CN115011488B - Penicillium purpureum C1-GP and application thereof in cultivation of blueberry mycorrhizal seedlings - Google Patents
Penicillium purpureum C1-GP and application thereof in cultivation of blueberry mycorrhizal seedlings Download PDFInfo
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Abstract
The invention relates to the technical field of blueberry culture, in particular to penicillium purpureum (Penicillium janthinellum) C1-GP and application thereof in cultivation of blueberry mycorrhizal seedlings. The preservation number of the penicillium violaceum C1-GP is GDMCCNO:62424. compared with seedlings cultivated by the prior seedling cultivation technology, the blueberry mycorrhizal seedlings cultivated by the penicillium violaceum C1-GP have the advantages that the mycorrhizal infection rate is improved by 280% -340%, the height of the blueberry mycorrhizal seedlings is increased by 29.9% -41.8%, the ground diameter growth is increased by 11.7% -16.0%, the root activity is improved by 1.38-1.55 times, the disease and pest resistance is enhanced, and the seedling quality and later-stage comprehensive benefits are obviously improved.
Description
Technical Field
The invention relates to the technical field of blueberry cultivation, in particular to penicillium purpureum C1-GP and application thereof in cultivation of blueberry mycorrhizal seedlings.
Background
As a new small berry tree seed with higher economic value and wide development prospect, the blueberry has fine and smooth fruit meat, extremely small seeds, agreeable sweetness and sourness and fresh and pleasant fragrance, and is rich in various vitamins, trace elements and other nutrient substances. The fresh blueberry fruit can be eaten as raw material, and can also be used as raw material for processing fruit juice, fruit wine, jam, etc., thus having higher economic value.
Under the big healthy background, the blueberry becomes the fine fruit that is favored by the consumer gradually, and its planting area increases rapidly, in recent years, and the demand to the high-quality seedling of blueberry is constantly increased, and the blueberry nursery stock that uses at present is mostly the nursery stock of tradition cultivation, dies easily after transplanting, grows slowly, still accompanies the phenomenon emergence of blooming and result delay, serious influence the development of blueberry industry.
Disclosure of Invention
In order to solve the problems, the invention provides penicillium violaceum C1-GP and application thereof in cultivation of blueberry mycorrhizal seedlings. Compared with nursery stocks cultivated by the prior seedling cultivation technology, the blueberry mycorrhiza seedlings cultivated by the penicillium violaceum C1-GP provided by the invention are high in mycorrhiza infection rate, remarkable in growth promotion effect and strong in disease resistance.
In order to achieve the above purpose, the invention provides the following technical scheme:
the invention provides Penicillium janthinillum (Penicillium janthinellum) C1-GP with disease-resistant and growth-promoting effects, wherein the preservation number of the Penicillium purillum C1-GP is GDMCC NO:62424.
the invention also provides a microbial agent with disease-resistant and growth-promoting effects, wherein the microbial agent comprises the penicillium violaceum C1-GP;
the number of spores of the penicillium chrysogenum C1-GP is more than or equal to 1 multiplied by 10 9 one/mL.
The invention also provides application of the penicillium purpureum C1-GP or the microbial agent in promoting plant growth and/or improving plant disease and insect resistance.
Preferably, the plant comprises blueberry.
The invention also provides a cultivation method of the blueberry mycorrhizal seedlings, which comprises the following steps:
transplanting the rooted seedlings into a composite matrix; the composite matrix comprises the microbial inoculum and a basic matrix.
Preferably, the volume ratio of the microbial inoculum to the basic matrix is 1: (100-150).
Preferably, the cultivation method further comprises: after transplanting for 30-40 d, adding the microbial inoculum into a composite matrix; the addition amount is 50-60 mL/plant in terms of the number of rooted seedlings.
Preferably, the base matrix comprises peat and perlite.
Preferably, the rooting culture medium for culturing the rooting seedlings comprises perlite, vermiculite and turf; the volume ratio of the perlite to the vermiculite to the turf is 1:1:3.
preferably, the blueberries comprise brilliant, pink blue or lychee.
Has the beneficial effects that:
the invention provides Penicillium janthinillum (Penicillium janthinellum) C1-GP with disease-resistant and growth-promoting effects, wherein the preservation number of the Penicillium purillum C1-GP is GDMCC NO:62424. the penicillium violaceum C1-GP provided by the invention has the functions of promoting plant growth and/or improving plant disease and pest resistance, compared with seedlings cultivated by the prior seedling cultivation technology, the blueberry mycorrhiza seedlings cultivated by the penicillium violaceum C1-GP have the mycorrhiza infection rate improved by 280% -340%, and the blueberry mycorrhiza seedling height is increased by 29.9% -41.8%. The growth of the ground diameter is increased by 11.7-16.0%, the root activity is improved by 1.38-1.55 times, the disease and pest resistance is enhanced, and the quality of the nursery stock and the later-stage comprehensive benefit are obviously improved.
Biological preservation information
Penicillium purpureum C1-GP, named Penicillium janthinellum, was deposited at the Guangdong province microbial culture Collection (GDMCC) 26.04.2022 with the deposition address of Dazhou large laboratory building No. 100, ministry of Youqing, guangdong province, with the deposition number GDMCC NO:62424.
Detailed Description
The invention provides Penicillium janthinillum (Penicillium janthinellum) C1-GP with disease-resistant and growth-promoting effects, wherein the preservation number of the Penicillium purillum C1-GP is GDMCC NO:62424.
the penicillium violaceum C1-GP provided by the invention has the functions of promoting plant growth and/or improving plant disease and pest resistance.
The invention also provides a microbial agent with disease-resistant and growth-promoting effects, wherein the microbial agent comprises the penicillium janthinellum C1-GP;
the number of spores of the penicillium chrysogenum C1-GP is more than or equal to 1 multiplied by 10 9 one/mL, preferably 1X 10 9 one/mL.
The invention also provides application of the penicillium purpureum C1-GP or the microbial agent in promoting plant growth and/or improving plant disease and insect resistance.
In the present invention, the plant preferably includes blueberry; the blueberries preferably comprise one or more of brilliant, pink blue or lycra.
The invention also provides a cultivation method of the blueberry mycorrhizal seedlings, which comprises the following steps:
transplanting the rooted seedlings into the composite matrix.
In the present invention, the blueberries preferably comprise brilliant, pink blue or lychee.
In the present invention, the method for culturing rooted seedlings preferably comprises: carrying out rooting culture on the tissue culture seedlings or the cutting seedlings in a rooting culture medium to obtain the rooting seedlings; the rooting medium preferably comprises perlite, vermiculite and turf; the volume ratio of the perlite to the vermiculite to the turf is preferably 1:1:3. the invention has no special requirement on the source of the tissue culture seedling or the cutting seedling, and the tissue culture seedling or the cutting seedling is obtained by adopting a method well known by the technical personnel in the field. The invention preferably refers to the method for cultivating the rabbit seedlings or the seedlings cultured by the method described in the literature [ small house crystal, li Yongxia, wen Guangqin, nie Fei ] blueberry tissue culture and rapid propagation technology research [ J ]. Seed, 2014,33 (09): 118-120 ], [ Nie Fei, liao Youjiang, he Jian, dan Jianggong, zhu Zhongrong ] American blueberry eye propagation technology research [ J ]. Subtropical plant science, 2004 (04): 39-41 ], or [ Yang Ling, zou Tiancai, nie Fei, liu Haiyan, zhou Hongying, liao Youjiang ] rabbit eye blueberry seedling breeding test and application technology research [ J ]. Seed, 2008 (01): 91-96 ]. The invention can improve the rooting rate by culturing the tissue culture seedling or the cutting seedling in a proper rooting culture medium.
In the invention, the composite matrix comprises the microbial inoculum and a basic matrix; the volume ratio of the microbial inoculum to the basic matrix is preferably 1: (100 to 150), more preferably 1: (100 to 130), more preferably 1:100, respectively; the base matrix preferably comprises turf and perlite; the volume ratio of the turf to the perlite is preferably 3:1.
in the present invention, the method for preparing the composite matrix preferably includes: sterilizing a basic matrix, and mixing the sterilized basic matrix with the microbial inoculum to obtain the composite matrix; the method of sterilization treatment preferably comprises radiation sterilization, more preferably irradiation at 12kGy for 7d.
In the present invention, the cultivation method preferably further comprises: after transplanting for 30-40 days, adding the microbial inoculum into a composite matrix; the addition amount is 50-60 mL/plant in terms of the number of rooted seedlings, and the microbial inoculum is added into the composite matrix after the transplanting is carried out for 30 more preferably; the addition amount is 50 mL/plant based on the number of rooted seedlings.
In the present invention, the adding method preferably comprises irrigating to the transplanting Miao Genbu, and the irrigating is preferably covered with turf.
In order to further illustrate the present invention, the following examples are provided to describe the penicillium microphyllum C1-GP of the present invention and its application in cultivating blueberry mycorrhizal seedlings in detail, but they should not be construed as limiting the scope of the present invention.
Example 1
Screening of Penicillium violaceum C1-GP
And in 2019, in 10 months, digging a material of a brilliant variety blueberry root in Huaxi high slope in Guiyang city of Guizhou province, slightly washing soil with tap water, and picking healthy living fine roots. After cleaning, washing with 75% alcohol for 1min, then washing with 10% 84 disinfectant for 20min, and after washing, washing with sterile water for 1min for 5 times.
Then, the roots were cut into 2 to 3mm root segments, 3 root segments were placed in each dish, and then placed in an incubator at 25 ℃ for dark culture. Picking single colony with inoculating loop and inoculating to PDA culture medium with pH 5.6 for purification, each colony has 5 replicates; carrying out blueberry infection experiments on the purified strains, and screening strains with high infection rate and good growth promoting effect, wherein the infection experiments of different strains refer to example 3, the screening indexes and methods refer to comparative application example 1, and finally screening the strains with the best infection rate and growth promoting effect as the strains with the numbers of C1-GP; the colony growth speed of the C1-GP strain is high, the edge is approximately regular, the front surface is grayish purple, and the back surface is light yellow;
the C1-GP strain is sent to Chongqing Ongqing biology company for ITS molecular identification and preservation.
The identified C1-GP strain is Penicillium violaceum, the latin name is Penicillium janthinellum, the strain is preserved in Guangdong province microorganism culture collection center (GDMCC) at 26.04.2022, and the preservation number is GDMCC NO:62424.
example 2
A microbial agent with disease-resistant and growth-promoting effects is prepared by the following steps:
inoculating the penicillium violaceum C1-GP screened in the example 1 into a PDB culture medium with the pH value of 5.6, wherein the inoculation amount is 0.1%; culturing at 25 deg.C and 200rpm for 7d to obtain the microbial agent (the number of spores of Penicillium microphyllum C1-GP in the microbial agent is 1 × 10) 9 one/mL).
Example 3
A cultivation method of blueberry mycorrhizal seedlings comprises the following steps:
1) Cutting green branch and single bud of 1-1.5 cm of half lignified blueberry, washing with saturated washing powder for 10min, washing with tap water for 10min, soaking in 70% ethanol under aseptic condition for 5min, and adding 0.1% of HgCl 2 Soaking green branch in the solution for 5min, and washing with sterile water for 6 times;
2) Inoculating single bud on improved WPM medium (WPM medium + zeatin 1.5 mg/L) to induce cluster bud, hardening seedling in greenhouse with transmittance of 30%, hardening seedling time of 12d, maintaining indoor and greenhouse temperature at about 15 deg.C and humidity of about 80%, and spraying water with sprayer to make the seedling have water drop;
3) After acclimatization, the culture medium was cut with tweezers, 2cm shoots were removed, the medium was washed out in clean water, the plants washed out of the medium were treated with 2000mg/L IBA quick dip, and were cut in a medium containing prepared rooting medium (perlite: vermiculite: the volume ratio of the turf is 1:1: 3) Placing the seedling in a greenhouse, covering a small arched greenhouse on a culture disc, keeping the temperature and moisture of the small arched greenhouse, keeping the temperature in the arched greenhouse at 15-25 ℃ in winter, and performing rooting culture under the condition that the temperature is controlled not to exceed 28 ℃ in summer to obtain rooted seedlings;
4) Mixing turf and perlite 3:1 (volume ratio) to obtain a transplanting matrix; performing radiation sterilization (12kGy, 7d) treatment on the transplanting matrix, and inoculating a microbial inoculum after the sterilization treatment: inoculating 10L of the microbial agent prepared in the example 2 to each cubic meter of transplanting matrix, uniformly stirring for later use, and filling into a plastic nutrition pot with the specification of 14-16 cm;
5) Transplanting the rooted seedlings cultured in the step 3) into the plastic nutrition pots filled with the transplanting substrates in the step 4), leveling, and then closely and uniformly placing the seedlings on prepared bed compartments, wherein the bed compartments are 20cm high, 200cm wide and different in length according to terrains; performing conventional management on seedlings according to the seedling management in DB 5226/T60-2016, and performing topdressing and sprinkling irrigation at proper time;
6) After seedlings are transplanted for one month, the microbial inoculum prepared in example 2 is poured to the roots of the transplanted seedlings, 50mL of each seedling is poured, and the seedlings are covered with turf after being poured.
Example 4
A cultivation method of blueberry mycorrhizal seedlings comprises the following steps:
1) Cultivating cuttage rooting seedlings: cutting current-year-old strong branches of blueberries (brilliant), cutting the branches into cutting slips with the length of 5-8 cm, removing leaves at the bottom of the cutting slips, only keeping one leaf at the top end, immersing the roots of the cutting slips below 2cm in IBA with the concentration of 2000mg/L, and dipping for 1min;
2) Mixing turf, perlite and vermiculite 3:1:1 (volume ratio), mixing, performing radiation sterilization (12kGy, 7 d) to obtain a cutting matrix, and spreading and constructing a cutting bed with the depth of 15cm in a nutrient disc sterilized by potassium permanganate; at a line spacing of 5cm × 5cm cutting the cutting slips into a cutting medium; after cutting, placing the nutrition disc in a greenhouse for seedling culture, wherein the transmittance of the greenhouse is 70%, moisturizing by adopting intermittent spraying, and culturing at the temperature of 20-25 ℃ in the greenhouse for 30d to obtain rooting cutting seedlings;
3) Mixing turf and perlite 3:1 (volume ratio) to obtain a transplanting matrix; performing radiation sterilization (12kGy, 7d) treatment on the transplanting matrix, and inoculating a microbial inoculum after the sterilization treatment: inoculating 10L of the microbial agent prepared in the example 2 to each cubic meter of transplanting matrix, uniformly stirring for later use, and filling into a plastic nutrition pot with the specification of 14-16 cm;
4) Transplanting the rooted seedlings cultured in the step 2) into the plastic nutrition pot filled with the transplanting matrix in the step 3), leveling, and then closely and uniformly placing the rooted seedlings on a prepared bed compartment, wherein the bed compartment is 20cm high and 200cm wide, and the length is different according to the terrain; performing conventional management, timely topdressing and timely sprinkling irrigation on seedlings according to the seedling management in DB 5226/T60-2016;
5) After seedlings are transplanted for one month, the microbial inoculum prepared in example 2 is poured to the roots of the transplanted seedlings, 50mL of each seedling is poured, and the seedlings are covered with turf after being poured.
Example 5
A cultivation method similar to that of example 3, the only difference being that blueberries were replaced with brilliant blueberries.
Example 6
A cultivation method similar to that of example 3, except that the brilliant blueberries were replaced with lecy blueberries.
Comparative example 1
A cultivation process similar to that of example 3, with the only difference that the rooting medium in step 3) is replaced by peat and perlite in a ratio of 3:1 by volume ratio.
Comparative example 2
A cultivation process similar to that of example 3, with the only difference that the rooting medium in step 3) is replaced with humus and perlite in a ratio of 3:2 by volume ratio.
The rooting rates after the rooting culture of example 3, comparative example 1 and comparative example 2 were counted, 30 seedlings were treated each, and the results are averaged out and shown in table 1.
TABLE 1 rooting percentage after cultivation in different rooting media
| Rooting culture medium | The rooting percentage is% |
| Peat + perlite (3:1) | 92.3 |
| Humus soil + perlite (3:2) | 91.5 |
| Peat + perlite + vermiculite (3 | 93.6 |
As can be seen from Table 1, the rooting medium provided by the invention has the highest rooting rate.
Comparative example 3
A cultivation method similar to that of example 3, with the only difference that the inoculum in steps 4) and 6) was replaced with an equal volume of clear water.
Comparative application example 1
After two months of cultivation, the infection rate, seedling height, ground diameter, total biomass, chlorophyll content and root activity of 4 treatment groups in examples 3-6 and comparative example 3 are counted; after the seedlings are cultivated for eight months, the health conditions of the seedlings in a natural state are checked, the infection rates of the seedlings infected with powdery mildew and gray mold are counted, 30 seedlings in each treatment group are subjected to three repetitions, and the results are shown in table 2.
TABLE 2 measurement results of different incubation methods
As can be seen from Table 2, compared with the seedlings cultivated by the prior seedling cultivation technology, the mycorrhiza infection rate of the blueberry mycorrhiza seedlings cultivated by the penicillium violaceum C1-GP is improved by 280% -340%, and the blueberry mycorrhiza seedling height is increased by 29.9% -41.8%. The growth of the ground diameter is increased by 11.7-16.0%, the root activity is improved by 1.38-1.55 times, the disease and pest resistance is enhanced, and the quality of the nursery stock and the later-stage comprehensive benefit are obviously improved.
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
Claims (6)
1. The penicillium purpureum (Penicillium janthinellum) C1-GP has the effects of resisting diseases and promoting growth, and is characterized in that the penicillium purpureum C1-GP is preserved in the Guangdong province microorganism strain preservation center at 26.04.2022 years, and the preservation number is GDMCC NO:62424.
2. a microbial agent having an anti-disease and growth-promoting effect, wherein the microbial agent comprises penicillium violaceum C1-GP according to claim 1;
the number of spores of the penicillium chrysogenum C1-GP is more than or equal to 1 multiplied by 10 9 one/mL.
3. Use of penicillium violaceum C1-GP according to claim 1 or a microbial inoculant according to claim 2 for promoting plant growth and/or improving plant disease and pest resistance; the plant is blueberry; the blueberries comprise one or more of brilliant, pink blue and lychee.
4. A cultivation method of blueberry mycorrhizal seedlings is characterized by comprising the following steps:
transplanting the rooted seedlings into a composite matrix; the composite matrix comprises the microbial inoculum of claim 2 and a base matrix;
the rooting culture medium for culturing the rooting seedlings comprises perlite, vermiculite and turf; the volume ratio of the perlite to the vermiculite to the turf is 1:1:3;
the basic matrix comprises turf and perlite;
the blueberries comprise one or more of brilliant, pink blue and lychee.
5. A cultivation method according to claim 4, wherein the volume ratio of the microbial inoculum to the base substrate is 1: (100-150).
6. The cultivation method as claimed in claim 4 or 5, further comprising: after 30-40 days of transplanting, adding the microbial inoculum of claim 2 into a composite matrix; the addition amount is 50-60 mL/plant in terms of the number of rooted seedlings.
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| ZA2022/09657A ZA202209657B (en) | 2022-05-31 | 2022-08-30 | Method for cultivating mountainous blueberry mycorrhiza seedlings |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102925363A (en) * | 2012-09-17 | 2013-02-13 | 中国农业科学院农业环境与可持续发展研究所 | Screening method of penicillium janthinillum and application thereof |
| CN106916765A (en) * | 2017-02-21 | 2017-07-04 | 江南大学 | A kind of method that utilization penicillium janthinellum adsorbs heavy metal in waste water zinc |
| CN113502228A (en) * | 2021-06-22 | 2021-10-15 | 浙江大学 | Penicillium janthinellum strain, biocontrol agent and application |
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| WO2016210238A1 (en) * | 2015-06-26 | 2016-12-29 | Indigo Agriculture, Inc | Penicillium endophyte compositions and methods for improved agronomic traits in plants |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102925363A (en) * | 2012-09-17 | 2013-02-13 | 中国农业科学院农业环境与可持续发展研究所 | Screening method of penicillium janthinillum and application thereof |
| CN106916765A (en) * | 2017-02-21 | 2017-07-04 | 江南大学 | A kind of method that utilization penicillium janthinellum adsorbs heavy metal in waste water zinc |
| CN113502228A (en) * | 2021-06-22 | 2021-10-15 | 浙江大学 | Penicillium janthinellum strain, biocontrol agent and application |
Non-Patent Citations (2)
| Title |
|---|
| 一株锌抗性菌株强化印度芥菜修复锌污染土壤的可行性研究;杜宪正 等;《食品与生物技术学报》;20191015(第10期);57-64 * |
| 细辛叶枯病生防真菌的筛选、鉴定及拮抗效果评价;刘亚苓 等;《中国植保导刊》;20200325(第03期);13-19 * |
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| ZA202209657B (en) | 2023-07-26 |
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