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Chamaecrista Bradyrhizobium sp. JYN6 and application thereof

A technology of JYN6 and Cassia genus is applied to the rhizobium of the genus Cassia JYN6 and its application field, which can solve the problems affecting planting and popularization, slow nodulation, low nitrogen fixation efficiency, etc., and achieves strong nitrogen fixation ability and high nodulation rate. , the effect of increasing the total nitrogen content

Inactive Publication Date: 2019-02-22
FUJIAN AGRI & FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, under natural conditions, Cassia rotifera forms nodules slowly and rarely, and its nitrogen fixation efficiency is low, which seriously affects its planting and popularization and application.

Method used

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  • Chamaecrista Bradyrhizobium sp. JYN6 and application thereof
  • Chamaecrista Bradyrhizobium sp. JYN6 and application thereof
  • Chamaecrista Bradyrhizobium sp. JYN6 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Isolation and purification of rhizobia JYN6

[0045] 1 Isolation of rhizobia

[0046] Take fresh, mature and large nodules of Cassia pinnata, rinse them with water, and dry the surface moisture with filter paper. Put it in 95% (w / v) ethanol for 3-5 minutes, take it out and rinse it with sterile water for 5-6 times, then put it in 0.1% (w / v) HgCl 2 Sterilize for 3-5 minutes, take it out and rinse with sterile water 5-6 times. Then cut in half on a flame-sterilized glass slide, clamp half of the tumor with sterile tweezers, make the incision face the surface of YMA (Table 1) culture medium, and incubate at 28°C after being upside down.

[0047] 2 purification

[0048] After the growth of bacteria, a small amount of rhizobia colonies were scraped from the plate with the tip of a pipette, diluted with 1 mL of sterile water, and then streaked on the plate for culture again. After 3 days, the colonies were observed until 15 days (slow-growing It takes 7-15 days...

Embodiment 2

[0054] Example 2 Identification of 16S rDNA Molecular Biology of Rhizobium JYN6

[0055] Perform PCR-specific amplification of 16S rDNA on rhizobia monoclonal liquid, the forward primer is SEQ ID NO.3: V4V5515-F 5'-GTGCCAGCMGCCGCGGTAA-3'; the reverse primer is SEQ ID NO.4: V4V5907- R 5'-CCGTCAATTCCTTTGAGTTT-3'. The enzyme is 2×star Mix. The PCR amplification products were detected by electrophoresis imaging technology to observe whether they had bands, and the remaining PCR amplification products were used for sequence determination. The sequencing result is shown in SEQ ID NO:5. The PCR reaction system is shown in Table 4.

[0056] Table 416S rDNA2×starMix enzyme reaction system

[0057]

[0058] Reaction conditions: 95°C for 5 min; (95°C for 20 s, 55°C for 20 s, 72°C for 50 s)×44 cycle; 70°C for 5 min.

[0059] Obtain 13 reference strain sequences from the NCBI (GenBank) database, use the software BioEdit and MEGA6 to analyze the 16S rDNA partial sequences of the iso...

Embodiment 3

[0060] Example 3 Tieback test

[0061] The purpose of the experiment: to screen out the rhizobia with high binding efficiency and strong nitrogen fixation ability with Cassia forage.

[0062] Test materials: Test plant: Minyu No. 1 Cassia rotifera; Test strain: isolated and purified Rhizobium.

[0063] Main test instruments and equipment: artificial climate growth chamber, ultra-clean workbench, high-pressure sterilization pot, constant temperature incubator, 25 potted pots of 15×15 cm, 2 beakers of 1 L, tweezers, Petri dishes, filter paper, glass Sticks, scissors and gauze etc.

[0064] Test drugs and reagents:

[0065] (1) Test drugs: mannitol, MgSO 4 ∙7H2O, NaCl, yeast powder, K 2 HPO 4 、KH 2 PO 4 , CaCO 3 , Ca(NO 3 ) 2 ∙4H 2 O, MgSO 4 ·7H 2 O, CaCl 2 2H 2 O, Na 2 HPO 4 12H 2 O, C 10 h 12 N 2 o 8 FeNa·3H 2 O, Na 2 MoO 4 , MnSO 4 、H 3 BO 3 、CuSO 4 ·5H 2 O and ZnSO 4 ·7H 2 O.

[0066] Test reagent:

[0067] 1) YMA (Yeast Mannitol Agar) liquid...

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Abstract

The invention discloses Chamaecrista Bradyrhizobium sp. JYN6 and application thereof, and belongs to the technical field of microorganisms. The strain is obtained by being separated and purified froma Chamaecrista chamaecrista forage fresh root nodule, and subjected to nodA gene detecting through a PCR and 16S rDNA molecular biological identification, and the strain is determined to be a new strain system of Bradyrhizobium and named as Anyu No.1; and the strain is preserved in the China Center for Type Culture Collection on April 10, 2018, and the preservation number is CCTCC NO: M2018192. Itproves that the Bradyrhizobium sp. JYN6 has the characteristics of efficient nodulation and high nitrogen fixation capacity through a laboratory potting tie-back test, by being inoculated with the Bradyrhizobium, the number of root nodules of Chamaecrista forage can be remarkably increased, the root nodule nitrogen fixation efficiency of the Chamaecrista forage can be remarkably improved, and thebiomass and the plant nitrogen content of the Chamaecrista forage can be remarkably increased, and thus the effect of soil fertility culturefertility improving and ecological environment improving are achieved.

Description

technical field [0001] The invention belongs to the field of microorganisms, and in particular relates to a Cassia rhizobia JYN6 and its application. Background technique [0002] my country has about 56.9 million hm 2 Acidic soil accounts for more than 42.2% of the total cultivated land area. In recent years, with the application of large amounts of chemical nitrogen fertilizers, soil acidification has become increasingly serious. The acidic soil in my country is mainly concentrated in the area south of the Yangtze River. This area is rich in light, heat, water and other resources, but the soil is thin, sticky, low in pH and nutrient availability, which seriously restricts the rapid development of agriculture in this area. In terms of production, crop yields are often increased by applying large amounts of chemical fertilizers. However, the input of a large amount of chemical fertilizers will not only lead to soil compaction and further acidification, but also cause eutr...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01G31/00A01G7/06C12R1/01
CPCA01G7/06A01G31/00C12N1/205C12R2001/01
Inventor 廖红杨庆李欣欣
Owner FUJIAN AGRI & FORESTRY UNIV
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