CN112772678B

CN112772678B - Application of Bacillus aryabhattai strain MB35-5 or microbial agent containing strain in regulation and control of plant stress resistance

Info

Publication number: CN112772678B
Application number: CN202110320433.5A
Authority: CN
Inventors: 康耀卫; 郭兴龙; 苑莹; 梁韵玲
Original assignee: Hebei Mengbang Biotechnology Co ltd
Current assignee: Hebei Mengbang Biotechnology Co ltd
Priority date: 2021-03-25
Filing date: 2021-03-25
Publication date: 2021-09-03
Anticipated expiration: 2041-03-25
Also published as: CN112772678A

Abstract

The invention provides an application of a bacillus aryabhattai strain MB35-5 or a microbial agent containing the strain in regulation and control of plant stress resistance, and relates to the technical field of microbial application. The invention discovers that the bacterial suspension of the Bacillus aryabhattai strain MB35-5 can promote the growth and development of monocotyledons or dicotyledons under the adverse environment, especially can improve the low temperature resistance and drought resistance of plants, and has important application value.

Description

Application of Bacillus aryabhattai strain MB35-5 or microbial agent containing strain in regulation and control of plant stress resistance

Technical Field

The invention relates to the technical field, in particular to application of a bacillus aryabhattai strain MB35-5 or a microbial agent containing the strain in regulation and control of plant stress resistance.

Background

Plant stress resistance refers to the resistance of a plant to stress or various stress factors. During the growth and development of plants, various biotic stresses and abiotic stresses are applied to the plants, and the growth and development, yield and quality of the plants are directly and seriously affected by the adverse circumstances. Abiotic stresses generally include drought, low temperature, rain, freezing, dry hot wind, typhoon, and the like, wherein low temperature and drought are the factors that have the greatest effect on crop yield among all abiotic stresses. The exploration of how to minimize the damage of adversity stress to plants becomes a key problem to be solved by research, researches the cold resistance and drought resistance of the plants and improves the tolerance of the plants, and has important significance for ecological environment protection and construction. The main methods for improving stress resistance of plants at present include genetic engineering methods, physical methods, methods using plant hormones (BR), methods using chemical agents, and methods using microbial agents, which have recently attracted much attention.

The microbial synergistic improvement of the stress resistance of the plants refers to the capability of promoting the growth of the plants, preventing and controlling plant pests and improving the stress resistance of the plants by partial beneficial microbes. The utilization of soil microorganisms to improve the stress tolerance of crops is a hot point of research in recent years. For example, Sunyan et al report that the growth-promoting rhizobacteria can enhance the resistance of plants against biotic stress (including pathogenic bacteria, pests, etc.) and abiotic stress (drought, salt damage, heavy metals, etc.). Arbuscular Mycorrhizal (AM) fungi can reduce stress to a certain extent and enhance the stress resistance of plants. It has also been found that certain endophytic fungi can enhance the tolerance of plants to biotic and abiotic stresses (drought, high temperature, mineral imbalance and high salt, etc.) (xu guang et al, 2006). The inventor reports that after the rape is inoculated with endophytic fungi Piriformospora indica, the expression of drought-resistant related genes BnGG2, BnD11, BnMPK3 and Bn-PKL is induced, the resistance of the rape to drought stress is enhanced, and the report also proves that the Piriformospora indica improves the drought-resistant capability of the corn by regulating the expression of ABA, NO and programmed cell death related genes, and can also improve the salt tolerance of crops such as barley, cotton, corn, tobacco, alfalfa and the like. (Duyun et al, 2015, Li Ling, 2016) report that dark color endophytic fungi are separated to improve the stress resistance of host plants, and Chaetomium globosum ND35 can obviously improve the stress resistance of crops.

Bacillus aryabhattai is a bacterium discovered at the beginning of the 21 st century, in 2009, a new Bacillus species in the stratosphere of 20-41 km was isolated by a hot air balloon with a low-temperature sampler emitted by the institute of Hadlahaba, India, and named as Bacillus aryabhattai (Bacillus aryabyabhattatai) under the name of Yellopoh, India astronaut, and the Bacillus aryabhattai is for short. Since the discovery of Bacillus aryabhattai, a great deal of research has been carried out by scholars at home and abroad. For example, Lizuhong and the like apply for a patent of Bacillus aryabhattai for preventing and treating tobacco black shank, while Jianchand applies for an invention patent of a compound microbial inoculum for preventing and treating wheat take-all, and the microbial inoculum comprises the Bacillus aryabhattai separated from soil. The researches show that the bacillus aryabhattai has obvious effects of resisting diseases and promoting growth of plants. However, the research on the plant stress resistance of bacillus aryabhattai is relatively few, and only some salt tolerance reports exist. Therefore, there is a need to further study and discover the functions and applications of Bacillus aryabhattai.

In view of this, the invention is particularly proposed.

Disclosure of Invention

In order to solve the technical problems in the prior art, the invention aims to provide a novel application of a Bacillus aryabhattai (Bacillus aryabhattai) strain MB 35-5.

The technical scheme provided by the invention is as follows:

in a first aspect, the invention provides the use of Bacillus aryabhattai (Bacillus aryabhattai) strain MB35-5 or a microbial agent containing the strain for regulating plant stress resistance.

The Bacillus aryabhattai strain MB35-5 related by the invention is already preserved in the common microorganism center of China Committee for culture Collection of microorganisms in 2019, 16.01.4.3 of the institute of microorganisms of China academy of sciences, the address of which is No. 1 Xilu-Beijing Hokko-Yang district, the preservation number is as follows: CGMCC No. 17204. The present invention aims to protect the new use of said known strains.

In one embodiment, said modulating plant stress resistance refers to increasing low/low temperature tolerance of a plant.

In one embodiment, said modulating plant stress resistance refers to increasing plant drought resistance.

In a specific embodiment, the plant comprises a monocot or a dicot; preferably, the plant is a crop plant of the class monocotyledonae or dicotyledonae.

In a specific embodiment, the plant is a graminaceous plant or a solanaceous plant under the class monocotyledonae.

It should be noted that the present invention does not limit the specific species of plants, and preferably, the plants in the present invention refer to crops; the crops are grain crops, economic crops or oil crops. Such plants include monocots including, but not limited to, e.g., rice, wheat, maize, sorghum, sugarcane, rye, shallot, garlic, and the like; the plant also includes monocots including, but not limited to, for example, tobacco, soybean, black soybean, cabbage, cotton, radish, tomato, watermelon, and the like. Alternatively, combinations of these plants are also included in some cases.

In one embodiment, the regulating plant stress resistance of the invention is improving drought resistance and improving low temperature resistance of a plant. For the plants involved in the invention, the Bacillus aryabhattai strain or the microbial agent containing the Bacillus aryabhattai strain can improve the drought resistance of the plants and can also improve the low-temperature resistance of the plants.

The improvement of the drought resistance of the plant refers to that the strain or the microbial agent containing the strain can promote the growth or development of the plant under the adverse conditions of dry stress, drought or water shortage, for example, the water loss of some plants is slowed down under the drought, and the yellowing and wilting degree of some plants is lighter. The improvement of the low temperature resistance of the plant means that the plant has stronger low temperature tolerance and stronger low temperature adaptability, so that the growth and development of the plant under low temperature stress and the physiological function of the plant are less influenced, for example, the germination and growth of seeds of the plant under low temperature stress are promoted. Therefore, the invention also provides the application of the strain or the microbial agent in promoting the germination of crops under low temperature stress.

"Low temperature" as referred to in the present invention means an environmental temperature above 0 ℃ and below the normal growth temperature of a plant, causing damage or even death of the plant, preferably an environmental temperature below 15 ℃, or more preferably below 10 ℃.

In a second aspect, the invention also provides the use of Bacillus aryabhattai (Bacillus aryabhattai) strain MB35-5 or a microbial agent containing the strain in the preparation of a product for improving stress resistance of plants.

In a particular embodiment, the product comprises a soil amendment, a seed coating agent or a microbial fertilizer.

In a third aspect, the present invention provides a method for improving plant stress resistance, comprising treating a plant with the aforementioned bacillus aryabhattai strain or a microbial agent comprising said strain, thereby improving plant stress resistance.

In a specific embodiment, said treating the plant comprises applying said strain to the plant, to the plant seed, or to the soil surrounding the plant or plant seed in a bacterial suspension; preferably, the bacterial suspension is used to coat plant seeds.

The preparation method of the bacterial suspension comprises the following steps: inoculating Bacillus aryabhattai to a bacterial culture medium and culturing to obtain OD_600nmThe bacterial liquid with the value of 0.5-1.0 is the bacterial suspension.

The microbial agent in the invention refers to a microbial agent containing the bacillus aryabhattai strain or a fermentation product thereof. The microbial agent may also comprise a solid carrier or a liquid carrier; the microbial agent may be in different forms, such as a liquid, an emulsion, a suspension, a powder, a granule, and the like.

Has the advantages that:

the invention discovers that the Bacillus aryabhattai strain MB35-5(Bacillus aryabhattai) also has the function of improving the stress resistance (low temperature resistance and drought resistance) of plants for the first time, and has extremely high commercial value and wide market prospect.

After the strain provided by the invention is prepared into bacterial suspension to treat seeds of monocotyledons and dicotyledons, the germination index of crop seeds is obviously improved compared with that of a control group under low-temperature adversity stress.

The strain is applied to the planting of monocotyledonous and dicotyledonous crops, has a great effect of improving the drought resistance of grain crops, particularly corn and economic crops, particularly tobacco, and is beneficial to ensuring the improvement of the yield and quality of the crops under the drought condition.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.

FIG. 1 is a graph showing the effect of various treatments on corn seed germination at different temperatures according to an embodiment of the present invention (where a is a room temperature condition of 25 ℃ to 30 ℃ and b is a low temperature condition of 10 ℃);

FIG. 2 is a graph showing the effect of various treatments on soybean seed germination at different temperatures according to an embodiment of the present invention (where c is a room temperature condition of 25 ℃ to 30 ℃ and d is a low temperature condition of 10 ℃);

FIG. 3 shows the effect of various treatments at different temperatures on the germination of black soybean seeds (where e is 25-30 ℃ at room temperature and f is 10 ℃ at low temperature) provided by an embodiment of the present invention;

FIG. 4 shows the leaf morphology of drought-stressed maize (Kyomyi No.: wherein CK represents a water-coated maize blank, 3 replicates; MB35-5 is a maize coated with MB35-5 Bacillus aryabhattai, 3 replicates);

FIG. 5 shows the leaf morphology of drought-stressed maize (Tahito 608) (where CK represents a water-coated maize blank, 3 replicates; MB35-5 represents a MB35-5 Bacillus aryabhattai coated maize, 3 replicates);

FIG. 6 shows the leaf morphology of tobacco under drought stress according to an embodiment of the present invention.

Detailed Description

The technical solutions of the present invention will be described clearly and completely with reference to the following embodiments, and it should be understood that the described embodiments are some, but not all, embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Example 1

1. Experimental Material

1.1 Strain Material

The strain materials used in the invention are:

bacillus aryabhattai (Bacillus aryabhattai) MB35-5, which is preserved in the China general microbiological culture Collection center of China Committee for culture Collection of microorganisms with the preservation number: CGMCC No. 17204;

a commercial growth-promoting strain Bacillus subtilis 92068, which is sourced from agricultural resource zoning institute of Chinese academy of agricultural sciences;

rhizobium japonicum (Brydyrhizobium japonicum) KY372, which is derived from Kangsheng (Zhaoqing) Biotech Co.

1.2 culture Medium

R2A medium: 0.50g of yeast powder, 0.50g of peptone, 0.50g of tryptone, 0.50g of glucose, 0.50g of soluble starch, 0.30g of dipotassium phosphate, 0.30g of sodium pyruvate, 0.05g of magnesium sulfate heptahydrate and 1 liter of water.

1.3 seed Material

Black beans are self-sustaining varieties of the unit; the soybean Jiyu 47, the rice Nanjing 9108, the corn Jing-Ming No. 1, the corn Jing-Ming-Suo 608 and the tobacco seeds are commercially available.

2. Experimental methods

2.1 Germination experiments

2.1.1 preparation of bacterial suspension

Respectively inoculating Bacillus aryabhattai MB35-5, Bacillus subtilis 92068 and Rhizobium sojae KY372 strains into 250mL conical flasks containing 50mL R2A liquid culture medium, culturing at 30 ℃ and 200rpm/min for 48 hours, and respectively preparing into OD_600nmThe bacterial solution was 0.1 for use.

2.1.2 seed coating

Spraying 300 μ L of the above different bacterial suspensions to each 100g of seeds, shaking for use, and setting the water-coated seeds as blank Control (CK).

2.1.3 preparation of agar 0.5% for germination test

The seeds which are full, uniform in size and not broken and have been subjected to the coating treatment are selected, inoculated into a culture medium with agar concentration of 0.5% (agar 5g and water 1000 ml) for germination test, 10 seeds are placed in each dish, each treatment is repeated for 3 times, and the seeds are sealed by a sealing film and cultured in a dark place.

2.1.4 germination percentage index calculation method:

a main root: 0 grade is no germination, 1 grade is bud length of 0-0.5 cm, 2 grade is bud length of 0.5-1.5 cm, and 3 grade is more than 2 cm;

lateral rooting: 1 is 1, 2 is 2, 3 is 3 … …; x total number of lateral roots ═ 1+2+3+ … ….

(Note: the research shows that the seed development is different from the main roots, the lateral roots of the main roots in the same grade are different, in order to further show the difference between the seed development, the invention adopts a main root measuring method and a main root plus lateral root measuring method, the main roots are well developed, the seeds with slow lateral root development are suitable for the main root measuring method such as leguminous plants, the main roots are well developed, the seeds with fast lateral root development are suitable for the main root plus lateral root measuring method such as corn, rice and the like.)

Measuring a main root:

the formula for calculating the germination index of leguminous seeds is as follows:

leguminous germination index G ═ 0 × N +1 × N +2 × N +3 × N (N denotes the number of grades);

measurement of the main root plus lateral root:

the germination index calculation formula of corn and rice seeds is as follows:

germination index G ═ N × (0+ X) + N × (1+ X) + N × (2+ X) + N × (3+ X) (N denotes the number of the grade and X denotes the total number of roots on the seed side of the grade).

2.2 method of drought resistance experiment

2.2.1 preparation of bacterial suspension

Inoculating Bacillus aryabhattai MB35-5 strain into 250mL conical flask containing 50mLR2A liquid culture medium, culturing at 30 deg.C and 200rmp/min for 48 hr to obtain OD_600nmThe bacterial solution was 0.1 for use.

2.2.2 seed coating

Spraying 300 μ L of the above bacterial liquid on 100g of corn or tobacco seed, shaking for use, and setting water-coated seed as Control (CK).

2.2.3 vermiculite potted plant (corn)

Sterilized vermiculite (sterilized at 121 ℃ for 3h) was placed in sterilized plastic pots (15 cm. times.10 cm) and each pot was drenched with vermiculite before planting. Selecting uniformly-sized and coated Jing-Nian I and sweet extract glutinous 608 corn seeds, planting 6 grains in each pot, setting 3 times of the seeds, setting the seeds coated with water as blank control, and randomly arranging the seeds. According to the growth requirements, a little fertilizer is applied after 2 weeks or so, the irrigation is stopped for drought stress about 3 weeks, and the data is recorded.

Calculating the leaf curl index and the aging index (the leaf curl index and the aging index are important morphological indexes for drought resistance identification, and the smaller the leaf curl index and the leaf aging index are, the stronger the drought resistance is)

Leaf curl index: the curling size of the blade is represented, and the value range of the curling degree of the blade is between 0 and 50.

The calculation formula is as follows:

(wherein, the number of the leaves refers to the number of the visible leaves of the corn, and the unfolded leaf width is the width of the middle part of the unfolded leaf at the top of the corn.)

Leaf senescence index: the index of aging and withering and yellowing of the leaves is represented by dividing the value range of 0-10 grade by 10 of the percentage of dead leaves (yellow leaves) to the whole plant leaves. The determination is carried out 1 time every 3d during the drought stress period, and the determination is carried out 2-3 times in total.

The calculation formula is as follows:

2.2.4 peat soil perlite Mixed substrate potted plant (tobacco)

The sterilized turfy soil and perlite mixed matrix is filled in a sterilized plastic flowerpot (15cm multiplied by 10cm), and each pot of the mixed matrix is thoroughly poured before planting. Selecting tobacco seeds with uniform sizes and subjected to coating treatment, planting 6 tobacco seeds in each pot, setting 3 times of repetition, setting seeds coated with water as blank control, and randomly arranging.

According to the growth requirement, irrigating properly. And (4) carrying out transplanting culture in a pot when the tobacco seedlings grow to five leaves and one heart, stopping watering for drought stress about 60 days, and recording data.

3. Results of the experiment

Results in terms of low temperature stress resistance:

3.1 MB35-5 has obvious promotion effect on corn seed germination under low temperature adverse conditions

The germination experiments of the corn seeds coated with the MB35-5 bacterial liquid are respectively carried out at 10 ℃ (low temperature) and 25 ℃ -30 ℃ (the optimum temperature for corn growth), the corn seeds coated with the commercial growth promoting bacterium bacillus subtilis 92068 are set as positive controls, the corn seeds coated with the rhizobium japonicum strain KY372 are set as negative controls, and the corn seeds coated with water are set as blank controls.

The germination condition of soybean seeds is analyzed by adopting a germination index, test data are processed by adopting SPSS software, data significance comparison is carried out by using a Ducan's new double-pole difference method, and the influence of different strains on seed germination is analyzed.

TABLE 1 influence of different temperature treatments on germination of corn seeds (Kyoto-Nippon-Hakko-No.)

Note that the difference between treatments (P < 0.05) is marked by different lower case letters in the same column of the table

FIG. 1 is a graph showing the effect of various treatments on corn seed germination at different temperatures. As can be seen from FIG. 1 (a), under optimum temperature conditions for maize (25 ℃ C. to 30 ℃ C.), the length of the roots and the number of lateral roots were also significantly greater for maize seeds coated with Bacillus aryabhattai MB35-5 than for the control (maize seeds coated with Bacillus subtilis 92068), whereas maize seeds coated with Rhizobium KY372 and maize seeds coated with water both germinated shorter and without lateral roots. And the data analysis in the table 1 shows that the germination index of the corn seeds coated with MB35-5 is significantly different from that of the corn seeds coated with 92068 at the optimum temperature (25 ℃ -30 ℃) of the corn, compared with other treatment germination indexes, so that the Bacillus aryabhattai MB35-5 has the effect of promoting the germination and development of the corn and has the most significant effect. As can be seen from FIG. 1 (b), the germination length of the corn seeds coated with the commercial growth promoting strain Bacillus subtilis 92068 was almost not different from the germination length of the corn seeds coated with Rhizobium sojae and water when the germination temperature was decreased to 10 ℃, but the corn seeds coated with Bacillus aryabhattai strain MB35-5 still developed well, the germination length was significantly higher than that of the other treatments in the experiment, and the data analysis in Table 1 also shows that the germination index of the coated corn seeds MB35-5 was significantly different from that of the corn seeds coated with the other treatments under the low temperature condition.

The result shows that the MB35-5 strain has obvious promotion effect on the germination of corn seeds under the low-temperature adversity condition, the positive control strain 92068 has good promotion activity on the germination of the corn at room temperature, and the promotion activity on the germination of the corn completely disappears under the low-temperature adversity condition. Under cold stress conditions, the germination index of MB35-5 strain was significantly higher compared to control strain 92068.

3.2 MB35-5 has obvious promotion effect on germination of rice seeds of temperature-preference crops under the condition of low-temperature adversity

The germination experiments of the rice seeds coated with the MB35-5 bacterial liquid are respectively carried out at 10 ℃ and 25 ℃ -30 ℃ (the optimum temperature for rice growth), the rice seeds coated with the commercial growth promoting bacterium bacillus subtilis 92068 are set as positive controls, the rice seeds coated with the rhizobium japonicum strain KY372 are set as negative controls, and the rice seeds coated with water are set as blank controls.

The germination condition of rice seeds is analyzed by adopting a germination index, test data are processed by adopting SPSS software, data significance comparison is carried out by using a Ducan's new double-pole difference method, and the influence of different strains on the germination of the seeds is analyzed. The result shows that MB35-5 has obvious promotion effect on rice seed germination under the condition of low temperature adversity.

TABLE 2 Effect of treatments at different temperatures on the germination of Rice seeds (Nanjing 9108)

Analysis of the data from table 2 shows: under the condition of 25-30 ℃ (the optimum temperature for rice growth), the germination index of rice seeds coated with MB35-5 is significantly different from that of negative control and blank control coating treatment, while the commercial growth-promoting bacterium bacillus subtilis 92068 only has a weak growth-promoting effect on the rice seeds of the temperature-favored crops, but the effect is almost negligible. Therefore, MB35-5 has the function of promoting the germination and development of the rice seeds which are the warm-loving crops.

When the germination temperature is reduced to 15 ℃, the commercial growth-promoting bacterium bacillus subtilis 92068 has no effect of promoting the germination and development of the rice seeds of the temperature-favored crops, and the germination index of the rice seeds coated with MB35-5 still has a significant difference compared with the germination index of the rice seeds coated with other controls.

In conclusion, the experimental results fully indicate that the arabidopsis strain MB35-5 still has the activity of promoting the germination of rice seeds under the condition of low-temperature stress.

3.3 MB35-5 has obvious promoting effect on the germination of dicotyledonous crop soybean seeds under the condition of low temperature adversity

Carrying out germination experiments on soybean seeds coated with MB35-5 bacterial liquid at 10 ℃ and 25-30 ℃ (the optimal temperature for soybean growth), setting the soybean seeds coated with commercial growth-promoting bacterium bacillus subtilis 92068 as a positive control, setting the soybean seeds coated with rhizobium japonicum strain KY372 as a negative control, and setting the soybean seeds coated with water as a blank control.

The germination condition of soybean seeds is analyzed by adopting a germination index, test data are processed by adopting SPSS software, data significance comparison is carried out by using a Ducan's new double-pole difference method, and the influence of different strains on seed germination is analyzed. The result shows that MB35-5 has obvious promotion effect on the germination of soybean seeds under the condition of low temperature adversity.

TABLE 3 Effect of treatments at different temperatures on the Germination of Soybean seeds (Jiyu 47)

FIG. 2 is a graph showing the effect of various treatments on soybean seed germination at different temperatures. As can be seen from FIG. 2 (c), under conditions of optimum temperature for soybean growth (25 ℃ C. to 30 ℃ C.), the shoot length of the soybean seeds coated with Bacillus aryabhattai MB35-5 was longer than that of the control (corn seeds coated with Bacillus subtilis 92068), while the shoot lengths of the soybean seeds coated with Rhizobium KY372 and the soybean seeds coated with water were shorter. Analysis of the data in Table 3 also shows that the germination index of the soybean seeds coated with MB35-5 is significantly different from that of the soybean seeds coated with Rhizobium japonicum KY372 and the soybean seeds coated with water, and thus it can be seen that Bacillus aryabhattai MB35-5 has the effect of promoting the germination and development of soybeans, and the effect is most significant.

As can be seen from FIG. 2 (d), when the germination temperature was decreased to 10 ℃, the shoot length of the soybean seeds coated with Bacillus subtilis 92068 was short, with almost no difference from those of the seeds coated with Rhizobium japonicum and those of the seeds coated with water. The soybean seeds coated with MB35-5 still developed well, the bud length was significantly higher than that of other experimental groups, and the data analysis in Table 3 also shows that the germination indexes of the soybean seeds coated with MB35-5 were significantly different from those of other experimental groups.

In conclusion, the experimental results fully indicate that the arabidopsis strain MB35-5 still has the activity of promoting the germination of soybean seeds under the low-temperature adversity condition.

3.4 MB35-5 has obvious promotion effect on the germination of self-sustained black bean seeds of dicotyledon crops under the condition of low temperature adversity

The black bean seeds coated with MB35-5 were subjected to germination tests at 10 ℃ and 25 ℃ -30 ℃ (optimum temperature for black bean growth), and the black bean seeds coated with commercial growth-promoting bacterium Bacillus subtilis 92068 were set as positive control and the black bean seeds coated with water were set as blank control.

The germination condition of the black bean seeds is analyzed by adopting a germination index, test data are processed by adopting SPSS software, data significance comparison is carried out by using a Ducan's new double-pole difference method, and the influence of different strains on seed germination is analyzed.

TABLE 4 Effect of treatments at different temperatures on the Germination of Black Soybean seeds (self-sustaining species)

FIG. 3 shows the effect of various treatments on the germination of black soybean seeds at different temperatures. As can be seen from FIG. 3 (e), the bud length of the black soybean seeds coated with MB35-5 and 92068 under the condition of the optimum temperature for the growth of black soybean (25 ℃ -30 ℃) was superior to that of the control group. The data analysis in Table 4 also shows that the germination index of coated MB35-5 black bean seeds and the germination index of coated 92068 black bean seeds are significantly different from the germination index of coated water. Therefore, the bacillus aryabhattai MB35-5 has the effect of promoting the germination and development of the black beans and has the most remarkable effect.

As can be seen from (f) in FIG. 3, when the germination temperature was decreased to 10 ℃, the bud length of 92068-coated black soybean seeds was almost the same as that of coated black soybean seeds, and they were short and small. However, the black bean seeds coated with MB35-5 still developed well, and the bud length was significantly higher than that of the other groups. Table 4 data analysis also showed that there was a significant difference in the germination index of the soybean seeds coated with MB35-5 compared to the germination index of the black soybean seeds coated with other experimental groups.

In conclusion, the experimental results fully indicate that the Bacillus aryabhattai MB35-5 still has the activity of promoting the germination of the black soybean seeds under the condition of low-temperature stress.

Results in terms of drought resistance:

3.5 MB35-5 for improving drought resistance of corn

Vermiculite potting drought resistance experiments (experimental methods described in the experimental methods section above) were performed on two varieties of corn seeds (Jing Niao No. one and Jing sweet glutinous 608) coated with MB35-5, with the water-coated corn seeds set as a blank. And (4) after the irrigation is stopped and drought stress is carried out for 20 days, carrying out data analysis on the drought resistance of the corn by adopting the leaf curl index and the leaf senescence index.

TABLE 5 comparison of leaf morphology indexes of maize (Jing Niao I) under drought stress

TABLE 6 comparison of leaf morphology index of corn (extracted sweet waxy 608) under water stress

Treatment of	Index of blade curling	Leaf senescence index
			35-5	3.45^a	3.78^a
CK	6.23^b	4.67^b

FIG. 4 is a photograph of leaves of maize (Jing Niao No. one) under drought stress; FIG. 5 shows the leaf morphology of maize (sweet and waxy 608) under drought stress.

As can be seen from FIGS. 4 and 5, the number of curled leaves and yellow leaves of the two corn varieties coated with MB35-5 Bacillus aryabhattai is less than that of CK, and the growth vigor is better, and the data in tables 5 and 6 also show that the curling index and the senescence index of the two corn varieties coated with MB35-5 Bacillus aryabhattai are less than that of CK, and the difference is obvious. In conclusion, the experimental results fully show that the arabidopsis strain MB35-5 improves the drought resistance of the corn.

3.6 MB35-5 improves the anti-yellowing capacity of tobacco leaves under drought conditions

A mixed matrix potting experiment was performed on tobacco seeds coated with MB35-5, with water-coated tobacco seeds as a blank control. After the tobacco grows to 3-4 leaves, all the tobacco is stopped watering for 3 weeks, and the development condition, the yellowing condition and the wilting condition of the leaves are observed, and the specific result is shown in fig. 6.

As can be seen in FIG. 6, the tobacco coated with MB35-5 grew well compared to the control, and the number of yellow leaves was significantly less for the tobacco coated with MB35-5 than for the control. In addition, the tobacco treated with MB35-5 was significantly greener in leaf color and less wrinkled than the control, thus indicating that the tobacco coated with MB35-5 had greater drought resistance.

The research of the invention shows that the temperature range of the strain MB35-5 for promoting the germination and the development of the seeds is wide, the strain MB35-5 has the activity for promoting the germination and the development of the seeds under the conditions of normal temperature and low temperature adversity, and the promoting effect of the strain MB35-5 is more obvious under the conditions of the low temperature adversity; under drought conditions, the MB35-5 strain has obvious difference compared with a blank control and strong drought resistance, so the MB35-5 strain has the function of improving the stress resistance of plants. And because it is harmless to the environment, can replace the use of chemical agent, solve the problem such as chemical agent, etc. other methods are not good in effect or have residue, etc., protect the environment, promote the sustainable development of agriculture.

Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. The application of the Bacillus aryabhattai strain MB35-5 or a microbial agent containing the strain in regulating and controlling the stress resistance of plants; the regulation and control of the plant stress resistance is to improve the low temperature resistance and/or the drought resistance of the plant.

2. The use of claim 1, wherein the plant comprises a monocot or a dicot.

3. Use according to claim 1, wherein the plant is a crop plant of the class monocotyledonae or dicotyledonae.

4. The use of claim 1, wherein when said modulating plant stress resistance is increasing plant drought resistance, said plant is maize or tobacco.

5. The application of the Bacillus aryabhattai strain MB35-5 or a microbial agent containing the strain in preparing a product for improving the stress resistance of plants; the improvement of the plant stress resistance is the improvement of the low temperature resistance of the plant and/or the improvement of the drought resistance of the plant.

6. Use according to claim 5, wherein the product comprises a soil amendment, a seed coating agent or a microbial fertilizer.

7. A method of increasing the low temperature resistance and/or increasing the drought resistance of a plant comprising treating the plant with the bacillus aryabhattai strain of claim 1 or a microbial agent comprising said strain.

8. The method of claim 7, wherein treating the plant comprises applying the strain to the plant, to plant seeds, or to soil surrounding the plant or plant seeds in an antimicrobial suspension.

9. The method of claim 8, wherein treating the plant comprises coating the plant seed with the bacterial suspension.