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Root nodule azotobacter strain RY1 bacterial strain and application thereof

A strain and nitrogen fixation technology, applied in bacteria, organic fertilizers, microorganisms, etc., can solve problems such as failure to achieve high-yield effects

Inactive Publication Date: 2010-07-21
TROPICAL CORP STRAIN RESOURCE INST CHINESE ACAD OF TROPICAL AGRI SCI +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because Australia's rhizobia agent is a high-efficiency strain selected based on Australia's soil and climate environment, the climate and soil environment in our country can only slightly increase the yield of stylophyllum, and cannot achieve the high-yield effect of Australia.
At present, no strains suitable for the climate and soil conditions of our country have been screened, so it is of great practical significance to screen the high-efficiency nitrogen-fixing rhizobia of Stylophyllum from the indigenous bacteria of different types of soil conditions for planting Stylophyllum in a specific area.

Method used

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  • Root nodule azotobacter strain RY1 bacterial strain and application thereof
  • Root nodule azotobacter strain RY1 bacterial strain and application thereof
  • Root nodule azotobacter strain RY1 bacterial strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The acquisition and cultivation of embodiment 1 rhizobia RY1

[0030] On February 27, 2009, the inventor collected a root nodule of a large and luxuriant Stylophyllum in Tianyang farmland wasteland (E10652.483 N23 47.971 H146) in Baise City, Guangxi Province. Put it into a fresh-keeping bag together with the root nodules and store it in an ice pack at low temperature.

[0031] (1) Take out the root system of Stylophyllum from the ice pack and rinse it 2 to 3 times under tap water to clean the root system. Use scissors to cut off the fresh, complete, dark red root nodule with 2mm root, and put it in a petri dish with grade 2 Water rinses the surface clean; this process ensures the integrity of the nodule surface.

[0032] (2) Transfer the rinsed root nodules to a sterilized petri dish, add 95% (volume ratio concentration) alcohol to soak for 3 minutes, pour out the alcohol, rinse 4 to 5 times with sterile water, add 0.1 % (mass volume concentration, 0.1g HgCl 2 / 100ml...

Embodiment 2

[0054] Example 2 The 16S rDNA sequence sequencing of Rhizobium RY1 and the determination of its category

[0055] In order to determine the phylogenetic status of Rhizobium RY1, the 16SDNA series of the isolated strains were sequenced. Firstly, total DNA was extracted using a kit from Omega, and PCR-specific amplification was performed using primers.

[0056] Upstream primer 35fc: CTKAAGAGTTTGATCMTGGCTCAGATTGAAC;

[0057] Downstream primer 1492r: TACGGYTACCTTGTTACGACTT).

[0058] The reaction conditions are as follows:

[0059] PCR reaction

[0060] Primer: 35fc, 1492r

[0061] PCR recipe:

[0062] 10×Reaction Buffer 5.0μL

[0063] dNTPs (10mM) 1.0μL

[0064] P35fc (25Pmol) primer 0.5μL

[0065] P1492r (25Pmol) primer 0.5μL

[0066] Taq DNA polymerase (5u / μL) 1.0μL

[0067] Template DNA 1.0 μL

[0068] Make up to 41 μL with ultrapure water

[0069] PCR procedure

[0070] Pre-denaturation at 94°C: 3min

[0071] Denaturation at 94°C for 50s

[0072] 56℃ annealing ...

Embodiment 3

[0077] Example 3 Application experiment of rhizobia RY1

[0078] In order to confirm the effect of RY1 on Stylophyllum, two main Stylophyllum varieties (TPRC2, TPRC5) were used to reconnect root nodules RY1 to the roots of Stylophyllum by sand culture method, with 4 replicates for each treatment (bacteria, sand and seed For the treatment of seedlings, refer to the backgrafting experiment), and nodule inoculation was used as the control (CK) for backgrafting comparison. The whole process is irrigated with low-nitrogen nutrient solution. See Table 2 and attached Figure 8 . attached Figure 8 The middle left square column (a) is TPRC5, and the right square column (b) is TPRC2 (attached Figure 8 Other square bar graphs in the middle are existing rhizobia control results, which are not directly related to the technology of the present invention, and are not marked).

[0079] It can be seen from Table 2 that the number of nodules, dry weight of nodules, plant height, fresh we...

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Abstract

The invention discloses a root nodule azotobacter strain RY1 bacterial strain and an application thereof. The bacterial strain (Bradyhizobium sp.RY1) is obtained from gathering fresh root nodules in main Stylosanthes guianensis planting areas in China and separating and purifying in a laboratory, the partial sequence of 16S rDNA of the bacterial strain is measured, and the bacterial strain is determined to be a new bacterial strain of Bradyrhizobium by the phylogenesis analysis and is named as Re Yan 1 (RY1). By the potted planting and the inoculation test in the laboratory, the nodule azotobacter strain RY1 bacterial strain is proven to have the characteristic of enabling the two main Stylosanthes guianensis varieties (TPRC2 and TPRC5) in China to realize the high-efficiency nodulation and achieve the high yield. Compared with the inoculation-free process, the inoculation with the root nodule can improve the yield of the Stylosanthes guianensis by more than 2 times, and can be used as one of conventional Stylosanthes guianensis culturing measures for planting the Stylosanthes guianensis.

Description

technical field [0001] The invention belongs to the technical field of plant nutrition and forage cultivation, and in particular relates to a biological nitrogen fixation technology, in particular to a rhizobia capable of effectively forming nodulation and high yield of stylo. Background technique [0002] Stylosanthes guianensis SW., also known as tropical alfalfa and Brazilian alfalfa, is native to South America. In 1982, it was introduced from the International Center for Tropical Agriculture (CIAT) by the Chinese Academy of Tropical Agricultural Sciences, and it was successfully planted in Hainan. Now it has been widely promoted in various provinces (regions) in South China. The dry matter of Stylophyllum in full bloom stage contains 16.0-20.0% of crude protein, 1.8-2.5% of crude fat, 17.0-19.0% of fiber, 38.0-44.0% of nitrogen-free extract, and 8.0-10.0% of crude ash. Excellent tropical legume. South China, such as Guangdong, Guangxi and other provinces (regions), int...

Claims

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Application Information

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IPC IPC(8): C12N1/20C05F11/08C12R1/01
Inventor 刘国道董荣书黄艳霞
Owner TROPICAL CORP STRAIN RESOURCE INST CHINESE ACAD OF TROPICAL AGRI SCI
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