New application of multifunctional talaromyces sp. Fungus W10

Abstract

The invention discloses a novel application of a multifunctional talaromyces sp. Fungus W10, and belongs to the technical field of agricultural microorganisms. In order to solve the technical problems that the soybean yield needs to be improved and the research on the effect of talaromyces in the aspect of promoting the growth and development of soybeans is less, the talaromyces fungus W10 with the capacity of dissolving inorganic phosphorus is further identified, and the growth promoting effect of the strain is explored; the bacterial strain has the capabilities of dissolving organophosphorus, degrading cellulose and generating indoleacetic acid, in addition, a seed germination experiment and a pot experiment find that fermentation liquor and fermentation supernate of the bacterial strain can promote germination of soybean seeds and growth of seedlings, and the content of chlorophyll and carotenoid in soybean seedling leaves is increased. Therefore, the talaromyces sp. Fungus W10 has the potential of being developed into the microbial fertilizer, and has a wide application prospect in agricultural production.

Description

Technical Field

[0001] The invention belongs to the technical field of agricultural microorganisms, and specifically 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's soybean imports in 2022 will be 91.08 million tons, 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 needs. Therefore, reasonably increasing the unit yield of soybean is a strategic measure with far-reaching significance. However, in the process of soybean planting, farmers usually apply excessive chemical fertilizers in order to pursue increased yields. 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 driving beneficial microorganisms in the soil through the addition of microorganisms, and promote the development of a benign cycle of soil structure. Some microorganisms can activate the insoluble phosphorus and insoluble potassium fixed in the soil into fast-acting phosphorus and fast-acting potassium that can be absorbed and utilized by plants, and some microorganisms can secrete plant hormones (such as indoleacetic acid, gibberellin, etc.) to directly promote plant growth. Microbial fertilizers have ecological effects 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 good 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]. Fungal 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 used sheath blight as bait and screened 4 strains of Talaromyces from rice soil. One of them, 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 T.flavus MT-06 from strawberry anthracnose samples and found that it had inhibitory effects on a variety of plant pathogenic fungi, with an inhibition rate of 71.75% against strawberry anthracnose (Mao Xueqin, Wei Caiyan, Chai Rongyao, et al. Determination of the efficacy and colonization ability of biocontrol strain MT-06 against 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 cucumber growth and increase 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 [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 for calcium phosphate and has the potential to be developed as a microbial fertilizer (Yin Xiaoman, Tan Jiajin, Fang Aiqin. Determination of phosphorus solubilization characteristics of a strain of Talaromyces flavus SH16 [J]. Journal of Nanjing Forestry University (Natural Science Edition), 2017, 41(05):169-174.); The golden yellow basket fungus (T. aurantiacus) JXBR04 isolated from the rhizosphere soil of bamboo by Yang Dou et al. can promote the absorption and utilization of soil available phosphorus and biomass growth of bamboo (Yang Dou, Shi Fuxi, Wan Songze, et al. Effects of golden yellow blue fungus on phosphorus components in bamboo soil and seedling biomass [J]. Forestry Science Research, 2021, 34(03):145-151.). In summary, Basileus fungi have demonstrated significant antagonistic effects on a variety of plant pathogens at the biological control level, and have 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 Basileus fungi in the agricultural field is still in its infancy, and there are very few studies on the efficacy of Basileus fungi in promoting soybean growth and development. Based on this, the applicant intends to further identify a strain of Basileus fungi W10 that has the function of dissolving inorganic phosphorus, and explore the growth-promoting effect of this Basileus fungi, in order to provide a theoretical basis for the production and application of this Basileus fungi in soybeans. Summary of the invention

[0006] In order to solve the problem that soybean yield needs to be improved and there is little research on the efficacy of Talaromyces in promoting soybean growth and development, the present invention further identified a Talaromyces fungus W10 that has the ability to dissolve inorganic phosphorus, and explored the growth-promoting effect of the strain. It was found that the 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 the strain can promote the germination of soybean seeds and seedling growth, and increase the chlorophyll and carotenoid content in the leaves of soybean seedlings. The specific technical scheme of the present invention is as follows:

[0007] The first objective of the present invention is to provide a strain of Talaromyces sp. fungus W10 for use in dissolving organic phosphorus. The deposit 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 Bacillaceae 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 an 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 an 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 treat the soil for planting soybeans with the fermentation liquid or fermentation supernatant of the Talaromyces fungus W10.

[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, and combined the results of the phylogenetic tree based on homologous genes and the gene family contraction and expansion evolutionary tree to identify the strain W10 as Talaromyces amestolkiae.

[0019] The present invention inoculates strain W10 in a culture medium with lecithin as the only phosphorus source, and finds that strain W10 not only has the ability to dissolve inorganic phosphorus (found in previous studies), but also has the function of degrading organic phosphorus, and its maximum phosphorus dissolving capacity for lecithin can reach 169.32 mg / L. Studies have shown that only a small amount of organic phosphorus can be directly absorbed and utilized by plants during plant growth, and most organic phosphorus needs to be converted into soluble inorganic phosphorus before it can be used by plants, and the content of organic phosphorus in the 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 Soil Organic Phosphorus [J / OL]. Acta Pedologica Sinica, 1-13 [2024-12-03].), so organic phosphorus is a very important potential phosphorus source in the process of plant growth. In general, the dual role of T.amestolkiae W10 in dissolving inorganic phosphorus and organic phosphorus can continuously provide stable phosphorus elements for plant growth, and has important 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 high-IAA-producing fungi, such as the high-IAA-producing Trichoderma harzianum mutant obtained by high-energy ray 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 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 Magazine, 1986, (09): 43-47+82.), it can be seen 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 after inoculation was able to significantly promote the growth of soybean germ roots and stems. During the same period of time, the production of IAA in the supernatant reached the highest value, which also confirmed the efficacy of IAA.

[0021] The present invention also found that T.amestolkiae W10 has the ability to produce cellulase activity. The cellulase activity in the culture fluid on the second day of fermentation reaches the highest value of 24.52U / ml. Although it does not reach the 70U / mL organic material maturity standard in the "General Technical Requirements for Quality Evaluation of Microbial Fertilizer Production Strains" stipulated by the state, the strain W10 has the function of promoting seed germination as a microbial agent. In the old seed germination experiment, the fermentation liquid and the fermentation supernatant of the strain W10 on the first day of fermentation showed different effects on the germination of soybean seeds. Since the cellulase activity content in the supernatant on the first day was low, the diluted supernatant could not make the old seeds germinate like the blank treatment. However, since soybean skin is rich in 40% cellulose, the fermentation liquid containing hyphae can promote the decomposition of cellulose and then promote the germination of old seeds. The seed germination effect of the fermentation supernatant of the strain W10 on the second and third days of fermentation also confirmed this point, so the strain W10 has the potential to be developed into 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 content in the leaves in the fermentation liquid treatment on the second day were significantly higher than those in CK. Although the various indicators of soybeans in the fermentation supernatant were also higher than those in CK, they did not show significant differences. It can be seen 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 is 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 the effective phosphorus content in the medium after strain W10 was inoculated into an organic phosphorus liquid medium with lecithin as the only phosphorus source and cultured for different periods of time;

[0026] Figure 4 This is the result graph showing the change of cellulase activity with 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 with culture time after strain W10 was inoculated into PDA liquid culture medium;

[0028] Figure 6 The results of the fermentation liquid and fermentation supernatant of strain W10 on the germination of fresh soybean seeds and old soybean seeds in the season 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 B is the result of the effect of the fermentation liquid and supernatant of strain W10 on the length of the embryonic axis during the germination of fresh soybean seeds in that season. Figure 6 C in the figure is 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 hypocotyl length during the germination of old soybean seeds;

[0029] Figure 7The results of the effects of the fermentation liquid and fermentation supernatant of strain W10 on the chlorophyll content and carotenoid content in soybean seedling leaves are shown in the figure; 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 is the result 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 is the effect of the fermentation liquid and fermentation supernatant of strain W10 on the total chlorophyll content in soybean seedling leaves. Figure 7 D in the figure is 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 scheme and advantages of the present invention more clear, the present invention is further described in detail below in conjunction with specific implementation methods and the accompanying drawings of the specification. 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 part of the embodiments of the present invention rather than all the 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 all conventional methods unless otherwise specified, and the materials, reagents, culture media and instruments used are all conventional materials, reagents, culture media and instruments in the art unless otherwise specified, and can be obtained by those skilled in the art through commercial channels.

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

[0033] The fresh soybean variety provided by the present invention for testing in the current season 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 involved in the present invention is as follows:

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

[0037] The organophosphorus culture medium consists 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 has been 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. According to the results of homologous gene and pan-genome analysis, different phylogenetic trees were constructed based on single-copy core gene data sets in a neighbor-joining clustering manner. Combining the results of phylogenetic trees based on homologous genes and gene family contraction and expansion evolutionary trees (see Figure 1 and Figure 2 ), strain W10 was identified as Talaromyces amestolkiae.

[0040] Example 2: Detection of the phosphate solubilizing activity of Bacillaceae 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 the 1st, 2nd, 3rd, 4th, 5th, 6th and 7th days respectively, and the effective phosphorus content in the supernatant was detected by the molybdenum antimony colorimetric method.

[0042] The strain W10 was inoculated into a Montana 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 in organic phosphorus. In the organic phosphorus culture medium with lecithin as the only phosphorus source, the effective 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 Bacillaceae W10 to degrade cellulose

[0044] Drawing of standard curve: Accurately prepare 400μg / mL, 600μg / mL, 800μg / mL, 1200μg / mL, 1600μg / mL, and 2000μg / mL glucose standard solutions, draw 1mL of each standard solution into a 20mL stoppered colorimetric tube, add 2mL of water and 2mL of DNS reagent, boil in a water bath for 5min, and dilute to 20mL with water after cooling, adjust to zero with a blank control, and detect its absorbance at a wavelength of 540nm. With absorbance as the horizontal coordinate and glucose concentration as the vertical coordinate, calculate the regression equation: y=1176.9x+69.233, and the 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 liquid was taken into a conical flask, diluted 3 times with citrate buffer, placed on a shaker for 30 min, centrifuged at 5000 r / min for 10 min, and the supernatant was collected as the enzyme solution to be tested.

[0046] Cellulase activity detection: Pipette 2mL of 0.51% sodium carboxymethyl cellulose solution into a 20mL stoppered colorimetric tube, place in a 50℃ water bath for 5min, add 0.5mL of preheated enzyme solution to be tested, shake well, place in a 50℃ water bath for 30min, add 2mL of DNS solution, cool to room temperature after a 5min boiling water bath, dilute to 20mL with distilled water, detect its absorbance at a wavelength of 540nm, set 3 replicates, and calculate the average value. Use a colorimetric tube that has been added with enzyme solution and DNS solution in advance as a blank control. Define the amount of enzyme that degrades sodium carboxymethyl cellulose to produce 1μg of glucose in 1mL of sample within 1min as 1 enzyme activity unit (U / mL).

[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 Bacillaceae W10 to produce indoleacetic acid

[0049] The content of indoleacetic acid (IAA) in the culture medium was determined by enzyme-linked immunosorbent assay. Add 50 μL of appropriately diluted sample to the microplate coated with indoleacetic acid antibody, add 50 μL of enzyme-labeled reagent, incubate at 37°C for 60 min, wash with washing solution 3-5 times, dry the water, add 100 μL of TMB and color for 15 min in the dark, add 50 μL of stop solution to terminate the reaction, use blank sample as control, and detect the absorbance of each well at 450 nm wavelength. 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 of the strain 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, the 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] The strain W10 was inoculated into PDA liquid culture medium for fermentation culture. The obtained fermentation broth and fermentation supernatant were diluted 1000 times with sterile water, respectively. The sterilized PDA liquid culture medium dilution was used as a blank control. Five sterilized soybean seeds were placed in each culture dish padded 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 added to each group and the dishes were 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 liquid and supernatant of strain W10 can significantly promote the germination of fresh soybean seeds (Williams 82) of the season, and the effect is most obvious on the second day. The fermentation supernatant on the second day has a significant difference in effect on the growth of soybean radicle length and axis length compared with CK, and the fermentation liquid on the second day has a significant difference in effect on the growth of soybean hypocotyl length compared with CK. In the experiment to detect the effects of the fermentation liquid 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 liquid groups 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 second day treatment group 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 ordinary black soil in the Northeast field. After collecting the soil, it was divided into seedling pots (upper diameter 11.6 cm, height 10.5 cm), 1.5 kg of black soil per pot, and the test seeds were Williams 82. The strain W10 was inoculated into PDA liquid culture medium and cultured with shaking at 28°C and 170 rpm. On the second day of culture, the fermentation liquid was aspirated, and a part of the fermentation liquid was centrifuged at 10000 rpm and 25°C for 10 min, and the fermentation 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] Uniform and relatively full soybean seeds were selected and disinfected before being sown in seedling pots, with one seed per pot and five treatments per group. 2 mL of strain W10 fermentation liquid or fermentation supernatant was evenly injected with a syringe 1-2 cm near the seed sowing. CK had no special treatment. Water was poured every 5 days after sowing. Plant height, fresh weight, dry weight, root length, root weight and number of nodules of soybean seedlings were tested 30 days later (see Table 1).

[0058] As shown in Table 1, when the soybean seedlings treated with the fermentation liquid and the fermentation supernatant of the strain grew to 30 days, their plant height, fresh weight, dry weight, root length, root weight and number of nodules were significantly higher than those of the blank control group. Only the plant height of the two treatment groups was significantly different from that of CK, and there was no significant difference in the root length between the two treatment groups and CK. Among the other indicators, only the fermentation liquid treatment group was significantly different from that of 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 liquid and fermentation supernatant of strain W10 on the growth of soybean seedlings

[0060]

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

[0062] The test soil was ordinary black soil in the Northeast field. After collecting the soil, it was divided into seedling pots (upper diameter 11.6 cm, height 10.5 cm), 1.5 kg of black soil per pot, and the test seeds were Williams 82. The strain W10 was inoculated into PDA liquid culture medium and cultured with shaking at 28°C and 170 rpm. On the second day of culture, the fermentation liquid was aspirated, and a part of the fermentation liquid was centrifuged at 10000 rpm and 25°C for 10 min, and the fermentation 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] Select uniform and relatively full soybean seeds and sow them in seedling pots after disinfection. One seed per pot, five treatments per group. Use a syringe to evenly inject 2 mL of strain W10 fermentation liquid or fermentation supernatant 1-2 cm near the seed sowing. CK has no special treatment. Water once every 5 days after sowing. After 30 days, test the chlorophyll content and carotenoid content in the leaves of soybean seedlings (see Figure 7 ).

[0065] Chlorophyll content detection: accurately weigh 0.1g of fresh leaves, cut them into pieces and put them into a 2mL centrifuge tube, add 1mL of distilled water, put them into a tissue homogenizer and crush them for 1min, draw the extract (absolute ethanol: acetone volume ratio = 1:2) and rinse the crushed tissue into a 10mL centrifuge tube, make the volume up to 10mL with the extract, extract for 3h in a dark environment, observe the color of the tissue residue at the bottom, if it is close to white, it is completely extracted. Take the extract as the blank control, and detect its absorbance at 645nm and 663nm wavelengths respectively.

[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 detection: accurately weigh 0.1g of fresh leaves, cut them into pieces and put them into a 2mL centrifuge tube, add 1mL of distilled water, put them into a tissue homogenizer and crush them for 1min, draw the extract (distilled water: acetone volume ratio = 1:4) and rinse the crushed tissue into a 10mL centrifuge tube, make the volume up to 10mL with the extract, extract for 3h in a dark environment, observe the color of the tissue residue at the bottom, and if it is close to white, it is completely extracted. Use the extract as a blank control, and detect its absorbance at wavelengths of 646nm, 663nm and 470nm respectively.

[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 potted 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 as above in the form of preferred embodiments, it 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. Use of a fungus W10 of the genus Talaromyces in dissolving organic phosphorus, characterized in that: The preservation number of the Talaromyces fungus W10 is CGMCC NO.40621.

2. The use according to claim 1, characterized in that: The organic phosphorus is lecithin.

3. Use of a phosphate-dissolving bacterial agent containing the Tularemella fungus W10 described in claim 1 in dissolving organic phosphorus in soil.

4. The use according to claim 3, characterized in that: The organic phosphorus is lecithin.

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

6. The use of the Talaromyces fungus W10 described in 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.

7. The use according to claim 6, characterized in that: The soybean seeds are fresh seeds of the season or old seeds.

8. The use of the Talaromyces fungus W10 in 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.

9. Use of the Talaromyces fungus W10 as claimed in 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.

10. Use of the Talaromyces fungus W10 as claimed in 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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