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Saline-alkali tolerant growth-promoting bacterium YS-AT1 capable of secreting IAA and application thereof

A YS-AT1, salt-alkali-resistant technology, applied in the field of microorganisms, to achieve the effect of promoting alkali reduction, promoting plant growth, and mitigating environmental hazards

Pending Publication Date: 2022-01-11
中国科学院东北地理与农业生态研究所农业技术中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The IAA-secreting bacteria currently found in the soil environment or inside plants are mainly Azospirillum, Pseudomonas, Bacillus, Microbacterium and Rhizobium. However, Klebslella (Klebslella), which has the characteristics of salt and alkali tolerance and can secrete IAA, has not yet been reported.

Method used

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  • Saline-alkali tolerant growth-promoting bacterium YS-AT1 capable of secreting IAA and application thereof
  • Saline-alkali tolerant growth-promoting bacterium YS-AT1 capable of secreting IAA and application thereof
  • Saline-alkali tolerant growth-promoting bacterium YS-AT1 capable of secreting IAA and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Isolation and purification of salt-alkali-resistant growth-promoting bacteria YS-AT1

[0032] Sample source: Roots of Suaeda salsa in tidal flats of Dongying City, Shandong Province, China

[0033] LB medium (solid): Tryptone 10g, Yeast extract 5g, NaCl 10g, agar 15g, H 2 O 1000mL, pH 7.0-7.2, sterilized at 121°C for 30min.

[0034] LB medium (liquid): Tryptone 10g, Yeast extract 5g, NaCl 10g, H 2 O 1000mL, pH7.0-7.2, sterilized at 121°C for 30min.

[0035] Separation and purification steps: collect Suaeda salsa growing on tidal flats, wash the soil particles on the surface of plant roots with tap water, cut the roots into several sections, place them in 70% ethanol, sodium hypochlorite and 70% ethanol for 7 minutes each, and then wash them with sterilized water 4 times, and then grind the roots with a sterilized mortar, and make a series of diluted bacterial solutions with sterile water, and choose a concentration gradient of 10 -2 and 10 -3 The bacteri...

Embodiment 2

[0036] Example 2: Identification of salt-alkali-tolerant growth-promoting bacteria YS-AT1

[0037] (1) Biological identification

[0038] Salt-alkali-resistant growth-promoting bacteria YS-AT1 was cultured on LB solid medium plate at 30°C for 2 days, the colony was round, with neat edges, smooth surface, raised, opaque, semi-liquid, bright chrysanthemum yellow, such as figure 1 shown.

[0039] (2) System classification identification

[0040] Using the colony PCR identification method, use an inoculation loop to pick up the colony of the saline-alkaline growth-promoting bacteria YS-AT1 on the LB solid medium plate and place it in 15 μL of sterile water, centrifuge at 12000 rpm for 5 minutes, discard the supernatant, and then suspend with 15 μL of sterile water, repeat Once, in a water bath at 95° C. for 5 min, placed on ice, using the treated bacterial suspension as a template for PCR identification, using bacterial universal primers 27F (5'-AGAGTTTGATCCTGGCTCAG-3', as shown...

Embodiment 3

[0042] Example 3: Salt and Alkali Resistance Evaluation of Salt-Alkali-Resistant Growth-promoting Bacteria YS-AT1

[0043] (1) Salt tolerance evaluation

[0044] Inoculate seed bacteria solution in LB medium containing 0, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9% (w / v) NaCl, 1% inoculum , cultured with shaking at 30°C for 48h, and measured the absorbance value (OD 600 ), draw the growth curve ( image 3 ). The culture medium with different NaCl concentration without inoculation was the control group. The results showed that the strain YS-AT1 could grow in LB medium with NaCl concentration of 0%-9% (w / v).

[0045] (2) Alkali resistance evaluation

[0046] Set the following pH gradient: 8.0, 8.5, 9.0, 9.5, 10.0, 10.5, 11.0, 11.5, 12.0, 12.5, 13.0, 13.5, 14.0, adjust the pH with 1N NaOH or 1N HCl, 1% inoculum size, shake at 30°C After culturing for 48 h, the absorbance value (OD 600 ), draw the growth curve, such as Figure 4 As shown, different pH gradient medium without inocul...

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Abstract

The invention discloses a saline-alkaline tolerant growth-promoting bacterium YS-AT1 capable of secreting IAA and an application thereof. The bacterium belongs to Klebslella and is preserved in the China Center for Type Culture Collection on March 23, 2021, and the preservation number is M2021237. The bacterium has strong salt resistance and alkali resistance, the growth tolerant NaCl concentration of the bacterium is larger than 9% (w / v), the tolerant pH value of the bacterium is 8.0-12.0, the bacterium has certain alkali reduction capacity under different pH growth conditions, and the alkali reduction range under the different pH growth conditions is 2.7%-13.8%. Besides, the capability of the bacterium for secreting IAA under the induction of L-tryptophan can reach 5.38 [mu] g.mL<-1>. In addition, the bacterium has an obvious promotion effect on wheat seed germination and arabidopsis thaliana seedling growth under saline and alkaline conditions. Therefore, the bacterium has important application value in the aspects of preparation of growth-promoting microbial bacterium for saline-alkali areas and the like.

Description

technical field [0001] The invention belongs to the field of microorganisms, and in particular relates to a salt-alkali-resistant growth-promoting bacterium YS-AT1 capable of secreting IAA and its application. Background technique [0002] Soil salinization is one of the serious ecological environmental problems facing the world. The area of ​​land salinization in my country has been increasing in recent years. The Northeast is an important area for grain production in my country, and it is also one of the areas where soil salinization is relatively serious, which seriously restricts animal husbandry. and agricultural development. [0003] Microbial strains are important resources for promoting plant growth and development, and growth-promoting bacteria can improve plant tolerance to abiotic stresses such as high salt and high alkali to a certain extent. At present, most of the research on the growth-promoting effect of microorganisms is the ability to secrete plant hormones...

Claims

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Application Information

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IPC IPC(8): C12N1/20C05F11/08C05G3/80A01N63/20A01P21/00C09K17/14C12R1/22C09K101/00
CPCC05F11/08C05G3/80A01N63/20C09K17/14C09K2101/00
Inventor 于镇华喻江李彦生王光华姚钦高志颖龙永
Owner 中国科学院东北地理与农业生态研究所农业技术中心
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