CN113801807A

CN113801807A - Growth-promoting microbial inoculum for inducing systemic resistance of processed tomatoes and application thereof

Info

Publication number: CN113801807A
Application number: CN202110765918.5A
Authority: CN
Inventors: 林青; 娄恺; 史应武; 霍向东; 杨红梅; 楚敏; 高雁; 曾军; 包慧芳; 王宁
Original assignee: Institute Of Microbial Applications Xinjiang Academy Of Agricultural Sciences (china Xinjiang-Armenia Bioengineering Research And Development Center)
Current assignee: Institute Of Microbial Applications Xinjiang Academy Of Agricultural Sciences (china Xinjiang-Armenia Bioengineering Research And Development Center)
Priority date: 2021-07-06
Filing date: 2021-07-06
Publication date: 2021-12-17
Anticipated expiration: 2041-07-06
Also published as: CN113801807B

Abstract

The invention discloses a growth-promoting bacterium Paenarthrobacter nitroguajacolicusD8 which can mediate processed tomatoes to generate induction system resistance and has a good disease prevention effect on early blight, gray mold and bacterial wilt of the tomatoes. The bacterial liquid of the strain D8 can obviously increase the jasmonic acid content of the processed tomatoes and maintain the jasmonic acid content for a certain time, so that the processed tomatoes have systemic resistance, and the disease resistance of the processed tomatoes to 3 pathogenic bacteria is enhanced. The novel growth-promoting microbial inoculum provided by the invention is used for processing tomato epidemic prevention and control, is green and environment-friendly, has low cost, and has wide application value in the technical field of expanding the application of microbial growth-promoting microbial strains.

Description

Growth-promoting microbial inoculum for inducing systemic resistance of processed tomatoes and application thereof

Technical Field

The invention relates to the technical field of microbial agents and application thereof, in particular to a Paenarthrobacter nitroguajacolicus D8 microbial growth promoting agent for inducing processed tomatoes to generate system resistance and the technical field of application thereof.

Background

The tomato processing is the dominant industry of Xinjiang-style economy, is one of the three major industries of Xinjiang, accounts for 18 percent of the total world production, and has important position in China and even the world. Early blight, bacterial wilt and gray mold are 3 serious diseases in the growth process of processed tomatoes, and the yield of the tomatoes is reduced by 10-30% every year. In recent years, diseases are increasingly generated with the increase of planting area of processed tomatoes, however, continuous cropping phenomena are commonly generated in the aspects of regional layout and industrial production, and in recent years, the types of diseases are increasingly increased and the diseases are more serious. Severely restricting the sustainable development of the local processing tomato planting industry.

However, no immune or highly resistant varieties are currently found against early blight, phytophthora, viral diseases or bacterial diseases of processed tomatoes. The disease control is mainly a method of applying chemical pesticides, the chemical pesticides need to adopt a control mode of applying different types of protective agents and therapeutic agents before and during the disease attack period, so that the production cost is increased, the problems of environmental pollution, pesticide residue and the like are easily caused, even beneficial bacteria in soil are harmed, and the ecological balance of planting soil is damaged. The identification of pathogenic bacteria and the biocontrol effect of antagonistic bacteria have become the focus of research in recent years. Many scholars have already clarified pathogenic bacteria of various diseases of tomato processed in Xinjiang, and successfully screen antagonistic bacteria with better antibacterial performance, and have certain control effect on the diseases. However, the antagonistic bacteria depends on stable and long-term colonization, is influenced by factors such as plant varieties, cultivation modes, pesticide application, field ecological environment and the like, and has unstable prevention and treatment effects.

The induction of plant resistance is one of the important means for biological control of certain endophytic bacillus, and some microbes can be used as plant activators to induce plants to generate systemic resistance, so that the plant growth regulator has a good disease prevention effect. After the biocontrol bacterium is used for treating the plants, a series of physiological reaction phenomena beneficial to enhancing the disease resistance of the plants are induced in the plants, which has important significance for the healthy growth of crops and the prevention and treatment of plant diseases. It is generally considered as a new disease control strategy for the development of the intrinsic resistance mechanism of plants.

At present, the research aiming at the tomato systemic resistance mainly focuses on the research of growth-promoting bacteria such as bacillus, photosynthetic bacteria and the like in inducing the tomato to have the systemic resistance to gray mold or early blight, the application effect needs to be improved, fewer growth-promoting bacteria suitable for inducing the tomato to have the systemic resistance to gray mold, early blight and bacterial wilt are needed to be processed, and the growth-promoting bacteria with obvious effect need to be screened urgently, and documents and patent reports about the application of a Paenarthrobacter nitroguajacolicus strain in inducing the tomato to have the systemic resistance to gray mold, early blight and bacterial wilt are not found, so that the growth-promoting bacteria capable of inducing the tomato to have the systemic resistance to gray mold, early blight and bacterial wilt are screened, and the method has a wide application prospect in eco-friendliness and cost reduction of the tomato disease.

Disclosure of Invention

Aiming at the problems that the existing processed tomatoes have various diseases and are mainly controlled by chemical pesticides, the application effect of antagonistic bacteria for biological control is unstable, and the like, and the application of the Paenarthrobacter nitroguajacolicus strain in inducing the systemic resistance of tomatoes to gray mold, early blight and bacterial wilt is not reported in documents and patents. The invention aims to provide a growth-promoting bacterium Paenarthrobacter nitroguajacolicus D8 which can mediate processed tomatoes to generate induction system resistance and has better control effect on early blight, gray mold and bacterial wilt. The influence of the strain on the systemic resistance of the induced and processed tomatoes to the tomato gray mold, the early blight and the bacterial wilt is further considered, and the result shows that the Paenarthrobacter nitirguajacolicus D8 bacterial liquid can obviously improve the jasmonic acid content of the processed tomatoes, so that the processed tomato plants generate the induced systemic resistance, and the mechanism of the strain is verified to find that the Paenarthrobacter nitirguajacolicus D8 strain has no antagonistic resistance to tomato gray mold pathogenic bacteria Botrytis cinerea (Botrytis cinerea), tomato early blight pathogenic bacteria Alternaria solani (Alternaria solani) and tomato bacterial wilt (Ralstonia solanacearum), which shows that the inhibition of the strain D8 to the tomato pathogenic bacteria does not pass the antagonistic action among the strains, but generates the induced systemic resistance through the induced and processed tomatoes, and achieves the control effect on the three types of pathogenic bacteria. The strain D8 is applied to the control of tomato epidemic situation, is green and environment-friendly, has low cost, provides a new growth-promoting bacterium for the development of tomato intrinsic resistance mechanism, and has wide application value in the technical field of expanding the application of microorganism growth-promoting bacterium strains.

In order to achieve the technical effects, the invention adopts the following technical scheme:

the invention provides a microbial agent of Paenarthrobacter nitroguajacolicus D8 for inducing systemic resistance of processed tomatoes, which is obtained by fermenting Paenarthrobacter nitroguajacolicus D8.

In the invention, a strain Paenarthrobacter nitroguajacolicus D8 is screened and separated from the rhizosphere soil of a vigna unguiculata field vigna unguiculata in West mountain farm of Wulu wood of Xinjiang, and the molecular level identification of a well-known and accepted strain system in the field is carried out to confirm the category of the obtained Paenarthrobacter, wherein the strain Paenarthrobacter nitroguajacolicus D8 is preserved in the China general microbiological culture Collection center, and the preservation number is as follows: CGMCC No.21584, preservation date: 2021, 01 month 04.

The gene sequence of the Paenarthrobacter nitroguajacolicus D8 CGMCC No.21584 strain is shown in SEQ ID NO. 1.

In the invention, the Paenarthrobacter nitroguajacolicus D8 in the microbial agent is separated and purified in a culture medium and culture conditions: 3g of beef extract powder, 10g of peptone, 5g of sodium chloride, 15g of agar and 1L of distilled water, wherein the pH value is 7.3 +/-0.1, and the beef extract powder is sterilized at 121 ℃ for 15 min.

The isolated and purified culture medium of Paenarthrobacter nitroguajacolius D8 provided above is suitable for use in the preparation of a corresponding plate medium, slant medium or liquid medium.

Meanwhile, the invention provides a preparation method of the microbial growth promoting microbial inoculum of Paenarthrobacter nitroguajacolicus D8 for inducing and processing systemic resistance of tomatoes, which comprises the following steps:

(1) activating the Paenarthrobacter nitroguajacolicus D8 preserved at low temperature on a plate culture medium, selecting a single colony on a slant culture medium, and culturing for 24h in an incubator at 37 ℃ to obtain the activated Paenarthrobacter nitroguajacolicus D8 strain.

(2) And (2) scraping a ring of the activated strain Paenarthrobacter nitroguajacolius D8 in the step (1) by using a sterile inoculating ring, inoculating the strain into 100mL of liquid culture medium, and performing shake culture for 24h at 37 ℃ and the rotating speed of a shaking table of 180r/min to obtain the Paenarthrobacter nitroguajacolius D8 microbial growth promoting bacteria liquid.

Further, the invention provides an application of the microorganism of Paenarthrobacter nitroguajacolius D8 for inducing the systemic resistance of processed tomatoes in preparing a medicament for preventing and treating the botrytis cinerea, the early blight and the bacterial wilt of the tomatoes. By using the Paenarthrobacter nitroguajacolicus D8 bacterial solution prepared by the invention to treat and process the tomato seeds, the bud length, the germination rate and the germination vigor of the tomato seeds are all obviously increased, and the root length, the fresh weight and the plant height of the tomatoes are also obviously improved.

Further, the invention provides an application of the microbial growth promoting agent of Paenarthrobacter nitroguajacolicus D8 for inducing the systemic resistance of processed tomatoes in preparing a medicament for preventing and treating the botrytis cinerea, the early blight and the bacterial wilt of the tomatoes. The bacterium solution of Paenarthrobacter nitroguajacolius D8 can significantly improve jasmonic acid content of processed tomato, so that the processed tomato plant generates induced system resistance, the induced resistance effect on three bacteria can reach more than 28%, and the epidemic situation index of the three tomato bacteria is reduced by 15-43%

By implementing the specific technical scheme provided by the invention, the following beneficial effects can be achieved by implementing the content of the invention:

(1) the invention provides a strain Paenarthhromobacter nitroguajacolicus D8, which is verified to belong to a new application of Paenarthhromobacter nitroguajacolicus with a strain number D8 in the category of the obtained Paenarthhromobacter by the molecular level identification of a well-known and accepted strain system in the field, and further needs to be preserved according to legal requirements.

(2) The invention provides a Paenarthrobacter nitroguajacolicus D8 strain which has no antagonism to Botrytis cinerea (Botrytis cinerea), tomato early blight pathogen Alternaria solani (Alternaria solani) and tomato bacterial wilt pathogen Ralstonia solani (Ralstonia solanacearum), and the inhibition of the strain D8 on tomato pathogenic bacteria is not through antagonistic action among strains, but through induction processing of systemic resistance of tomato to three pathogenic bacteria, the prevention and treatment effect is achieved. The strain D8 is applied to the control of tomato epidemic situation, is green and environment-friendly, has low cost, provides a new growth-promoting bacterium for the development of an intrinsic resistance mechanism of tomato, and has wide application value in the technical field of expanding the application of microbial growth-promoting bacterium strains.

(2) The application of the Paenarthrobacter nitroguajacolicus D8 in the prevention and treatment of the gray mold, the early blight and the bacterial wilt of the tomatoes, the prepared Paenarthrobacter nitroguajacolicus D8 bacterial liquid treats and processes the tomato seeds, the bud length, the germination rate and the germination vigor of the tomato seeds are all obviously increased, and the root length, the fresh weight and the plant height of the tomatoes are also obviously improved. The strain D8 does not inhibit tomato pathogenic bacteria through antagonistic action among strains, and the Paenarthrobacter nitroguajacolicus D8 bacterial solution can improve the jasmonic acid content of processed tomatoes, so that the processed tomato plants generate induction system resistance, the induction effect on the three pathogenic bacteria can reach more than 28%, and the epidemic situation index of the three tomato pathogenic bacteria is reduced by 15-43%. The invention provides a new growth-promoting bacterium for discovering and processing the internal resistance mechanism of tomatoes, and the strain D8 is applied to the control of tomato epidemic situation, is not only green and environment-friendly, but also has low cost, and has wide application value in the technical field of expanding the application of microbial growth-promoting bacterium strains.

Drawings

FIG. 1 is a diagram of a phylogenetic tree constructed by strain D8 based on the 16SrDNA sequence.

FIG. 2 is a colony morphology of strain D8.

FIG. 3 is a graph showing the change of jasmonic acid content of processed tomato leaves with time after D8 bacterial liquid root irrigation treatment.

FIG. 4 is an experimental graph showing the antagonism of strain D8 with tomato gray mold, early blight and bacterial wilt pathogens, wherein FIG. 4A is an antagonistic graph of strain D8 with tomato gray mold pathogen Botrytis cinerea (Botrytis cinerea); FIG. 4B is a graph showing the antagonistic activity of strain D8 against the tomato early blight pathogen Alternaria solani (Alternaria solani); FIG. 4C is a graph showing the antagonism of strain D8 against the bacterial wilt bacterium, Ralstonia solanacearum.

Detailed Description

The present invention will be described below by way of examples, but the present invention is not limited to the following examples. All the raw and auxiliary materials selected in the present invention, as well as the selected culture method of the strain and the selected DNA extraction kit are well known and selected in the art, and the% referred to in the present invention are weight percentages unless otherwise indicated. DNA extraction kit: biometrics (Shanghai) Ltd.

Example 1: isolation, screening and identification of Paenarthrobacteroidnigujacolicus D8 CGMCC No.21584

(I) separation and purification

In 2020, the soil of the rhizosphere of the vigna unguiculata field vigna unguiculata plants collected from farm vigna unguiculata in Xinjiang, and the root surface soil refers to a small amount of soil adhered to the surface of roots after a large amount of soil adhered to the root system is shaken off. Cutting off roots, weighing the roots, placing the roots into a triangular flask filled with 45mL of sterile normal saline, shaking for 1-2 min by hand until the soil adhered to the surfaces of the roots is washed, taking out the roots, and drying the water on the surfaces of the roots by using absorbent paper to obtain the roots, weighing the roots by weight (W2), wherein the weight of the roots is W1-W2. Diluting root surface soil to 10-4 times, taking 10-1,10-2,10-3 and 10-4, sucking 100 mu L of diluent, respectively coating the diluent on NA culture medium plates, and culturing in an incubator at 30 ℃. Picking single colony from plate, inoculating to new separation plate for purification, repeating for 3-4 times for purification. The purified strain is inoculated on a culture medium, cultured for 24 hours at the constant temperature of an incubator at 37 ℃, and stored at 4 ℃ for later use, see the attached figure 2.

(II) Classification and identification

Sequencing and analysis of the 16Sr RNA gene of Paenarthrobacter nitroguajacolicus D8 (hereinafter referred to as "Strain D8"):

(1) extraction of PCR template DNA

Inoculating the strain D8 into a culture medium, carrying out shake culture at 37 ℃ for 24h at the rotation speed of 180r/min, centrifugally collecting thalli, and extracting the genome DNA by adopting a genome DNA extraction kit of an Ezup-type bacteria.

(2) PCR amplification

Inoculating the strain BJT-7 into a culture medium, carrying out shake culture at the rotation speed of 3000r/min and the temperature of 37 ℃ for 24h, centrifugally collecting the thallus, and extracting the genome DNA by adopting a novel plant genome DNA rapid extraction kit. The total DNA of the strain D8 was extracted as a template for PCR amplification with primers 27F (5'-AGAGTTTGATCCTGGCTCAG-3') and 1492R (5'-GGTTACCTTGTTACGACTT-3'). Carrying out PCR amplification on a target fragment by adopting a 16S rDNA specific primer, wherein the total volume of a PCR reaction system is 25 mu L, and the PCR amplification condition is 94 ℃ for 5 min; 30s at 94 ℃, 30s at 54 ℃, 1min and 30s at 72 ℃, 35 cycles, 10min at 72 ℃ and 4 ℃ for storage.

(3) Sequence determination

The PCR amplification product is subjected to electrophoresis detection and purification and then sequenced, the 16SrDNA sequence of the strain D8 is sequenced, the partial ordered sequence of the 16SrDNA is obtained by DNAstar software splicing, the length is 1344bp, the sequence is shown as SEQ ID NO:1, BLAST homologous sequence retrieval is carried out in GenBank, a phylogenetic tree (repeated sampling is carried out for 1000 times) is established by utilizing MEGA 5.0 software commonly adopted in the field through a Neighbor-Joining method, and the result is shown in figure 1. The 16Sr DNA gene sequence of the strain D8 has the highest homology with the Paenarthrobacter nitroguajacolicus and the similarity is 100 percent, in a phylogenetic tree constructed by the 16Sr DNA gene sequence, the 16Sr DNA sequence of the strain D8 has the closest genetic relationship with the Paenarthrobacter nitroguacolicus (the accession number is NR _027199) and the confidence is 99.93 percent, and the strain D8 is determined to be a strain in the category of the Paenarthrobacter through the molecular level identification of series strains and is tentatively named as the Paenarthrobacter nitroguajacolicus D8 from the taxonomic angle.

The above-mentioned strain, Paenarthrobacter nitroguajacolicus D8, has been deposited with the International Collection of microorganisms under the Budapest treaty: china general microbiological culture Collection center (CGMCC). Address: west road No. 1, north west of the republic of kyo, yang, institute of microbiology, academy of sciences of china, zip code: 100101, preservation date of 2021, 01 month and 04 days, preservation number of CGMCC No. 21584.

Example 2: preparation of microbial growth promoting agent of Paenarthrobacter nitroguajacolicus D8 CGMCC No.21584

The embodiment provides a preparation method of a Paenarthrobacter nitroguajacolicus D8 microbial agent for inducing systemic resistance of processed tomatoes, which comprises the following specific steps:

The Paenarthrobacter nitroguajacolicus D8 separation and purification culture medium and culture conditions are as follows: 3g of beef extract powder, 10g of peptone, 5g of sodium chloride, 15g of agar and 1L of distilled water, wherein the pH value is 7.3 +/-0.1, and the beef extract powder is sterilized at 121 ℃ for 15 min.

Example 3: application of Paenarthrobacter nitroguajacolicus D8 CGMCC No.21584

(1) Effect of Paenarthrobacter nitroguajacolicus D8 Strain on germination of processed tomato seeds

Shake-culturing strain D8 in liquid culture medium at 37 deg.C under 180rpm for 24h, diluting with physiological saline to 10%⁴Multiple, bacterial liquid concentration 5.9X 10⁴cfu/ml, used for processing the seed dip of tomato seeds. Treating with clear water as a blank control, soaking the seeds in 2% sodium hypochlorite for 10min, rinsing with clear water for 5 times, soaking in 75% alcohol for 30s, rinsing with clear water for 5 times, and performing surface disinfection. The disinfected seeds are respectively soaked in diluted bacterial liquid and clear water, the temperature is kept at 37 ℃ for 4 hours, 10 seeds are sown on a culture dish paved with sterile water-wet filter paper in a spot mode, each treatment is repeated for 3 times, the culture dish is placed in a 28 ℃ incubator to be cultured in a dark place, the germination rate of the seeds is investigated after 3 days, the bud length is measured, the germination potential of the seeds is investigated after 5 days, and specific results are shown in table 1.

Table 1: the present invention provides the effect of strains on processing tomato seeds

	Bud length (cm)	Germination rate	Vigor of germination
				Control	1.73±0.68a	0.60±0.12a	0.70±0.13a
D8	3.09±0.56b	0.83±0.06b	0.97±0.06b

Note: after the data in the same column are marked with different letters, the new repolarization test is remarkable, and the lower case letter is P which is 0.05.

As can be seen from the data in Table 1, the tomato seeds processed by the bacterial liquid of the strain D8 provided by the invention have obviously improved bud length, germination rate and germination potential, wherein the bud length is increased by nearly two times, the germination rate is increased by 20%, and the germination potential is also increased by nearly 30%, which indicates that the strain provided by the invention has growth promoting effect on the tomato seeds processed, and the effect is obvious.

(2) Test of the Effect of the different treatment methods of the bacterium paenrobacter nitroguajacolicus D8 on the processing of tomatoes

In this experiment, on the basis of the test (1), the strain D8 was subjected to shake flask culture at 180rpm and 37 ℃ in a liquid medium for 24 hours. The bacterial liquid is subjected to two treatments: a is diluted to 10 with normal saline⁴Multiple, the concentration of the bacteria liquid is 10⁴cfu/ml, soaking the processed tomato seeds; b, centrifuging the bacterial liquid of the strain D8 at the rotating speed of 9000rpm for 10min, discarding the supernatant, and resuspending the precipitated thallus into 10-degree bacteria with clear water⁹cfu/ml bacterial solution, and performing root irrigation treatment on the processed tomato seeds. The control group and the root irrigation treatment group are soaked in normal saline, the seed soaking treatment group is soaked in diluted bacterial liquid, and the temperature is kept at 37 ℃ for 4 hours. Using clear water treatment as a blank control, and respectively adding the following components in percentage by weight: and (3) dibbling the seeds after the seeds are soaked in the small flowerpot with the vermiculite-2: 1 matrix, pouring 100ml of clear water into each pot of the control group and the seed soaking group, adding 15ml of heavy suspension bacteria liquid into each pot of the root irrigation group, and adding 75ml of clear water. Each treatment was repeated 3 times with 10 pots. The clear water is poured once every 5 days, and each time, 100ml of the clear water is poured in each basin. When the tomato grows to 3 leaves and 1 heart, the root length, plant height and fresh weight are measured, and specific results are shown in table 2.

Table 2: the invention provides the influence of different treatment modes of bacterial liquid of a strain D8 on tomato processing

	Root length	Plant height	Fresh weight
				Control	10.82±3.30a	18.38±2.10a	0.41±0.29a
D8 root irrigation	13.72±2.79b	20.99±2.91b	0.75±0.33b
				D8 soaking seeds	13.79±2.12b	20.50±2.70b	1.21±0.53c

Note: after the data in the same column are marked with different letters, the new repolarization test is obvious, and the lower case letter is P0.05

As can be seen from the data in Table 2, the strain D8 provided by the invention has a promoting effect on the growth of processed tomatoes regardless of seed soaking treatment or root irrigation treatment. The root length of the processed tomatoes treated by the strain D8 bacterial liquid provided by the invention is increased by nearly 3cm compared with a control group, the plant height of the processed tomatoes is increased by 3cm, and the fresh weight of the processed tomatoes is increased by 0.5g in the seed soaking treatment compared with the root soaking treatment and 0.8g in the control fresh weight. Therefore, the bacterial liquid of the strain D8 provided by the invention has obvious growth promotion effect on the germination of the processed tomatoes and the later growth process.

(3) Effect of bacterial liquid of strain D8 on the resistance of tomato processing system

In the experiment, based on the experiment 1-2, when the tomato seedlings to be processed grow to 3 leaves and 1 heart, the tomato seedlings with consistent growth vigor and plant height are selected for root irrigation treatment. The strain D8 was placed in a liquid medium and cultured in a shake flask at 37 ℃ and 180rpm for 24 h. Centrifuging the bacterial solution at 9000rpm for 10min, discarding the supernatant, and resuspending the precipitated bacterial solution with clear water to a concentration of 10⁹cfu/ml, adding 15ml of heavy suspension bacteria liquid into each processed tomato pot, and adding 75ml of clear water. Each treatment was repeated 3 times with 10 pots. Clear water treatment was used as a blank control, the watering amount was the same, and the substrate was kept moist. Collecting tomatoes continuously for 5d after root irrigationThe leaf of the tomato is measured by using a plant Jasmonic Acid (JA) enzyme-linked immunosorbent assay (ELISA) kit according to the instruction, and the specific result is shown in figure 3.

As can be seen from the data in FIG. 3, the jasmonic acid content in the control group of processed tomato leaves shows regular fluctuation along with watering, and the jasmonic acid content in the treatment group of processed tomato leaves obviously increases on day 3 after the root is irrigated with D8 bacterial liquid and is maintained for a period of time. The bacterium liquid of the strain D8 can obviously improve the jasmonic acid content of the processed tomatoes.

(4) Influence of bacterial liquid of strain D8 on disease resistance of processed tomatoes

Preparation of pathogenic bacteria: inoculating Botrytis cinerea (Botrytis cinerea) and Alternaria solani (Alternaria solani) which are pathogenic bacteria of Botrytis cinerea and early blight to a PDA culture medium plate, culturing at 28 deg.C for 7-10 days until spores grow, washing with sterile water under aseptic condition to remove spores, counting with a blood counting plate, and adjusting the concentration to 10⁶cfu/ml spore suspension. Culturing Ralstonia solanacearum in NB liquid medium at 37 deg.C under 180rpm for 24 hr to obtain 10% solution⁹cfu/ml of bacterial liquid.

Induction and inoculation: when the processed tomato seedlings grow to 3 leaves and 1 heart, selecting the tomato seedlings with consistent growth vigor and plant height, and carrying out induced inoculation on the tomato seedlings by irrigating the roots with the heavy suspension D8 bacterial liquid (the method is the same as the test (2)).

Challenge inoculation: after the induction treatment for 5 days, respectively spraying the prepared 3 pathogenic bacteria spore suspension/bacterial liquid on the leaf surfaces by using a spraying method, and preserving moisture for 24-48h after inoculation, wherein the temperature of a culture room is 25 ℃, and the humidity RH is more than or equal to 90%. The epidemic situation index and the resistance inducing effect are counted on the 5 th day and the 14 th day of the challenge inoculation, and the results are shown in tables 3-4.

Table 3: the invention provides the influence of the bacterial liquid of the strain D8 on the epidemic situation index of the processed tomatoes

Table 4: the invention provides the influence of the bacterial liquid of the strain D8 on the resistance inducing effect of processed tomatoes

	Gray mold	Early blight	Bacterial wilt
				Resistance inducing effect (%)	37.9±5.06	32.97±6.59	28.77±4.03

As can be seen from the data in tables 3-4, the tomato processed by the bacterial liquid of the strain D8 can effectively enhance the disease resistance of the plant to 3 pathogenic bacteria, and the disease degree is obviously lower than that of the control. When the bacterial solution of the strain D8 is treated on the 5 th day, the epidemic situation index of the tomato gray mold is reduced by 15% compared with that of a control group, the epidemic situation index is further reduced along with the prolonging of the treatment time, and when the bacterial solution is treated on the 14 th day, the epidemic situation index of the tomato gray mold is reduced by 22% compared with that of the control group; when the strain D8 is used for treating the tomato early blight on day 5, the epidemic index of the tomato early blight is reduced by 14% compared with that of a control group, and when the strain D8 is used for treating the tomato early blight on day 14, the epidemic index of the tomato early blight is reduced by 16% compared with that of the control group; the epidemic situation index of the tomato bacterial wilt is reduced by 43% compared with that of a control group on the 5 th day of the bacterial strain D8 bacterial liquid treatment, and the epidemic situation index of the tomato early blight is reduced by 25% compared with that of the control group on the 14 th day of the bacterial strain treatment. As can be seen from the data of the induced resistance effect of the strain D8 on the processed tomatoes in Table 4, the induced resistance effect of the strain D8 on gray mold, early blight and bacterial wilt is over 28%, the strain D8 has a remarkable growth promoting effect on seed germination and late growth of the processed tomatoes, and the epidemic index of the three main pathogenic bacteria of the processed tomatoes is remarkably reduced by the treatment of the strain D8. The invention provides a new growth-promoting bacterium for discovering and processing the internal resistance mechanism of tomatoes, and the strain D8 is applied to the control of tomato epidemic situation, so that the method is green and environment-friendly, has low cost, and has wide application value in the technical field of expanding the application of microbial growth-promoting bacterium strains.

(4) Antagonistic test of Paenarthhromobacter nitroguajacolicus D8 strain against tomato pathogenic bacteria

In order to further verify the inhibition mechanism of the strain D8 on tomato pathogenic bacteria, the test inspects the influence of the strain D8 on the main tomato processing pathogenic bacteria, the test selects tomato gray mold pathogenic bacteria Botrytis cinerea (Botrytis cinerea), tomato early blight pathogenic bacteria Alternaria solani (Alternaria solani) and tomato bacterial wilt pathogenic bacteria Ralstonia solanacearum to carry out the test, and the specific test result is shown in figure 4.

As can be seen from the results in FIG. 4, FIG. 4A shows the antagonistic effect of the strain D8 on Botrytis cinerea (Botrytis cinerea), a pathogen of tomato gray mold; 4B is the antagonistic effect of the strain D8 on tomato early blight pathogen Alternaria solani (Alternaria solani); 4B is the antagonistic effect of the strain D8 on the pathogen bacterial wilt of tomato bacterial wilt, Ralstonia solanacearum. As can be seen, the strain Paenarthrobacter nitroguajacolicus D8 has no antagonism against Staphylococcus griseus (Botrytis cinerea), tomato early blight pathogen Alternaria solani (Alternaria solani) and tomato bacterial wilt pathogen Ralstonia solani (Ralstonia solanacearum), indicating that the strain D8 exerts a function of enhancing disease resistance against three tomato pathogens by inducing and processing systemic resistance of tomato instead of antagonism. The strain Paenarthrobacter nitroguajacolicus D8 is applied to the control of tomato epidemic situation, is green and environment-friendly, has low cost, provides a new growth-promoting bacterium for the development of an internal resistance mechanism of processed tomatoes, and has wide application value in the technical field of application of expanding microbial growth-promoting bacterium strains.

The above examples are merely illustrative for clearly illustrating the present invention and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications can be made while remaining within the scope of the present invention.

Sequence listing

<110> institute of microorganism application of academy of agricultural sciences in Xinjiang (Xinjiang-Yameiya bioengineering research and development center, China)

<120> growth promoting microbial inoculum for inducing systemic resistance of processed tomatoes and application thereof

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<213> Paenarthrobacter nitroguajacolicus D8

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taggcggttt gtcgcgtctg ctgtgaaaga ccggggctca actccggttc tgcagtgggt 540

acgggcagac tagagtgcag taggggagac tggaattcct ggtgtagcgg tgaaatgcgc 600

agatatcagg aggaacaccg atggcgaagg caggtctctg ggctgtaact gacgctgagg 660

agcgaaagca tggggagcga acaggattag ataccctggt agtccatgcc gtaaacgttg 720

ggcactaggt gtgggggaca ttccacgttt tccgcgccgt agctaacgca ttaagtgccc 780

cgcctgggga gtacggccgc aaggctaaaa ctcaaaggaa ttgacggggg cccgcacaag 840

cggcggagca tgcggattaa ttcgatgcaa cgcgaagaac cttaccaagg cttgacatga 900

accggaaaga cctggaaaca ggtgccccgc ttgcggtcgg tttacaggtg gtgcatggtt 960

gtcgtcagct cgtgtcgtga gatgttgggt taagtcccgc aacgagcgca accctcgttc 1020

tatgttgcca gcgcgttatg gcggggactc ataggagact gccggggtca actcggagga 1080

aggtggggac gacgtcaaat catcatgccc cttatgtctt gggcttcacg catgctacaa 1140

tggccggtac aaagggttgc gatactgtga ggtggagcta atcccaaaaa gccggtctca 1200

gttcggattg gggtctgcaa ctcgacccca tgaagtcgga gtcgctagta atcgcagatc 1260

agcaacgctg cggtgaatac gttcccgggc cttgtacaca ccgcccgtca agtcacgaaa 1320

gttggtaaca cccgaagccg gtgg 1344

Claims

1. A microbial growth promoting agent of Paenarthhromobacter nitroguajacolicus D8 for inducing systemic resistance of processed tomatoes is characterized in that the microbial growth promoting agent is obtained by activated culture of the Paenarthhromobacter nitroguajacolicus D8.

2. The plant growth promoting microbial inoculant of paenrobacter nitroguajacolius D8 for inducing systemic resistance in processing tomatoes as claimed in claim 1, wherein the deposit number of the paenrobacter nitroguajacolius D8 is CGMCC No. 21584.

3. The pro-growth microbial inoculant for the microorganism, Paenarthrobacter nitroguajacolius D8 for inducing systemic resistance in processing tomato as claimed in claim 1, wherein the gene sequence of the Paenarthrobacter nitroguajacolius D8 is as shown in SEQ ID NO 1.

4. The plant growth promoting microbial inoculant of paenrobacter nitroguajaccicus D8 for inducing systemic resistance in tomato processing as claimed in claim 1, wherein the isolated and purified culture medium and culture conditions of paenrobacter nitroguajaccicus D8 are as follows: 3g of beef extract powder, 10g of peptone, 5g of sodium chloride, 15g of agar and 1L of distilled water, wherein the pH value is 7.3 +/-0.1, and the beef extract powder is sterilized at 121 ℃ for 15 min.

5. The plant growth promoting microorganism of Paenarthrobacter nitroguajaccius D8 for inducing systemic resistance in processing tomato according to claim 4, wherein the isolated and purified culture medium of Paenarthrobacter nitroguajaccius D8 can be used for preparing a plate culture medium, a slant culture medium or a liquid culture medium.

6. A preparation method of a Paenarthrobacter nitroguajacolicus D8 microbial growth promoting microbial inoculum for inducing and processing tomato systemic resistance is characterized by comprising the following steps:

7. An application of the microorganism Paenarthrobacter nitroguajacolicus D8 in preparing the medicines for preventing and treating the gray mould, early blight and bacterial wilt of tomato.

8. An application of the microbial growth-promoting microbial inoculum of Paenarthrobacter nitroguajacolicus D8 for inducing the systemic resistance of tomato in preparing the medicines for preventing and treating the gray mold, early blight and bacterial wilt of tomato.