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Bacterial strain of root nodule nitrogen-fixing strain series RY3 and application thereof

A strain and nodule technology, applied in bacteria, organic fertilizers, microorganisms, etc., can solve the problem of not being able to achieve high-yield effects, and achieve the effect of promoting nitrogen fixation by nodules

Inactive Publication Date: 2010-06-23
TROPICAL CORP STRAIN RESOURCE INST CHINESE ACAD OF TROPICAL AGRI SCI +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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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  • Bacterial strain of root nodule nitrogen-fixing strain series RY3 and application thereof
  • Bacterial strain of root nodule nitrogen-fixing strain series RY3 and application thereof
  • Bacterial strain of root nodule nitrogen-fixing strain series RY3 and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] The acquisition and cultivation of embodiment 1 rhizobia RY3

[0039]On February 27, 2009, the root nodules of a plant with large nodules and lush growth were collected from the Tropical Ecological Agriculture Research Institute of Yunnan Academy of Agricultural Sciences (E101.49N25.50H1084). Put them together in a fresh-keeping bag and store them in an ice pack at low temperature. After isolation and purification, a nitrogen-fixing bacteria strain RY3 (Bradyrhizobium sp. RY3) was obtained. It was preserved in the China Center for Type Culture Collection on November 13, 2009, with the accession number It is CCTCC NO: M 209267.

[0040] (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 clean; this process ensures the integrity of the ...

Embodiment 2

[0063] The 16S rDNA sequence sequencing of embodiment 2 Rhizobium RY3 and the determination of category

[0064] In order to determine the phylogenetic status of Rhizobium RY2, the 16SDNA series of the isolated strains were sequenced. First, use the kit from Omega to extract the total DNA (this method is not a conventional method. Extracting with a kit is an efficient and convenient method, but the cost is a bit higher compared with the conventional method), and then use the primers to perform PCR. specific amplification

[0065] Upstream primer 35fc: CTKAAGAGTTTGATCMTGGCTCAGATTGAAC;

[0066] Downstream primer 1492r: TACGGYTACCTTGTTACGACTT).

[0067] The reaction conditions are as follows:

[0068] PCR reaction

[0069] Primer: 35fc, 1492r

[0070] PCR recipe:

[0071] 10×Reaction Buffer 5.0μL

[0072] dNTPs (10mM) 1.0μL

[0073] P35fc (25Pmol) primer 0.5μL

[0074] P1492r (25Pmol) primer 0.5μL

[0075] Taq DNA polymerase (5u / μL) 1.0μL

[0076] Template DNA 1.0 μL ...

Embodiment 3

[0086] The application experiment of embodiment 3 rhizobia RY3

[0087]In order to confirm the effect of RY3 on Stylophyllum, two Stylophyllum varieties (TPRC2, TPRC5) were used, and the root nodules RY3 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 Stylosia TPRC2 and TPRC5 after inoculation with Rhizobium RY3 are shown in Table 2 and attached Figure 8 , attached Figure 9 . attached Figure 8 And attached Figure 9 In the figure, the left column (a) is TPRC2, and the right column (b) is TPRC5. (attached Figure 8 And attached Figure 9 Other square bar graphs in the middle are existing rhizobia control results, which are not directly related to ...

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Abstract

The invention discloses a bacterial strain of a root nodule nitrogen-fixing strain series RY3 and application thereof. The bacterial strain (Bradyrhizobium sp. RY3) is preserved in the China Center for Type Culture Collection on November 13th, 2009, and the collection number of the bacterial strain is CCTCC No: M 209267. The root nodule nitrogen-fixing strain series RY3 of the invention is grafted back to Stylosanthes guianensis TPRC2 and TPRC5 so that the fresh weight and dry weight of the Stylosanthes guianensis are increased by over triple. The RY3 is a highly-efficient bacterial strain which can ensure that the high yield of both the fresh weight and dry weight of the Stylosanthes guianensis can be achieved and the dissemination capacity of the Stylosanthes guianensis is stronger after the Stylosanthes guianensis is grafted with rhizobia; and after being released to the natural environment, the RY3 has no harm to human, animals and plants and does 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. 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 刘国道黄艳霞董荣书
Owner TROPICAL CORP STRAIN RESOURCE INST CHINESE ACAD OF TROPICAL AGRI SCI
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