Plant endophyte and application thereof

A technology of plant endophytic bacteria and bacterial liquid, applied in plant growth regulators, plant growth regulators, applications, etc., to achieve the effect of increasing zinc enrichment in grains, simple operation, and reduced investment

Active Publication Date: 2013-07-31
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are very few studies in this area at present. In the actual agricultural production, it can be considered to us

Method used

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  • Plant endophyte and application thereof
  • Plant endophyte and application thereof
  • Plant endophyte and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0034] Example 1 Isolation of Enterobacter CGMCC7381

[0035] (1) Medium

[0036] DF medium: each liter of culture medium contains KH 2 PO 4 4g, Na 2 HPO 4 6g, MgSO 4 ·7H 2 O0.2g, glucose 2g, sodium gluconate 2g, citric acid 2g, (NH 4 ) 2 SO 4 2g, 0.1mL each of component 1 and component 2, 15g agar, water, pH 7.2, autoclave sterilization (121°C, 20min).

[0037] Among them, component 1: H 3 BO 3 10mg, MnSO 4 ·H 2 O11.19mg, ZnSO 4 ·7H 2 O124.6mg, CuSO 4 ·5H 2 O78.22mg, MoO 3 10mg dissolved in 100mL sterilized distilled water, stored at -4℃;

[0038] Component 2: FeSO 4 ·7H 2 O100mg is dissolved in 10mL sterilized distilled water and stored at -4°C.

[0039] DF+ACC medium: Add 3mM ACC per liter of DF medium.

[0040] LB liquid medium: each liter of culture medium contains 10g tryptone, 5g yeast extract, 10g NaCl, solvent is water, pH7.0, autoclave sterilization (121℃, 20min).

[0041] (2) Separation and purification of strains

[0042] Use sterile scissors to cut the root system of the south...

Example Embodiment

[0045] Example 2 Identification of SaCS20 strain

[0046] (1) Observation of colony and cell morphology of SaCS20 strain

[0047] Morphological study of bacterial cells: Gram-stained bacterial cells were observed under a 16×100 oil lens under an optical microscope.

[0048] Study on the internal structure of the bacteria: Ultrathin sectioning of the bacteria and observation of the internal structure of the bacteria with a transmission electron microscope.

[0049] On the solid LB medium, the size of the SaCS20 strain is 2-5mm; it is round with neat edges; the surface is smooth, raised, and opaque.

[0050] (2) Molecular biological identification of SaCS20 strain

[0051] The preserved SaCS20 strain was inoculated into LB liquid medium and cultured overnight at 30°C and 180r / min on a constant temperature shaker. The resulting bacterial solution was extracted with the Shanghai Shenggong Biological Bacterial DNA Extraction Kit to amplify the 16S rDNA of the strain. The PCR products obtaine...

Example Embodiment

[0062] Example 3 Preparation of enterobacteria CGMCC7381 inoculum

[0063] (1) Strain activation

[0064] Inoculate the cold-preserved Enterobacter CGMCC7381 stock in LB slant medium for activation culture, and incubate in a constant temperature incubator at 37°C for 22-24 hours to obtain bacterial slant;

[0065] (2) Expanding cultivation

[0066] The activated strains were connected to LB liquid medium for propagation, cultured on a constant temperature shaker at 30°C, 180r / min, shaken to the logarithmic phase to obtain the bacterial solution, centrifuged the bacterial solution, discarded the supernatant, and used aseptic Wash the precipitate 3 times with water, and suspend the precipitate with pH 7.0 phosphate buffer to obtain a bacterial suspension containing Enterobacter CGMCC7381. The OD of the bacterial suspension must be removed before application 600 Adjust to 0.8~1.0.

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Abstract

The invention discloses a plant endophyte with a category name of Enterobacter sp. and a complete name of Enterobacter sp. SaCS20. The strain is preserved at China General Microbiological Culture Collection Center (CGMCC) on March 28th, 2013, and has a preservation number of CGMCC NO: 7381. The invention also discloses the application of the plant endophyte in promoting paddy rice growth, improving paddy rice yield, or promoting trace element zinc accumulation in paddy rice. The Enterobacter sp. CGMCC 7381 provided by the invention is separated from the root system of a zinc hyperaccumulator sedum alfredii. The plant endophyte has no pathogenicity upon paddy rice, and can be successfully colonized into paddy rice root systems. With the plant endophyte, paddy rice growth can be promoted, rhizosphere soil available zinc content can be increased, and grain zinc enrichment can be promoted. The application provided by the invention is simple to operate, and is safe and economical. With the plant endophyte and the application, soil micronutrient fertilizer investment can be reduced, and crop yield can be increased.

Description

technical field [0001] The invention belongs to the technical field of rice microelement bioaugmentation, and in particular relates to a plant endophyte and its application. Background technique [0002] Zinc is closely related to human production and life activities. Zinc is an essential micronutrient element for plant growth and an important nutrient element necessary for the human body. Zinc has important physiological functions and nutritional value for the human body. Under normal circumstances, there are nearly 200 kinds of enzyme activities in the body related to zinc, so many enzyme activities are affected when the human body is deficient in zinc, resulting in a variety of metabolic disorders, which in turn affect human health. Studies have shown that zinc deficiency can cause short stature and skin disorders, especially for pregnant women and children. [0003] Zinc in the edible part of food crops is the main way for the human body to obtain zinc. Due to insuffic...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01P21/00C12R1/01
CPCY02A40/10
Inventor 杨肖娥王玉艳张新成董兰学冯英韦燕燕高俊
Owner ZHEJIANG UNIV
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