CN110591957A - Salt-tolerant alfalfa rhizobium and application thereof - Google Patents
Salt-tolerant alfalfa rhizobium and application thereof Download PDFInfo
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Abstract
The invention discloses an alfalfa rhizobium salt-tolerant strain and application thereof, wherein the strain is obtained by separating rhizobium of alfalfa in Xinjiang. Determining 16S rDNA sequence of the rhizobium strain and phylogenetic analysis to determine the rhizobiumSinorhizobiumsp.) was named XGL 180. The rhizobia XGL180 has strong salt tolerance, and inoculation of the rhizobia can increase the salt tolerance of alfalfa, improve the biomass, increase the content of nitrogen, phosphorus and potassium in plants under the salt treatment condition, and is suitable for one of measures of alfalfa cultivation and production in saline soil such as Xinjiang and the like.
Description
Technical Field
The invention relates to a microbial strain resource, in particular to a salt-tolerant alfalfa rhizobium and application thereof.
Background
Rhizobia (Rhizobia) is a gram-negative bacillus-free bacterium that can symbiotically form nodules with legumes, converting free nitrogen in the air into a chemical nitrogen that can be utilized by plants. At the same time, the host plant will provide photosynthetic product carbohydrates to the rhizobia for normal reproductive growth thereof. The symbiotic nitrogen fixation effect of the rhizobia plays a role in continuously supplying nitrogen nutrition to leguminous plants, improving soil, improving and maintaining soil fertility and has important significance in agricultural production.
Alfalfa is a perennial herb of the family fabaceae, and is called the king of pasture due to its strong adaptability, high yield and rich protein. As the most important pasture plant in leguminous plants, the alfalfa has strong root system, has strong nitrogen fixation capacity with rhizobium symbiosis, not only is used as a good forage grass, but also can play a role in improving soil and fertility fields, maintaining water and soil and improving ecological environment, and is a preferred grass seed in western grassland construction and desertification transformation. The planting area of the alfalfa in China is over 130 kilohm2The main cultivation area is mostly concentrated in the northwest arid and semi-arid areas. Sinkiang is the main cultivation and production area of alfalfa in China, the alfalfa industrialization is rapidly developed along with the adjustment of the agricultural industrial structure in recent years, the planting area is rapidly enlarged, and the planting area in the whole Xinjiang is up to 30.1 kilohm2Expected to increase to 50 kilohm2. Certain technical measures are taken to improve the production level of the alfalfa, and a high-yield, high-efficiency and high-quality industrial system is established, which is the key for promoting the healthy and sustainable development of the alfalfa industry.
Research and production practices prove that the alfalfa inoculation rhizobium can promote early nodulation, increase the nitrogen fixation amount, improve the nitrogen nutrition, reduce the fertilizer input and improve the yield and the quality of the alfalfa. Before rhizobia invades leguminous plants to establish symbiotic relationship, various external adverse environmental factors inevitably influence the survival and the nitrogen fixation process of the rhizobia in soil, and the stress resistance of the rhizobia is an important index influencing the ecological adaptation range and the nitrogen fixation capacity of the rhizobia. The soil salinization is a remarkable characteristic of the ecological environment of Xinjiang and seriously restricts the infection and nitrogen fixation effect of rhizobia. The rhizobia strain which is suitable for the local soil environment and has strong salt tolerance and high-efficiency nitrogen fixation capacity is screened, and is inoculated and applied in alfalfa production, so that the nitrogen fixation function of the rhizobia strain can be fully exerted, the nitrogen fixation efficiency is improved, the application range of the rhizobia strain can be expanded, the stress resistance of the alfalfa is improved, and the rhizobia strain has important significance for improving the grassland productivity and promoting the high-efficiency development of the alfalfa industry.
Disclosure of Invention
The invention aims to provide an alfalfa rhizobium strain XGL180 which is used for identifying a new strain belonging to the genus Sinorhizobium and has the characteristics of strong salt tolerance and high nitrogen fixation activity. The strain has been deposited in the Budapest treaty International Collection of microorganisms: china general microbiological culture Collection center (CGMCC for short). And (4) storage address: the institute of microbiology, national academy of sciences No. 3, Xilu No.1, Beijing, Chaoyang, Beijing. The preservation date is 3 months and 21 days in 2019, and the preservation number is CGMCC No. 17425.
The invention also aims to provide the rhizobium strain XGL180 for planting alfalfa in a soil salt environment, which can improve the salt tolerance of alfalfa, obviously improve the biomass and increase the contents of nitrogen, phosphorus and potassium nutrients in plants.
The Sinorhizobium meliloti XGL180 is applied to the cultivation and production of the alfalfa artificial grassland in the saline soil of Xinjiang.
The technical purpose of the invention is realized by the following technical scheme:
specifically, the rhizobia XGL180 is separated and screened from alfalfa rhizobia in south Xinjiang and north Xinjiang. The biological characteristics of the strain are as follows: the optimal growth temperature is 28 ℃, and the aerobic culture is performed. On YMA plate medium, circular or nearly circular, viscous, smooth, prominent, off-white or milky-white colonies are formed. Under an optical microscope, the fiber is in a short rod shape, contains beta-hydroxybutyric acid, is in a ring segment shape, mostly moves, is periflagellar, has no spores, and is gram-negative bacteria with red gram stain. Sequencing the 16S rDNA gene sequence of the separated strain, utilizing BLAST program in GenBank data base to make similarity search of the sequencing result and making it obtain the invented productMultiple sequence comparison and phylogenetic tree construction are carried out through DNAStar and MEGA4.1 software, and the result shows that the 16S rDNA gene sequence of the rhizobium strain provided by the invention and the Chinese rhizobium (R) areSinorhizobium sp.) Model strain of (4)Sinorhizobiumsp. The sequence similarity of CCBAU 83431 (EF 549401) reaches 99.9 percent, so that the separated rhizobium XGL180 is determined to be the rhizobiumSinorhizobium sp.) The novel strain of (1).
Furthermore, salt tolerance determination is carried out on the rhizobium strain XGL180, and the isolate strain XGL180 is a rhizobium strain which has high salt tolerance, strong nodulation capacity and can produce auxin. The invention has the following beneficial effects:
1. the Sinorhizobium sinorhizobium strain XGL180 has the capability of secreting auxin (IAA).
2. The Sinorhizobium strain XGL180 has strong salt tolerance and is prepared from 2% ~ 10% NaCl or 2% ~ 20% Na2SO4Can grow in the range of salt ion concentration.
3. The Sinorhizobium sinorhizobium strain XGL180 disclosed by the invention is used for inoculating alfalfa in a soil salt environment, so that the salt tolerance of the alfalfa can be improved, the biomass is obviously improved, and the nitrogen, phosphorus and potassium nutrient contents of the overground part and the root of a plant are increased.
Drawings
FIG. 1 shows the morphological features of the strain Sinorhizobium XGL180 under a microscope after gram staining.
FIG. 2 shows a phylogenetic dendrogram of the 16S rDNA sequence of the strain Sinorhizobium XGL 80.
FIG. 3 shows the salt tolerance test of Sinorhizobium XGL80 strain.
FIG. 4 shows the effect of inoculation with Sinorhizobium meliloti XGL80 strain on alfalfa biomass. In the figure, ZM refers to alfalfa of alfalfa No. 3, GJ refers to alfalfa of Alkanjin, Ri refers to treatment without rhizobium inoculation, USDA1002 refers to treatment with rhizobium inoculation standard strain USDA1002, and XGL180 refers to treatment with Chinese rhizobium XGL 180.
FIG. 5 shows the effect of inoculation with Sinorhizobium strain XGL80 on the aerial, root nitrogen content of the plants. In the figure, ZM refers to alfalfa of alfalfa No. 3, GJ refers to alfalfa of Alkanjin, Ri refers to treatment without rhizobium inoculation, USDA1002 refers to treatment with rhizobium inoculation standard strain USDA1002, and XGL180 refers to treatment with Chinese rhizobium XGL 180.
FIG. 6 shows the effect of inoculation with Sinorhizobium strain XGL80 on the aerial, root potassium content of the plants. In the figure, ZM refers to alfalfa of alfalfa No. 3, GJ refers to alfalfa of Alkanjin, Ri refers to treatment without rhizobium inoculation, USDA1002 refers to treatment with rhizobium inoculation standard strain USDA1002, and XGL180 refers to treatment with Chinese rhizobium XGL 180.
FIG. 7 shows the effect of inoculation with Sinorhizobium strain XGL80 on the aerial, root phosphorus content of plants. In the figure, ZM refers to alfalfa of alfalfa No. 3, GJ refers to alfalfa of Alkanjin, Ri refers to treatment without rhizobium inoculation, USDA1002 refers to treatment with rhizobium inoculation standard strain USDA1002, and XGL180 refers to treatment with Chinese rhizobium XGL 180.
Detailed Description
The invention will be further illustrated with reference to the following specific examples.
It will be understood by those skilled in the art that these examples are only for illustrating the present invention and do not in any way limit the scope of the present invention. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
All reagents and equipment selected for use in the present invention are well known in the art and are not intended to limit the practice of the present invention, and other reagents and equipment well known in the art may be suitable for use in the practice of the following embodiments of the present invention.
The alfalfa varieties tested by the invention are as follows: new herding No.1 alfalfa (Medicago sativa Xinmu No. 1), Alokang alfalfa (Medicago sativa)Medicago sativa Algonquin), alfalfa 3 (alfalfa of Chinese alfalfa 3)Medicago sativa L.cv. ‘Zhongmu No.3’）。
The present invention is tested with reference rhizobia strains: the Sinorhizobium meliloti USDA1002 strain is provided by the U.S. department of agriculture, and the Sinorhizobium meliloti strains H1, H2 are strains isolated from Sinkiang university Triflon experimental farm Sinorhizobium meliloti.
Example 1 isolation, purification and characterization of Rhizobium meliloti XGL180 Strain
1. Isolation and purification of the strains
Collecting a large amount of root nodules from alfalfa cultivated in saline soil in south Xinjiang and north Xinjiang. Selecting large and plump red root nodule of main and lateral root of robust plant, washing the root nodule with sterile water, soaking in 75% ethanol for 5 min, and adding 0.1% mercuric chloride (HgCl)2) And after sterilizing for 5 min, washing with sterile water for 5-6 times, coating the sterile water cleaning solution for the last time on a YMA plate culture medium for 72 h for aseptic inspection, crushing a single nodule by using a sterile forceps, picking a bacterial suspension by using an inoculating loop, scribing on the YMA plate culture medium, culturing for 2 ~ 4 d under the constant temperature condition of 28 ℃, purifying the strain for multiple times, picking a single colony transfer inclined plane, storing at the low temperature of 4 ℃, and naming the obtained strain as XGL 180.
The composition of YMA solid medium (yeast juice mannose agar medium) is (g/L): 10 g of mannitol, 0.1 g of NaC1, 0.2 g of MgSO4·7H2O, 1.0 g yeast powder, 0.5 g K2HPO416 g of agar powder and 1000 mL of distilled water, and the pH value is 7.0-7.2.
2. Identification of strains
(1) Morphological and cultural characteristic Observation of Strain
Strain XGL180 was inoculated on YMA solid medium for culture and observed for recording. After the culture is carried out for 72 h at the optimal growth temperature of 28 ℃, the bacterial colony is circular or approximately circular, gray white or milky white, semitransparent, neat in edge, viscous and 0.8-2.0 mm in diameter. Under the optical microscope, the thallus is in a short rod shape, the intracellular staining is uneven and is in a ring joint shape, no spores exist, the thallus is single or paired, the gram staining is pink, and the thallus is a gram negative bacterium (see figure 1).
(2) Sequence determination and analysis of 16S rDNA gene of strain
And extracting the total DNA of the thalli. The 16S rDNA sequence was amplified using the bacterial universal primer pair P1 (5'-CGGGATCCAGAGTTTGATCCTGGCTCAGAACGAACGCT-3') and P6 (5'-CGGGATCCTACGGCTACCTTGTTACGACTTCACCCC-3'). PCR reaction (50 . mu.L) 10 XPCR Buffer 5.0 μ L, 2.5 mmol/L MgCl23.0 muL, 1.0 muL of each primer P1/P6 of 10 mumol/L, dNTP 8.0 muL of 2.5 mmol/L, 5 U/muLTaqDNA polymerase 0.5 μ L, template 2.5 μ L, sterilized ddH2O29 μ L. Reaction conditions are as follows: pre-denaturation at 95 deg.C for 5 min; denaturation at 94 deg.C for 1 min, annealing at 56 deg.C for 1 min, extension at 72 deg.C for 2 min, and amplification for 32 cycles; 72 ℃ compensation extension 6 min. After the amplification product is detected by 10 g/L agarose gel electrophoresis, the PCR product is subjected to clone sequencing by Shanghai biological engineering Co., Ltd, and the sequencing result is shown in a nucleotide sequence table.
The sequence results obtained were subjected to BLAST search at the National Center for Biotechnology Information (NCBI) of the United states, and model strains having high similarity were selected as reference strains to construct a 16S rDNA gene phylogenetic tree by the Neighbor-joining method (Neighbor-joining) in MEGA4.1 software. From the alignment and the phylogenetic tree (see FIG. 2), the sequence of the rDNA gene of Rhizobium XGL 18016S and Sinorhizobium (see FIG.: 2)Sinorhizobium sp.) Model strain of (4)Sinorhizobium sp. CCBAU 83431 has a sequence similarity of 99.9% and is located in the same clade. Thereby determining that the separated rhizobia XGL180 is the genus Sinorhizobium (A)Sinorhizobium sp.) The novel strain of (1).
Based on the above biological characteristics, strain XGL180 was identified as Sinorhizobium (R) ((R))Sinorhizobium sp.). The strain has been deposited in the Budapest treaty International Collection of microorganisms: china general microbiological culture Collection center (CGMCC for short). And (4) storage address: the institute of microbiology, national academy of sciences No. 3, Xilu No.1, Beijing, Chaoyang, Beijing. The preservation date is 3 months and 21 days in 2019, and the preservation number is CGMCC No. 17425.
Example 2 Bacteria meliloti XGL180 Back-joining test
The tieback test of the sinorhizobium XGL180 adopts an agar tube method: selecting big, plump and undamaged alfalfa seeds, wherein the variety is New Muge No. I, Algang jin. Soaking in 95% ethanol for 5 min, removing ethanol, adding 0.1% mercuric chloride solution, sterilizing for 5 min, and cleaning with sterile water for 5 min for 4-6 times. Placing the disinfected seeds on an agar plate, accelerating germination at 28 ℃ for 2 d, placing the accelerating germination seeds with consistent growth on a test tube inclined plane of a 1% agar nitrogen-free culture solution, inoculating when a first piece of true leaves appears, inoculating 1mL of the culture solution per tube, repeating 4 tubes, additionally arranging a blank non-inoculation control, culturing in a lighting incubator, and observing and recording nodulation results 10-30 d after emergence of seedlings. The result of a back-grafting test shows that the strain XGL180 shows good nodulation capacity, can efficiently nodulate on 2 alfalfa varieties of Argang gold in New Murray I, the average nodulation rate is 83.3%, the average nodulation number reaches 2.1/strain, and the nodulation plants grow well.
Example 3 salt tolerance test of Rhizobium meliloti XGL180 Strain
Preparing YMA culture medium with NaCl content of 0%, 2%, 4%, 6%, and 8% and 5 salt concentration gradients. 2 mL of each of the purified 4 rhizobia XGL180, USDA1002, H1 and H2 were aspirated in a clean bench, inoculated in 15 mL of YMA liquid medium, and shake-cultured for 48H in a constant-temperature shake incubator at 28 ℃. Meanwhile, a sheet of a4 paper (square 8.3mm long and wide) was prepared, on which small squares were drawn. And measuring the light absorption value of the bacterial liquid under the OD600 light absorption value after oscillating for 48 hours, wherein the bacterial liquid can be used only when the light absorption value is more than 1. In an ultraclean workbench, A4 paper with squares is drawn on the lower pad of a culture dish filled with YMA solid culture medium, cultured bacterium liquid is sucked by a liquid transfer gun to be 10 mu L and is beaten on the culture medium, one square is beaten, each strain is beaten into one line, four lines of each dish are respectively made into 4 dishes by each NaCl concentration gradient, and 20 dishes are counted. After the bacterial liquid in the YMA solid culture medium is dried in an ultra-clean workbench, a cover is covered, and the YMA solid culture medium is filled into a freshness protection package and is placed in a constant temperature incubator at 28 ℃ for culturing for 24 hours. The YMA solid medium cultured for 24 hours is taken out from the incubator, and the growth of colonies of four kinds of rhizobia including XGL180, USDA1002, H1 and H2 in YMA medium with 0%, 2%, 4%, 6%, 8% and 5 NaCl concentrations is observed.
As shown in FIG. 3, the salt tolerance of the Rhizobium USDA1002 is the worst, and the Rhizobium H1 can tolerate 6% of NaCl culture concentration at the highest, the Rhizobium H2 can tolerate 2% of NaCl culture concentration at the highest, and the salt tolerance of the Rhizobium XGL180 is the strongest, and the Rhizobium USDA can grow in YMA culture medium with 8% of NaCl concentration.
The salt tolerance of the XGL180 strain is further determined, and the strain can contain 10% of NaCl and 20% of Na at most2SO4The growth of the cells is maintained in the culture medium, and a certain growth amount is maintained.
Example 4 determination of auxin-secreting ability of Rhizobium meliloti XGL180 Strain
The ability of rhizobia to secrete auxin (IAA) was determined colorimetrically. Inoculating the activated rhizobium strain XGL180 into a triangular flask filled with 50 mL of YMA culture solution, culturing at 28 ℃ at 120 r/min for 3 times, after culturing for 12 d, taking 100 muL rhizobium suspension, dripping the rhizobium suspension on a white ceramic colorimetric plate, and simultaneously adding 100 muL colorimetric solution (the composition of the colorimetric solution: 4.5 g FeCl)3，H2SO430 mL), and 50 μ L of auxin (IAA) was added to the colorimetric solution alone for control. The ceramic color plate was placed at room temperature for 15 min and the color change was observed. The color is changed into pink to be positive, which indicates that the strain can secrete IAA, and the darker the color indicates that the intensity of IAA secretion is higher; no color change was negative, indicating that the strain was unable to secrete IAA. The experimental result shows that the color of the culture solution for the rhizobium strain XGL180 to grow is changed into dark pink, which indicates that the capability of the Sinorhizobium meliloti XGL180 for secreting IAA is strong.
Example 5 Effect of alfalfa inoculation with Rhizobium XGL180 Strain on alfalfa salt tolerance
Alfalfa varieties are medium alfalfa 3 (ZM) and Algang Gold (GJ) used in the experiment, alfalfa 3 screened out according to the water culture experiment is a salt-tolerant variety in the vegetative growth period, and Algang gold is a salt-sensitive variety and is used as a salt-tolerant alfalfa tieback plant material for rhizobia. Three treatments of rhizobium not-inoculated (-Ri), rhizobium inoculated standard strains USDA1002 and XGL180 were set, and NaCl salt concentrations were 0mM and 100 mM. The experiment was in a completely randomized block design with 4 replicates per treatment.
Uniformly mixing cleaned quartz sand according to a ratio of coarse sand to medium sand to fine sand according to a ratio of 1: 2: 1, sterilizing at high temperature, and filling into a basin with a diameter of 5 cm and a height of 15 cm. Alfalfa seeds were surface sterilized as described in example 2, and placed in 1/4 Hoagland (Hoagland) nutrient-wetted filter paper petri dishes for dark culture for 12h, and viable seeds with exposed root tips were transferred to pots with 4 seeds per pot. Thinning seedlings after one week, inoculating bacteria, wherein the OD value of the bacteria liquid is required to be more than 1, and taking 1mL of the bacteria liquid to be inoculated to the periphery of the root of the alfalfa. After one week of inoculation, salt treatment is carried out, NaCl treatment with the concentration of 0mM and 100 mM is set, NaCl with the corresponding concentration is weighed and added into 1/4 Hoagland (Hoagland) low-nitrogen nutrient solution, the pH is adjusted to be between 5.8 and 6.0, and the treatment is carried out once in 3 days.
The plants were harvested after 4 weeks of growth and the various indices were determined. The results of the measurement of the biomass of alfalfa treated differently (see FIG. 4) show that inoculation of both the USDA1002 and XGL180 standard strains increased alfalfa biomass compared to plants without inoculation without salt treatment. Under the salt treatment, the effect of increasing biomass by inoculating the XGL180 strain Alokang gold plant is better than that of a standard strain USDA1002, and is obviously better than that of non-inoculation treatment.
The results of the nitrogen content measurement of the aerial parts and roots of the alfalfa treated differently (see FIG. 5) show that inoculation of the standard strains USDA1002 and XGL180 increased the aerial parts and root nitrogen content of the alfalfa compared to the plants without inoculation, both with and without salt treatment.
The results of the measurement of potassium content in the overground part and the root part of the alfalfa processed differently (see fig. 6) show that compared with the plants without inoculated strains, the standard strains USDA1002 and XGL180 are inoculated to increase the potassium content in the overground part of the alfalfa under the condition of not adding salt, the effect of the standard strains USDA1002 and XGL180 is the same in the No. 3 alfalfa of salt-tolerant varieties, and the effect of inoculating the standard strain USDA1002 in the Alkanjin of salt-sensitive varieties is better than that of the XGL180 strains. Under the salt treatment, the effect of increasing the potassium content of roots by inoculating the XGL180 strain is obviously better than that of a standard strain USDA1002 and is obviously better than that of non-inoculation treatment.
The determination results of phosphorus content in the overground part and the root part of the alfalfa treated differently (see fig. 7) show that compared with the plant without inoculated strain, the inoculated standard strains USDA1002 and XGL180 can increase the phosphorus content in the overground part of the alfalfa under the condition of not adding salt, the effect of the inoculated standard strains USDA1002 and XGL180 is the same in the alfalfa No. 3 in a salt-tolerant variety, and the effect of the phosphorus content in the overground part in the inoculated standard strain USDA1002 is better than that of the XGL180 in a salt-sensitive variety Algang gold. However, inoculation of the XGL180 strain was significantly more effective in increasing the phosphorus content in the aerial and root sections under salt treatment, and the XGL180 strain was more effective than the USDA1002 standard strain and significantly better than the treatment without inoculation in view of the phosphorus content in the root section.
The research shows that under the condition of salt treatment, the inoculation of the rhizobium XGL180 has a relatively obvious yield increase effect, and particularly can effectively relieve salt damage when being used for salt-sensitive alfalfa varieties. The indexes of biomass, nitrogen content, potassium content, phosphorus content and the like of alfalfa inoculated with the rhizobium XGL180 strain are all higher than those of plants which are not inoculated, and part of indexes are better than those of inoculated standard strain USDA 1002.
Experiments show that the rhizobium XGL180 strain has stronger salt tolerance than a reference strain, alfalfa inoculation can effectively nodulate and fix nitrogen, plant nitrogen nutrition is increased, the content of nitrogen, phosphorus and potassium nutrient elements in a plant body is improved, plant growth is promoted, and the dry weight of biomass is improved. Therefore, the XGL180 strain can be popularized and applied to alfalfa production in saline soil areas in Xinjiang, the adaptability of alfalfa to the saline soil environment is enhanced, and the yield of pasture is increased.
Sequence listing
<110> Sinkiang university of agriculture
South China university of agriculture
<120> salt-tolerant alfalfa rhizobium and application thereof
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cggtagggtc aggccggagc cta 1403
Claims (2)
1. A salt-tolerant alfalfa rhizobium and application thereof are characterized in that the preservation name of the strain is Chinese rhizobium (Sinorhizobium sp) XGL180 which is preserved in China general microbiological culture Collection center (CGMCC for short), the preservation address is the microbiological research institute of China academy of sciences No. 3, West Lu No.1 Hopkins, Xingyang area, Beijing city, the preservation date is 3 and 21 days in 2019, and the preservation number is CGMCC NO. 17425.
2. Use of a Sinorhizobium sp XGL180 as claimed in claim 1, in alfalfa cultivation under saline soil conditions in Xinjiang.
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| CN111466282A (en) * | 2020-02-28 | 2020-07-31 | 甘肃农业大学 | Determining the growth promoting effect of alfalfa and rhizobia specific symbiosis nodulation by biomass dry weight |
| CN113088510B (en) * | 2021-04-14 | 2022-11-18 | 上海健康医学院 | Agrobacterium tumefaciens, immobilized bacterium thereof and method for degrading azithromycin |
| CN115261245A (en) * | 2021-04-29 | 2022-11-01 | 中国农业科学院烟草研究所(中国烟草总公司青州烟草研究所) | Root nodule bacterial strain for inducing plant to have salt tolerance and application thereof |
| CN115261245B (en) * | 2021-04-29 | 2023-09-19 | 中国农业科学院烟草研究所(中国烟草总公司青州烟草研究所) | Root nodule strain capable of inducing plants to have salt tolerance and application thereof |
| CN114480171A (en) * | 2021-12-24 | 2022-05-13 | 山东省科学院生物研究所 | Rhizobium japonicum and application thereof |
| CN114480171B (en) * | 2021-12-24 | 2023-06-13 | 山东省科学院生物研究所 | Rhizobium sojae strain and application thereof |
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