New application of a multifunctional Talaromyces W10

Abstract

The new application of a multifunctional Talaromyces fungus W10 belongs to the field of agricultural microbial technology. In order to solve the technical problems that the soybean yield needs to be improved and the research on the growth and development of Talaromyces in promoting soybean is less, the present application further identifies a Talaromyces fungus W10 with the ability of dissolving inorganic phosphorus, and explores the growth promoting effect of the strain. It is found that the strain has the ability of dissolving organic phosphorus, degrading cellulose and producing indole acetic acid. In addition, through seed germination experiment and pot experiment, it is found that the fermentation liquid and fermentation supernatant of the strain can promote the germination of soybean seeds and the growth of seedlings, and improve the chlorophyll and carotenoid content in soybean seedling leaves. It can be seen that the Talaromyces fungus W10 in the present application has the potential to be developed into a microbial fertilizer, and has a broad application prospect in agricultural production.

Description

Technical Field

[0001] The present invention belongs to the technical field of agricultural microorganisms, and in particular relates to a new application of a multifunctional Talaromyces fungus W10. Background Art

[0002] Soybean is an important food crop in my country and has strong application value in the food, health care, pharmaceutical and feed industries (Wang Liaowei. Analysis of my country's soybean production situation and development suggestions [J]. Soybean Science and Technology, 2024, (04): 1-6.). With the improvement of people's living standards, the demand for soybean food continues to expand. According to statistics, China imported 91.08 million tons of soybeans in 2022, accounting for 58.3% of the global trade volume (Ma Cuiping, Yang Shuiqing. Research on soybean trade under the evolution of Sino-US economic and trade relations since joining the WTO [J]. World Agriculture, 2024, (05): 69-80.). The current soybean production is far from meeting national demand. Therefore, reasonably increasing soybean unit yield is a strategic measure with far-reaching significance. However, in the process of soybean cultivation, farmers usually apply excessive chemical fertilizers in order to increase yield. This not only brings additional economic investment to farmers, but also destroys soil structure and damages soil fertility, which is not conducive to the sustainable development of agriculture.

[0003] Microbial fertilizers can improve the soil environment by promoting the growth of beneficial microorganisms in the soil through the addition of microorganisms, promoting the development of a virtuous cycle of soil structure. Some microorganisms can activate insoluble phosphorus and potassium fixed in the soil into fast-acting phosphorus and potassium that can be absorbed and utilized by plants. Other microorganisms can secrete plant hormones (such as indoleacetic acid and gibberellins) to directly promote plant growth. Microbial fertilizers have ecological benefits that traditional chemical fertilizers do not have, such as activating soil nutrients, improving soil structure, and promoting nutrient absorption (Wang Yikun, Gao Fei, Li Changwei, et al. Effects of microbial fertilizers on soil and crop growth [J]. Agricultural Engineering, 2024, 14(09): 68-72.), and have great development significance.

[0004] Talaromyces sp. is an endophytic fungus widely distributed in soil, plants, and oceans. Its research in the fields of medical antibiotics, enzyme preparations, natural pigments, and agriculture is relatively common (Sun Jianqiu, Ruan Yongming, Jin Shiyu, et al. The importance of the genus Talaromyces and an overview of its taxonomic research [J]. Mycological Research, 2021, 19(2): 83-93.).In the process of agricultural research, Abbas A et al. used microbiome analysis technology to find that the species abundance of Talaromyces in rice soil was much higher than that in fallow soil, indicating the dominant position of Talaromyces in rice soil. They also screened four strains of Talaromyces from rice soil using rice sheath blight as bait, one of which, Talaromyces TF-04, could significantly promote rice growth, reduce rice sheath blight, and increase rice yield under outdoor conditions (Aqleem, Abbas, Yanping, Fu, Zheng, Qu et al. Isolation and evaluation of the biocontrol potential of Talaromyces pp. against rice sheath blight guided by soil microbiome.[J].EnvironMicrobiol,2021,23:0.); Mao Xueqin et al. isolated the yellow basket fungus (T.flavus) MT-06 from the strawberry anthracnose standard sample and found that it has an inhibitory effect on a variety of plant pathogenic fungi, and the inhibition rate on strawberry anthracnose fungi reached 71.75% (Mao Xueqin, Wei Caiyan, Chai Rongyao, et al. Determination of the control effect and colonization ability of the biocontrol strain MT-06 on strawberry anthracnose [J]. Jiangsu Agricultural Science, 2011,39(02):193-194+281.); Yin Chenglin et al. added the purple basket fungus Q2 strain to the fumigated soil to effectively promote the growth of cucumbers, increase the plant photosynthetic rate and root activity (Yin Chenglin, Xu Peipei, Jin Yirong, et al. Study on the effect of purple basket fungus on cucumber growth after soil fumigation treatment [J]. Anhui Agricultural Science, 2024,52(1 2):13-15+21.); The growth-promoting fungus T. flavus isolated from the roots of poplar by Yin Xiaoman et al. showed high solubility activity on calcium phosphate and has the potential to be developed as a microbial fertilizer (Yin Xiaoman, Tan Jiajin, Fang Aiqin. Determination of phosphorus solubility characteristics of a yellow-blue fungus (Talaromyces flavus) SH16 [J]. Journal of Nanjing Forestry University (Natural Science Edition), 2017, 41(05):169-174.); The golden-yellow ... In summary, Basileus exhibits significant antagonistic effects against a variety of plant pathogens at the biological control level, and has an outstanding ability to efficiently convert insoluble phosphorus in terms of phosphorus solubilization. These characteristics give it excellent development potential and broad prospects in the field of agricultural microbial fertilizers.

[0005] At present, domestic research on basket fungi in the agricultural field is still in its infancy, and there are very few studies on the efficacy of basket fungi in promoting soybean growth and development. Based on this, the applicant intends to further identify a strain of basket fungi W10 that has the function of dissolving inorganic phosphorus, and explore the growth-promoting effect of the basket fungi, in order to provide a theoretical basis for the production and application of the basket fungi in soybeans. Summary of the Invention

[0006] To address the problem of soybean yields needing to be increased and the lack of research on the efficacy of Talaromyces in promoting soybean growth and development, the present invention further identified a Talaromyces fungus W10 with the ability to dissolve inorganic phosphorus and explored the growth-promoting efficacy of this strain. It was found that this strain has the ability to dissolve organic phosphorus, degrade cellulose, and produce indoleacetic acid. In addition, through seed germination experiments and potted experiments, it was found that the fermentation liquid and fermentation supernatant of this strain can promote soybean seed germination and seedling growth, and increase the chlorophyll and carotenoid content in soybean seedling leaves. The specific technical solutions of the present invention are as follows:

[0007] The first object of the present invention is to provide a Talaromyces sp. fungus W10 for use in dissolving organic phosphorus. The preservation number of the Talaromyces sp. fungus W10 is CGMCC NO.40621.

[0008] In one embodiment of the present invention, the organic phosphorus is lecithin.

[0009] The second object of the present invention is to provide a phosphate-dissolving bacterial agent containing the above-mentioned Tularemia fungus W10 for use in dissolving organic phosphorus in soil.

[0010] In one embodiment of the present invention, the organic phosphorus is lecithin.

[0011] The third object of the present invention is to provide the use of the above-mentioned Talaromyces fungus W10 in degrading cellulose.

[0012] The fourth object of the present invention is to provide the application of the above-mentioned Talaromyces fungus W10 in promoting soybean seed germination, wherein the application is to treat soybean seeds with the fermentation liquid or fermentation supernatant of the Talaromyces fungus W10.

[0013] In one embodiment of the present invention, the soybean seeds are fresh seeds of the season or old seeds.

[0014] The fifth object of the present invention is to provide the application of the above-mentioned Talaromyces fungus W10 in promoting the growth of soybean seedlings, wherein the application is to use the fermentation liquid or fermentation supernatant of the Talaromyces fungus W10 to treat the soil for planting soybeans.

[0015] The sixth object of the present invention is to provide the application of the above-mentioned Talaromyces fungus W10 in increasing the chlorophyll content in soybean seedling leaves, wherein the application is to use the fermentation liquid or fermentation supernatant of the Talaromyces fungus W10 to treat the soil for planting soybeans.

[0016] The seventh object of the present invention is to provide the application of the above-mentioned Talaromyces fungus W10 in increasing the carotenoid content in soybean seedling leaves, wherein the application is to use the fermentation liquid or fermentation supernatant of the Talaromyces fungus W10 to treat the soil for planting soybeans.

[0017] Beneficial effects of the present invention:

[0018] The present invention further identified the Talaromyces fungus W10 disclosed in the Chinese patent application number 202310752176.1. Combining the results of the phylogenetic tree based on homologous genes and the gene family contraction and expansion evolutionary tree, the strain W10 was identified as Talaromyces amestolkiae.

[0019] The present invention inoculated strain W10 into a culture medium containing lecithin as the sole phosphorus source. The strain was found to not only dissolve inorganic phosphorus (as previously discovered) but also degrade organic phosphorus, achieving a maximum phosphorus solubility of 169.32 mg / L on lecithin. Studies have shown that only a small amount of organic phosphorus can be directly absorbed and utilized by plants during growth; the majority must be converted into soluble inorganic phosphorus before it can be utilized by plants. Organic phosphorus in soil accounts for 30-65% of the total phosphorus content (Zhang Wannian, Yang Zi, Yan Yupeng, et al. Research Progress on Mineralization and Regulation of Organic Phosphorus in Soil [J / OL]. Acta Pedologica Sinica, 1-13 [2024-12-03].), making organic phosphorus a very important potential phosphorus source for plant growth. Overall, the dual role of T. amestolkiae W10 in dissolving both inorganic and organic phosphorus allows for a continuous and stable supply of phosphorus for plant growth, possessing significant research and application value.

[0020] In this study, the ELISA detection kit was used to accurately detect the presence of indoleacetic acid (IAA) in the fermentation broth of W10, with the highest content being 59.29 μg / L. Compared with fungi with high IAA production, such as the high IAA-producing Trichoderma harzianum mutant strain obtained by high-energy irradiation screening, its maximum yield is 456.81 μg / L (Gao Yaxin, Wang Jiaguo, Zhang Xiangkai, et al. Screening of high-IAA-producing Trichoderma harzianum mutant strain M95 and its growth-promoting effect on cucumber [J]. Journal of Nanjing Agricultural University, 2023, 46(03): 499-509.), although the IAA content produced by strain W10 is low, it is well known that excessive concentration of IAA will inhibit plant growth. The optimal concentration range of IAA for plants is 0.01-10 mg / L (Zhang Zhiyong, Zhou Jiahuai. Hormone regulation of plant root growth [J]. Nature Journal, 1986, (09): 43-47+82.), which shows that the IAA concentration produced by strain W10 can effectively promote plant growth. In the seed germination experiment, the fermentation supernatant of strain W10 on the second day of inoculation was able to significantly promote the growth of soybean embryo roots and stems. During the same period, the IAA production in the supernatant reached the highest value, which also confirmed the efficacy of IAA.

[0021] The present invention also discovered that T. amestolkiae W10 has the ability to produce cellulase activity. The cellulase activity in the culture broth reached a maximum of 24.52 U / mL on the second day of fermentation. Although this does not meet the organic material maturity standard of 70 U / mL specified in the national "General Technical Requirements for Quality Evaluation of Microbial Fertilizer Production Strains," strain W10, as a microbial agent, does promote seed germination. In a stale seed germination experiment, the fermentation broth and supernatant of strain W10 on the first day of fermentation exhibited different effects on soybean seed germination. Due to the low cellulase activity content in the supernatant on the first day, the diluted supernatant, like the blank treatment, failed to germinate the stale seeds. However, since soybean hulls are rich in 40% cellulose, the fermentation broth containing mycelium can promote cellulose decomposition and, consequently, promote germination of stale seeds. This was also confirmed by the seed germination effects of the supernatants of strain W10 on days 2 and 3 of fermentation, indicating that strain W10 has the potential to be developed as a seed coating agent.

[0022] In the potted experiment, the present invention selected the fermentation liquid and the fermentation supernatant on the second day of fermentation to carry out the soybean seedling experiment. The results showed that the soybean plant height, fresh weight, dry weight, root weight, number of nodules, and chlorophyll and carotenoid contents in the fermentation liquid treatment on the second day were significantly higher than those in the CK. Although the various soybean indicators in the fermentation supernatant were also higher than those in the CK, they did not show significant differences. This shows that the effect of a one-time application of the supernatant is not as long-lasting as the effect of applying the fermentation liquid containing mycelium. It is worth mentioning that the number of rhizobia in the soybean root system treated with the fermentation liquid was also significantly higher than that of the CK group, indicating that strain W10 may be able to recruit beneficial microorganisms (such as rhizobia, etc.) in the soil to enrich in the root system, further showing the application potential of strain W10 in soybean crop cultivation. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 This is the phylogenetic tree analysis result of strain W10 based on the single-copy gene set;

[0024] Figure 2 This is the result of the gene family contraction and expansion evolutionary tree analysis of strain W10;

[0025] Figure 3 This is a graph showing the changes in available phosphorus content in the culture medium after strain W10 was inoculated into an organophosphorus liquid culture medium with lecithin as the sole phosphorus source and cultured for different periods of time;

[0026] Figure 4 This is a graph showing the changes in cellulase activity over culture time after strain W10 was inoculated into PDA liquid culture medium;

[0027] Figure 5 This is a graph showing the change in indoleacetic acid content in the culture medium over culture time after strain W10 was inoculated into PDA liquid culture medium;

[0028] Figure 6 The results of the effects of the fermentation liquid and supernatant of strain W10 on the germination of fresh soybean seeds and old soybean seeds are shown in the figure. Figure 6 A in the figure is the effect of the fermentation liquid and supernatant of strain W10 on the radicle length during the germination of fresh soybean seeds in that season. Figure 6 Figure B shows the effect of the fermentation liquid and supernatant of strain W10 on the embryonic axis length during the germination of fresh soybean seeds in that season. Figure 6 Figure C shows the effect of the fermentation liquid and supernatant of strain W10 on the radicle length during the germination of old soybean seeds. Figure 6 D in the figure is the effect of the fermentation liquid and fermentation supernatant of strain W10 on the embryonic axis length during the germination of old soybean seeds;

[0029] Figure 7The figure shows the effect of the fermentation liquid and supernatant of strain W10 on the chlorophyll content and carotenoid content in soybean seedling leaves; Figure 7 A in the figure is the effect of the fermentation liquid and fermentation supernatant of strain W10 on the chlorophyll a content in soybean seedling leaves. Figure 7 B in the figure is the effect of the fermentation liquid and fermentation supernatant of strain W10 on the chlorophyll b content in soybean seedling leaves. Figure 7 C in the figure shows the effect of the fermentation liquid and supernatant of strain W10 on the total chlorophyll content in soybean seedling leaves. Figure 7 D in the figure shows the effect of the fermentation liquid and fermentation supernatant of strain W10 on the carotenoid content in soybean seedling leaves. DETAILED DESCRIPTION

[0030] In order to make the purpose, technical solutions and advantages of the present invention more clear, the present invention is further described in detail below with reference to specific embodiments and the accompanying drawings. It should be noted that the embodiments mentioned below are only applicable to explaining the present invention, but are not intended to limit the scope of the present invention. The embodiments mentioned below are only some embodiments of the present invention and not all embodiments. In this field, if other technicians do not make creative work, the embodiments they obtain are protected by the present invention.

[0031] The experimental methods used in the following examples are conventional methods unless otherwise specified. The materials, reagents, culture media and instruments used are conventional materials, reagents, culture media and instruments in the art unless otherwise specified, and can be obtained from commercial channels by those skilled in the art.

[0032] The Talaromyces sp. fungus W10 in the present invention is deposited in the General Microbiology Center of the China Culture Collection Administration, with a deposit number of CGMCC NO.40621 and a deposit date of May 10, 2023. The depository address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences. It is disclosed in the Chinese patent application No. 202310752176.1 and is isolated and preserved by the Institute of Microbiology, Heilongjiang Academy of Sciences.

[0033] The fresh soybean variety used in the test season provided by the present invention is Williams 82; the old seeds are Heihe 43.

[0034] The test soil provided by the present invention has the following composition: organic matter content of 26.52 g / kg, total nitrogen content of 106.0 mg / kg, available phosphorus content of 25.0 mg / kg, available potassium content of 83.7 mg / kg, and pH value of 6.4.

[0035] The culture medium of the present invention is as follows:

[0036] PDA medium consisted of the following ingredients: 1000 mL of potato juice, 20 g of glucose, 2 g of dipotassium hydrogen phosphate, 1 g of magnesium sulfate, pH 7.0.

[0037] The organophosphorus culture medium is composed of the following ingredients: glucose 10 g, ferrous sulfate 0.03 g, magnesium sulfate 0.3 g, sodium chloride 0.3 g, potassium chloride 0.3 g, ammonium sulfate 0.5 g, manganese sulfate 0.03 g, lecithin 5 g, distilled water 1000 mL, pH 7.2-7.4.

[0038] Example 1: Further identification of Tularemia W10

[0039] In the Chinese patent application number 202310752176.1, strain W10 was identified as a fungus of the genus Talaromyces sp. based on its colony characteristics, microscopic morphological description and ITS rDNA sequence analysis results. In order to further identify strain W10, strain W10 was sent to Sangon Biotech (Shanghai) Co., Ltd. for whole genome sequencing. Based on the results of homologous gene and pan-genome analysis, different phylogenetic trees were constructed using neighbor-joining clustering based on single-copy core gene datasets. Combining the results of the phylogenetic tree based on homologous genes and the gene family contraction and expansion evolutionary tree (see Figure 1 and Figure 2 ), strain W10 was identified as Talaromyces amestolkiae.

[0040] Example 2: Detection of phosphate solubilization activity of Tularemia spp. W10 on organic phosphorus

[0041] Add 2 mL of the prepared spore suspension of strain W10 to 200 mL of Montana liquid medium with lecithin as the only phosphorus source. The concentration of the spore suspension is 1 × 10 7 cfu / mL, and cultured in a shaker at 28°C. The supernatant was aspirated on days 1, 2, 3, 4, 5, 6, and 7, and the available phosphorus content in the supernatant was detected using the molybdenum antimony colorimetric method.

[0042] The strain W10 was inoculated into a Montgina liquid medium with lecithin as the only phosphorus source, and the changes in the available phosphorus content in the medium were detected every other day. Figure 3 It can be seen that strain W10 has a certain solubility for organic phosphorus. In the organic phosphorus culture medium with lecithin as the only phosphorus source, the available phosphorus content increases with the increase of culture time. The value on the last day of detection is 169.32 mg / L and has not reached the maximum limit.

[0043] Example 3: Determination of the ability of Tularemia spp. W10 to degrade cellulose

[0044] To create a standard curve: Accurately prepare glucose standard solutions at 400 μg / mL, 600 μg / mL, 800 μg / mL, 1200 μg / mL, 1600 μg / mL, and 2000 μg / mL. Pipette 1 mL of each standard solution into a 20 mL stoppered colorimetric tube. Add 2 mL of water and 2 mL of DNS reagent to each tube. Boil in a boiling water bath for 5 minutes. After cooling, dilute to 20 mL with water. Zero the solution with a blank. Measure the absorbance at a wavelength of 540 nm. Using absorbance as the horizontal axis and glucose concentration as the vertical axis, calculate the regression equation: y = 1176.9x + 69.233, with a correlation coefficient R. 2 =0.9955.

[0045] Preparation of cellulase solution: After strain W10 was cultured in PDA liquid medium for a certain period of time, 5 mL of fermentation broth was aspirated into a conical flask, diluted 3 times with citrate buffer, placed on a shaker for 30 minutes, centrifuged at 5000 rpm for 10 minutes, and the supernatant was collected as the enzyme solution to be tested.

[0046] Cellulase activity assay: Pipette 2 mL of 0.51% sodium carboxymethyl cellulose solution into a 20 mL stoppered colorimetric tube. Incubate at 50°C for 5 minutes. Add 0.5 mL of preheated enzyme solution to be tested, shake thoroughly, and incubate at 50°C for 30 minutes. Add 2 mL of DNS solution, boil in a water bath for 5 minutes, and cool to room temperature. Add distilled water to 20 mL. Measure the absorbance at 540 nm for three replicates, and calculate the average. A colorimetric tube containing enzyme solution and DNS solution pre-added for reaction serves as a blank control. One unit of enzyme activity (U / mL) is defined as the amount of enzyme that degrades sodium carboxymethyl cellulose to produce 1 μg of glucose in 1 minute per 1 mL of sample.

[0047] The spore suspension of strain W10 was inoculated into PDA liquid culture medium, and the cellulase activity in the culture medium was detected after 1, 2, 3, and 4 days. Figure 4 As shown in the figure, it can be seen that the cellulase activities of strain W10 at 1, 2, 3, and 4 days after inoculation were 15.1 U / mL, 24.52 U / mL, 16.75 U / mL, and 7.57 U / mL, respectively, showing a trend of first increasing and then decreasing.

[0048] Example 4: Determination of the ability of Tularemia spp. W10 to produce indoleacetic acid

[0049] The enzyme-linked immunosorbent assay (ELISA) was used to determine the indoleacetic acid (IAA) content in the culture medium. Add 50 μL of appropriately diluted sample to a microplate coated with an IAA antibody, add 50 μL of enzyme-labeled reagent, incubate at 37°C for 60 minutes, wash 3-5 times with detergent, spin dry, add 100 μL of TMB, develop in the dark for 15 minutes, and terminate the reaction by adding 50 μL of stop solution. Use a blank sample as a control and measure the absorbance of each well at a wavelength of 450 nm. Standard curve: y = 111.92-93.974x, correlation coefficient R 2 =0.9925.

[0050] The spore suspension of strain W10 was inoculated into PDA liquid culture medium, and the culture medium was collected every other day. The changes in the indoleacetic acid content in the culture medium were detected by ELISA kit. Figure 5 As shown in the figure, the change of indoleacetic acid content in the culture medium showed a trend of slowly increasing and then decreasing. The indoleacetic acid content reached the highest value of 59.29 μg / L on the second day of culture.

[0051] Example 5: Application of strain W10 in promoting soybean seed germination

[0052] Strain W10 was inoculated into PDA liquid medium for fermentation. The obtained fermentation broth and fermentation supernatant were diluted 1000-fold with sterile water, respectively. The sterilized PDA liquid medium dilution was used as a blank control. Five sterilized soybean seeds were placed in each culture dish lined with two layers of filter paper. 10 mL of the diluted fermentation broth or fermentation supernatant was added to each culture dish. Three replicates were placed in each group and placed in a 28°C incubator. After culturing for 3 days, the radicle and hypocotyl lengths of the germinated seeds were detected.

[0053] The results are as follows Figure 6 As shown in the results, both the fermentation broth and supernatant of strain W10 can significantly promote the germination of fresh soybean seeds (Williams 82) of the season, and the effects are most obvious on the second day. The effects of the fermentation supernatant on the second day on the growth of soybean radicle length and axis length are significantly different from those of CK, and the effects of the fermentation broth on the second day on the growth of soybean hypocotyl length are significantly different from those of CK. In the experiment to detect the effects of the fermentation broth and supernatant of strain W10 on the germination of old soybean seeds, the seeds in the blank control group and the fermentation supernatant on the first day did not germinate, and some seeds in the fermentation broth treatment groups also did not germinate, while the seeds in the fermentation supernatant on the second and third days germinated normally, and the radicle length and axis length of the group treated on the second day were the highest among all groups.

[0054] Example 6: Application of strain W10 in promoting the growth of soybean seedlings

[0055] The test soil was common black soil from Northeast China. The soil was collected and distributed into seedling pots (11.6 cm top diameter, 10.5 cm height), with 1.5 kg of black soil per pot. Williams 82 seeds were used. Strain W10 was inoculated into PDA liquid medium and cultured with shaking at 28°C and 170 rpm. On the second day of culture, the fermentation broth was aspirated and a portion was centrifuged at 10,000 rpm and 25°C for 10 min. The supernatant was collected.

[0056] Experimental groups: (1) CK: ordinary soil; (2) fermentation liquid: fermentation liquid on the second day after inoculation with W10; (3) fermentation supernatant: fermentation supernatant on the second day after inoculation with W10.

[0057] Uniformly sized and relatively plump soybean seeds were selected and sterilized before being sown in seedling pots. One seed was placed in each pot, and five treatments were assigned to each group. 2 mL of the fermentation liquid or fermentation supernatant of strain W10 was evenly injected with a syringe 1-2 cm from the seed sowing point. CK was not specially treated. Water was applied every 5 days after sowing. After 30 days, the height, fresh weight, dry weight, root length, root weight, and number of nodules of soybean seedlings were tested (see Table 1).

[0058] As shown in Table 1, when the soybean seedlings treated with the fermentation liquid and fermentation supernatant of the strain grew to 30 days, their plant height, fresh weight, dry weight, root length, root weight and nodule number were significantly higher than those of the blank control group. In both treatment groups, only the plant height showed significant difference compared with the CK, and there was no significant difference in root length between the two treatment groups and the CK. Among the other indicators, only the fermentation liquid treatment group showed significant differences compared with the CK. Compared with CK, the plant height, fresh weight, dry weight, root length, root weight and number of nodules in the fermentation liquid treatment group increased by 4.8 cm, 0.3175 g, 0.0625 g, 1.025 cm, 0.0425 g and 7, respectively, with an increase of 14.48%, 16.26%, 36.23%, 6.91%, 13.08% and 73.68%, respectively. Compared with CK, the plant height, fresh weight, dry weight, root length and number of nodules in the fermentation supernatant treatment group increased by 3.9 cm, 0.1525 g, 0.0425 g, 0.825 cm and 1.5, respectively, with an increase of 11.76%, 7.81%, 24.64%, 5.56% and 15.79%, respectively.

[0059] Table 1 Effects of the fermentation broth and supernatant of strain W10 on the growth of soybean seedlings

[0060]

[0061] Example 7: Application of strain W10 in increasing chlorophyll and carotenoid content in soybean seedling leaves

[0062] The test soil was common black soil from Northeast China. The soil was collected and distributed into seedling pots (11.6 cm top diameter, 10.5 cm height), with 1.5 kg of black soil per pot. Williams 82 seeds were used. Strain W10 was inoculated into PDA liquid medium and cultured with shaking at 28°C and 170 rpm. On the second day of culture, the fermentation broth was aspirated and a portion was centrifuged at 10,000 rpm and 25°C for 10 min. The supernatant was collected.

[0063] Experimental groups: (1) CK: ordinary soil; (2) fermentation liquid: fermentation liquid on the second day after inoculation with W10; (3) fermentation supernatant: fermentation supernatant on the second day after inoculation with W10.

[0064] Soybean seeds of uniform size and relatively full size were selected and sterilized before sowing in seedling pots. One seed was placed in each pot and five treatments were applied in each group. 2 mL of the fermentation liquid or fermentation supernatant of strain W10 was evenly injected with a syringe 1-2 cm from the seed sowing point. CK was not specially treated. Water was applied every 5 days after sowing. The chlorophyll content and carotenoid content in the leaves of soybean seedlings were tested 30 days later (see Figure 7 ).

[0065] Chlorophyll content assay: Accurately weigh 0.1g of fresh leaves, chop them into pieces, and place them in a 2mL centrifuge tube. Add 1mL of distilled water and homogenize the leaves in a tissue homogenizer for 1 minute. Rinse the broken tissue into a 10mL centrifuge tube using the extract (ethanol:acetone volume ratio = 1:2). Dose the extract to 10mL. Extract in the dark for 3 hours. Observe the color of the tissue residue at the bottom; if it approaches white, complete extraction is indicated. Measure the absorbance of the extract at 645nm and 663nm, respectively, using the extract as a blank control.

[0066] Chlorophyll a content (mg / g) = (21.2×A663-4.48×A645)×V 提 ÷0.1g÷1000;

[0067] Chlorophyll b content (mg / g) = (38.2×A645-7.8×A663)×V 提 ÷0.1g÷1000;

[0068] Total chlorophyll content (mg / g) = (13.4×A663+33.7×A645)×V 提 ÷0.1g÷1000.

[0069] Carotenoid content assay: Accurately weigh 0.1g of fresh leaves, mince, and place in a 2mL centrifuge tube. Add 1mL of distilled water and homogenize in a tissue homogenizer for 1 minute. Aspirate the extract (distilled water:acetone volume ratio = 1:4) and rinse the broken tissue into a 10mL centrifuge tube. Dose the extract to 10mL. Extract in the dark for 3 hours. Observe the color of the tissue residue at the bottom; complete extraction is indicated if it approaches white. Measure the absorbance of the extract at 646nm, 663nm, and 470nm, using the extract as a blank control.

[0070] Carotenoid content (mg / g) = [4.367×A470-0.014×(12.21×A663-2.81×A646)-0.454×(20.13×A646-5.03×A663)]×V 提 ÷0.1g.

[0071] Depend on Figure 7 It can be seen that in the soybean pot experiment, the chlorophyll a, chlorophyll b, total chlorophyll content and carotenoid content in the leaves of soybean seedlings treated with strain fermentation liquid and fermentation supernatant were significantly higher than those in the blank control group. Only the fermentation liquid treatment group showed significant differences compared with CK. The chlorophyll a, chlorophyll b, total chlorophyll content and carotenoid content in the fermentation liquid treatment group increased by 0.1325 mg / g, 0.0695 mg / g, 0.2020 mg / g and 0.0154 mg / g respectively compared with CK, with an increase of 32.11%, 38.21%, 33.98% and 56.20% respectively. There was no significant difference between the fermentation supernatant treatment group and CK.

[0072] Although the present invention has been disclosed above with reference to preferred embodiments, this is not intended to limit the present invention. Anyone familiar with this technology can make various changes and modifications without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be based on the definition of the claims.

Claims

1. A strain of the genus Talaromyces ( Talaromyces sp.) fungus W10 in dissolving lecithin for purposes other than disease diagnosis or treatment, characterized in that: The preservation number of the Talaromyces fungus W10 is CGMCC NO. 40621.

2. Use of a phosphate-dissolving bacterial agent containing the Talaromyces fungus W10 described in claim 1 in dissolving lecithin in soil.

3. Use of the Talaromyces fungus W10 described in claim 1 in degrading cellulose.

4. The use of the Talaromyces fungus W10 according to claim 1 in promoting soybean seed germination, characterized in that: The application is to use the fermentation liquid or fermentation supernatant of the Talaromyces fungus W10 to treat soybean seeds.

5. The use according to claim 4, characterized in that The soybean seeds are fresh seeds of the season or old seeds.

6. Use of the Talaromyces fungus W10 according to claim 1 in promoting the growth of soybean seedlings, characterized in that: The application is to use the fermentation liquid or fermentation supernatant of the Talaromyces fungus W10 to treat the soil for planting soybeans.

7. Use of the Talaromyces fungus W10 according to claim 1 for increasing the chlorophyll content in soybean seedling leaves, characterized in that: The application is to use the fermentation liquid or fermentation supernatant of the Talaromyces fungus W10 to treat the soil for planting soybeans.

8. Use of the Talaromyces fungus W10 according to claim 1 for increasing the carotenoid content in soybean seedling leaves, characterized in that: The application is to use the fermentation liquid or fermentation supernatant of the Talaromyces fungus W10 to treat the soil for planting soybeans.

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