Salt-tolerant rice variety root system Exiguobacterium indicum SaltT2 and application thereof
By using the bacterial suspension of the root system of the salt-resistant rice variety Exiguobacterium indicum SaltT2, the problem of restricted growth of rice in saline-alkali land was solved, and the effect of improving the salt tolerance and yield of rice was achieved, while also having the advantages of environmental protection.
Patent Information
- Application Number
- CN202510075601.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-01-17
- Publication Date
- 2025-04-25
AI Technical Summary
The growth of rice in saline-alkali land is severely restricted by soil salinization. The prior art is difficult to effectively improve the salt tolerance of rice, and traditional methods have problems of water resource waste and environmental pollution.
The root system of the salt-resistant rice variety Exiguobacterium indicum SaltT2 and its application are provided. By soaking the bacterial suspension of this strain on rice seeds, the salt tolerance of rice during the bud stage is improved.
It significantly improves the salt tolerance of rice in saline-alkali land, reduces damage under salt stress, improves rice yield and quality, and avoids environmental pollution and waste of water resources.
Smart Images
Figure CN119875915A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of biological pesticides, and in particular to a salt-tolerant rice variety root system Exiguobacterium indicum SaltT2 and an application thereof. Background Art
[0002] As one of the main food crops, rational use of saline-alkali land has become one of the effective measures. However, soil salinization seriously restricts the growth and development of rice and affects its yield and quality, and even leads to the death of rice plants.
[0003] Improving soil physical and chemical properties through some physical (irrigation to wash salt and drainage) and chemical (adding soil conditioners such as gypsum) methods often causes problems such as water waste or environmental pollution. In addition, although screening salt-tolerant rice germplasm resources, exploring rice salt-tolerant genes, and creating and cultivating new salt-tolerant rice germplasm and new varieties are the most economical and effective, they are limited by the few existing strong salt-tolerant germplasm resources, the few cloned rice salt-tolerant genes, and the long breeding cycle.
[0004] At present, the types of rhizosphere microorganisms that enhance rice salt tolerance include arbuscular mycorrhizal fungi (AMF), Serendipita indica, Bacillus spp., and Enterobacter spp. The physiological mechanisms by which these microorganisms enhance rice salt tolerance include alleviating Na+ ion toxicity, reducing the content of reactive oxygen in cells, and inducing hormone regulation in rice plants. Wang et al. identified the strain Rossellomorea vietnamensis RL-WG62 in the root system of Haidao 86 that can enhance its salt tolerance, proving that beneficial rhizosphere microbiomes are also enriched in the root system of rice to reduce the damage of rice under salt stress.
[0005] To this end, the present invention aims to provide a rice root Exiguobacterium indicumSaltT2 and its application to solve the above problems. Summary of the invention
[0006] The purpose of the present invention is to solve the above problems and provide a salt-tolerant rice variety root system Exiguobacterium indicum SaltT2 and its application to enhance the salt tolerance of rice, reduce the damage under salt stress, improve the yield and quality of rice in saline-alkali land, thereby providing guarantee for food security.
[0007] The technical scheme of the present invention is as follows: a salt-tolerant rice variety root system Exiguobacterium indicum SaltT2, the Exiguobacterium indicum SaltT2 is derived from the root system of the salt-tolerant rice variety Guanghong No. 3 planted in the Taishan Experimental Base of the Salt-Alkali Tolerant Rice Research Center of the Rice Research Institute of Guangdong Academy of Agricultural Sciences, Beidou Town, Taishan City, Guangdong Province, and the strain was deposited in the Guangdong Provincial Microbiological Culture Collection Center on September 23, 2024, with a deposit number of GDMCC No: 65178; the 16S rRNA gene sequence of the strain is shown in Seq ID No.1.
[0008] The present invention is further configured as follows: the homology of the 16S rRNA sequence of the strain to the 16S rRNA sequence of the Exiguobacterium indicum BDNA-E69 strain with the accession number of MN636679.1 in GenBank is 97.04%.
[0009] Another object of the present invention is to provide an application of Exiguobacterium indicum SaltT2 in the root system of a salt-tolerant rice variety, wherein the Exiguobacterium indicum SaltT2 in the root system of the rice is used to improve the salt tolerance of rice during the bud stage, and the rice variety is the salt-sensitive variety Zhefu 80.
[0010] The present invention is further configured as follows: the application includes utilizing the environmental protection and non-toxicity of the strain described in claim 1 to soak rice seeds for planting in saline-alkali land, which has little impact on the ecological environment and is safe and reliable for the consumption of rice and its processed products.
[0011] Another object of the present invention is to provide a method for preparing an Exiguobacterium indicum SaltT2 bacterial suspension, comprising inoculating into LB liquid culture medium for culturing, centrifuging, removing the supernatant, and resuspending with sterile pure water to obtain an Exiguobacterium indicum SaltT2 bacterial suspension, wherein the bacterial suspension is used to improve the salt tolerance of rice during the bud stage.
[0012] Another object of the present invention is to provide a method for improving the salt tolerance of rice by using Exiguobacterium indicumSaltT2, comprising soaking rice seeds with a bacterial suspension of Exiguobacterium indicumSaltT2 to resist salt stress during the germination period.
[0013] Another object of the present invention is to provide a rice planting method, comprising treating rice seeds with the Exiguobacterium indicum SaltT2 bacterial suspension according to claim 1, and then planting the treated seeds in saline-alkali land.
[0014] Compared with the prior art, the beneficial effects of this scheme are as follows: the present invention provides a salt-tolerant rice variety root system Exiguobacterium indicum SaltT2 and its application. First, this strain can effectively colonize the rice root system and significantly improve the salt tolerance of rice seeds in the bud stage, which is of great significance for growing rice in saline-alkali land because it can help rice grow in a high-salt environment and increase crop yield. Secondly, compared with traditional physical and chemical methods, the use of Exiguobacterium indicum SaltT2 as a biological method has less impact on the environment and will not cause waste of water resources or environmental pollution. In addition, due to the characteristics of its biological source, the strain is non-toxic and harmless to human health and the ecosystem, ensuring the edible safety of rice and the reliability of processed products. Through its physiological mechanism, including alleviating Na+ ion toxicity, reducing the content of active oxygen in cells and inducing the regulation of hormones in rice plants, the strain of the present invention promotes the growth and development of rice under salt stress and improves the adaptability and survival ability of rice.
[0015] The beneficial effect of the present invention is that the success rate of rice planting in saline-alkali land is increased, thereby improving the utilization rate of saline-alkali land, which is of great significance for alleviating the pressure on cultivated land and ensuring food security. At the same time, the salt-tolerant rice root microbial resources provided by the present invention provide a new option for the development of salt-tolerant rice germplasm resources, help overcome the limitations of existing salt-tolerant germplasm resources, and promote the creation and cultivation of new salt-tolerant rice germplasms and new varieties.
[0016] In summary, the beneficial effect of the present invention is to provide a new strain Exiguobacterium indicum SaltT2, which improves the salt tolerance of rice by using Exiguobacterium indicumSaltT2, reduces the restriction of saline-alkali land on rice growth, and has the advantages of environmental protection and sustainable development. This method not only helps to improve rice yield and quality, but also provides effective technical support. BRIEF DESCRIPTION OF THE DRAWINGS
[0017] Figure 1 This is a single colony image of Exiguobacterium indicum SaltT2 on LB solid culture medium in an embodiment of the present invention;
[0018] Figure 2 It is a diagram showing the effect of Exiguobacterium indicum SaltT2 on the salt tolerance of the salt-sensitive rice variety Zhefu 802 at the germination stage and a diagram showing the statistical results of the root length and bud length of the seeds during germination in an embodiment of the present invention. DETAILED DESCRIPTION
[0019] In order to enable those skilled in the art to better understand the solution of the present invention, the technical solution of the present invention will be further described in detail below in conjunction with the embodiments of the present invention and the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work should fall within the scope of protection of the present invention.
[0020] It should be noted that, in the absence of conflict, the embodiments and features in the embodiments of the present invention can be combined with each other. The present invention will be described in detail below in conjunction with the embodiments.
[0021] Example 1: Isolation, purification and preservation of Exiguobacterium indicum SaltT2 from rice roots
[0022] 1.1 Preparation of LB solid medium
[0023] The present embodiment first performs the configuration of LB solid culture medium. The required raw materials include 10g of tryptone, 5g of yeast extract, 10g of sodium chloride and 1000mL of deionized water. The above raw materials are added to deionized water and stirred with a stirring rod or a magnetic stirrer until completely dissolved to ensure that there are no particles or lumps. During the heating process, 20g of agar is added, and heating and stirring are continued until the agar is completely dissolved. Subsequently, the prepared LB solid culture medium is dispensed into culture dishes or test tubes to ensure that the culture medium thickness of each culture dish or test tube is uniform. Finally, the dispensed culture medium is placed in an autoclave and sterilized at 121°C for 20 minutes. After sterilization is completed, the culture medium is placed in an incubator to cool and store for standby use.
[0024] 1.2 Isolation and purification of Exiguobacterium indica from rice roots
[0025] This embodiment separates and purifies Microbacterium indica from rice roots. First, root samples of salt-tolerant rice variety Guanghong No. 3 were collected from the Taishan Experimental Base of Salt-Alkali Tolerant Rice Research Center, Rice Research Institute, Guangdong Academy of Agricultural Sciences, Beidou Town, Taishan City, Guangdong Province. 10g of root sample (including soil) was placed in a pre-sterilized triangular bottle, and an appropriate amount of sterile pure water was added to ensure that the root tissue and soil were completely immersed. Then, after the triangular bottle was sealed, it was placed in a shaker and shaken at a speed of 150rpm for 5min to help release the microorganisms in the root system. After shaking, the triangular bottle was placed in a clean bench and allowed to stand for 15min to allow the suspended microorganisms to settle. Next, 100μL of the upper root extract was drawn and evenly spread on the LB plate. Three replicates were set for each treatment, and the coated LB plate was placed in a constant temperature incubator at 28°C for 48 to 72 hours to observe the growth of the colony.
[0026] 1.3 Strain identification
[0027] In terms of strain identification, the strain isolated from the rice root system was subjected to 16S rRNA gene sequence analysis, and it was found that the 16S rRNA sequence homology of the strain was 97.04% with the Exiguobacterium indicum BDNA-E69 strain with accession number MN636679.1 in GenBank. Combined with the colony morphology characteristics of the bacteria on LB solid medium plates and related physiological and biochemical characteristics tests, it was confirmed that it was classified and named Exiguobacterium indicum, and named Exiguobacterium indicum SaltT2. After the strain was cultured on LB solid medium at 28°C for 24 hours, the colonies were orange-yellow, opaque, slightly convex, smooth and moist on the surface, with neat edges, as shown in the attached Figure 1 The sequence fragment is shown as SeqID No.1.
[0028] 1.4 Strain preservation
[0029] The Exiguobacterium indicum SaltT2 of this embodiment was deposited in the Guangdong Microbiological Culture Collection Center on September 23, 2024, with the address in Guangzhou, Guangdong Province, and the deposit number is GDMCC No: 65178, for future research and application.
[0030] Example 2: Determination of the effect of Exiguobacterium indicum SaltT2 on promoting salt tolerance in rice bud stage
[0031] 2.1 Preparation of LB liquid medium
[0032] The LB solid medium formula is the same as that in Example 1, and agar is not required for the preparation of the LB liquid medium. 10 g of tryptone, 5 g of yeast extract, and 10 g of sodium chloride are mixed with 1000 mL of deionized water and stirred thoroughly until the agent is completely dissolved. The prepared liquid medium is divided into 100 mL conical flasks, the bottle mouths are plugged and bandaged, and then sterilized at 121°C for 20 min. After sterilization, the medium is cooled and stored for later use.
[0033] 2.2 Preparation of single colonies of Exiguobacterium indica
[0034] Exiguobacterium indicum SaltT2 was streaked onto an LB solid culture medium plate, and after culturing for 24 hours, a single colony with good growth was picked for streaking preservation to ensure that a pure strain was obtained. Subsequently, the selected single colony was accessed into the LB liquid culture medium conical flask prepared in the above step, and placed in a shaking incubator at 28°C and cultured at a speed of 200rpm for 24 hours to obtain a fermentation broth of Exiguobacterium indicum. After the culturing was completed, the obtained fermentation broth was centrifuged at 12000rpm for 10min, the supernatant was discarded, and the bacteria were resuspended with sterile pure water to obtain an Exiguobacterium indicum bacterial suspension.
[0035] 2.3 Determination of the effect of Exiguobacterium indica on salt tolerance of rice buds
[0036] Take the same amount of the above-mentioned Exiguobacterium indica suspension and add an appropriate amount of NaCl to make the NaCl concentration of the bacterial suspension reach 0.7%. At the same time, sterile pure water and 0.7% NaCl solution were used as controls to treat the seeds of the salt-sensitive rice variety Zhefu 802. Under the same temperature and light conditions, germination tests were carried out. During the test, the length of the germinated roots and shoots was counted and photographed and recorded, as shown in the attached figure. Figure 2 Through these steps, this example will evaluate the effectiveness of Exiguobacterium indicum SaltT2 in improving the salt tolerance of rice during the budding period, and provide experimental basis for subsequent application research.
[0037] The above specific embodiments are merely explanations of the present invention and are not limitations of the present invention. After reading this specification, those skilled in the art may make modifications to the embodiments without any creative contribution as needed. However, such modifications are protected by the patent law as long as they are within the scope of the claims of the present invention.
Claims
1. A salt-tolerant rice variety root system Exiguobacterium indicum Salt T2, characterized by: The Exiguobacterium indicum SaltT2 is derived from the rice root system of the salt-tolerant rice variety Guanghong No. 3 planted at the Taishan Experimental Base of the Salt-Alkali Tolerant Rice Research Center, Rice Research Institute, Guangdong Academy of Agricultural Sciences, Beidou Town, Taishan City, Guangdong Province, and the strain was deposited in the Guangdong Provincial Microbiological Culture Collection Center on September 23, 2024, with the deposit number GDMCCNo: 65178; the 16S rRNA gene sequence of the strain is shown in Seq ID No.
1.
2. Exiguobacteriumindicum SaltT2 as claimed in claim 1, characterized in that: The homology of the 16S rRNA sequence of the strain and the 16S rRNA sequence of the Exiguobacterium indicum BDNA-E69 strain with the accession number of MN636679.1 in GenBank is 97.04%.
3. The use of Exiguobacterium indicum SaltT2 in rice roots as claimed in claim 1, characterized in that: The rice root system Exiguobacterium indicum SaltT2 is used to improve the salt tolerance of rice during the budding period, and the rice variety is the salt-sensitive variety Zhefu 80.
4. The use according to claim 1, characterized in that: The application includes utilizing the environmental protection and non-toxicity of the strain described in claim 1 to soak rice seeds for planting in saline-alkali land, which has little impact on the ecological environment and is safe and reliable for the consumption of rice and its processed products.
5. A method for preparing a bacterial suspension of Exiguobacterium indicum Salt T2, characterized in that: The method comprises the steps of inoculating into LB liquid culture medium for cultivation, centrifuging, removing supernatant, and resuspending with sterile pure water to obtain Exiguobacterium indicum SaltT2 bacterial suspension, and the bacterial suspension is used for improving salt tolerance of rice during bud stage.
6. A method for improving the salt tolerance of rice using Exiguobacterium indicum Salt T2, characterized in that: The method comprises soaking rice seeds with the Exiguobacteriumindicum SaltT2 bacterial suspension to resist salt stress during the germination period.
7. A rice planting method, characterized in that: The method comprises treating rice seeds with the Exiguobacterium indicum Salt T2 bacterial suspension according to claim 1, and then planting the treated seeds in saline-alkali land.