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Method for promoting nodulation and nitrogen fixation of root system of tumorous stem mustard by using azorhizobium caulinodans

A technology of nitrogen-fixing rhizobia and stem mustard, applied in horticultural methods, botany equipment and methods, applications, etc., to achieve great application value, increase crop yield, and reduce environmental pollution

Pending Publication Date: 2021-07-30
YANGTZE NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, although some progress has been made in the basic experiment of inducing nodulation of non-legume plants, the research is still in the basic stage. Before systematic and perfect application in actual agricultural production, more time and energy are needed to invest in research in this area.

Method used

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  • Method for promoting nodulation and nitrogen fixation of root system of tumorous stem mustard by using azorhizobium caulinodans
  • Method for promoting nodulation and nitrogen fixation of root system of tumorous stem mustard by using azorhizobium caulinodans
  • Method for promoting nodulation and nitrogen fixation of root system of tumorous stem mustard by using azorhizobium caulinodans

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Research on the Growth-promoting Effect of Rhizobium on Stem Mustard

[0034] 1. Seed treatment and bacterial solution configuration

[0035] Select uniform and plump mustard seeds with roughly the same particle size and place them in 2ml centrifuge tubes, place the centrifuge tubes in an ultra-clean workbench, soak the seeds with 50% NaClO for 10 minutes, shake the centrifuge tubes during soaking to ensure that all the seeds are soaked, pour out the cleaning solution Use a pipette gun to draw sterile water to rinse and disinfect the surface. The rinsing solution is replaced with a pipette gun with a sterilized 1ml tip. The surface-sterilized seeds were placed in a tissue culture bottle, and the surface of a high-temperature sterilized agar-water solid medium (hereinafter referred to as the medium) was poured into the tissue culture bottle for limited bacterial culture. Placed on a conventional culture rack for cultivation.

[0036]Activation and expansion...

Embodiment 2

[0041] Example 2 Different concentrations of rhizobia infecting the tuber mustard infection experiment

[0042] Disinfect the surface of the seeds of Fuza No. 2 in the same way as above, place them on agar-water solid medium for bacterial culture at the upper limit, and when the seeds germinate to the stage of two leaves and one heart, remove the ungerminated seeds with tweezers, leaving seedlings with roughly the same growth status. Put the rhizobia bacteria liquid in the logarithmic growth phase in a 50ml centrifuge tube, centrifuge at 4500r / m for 10min to collect the bacteria, and dilute them with PBS buffer to 1.0×10 9 cfu / mL, 1.0×10 8 cfu / mL, 1.0×10 7 cfu / mL, 1.0×10 6 Four different concentrations of cfu / mL are available for use. Pipette 1ml of rhizobia bacteria solution at different concentrations to the surface of the culture medium, set 4 replicates for each concentration, 10 grains per bottle, and connect the same amount of PBS for parallel control, set 4 replicate...

Embodiment 3

[0046] Example 3 Observation on the colonization of rhizobia in the root system of Stem mustard

[0047] Under indoor conditions, the plump seeds of Fuza No. 2 were used for surface disinfection, and the disinfection method was the same as above. Placed in agar-water solid medium for upper-limit bacterial culture, germinated to the stage of two leaves and one heart, each tissue culture bottle draws 1mL1.0×10 8 The cfu / mL PBS diluted bacterial solution was added to the surface of the agar-water solid medium to inoculate the tuber mustard seedlings. After inoculation, seal the tissue culture vials with parafilm to avoid contamination. Place conventional intelligent culture racks to limit bacterial culture. Set time gradients on the 3d, 6d, 9d, 12d, 15d, 18d, and 21d of inoculation, and regularly use a fluorescence microscope to detect the colonization of GFP-labeled A. caulinodans in the roots of the stem mustard seedlings.

[0048] Pick representative tuber mustard seedlings...

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Abstract

The invention discloses a method for promoting nodulation and nitrogen fixation of a root system of a tumorous stem mustard by utilizing azorhizobium caulinodans. The method comprises the following steps that in the culture or cultivation process of the tumorous stem mustard, the azorhizobium caulinodans is applied to a culture medium or soil of the tumorous stem mustard, or the roots of the tumorous stem mustard are soaked with the azorhizobium caulinodans. Researches show that the rhizobium can infect leguminous plants to establish a symbiotic relationship, and can infect the tumorous stem mustard through an artificial induction method and promote the tumorous stem mustard to nodulate and fix nitrogen. Therefore, the nitrogen fixation capability of the root nodules can be utilized to reduce the use amount of chemical nitrogen fertilizer in cultivation, relieve the dependence on the chemical nitrogen fertilizer in the agricultural process, reduce environmental pollution and improve the crop yield.

Description

technical field [0001] The invention relates to the field of crop cultivation, in particular to a method for cultivating stem mustard, and more specifically to a method for promoting root nodulation and nitrogen fixation of stem mustard by utilizing the nitrogen-fixing rhizobia of the mustard. Background technique [0002] Tumida Tsenet Lee (Brassica junceavar.tumida Tsenet Lee) belongs to the Brassicaceae plant, belongs to the class of Dicotyledonous Plants, and belongs to the genus Brassica. The origin of mustard in China is the cold northern region, and then moved from the northern region to Sichuan for planting. The history of growing mustard in Sichuan has been more than 100 years, commonly known as "Qingcaitou". Yang Yigeng, an expert from the Chongqing Institute of Agricultural Sciences in my country, and others carried out the work of systematic classification and induction of mustard. Classify "stem mustard" as a class of mustard, and "Qing Caitou" was renamed "st...

Claims

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Application Information

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IPC IPC(8): A01G22/15A01G7/06
CPCA01G22/15A01G7/06
Inventor 王殿东常黎洁张卓廖静静蔡兆明罗路云王攀石家宇
Owner YANGTZE NORMAL UNIVERSITY
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