGTTTTCAGTTAATGCTTCTA

[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 is described above by way of example. It should be noted that, without departing from the core of the present invention, any simple deformation, modification or other equivalent replacement that can be made by those skilled in the art without inventive effort falls within the protection scope of the present invention.

Claims

1. A growth-promoting chlorella Chlorella vulgaris E1, characterized in that: It was deposited in the China Center for Type Culture Collection on December 20, 2024, and its deposit number is: CCTCC NO:M 20242875.

2. The present invention provides a Chlorella preparation, characterized in that: It comprises the growth-promoting Chlorella vulgaris E1 according to claim 1.

3. A microbial fertilizer, characterized in that: Contains at least one of the following: The growth-promoting chlorella Chlorella vulgaris E1, its culture or its processed product according to claim 1; The Chlorella preparation according to claim 2.

4. A soil conditioner, characterized in that: Contains at least one of the following: The growth-promoting chlorella Chlorella vulgaris E1, its culture or its processed product according to claim 1; The Chlorella preparation according to claim 2.

5. A plant growth regulator, characterized in that: Contains at least one of the following: The growth-promoting chlorella Chlorella vulgaris E1, its culture or its processed product according to claim 1; The Chlorella preparation according to claim 2.

6. A biostimulant, characterized in that: Contains at least one of the following: The growth-promoting chlorella Chlorella vulgaris E1, its culture or its processed product according to claim 1; The Chlorella preparation according to claim 2.

7. A biological pesticide, characterized in that: It comprises a carrier and an active ingredient; the active ingredient comprises: the growth-promoting chlorella Chlorella vulgaris E1, its culture or its processed product in the above technical solution.

8. Use of the growth-promoting agent for Chlorella vulgaris E1 according to claim 1, the Chlorella preparation according to claim 2 or the microbial fertilizer according to claim 3 in at least one of the following: Improve crop stress resistance, especially salt resistance; Promote crop growth; Soil improvement.

9. The use according to claim 8, characterized in that: The growth-promoting Chlorella vulgaris E1 of claim 1, the Chlorella preparation of claim 2 or the microbial fertilizer of claim 3 is added to the soil for growing crops.

10. A method for preparing a biological pesticide, characterized in that: The biopesticide is prepared by using the growth-promoting chlorella Chlorella vulgaris E1, a culture thereof or a processed product thereof.

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