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Vicia sativa rhizobium strain VS21-2 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

Inactive 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 VS21-2 and application thereof
  • Vicia sativa rhizobium strain VS21-2 and application thereof
  • Vicia sativa rhizobium strain VS21-2 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Example 1 Isolation, purification and storage of Rhizobium vulgaris VS21-2

[0016] From wild arrow pea collected from Xiangshui Village, Yanjiang Town, Yanjiang District, Ziyang City, Sichuan Province, choose strong plants with large and full red nodules on the main roots, wash the nodules, take part of the root bark off, and perform surface disinfection: use 95% Soak in ethanol for 5 minutes to eliminate the surface tension, then disinfect the surface with mercury (0.1% m / v) for 5 minutes, and finally rinse with sterile water 6-8 times. Put the nodules on a sterilized white porcelain plate in a sterile environment, use bamboo sticks to break the nodules and smear them on a plate of YMA medium, carry out streaking purification, and cultivate 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 O0.1g, NaCl 0.1g, ammonium molybdate (1%) 2mL, boric acid (1%) 2mL, Congo red (1%) 2.5mL, mannitol 10g, yeast powder 0.8g, agar 18-20g, water...

Embodiment 2

[0019] Example 2 Rhizobium back joint test

[0020] The back inoculation test of rhizobia used sand culture method. The arrow pea species used in the experiment was Northern Sichuan arrow pea, which originated from Xiangshui Village, Yanjiang Town, Yanjiang District, Ziyang City, Sichuan. It was carried out in a light room (temperature controlled 22-24°C, light intensity about 2800 lux, sunshine time 14h), planted 46d and harvested. Regularly supplement the sterilized nitrogen-free nutrient solution, and inoculate Rhizobium versicolor VS21-2 into the sand culture vessel. The sand culture device uses a 300mL plastic cup, and the substrate is vermiculite, so that Rhizobium versicolor VS21-2 is not inoculated. The plant of the same product was the control CK, and each treatment was repeated three times. After harvest, the number of nodules and the dry weight of the pea plant were used to evaluate the inoculation effect of Rhizobium vulgare VS21-2.

[0021] (1) Bacterial liquid cultu...

Embodiment 3

[0031] Example 3 The resistance ability of Rhizobium sagittatum VS21-2

[0032] The resistance to stress of Rhizobium sagittatum VS21-2 was mainly tested for salt tolerance and acid and alkali tolerance. The YMA medium was used as the basic medium, and the YMA plates cultured at pH 7, 28°C for 7 days were used as positive control (CK). The above-mentioned YMA slant culture of Rhizobium sagittatum VS21-2 was scraped with sterile water to prepare a bacterial suspension for use. The spot inoculation method was adopted, and each treatment was repeated 3 times. The plates for acid and alkali resistance and salt resistance tests were all incubated at 28°C for 3 days and the recorded results were observed.

[0033] (1) Medium formula for acid and alkali resistance determination: use YMA medium as the basic medium, adjust the pH value with HC1 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 VS21-2 and an application thereof. A bacterial strain is obtained from fresh Vicia sativa root nodules by separation and purification, and belongs to a Rhizobium sophorae 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:M2017608. The Vicia sativa rhizobium strain VS21-2 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 Vicia sativa without application of nitrogenous fertilizers and other fertilizers and only with inoculation of rhizobium VS21-2 reaches 66.7% or above, and yield increase of fresh roots in the condition reaches 73%.

Description

Technical field [0001] The present invention relates to the field of microorganisms, in particular to a strain VS21-2 of the pea pea root nodule strain and its application. Background technique [0002] Arrow pea (Vicia sativa L.), also known as large vetch, belongs to a cultivar of Leguminosae (Leguminosae) Vicia genus. It has a large main root, shallow soil, many nodules, and arrow-shaped leaves, so it is named Arrow The origin of pea is in southern Europe and western Asia. It was introduced into my country for cultivation in the 1940s. The cultivation range of my country covers 29 provinces (regions) including Qinghai, Jiangsu, Hunan, and Taiwan (Hong Ruxing et al., 1985). As a kind of excellent feed and green manure, the pea has high nutritional value, tender stems and branches, more leaves, and good palatability. It is a commonly used green manure germplasm resource in production (Lu Binglin et al., 2015; Zhao Hejing, 2015 ). Farrow pea also has the characteristics of droug...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/41
CPCC12N1/205C12R2001/41
Inventor 陈远学徐开未彭丹吴永成周元胡斐杨云飞
Owner SICHUAN AGRI UNIV
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