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RY2 strain of root nodule nitrogen-fixing bacterial strain system and application thereof

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

Inactive Publication Date: 2010-09-08
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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  • RY2 strain of root nodule nitrogen-fixing bacterial strain system and application thereof
  • RY2 strain of root nodule nitrogen-fixing bacterial strain system and application thereof
  • RY2 strain of root nodule nitrogen-fixing bacterial strain system and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] The acquisition and cultivation of embodiment 1 rhizobia RY2

[0031] On November 14, 2008, a root nodule of a luxuriant Stylophyllum was collected in the experimental site of the Institute of Thermal Economics, Yunnan Academy of Agricultural Sciences (N=24°45.684′, E=98°55.546′, H=1023m). Put the whole root system together with the root nodules into the fresh-keeping bag for the aboveground part of the stigma, and put it in the ice pack for low temperature preservation. A nitrogen-fixing bacteria strain RY2 (Bradyrhizobium yuanmingense RY2) 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 209266.

[0032] (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...

Embodiment 2

[0055] The 16S rDNA sequence sequencing of embodiment 2 Rhizobium RY2 and the determination of category

[0056] In order to determine the phylogenetic status of Rhizobium RY2, the 16SDNA 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.

[0057] Upstream primer 35fc: CTKAAGAGTTTGATCMTGGCTCAGATTGAAC;

[0058] Downstream primer 1492r: TACGGYTACCTTGTTACGACTT.

[0059] The reaction conditions are as follows:

[0060] PCR reaction

[0061] Primer: 35fc, 1492r

[0062] PCR recipe:

[0063] 10×Reaction Buffer 5.0μL

[0064] dNTPs (10mM) 1.0μL

[0065] P35fc (25Pmol) primer 0.5μL

[0066] P1492r (25Pmol) primer 0.5μL

[0067] Taq DNA polymerase (5u / μL) 1.0μL

[0068] Template DNA 1.0 μL

[0069] Make up to 41 μL with ultrapure water

[0070] PCR procedure

[0071] Pre-denaturation at 94°C: 3min

[0072] Denaturation at 94°C for 50s

[0073] 56℃ a...

Embodiment 3

[0078] The application experiment of embodiment 3 rhizobia RY2

[0079] In order to confirm the effect of RY2 on Stylophyllum, two main Stylophyllum varieties (TPRC2, TPRC5) were used, and the root nodules RY2 were reattached 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 step of backgrafting experiment), and nodule inoculation was used as the control (CK) for backgrafting comparison. The whole process is irrigated with low-nitrogen nutrient solution. The physiological indicators of Stylosia TPRC2 and TPRC5 after inoculation with Rhizobium RY2 are shown in 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, an...

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Abstract

The invention discloses a RY2 strain of a root nodule nitrogen-fixing bacterial strain system and application thereof. The strain (Bradyrhizobium yuanmingense RY2) is preserved in the China Center for Type Culture Collection (CCTCC) on 13th November in 2009. The preservation number is CCTCC NO:M 209266. The RY2 strain of the root module nitrogen-fixing bacterial strain system can exclusively greatly improve the nitrogenase activity of TPRC2, so that the nitrogenase activity of the TPRC2 is over three times higher than that of TPRC5 and can achieve higher nitrogenase activity by using a few root nodules. The yield of stylosanthes guianensis can be improved by over 30 to 40 percent by inoculating the root nodule bacteria, and the root nodule bacteria can be suitable for intercropping of stylosanthes guianensis TPRC2 and provides a large number of nitrogen for the stylosanthes guianensis, crops and soil. Meanwhile, the RY2 has strong salt tolerance, can grow in the environment that the NaCl concentration is higher than 0.25mol / L, can be planted in the salinized soil and has better effects of improving the yield of the stylosanthes guianensis and utilizing and improving the salinized soil.

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 successfully planted in Hainan, and 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. In southern China such as Guangdong, Guangxi and other provinces (regions), interpla...

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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