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Vicia sativa rhizobium strain VS6-3 and application thereof

A technology of arrow pea and rhizobia, which is applied in the field of microorganisms to achieve the effect of obvious yield increase, strong nitrogen fixation ability and good affinity

Active Publication Date: 2018-05-15
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the research on high-efficiency rhizobia suitable for Sichuan ecological environment is almost blank

Method used

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  • Vicia sativa rhizobium strain VS6-3 and application thereof
  • Vicia sativa rhizobium strain VS6-3 and application thereof
  • Vicia sativa rhizobium strain VS6-3 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Example 1 Isolation, purification and preservation of Arrowhead pea rhizobia VS6-3

[0016] From the wild arrowhead peas collected in Zhongxiang Town, Renshou County, Meishan City, Sichuan Province, select the large and plump red root nodules on the main roots of robust plants, wash the root nodules, and take part of the root bark for surface disinfection: soak in ethanol (95%) Eliminate surface tension for 5 minutes, then soak in mercuric chloride (0.1% m / v) for 5 minutes, and finally rinse with sterile water for 6-8 times. Put the root nodules on a sterilized white porcelain plate under a sterile environment, clip the root nodules with a bamboo stick and smear them on the plate of YMA medium, perform streaking and purification, and culture in a 28°C incubator. YMA medium formula: KH 2 PO 4 0.25g, MgSO 4 ·7H 2 O 0.2g, CaCl 2 ·6H 2 O 0.1g, NaCl 0.1g, ammonium molybdate (1%) 2mL, boric acid (1%) 2mL, Congo red (1%) 2.5mL, mannitol 10g, yeast powder 0.8g, agar 18-2...

Embodiment 2

[0019] The back graft test of embodiment 2 rhizobia

[0020] The backgrafting test of rhizobia uses the sand culture method, and the arrowhead pea variety used in the test is North Sichuan arrowhead pea, which comes from Xiangshui Village, Yanjiang Town, Yanjiang District, Ziyang City, Sichuan. Carry out in the light room (temperature control 22~24 ℃, light intensity about 2800 lux, sunshine time 14h), plant and harvest 46 days. Regularly supplement the sterilized nitrogen-free nutrient solution, inoculate the rhizobia VS6-3 into the sand culture device, the sand culture device uses a 300mL plastic cup, and use vermiculite as the substrate, so as not to inoculate the rhizobia VS6-3 The plants of the same variety as the control CK, each treatment was repeated three times. After harvesting, the nodule number and dry weight of the plants were used to evaluate the inoculation effect of Rhizobia VS6-3.

[0021] (1) Bacterial culture: inoculate the Rhizobium spp. VS6-3 in YMA liqu...

Embodiment 3

[0031] Example 3 The Stress Resistance of Pea Arrowhead Rhizobium VS6-3

[0032] The stress resistance of Rhizobium spp. VS6-3 was mainly tested for salt tolerance, acid and alkali resistance. The YMA medium was used as the basal medium, and the YMA plate cultured at pH 7 and 28°C for 7 days was used as the positive control (CK). Scrape and wash the above-mentioned YMA slant culture of Rhizobium argentina VS6-3 with sterile water to make a bacterial suspension for use. The spot inoculation method was used, and each treatment was repeated 3 times. Plates for acid and alkali resistance and salt resistance tests were all cultured at 28°C for 3 days, and the results were observed and recorded.

[0033] (1) Medium formula for acid and alkali resistance determination: take YMA medium as the basic medium, adjust the pH value with HCl and NaOH, so that the pH value of the medium is 4.0, 5.0, 6.0, 8.0, 9.0, 10.0, 11.0 .

[0034] (2) Medium formula for salt tolerance determination: ...

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Abstract

The invention provides a Vicia sativa rhizobium strain VS6-3 and an application thereof. A bacterial strain is obtained from fresh Vicia sativa root nodules by separation and purification, and belongsto a Rhizobium anhuiense strain of Rhizobium. The bacterial strain is preserved in China Center for Type Culture Collection in Wuhan university at October 23th, 2017, and the preservation number is CCTCC No:M2017606. The Vicia sativa rhizobium VS6-3 is an excellent Vicia sativa rhizobium strain, which is applied to production of Vicia sativa in Sichuan, has good symbiotic nitrogen fixation ability and obvious yield increasing effects of Vicia sativa, has capabilities for secreting IAA, dissolving phosphorus and potassium, and good stress resistance. The yield increase of fresh grass of Viciasativa without application of nitrogenous fertilizers and other fertilizers and only with inoculation of rhizobium VS6-3 reaches 77.8% or above, and yield increase of fresh roots in the condition reaches 73.7%.

Description

technical field [0001] The present invention relates to the field of microbes, in particular to a strain VS6-3 of pea rhizobium and application thereof. Background technique [0002] Arrowhead pea (Vicia sativa L.), also known as Vicia sativa L., belongs to a cultivar of the genus Vetaceae of the family Leguminosae. It has a large main root, shallow penetration into the soil, many root nodules, and arrow-shaped leaves, so it is named Arrowhead Pea. Pea, which originated in southern Europe and western Asia, was introduced into my country for planting and cultivation in the 1940s. The planting range in my country reached 29 provinces (regions) including Qinghai, Jiangsu, Hunan, and Taiwan (Hong Ruxing et al., 1985). Arrowhead pea, as an excellent feed and green manure dual-purpose crop, has high nutritional value, tender stems and branches, more leaves, and good palatability. It is a relatively common green manure germplasm resource used in production (Lu Binglin et al., 2015; ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12R1/41
CPCC12N1/205C12R2001/41
Inventor 徐开未陈远学陈沁园蒋帆彭丹刘明吴永成
Owner SICHUAN AGRI UNIV
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