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Bradyrhizobium sp.RY4 strain and application thereof

A technology of strains and nodules, applied in the direction of bacteria, organic fertilizers, microorganisms, etc., can solve the problems of not achieving the effect of increasing yield, not being able to adapt well to the planting conditions of styli, and achieving the effect of increasing yield

Inactive Publication Date: 2010-06-16
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, the rhizobia introduced from Australia cannot be well adapted to the planting conditions of Stylophyllum in southern my country, and cannot achieve a significant increase in production after inoculation on Stylophyllum
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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  • Bradyrhizobium sp.RY4 strain and application thereof
  • Bradyrhizobium sp.RY4 strain and application thereof
  • Bradyrhizobium sp.RY4 strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 The acquisition and cultivation of Rhizobium RY4

[0037] In November 2008, the inventor collected a root nodule of a luxuriant flower and grass from the mango field (E101. 46.137'N26.25.276 H1100) in the sticky soil of Panzhihua, Sichuan, cut off the aboveground part of the stylophyllum, and packed the whole root system together with the root nodule. Put it in a fresh-keeping bag and store it in an ice pack at low temperature. A nitrogen-fixing bacteria strain RY4 (Bradyrhizobium sp. RY4) was obtained after isolation and purification, and was deposited in China Center for Type Culture Collection on November 13, 2009, with the preservation number CCTCC NO: M 209268.

[0038] (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 nodules with 2 mm in color and place them in a petri dish with grade 2 water. The surface is rinsed...

Embodiment 2

[0061] Example 2 The 16S rDNA sequence sequencing of Rhizobium RY4 and the determination of its category

[0062] In order to determine the phylogenetic status of Rhizobium RY4, the 16SrDNA series of the isolated strains were sequenced. Firstly, total DNA was extracted using a kit from Omega, and then PCR-specific amplification was performed using primers.

[0063] Upstream primer 35fc: CTKAAGAGTTTGATCMTGGCTCAGATTGAAC;

[0064] Downstream primer 1492r: TACGGYTACCTTGTTACGACTT.

[0065] The reaction conditions are as follows:

[0066] PCR reaction

[0067] Primer: 35fc, 1492r

[0068] PCR recipe:

[0069] 10×Reaction Buffer 5.0μL

[0070] dNTPs (10mM) 1.0μL

[0071] P35fc (25Pmol) primer 0.5μL

[0072] P1492r (25Pmol) primer 0.5μL

[0073] Taq DNA polymerase (5u / μL) 1.0μL

[0074] Template DNA 1.0 μL

[0075] Make up to 41 μL with ultrapure water

[0076] PCR procedure

[0077] Pre-denaturation at 94°C: 3min

[0078] Denaturation at 94°C for 50s

[0079] 56℃...

Embodiment 3

[0084] Example 3 Application experiment of rhizobia RY4

[0085] In order to confirm the effect of RY4 on Stylophyllum, two Stylophyllum varieties (TPRC2, TPRC5) were used, and the root nodules RY4 were reattached to the roots of Stylophyllum by sand culture method, with 4 replicates for each treatment (bacteria, sand and seedlings). For treatment, refer to the backgrain experiment procedure), and nodule inoculation was used as the control (CK) for backgrain comparison. The whole process is irrigated with low-nitrogen nutrient solution. The physiological indicators of Stylophyllum TPRC2 and TPRC5 back-inoculated with Rhizobium RY4 are shown in Table 2 and the appendix Figure 8 . attached Figure 8 The middle left square column (a) is TPRC2, and the left square column (b) is TPRC5 (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).

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Abstract

The invention discloses a Bradyrhizobium sp.RY4 strain and application thereof. The Bradyrhizobium sp.RY4 strain is preserved at the China center for type culture collection and the preservation number is CCTCC NO:M209268. When the Bradyrhizobium sp.RY4 is back-jointed to stylosanthes guianensis TPRC2 and TPRC5, the number of root nodules, the dry weight of the root nodule, the height of the strain, the fresh weight of the plant and the nitrogenase activity are greatly increased and obviously higher than those in a control group; and the effect of the RY4 back jointed to the stylosanthes guianensis TPRC2 is better. Thalli are released to the natural environment, the thalli increase the number of the groups of nodule bacteria in soil, contribute to the nodulation and nitrogen fixation of the stylosanthes guianensis, are harmless for human beings, animals and plants, and do not pollute the environment.

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. The main leguminous forage, there is a saying in the grass industry that "there is alfalfa in the nort...

Claims

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Application Information

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