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Method for identifying tropical resistance of different banana varieties to No. 4 physiological race of fusarium wilt by using tissue culture seedlings

A technology of physiological races and tissue culture seedlings, which is applied in biochemical equipment and methods, biological material analysis, microbial measurement/inspection, etc., and can solve problems such as large footprint, cross-infection of growth conditions, and influence of resistance identification

Active Publication Date: 2021-09-14
INST OF AGRI ENVIRONMENT & RESOURCES YUNNAN ACAD OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method has a long identification cycle and a large area, and there are other uncontrollable growth conditions and other organisms that may cause cross-infection and affect the results of resistance identification.
[0008] At present, there is no report on the method of identifying the resistance of different banana varieties to the tropical type of fusarium wilt No.

Method used

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  • Method for identifying tropical resistance of different banana varieties to No. 4 physiological race of fusarium wilt by using tissue culture seedlings
  • Method for identifying tropical resistance of different banana varieties to No. 4 physiological race of fusarium wilt by using tissue culture seedlings
  • Method for identifying tropical resistance of different banana varieties to No. 4 physiological race of fusarium wilt by using tissue culture seedlings

Examples

Experimental program
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Effect test

Embodiment 1

[0047] Example 1. A method of utilizing tissue culture seedlings to identify the resistance of different banana varieties to No. 4 physiological race tropical type of fusarium wilt

[0048] 1.1 Cultivation of rooted seedlings of different banana varieties

[0049] The tissue-cultured seedlings of different banana varieties were multiplied and cultivated to obtain clustered shoots with relatively consistent growth; the clustered shoots were cut into individual plants and transferred to the rooting medium, and each bottle of medium was transferred to 5 plants, a total of 40 bottles, 200 strain.

[0050] Culture conditions: the temperature of the culture room is 25-28°C, the photoperiod is 12h light and 12h dark, and the light intensity is 1600-2000Lux.

[0051] 1.2 Inoculation method of pathogenic bacteria in banana tissue culture seedlings

[0052] Preparation of banana fusarium wilt spore suspension: Activate the TR4 strain stored at -20°C on a PDA solid medium, culture at a...

Embodiment 2

[0069] Embodiment 2 utilizes tissue culture plantlet to identify the method for TR4 resistance of different banana varieties in greenhouse

[0070] Banana germplasm resources to be tested: Banana germplasm resources introduced from the International Biodiversity Research Center and preserved in the Yunnan Academy of Agricultural Sciences. Using the above-mentioned identification results of the same banana germplasm resources in the greenhouse for disease resistance, the inoculated strains were the same strain.

[0071] 2.1 Banana seedling cultivation

[0072] The above-mentioned banana germplasm resources were selected for tissue culture before inoculation. The seedlings were cultured in the matrix coconut peat in the greenhouse for about 6 weeks. After growing to 10cm, they were used for artificial inoculation of TR4. Each germplasm was inoculated with more than 20 plants per batch. July and July 2019 In August 2019, 2 batches were inoculated.

[0073] 2.2 Culture of TR4 sp...

Embodiment 3

[0086] Example 3 Inoculation of rooted tissue cultured seedlings wounded roots in tissue culture bottle with TR4 strain

[0087] Steps: (1) After the rooted seedlings are cultivated for 15-20 days, add 300 μL of 1×10 6 The number of spores / mL bacterial liquid is tested in a sterile workbench; (2) Reclose the tissue culture bottle that has been inserted into the strain, and place it at a temperature of 25-28°C, photoperiod: 12h light and 12h dark, and the light intensity is 1600-2000Lux tissue culture indoor culture observation, see the results Figure 4 .

[0088] Result: if Figure 4 As shown in (A), after 3 days of inoculating the TR4 strain, the hyphae were covered with the medium; as Figure 4 As shown in (B), the seedlings died after 5 days, but there was no obvious yellow leaf symptom, which could not be determined to be caused by TR4 infection. The results showed that rooted banana seedlings were not suitable for TR4 inoculation by this method.

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Abstract

The invention discloses a method for identifying tropical resistance of different banana varieties to No. 4 physiological race of fusarium wilt by using tissue culture seedlings, and belongs to the field of agricultural production and plant protection. The method comprises the following steps: preparing rooting seedlings, preparing an inoculation spore suspension, inoculating pathogenic bacteria, and investigating diseases. According to the method, the pathogenic bacteria are directly inoculated by puncturing leaves of the rooting seedlings in a tissue culture bottle, the tissue culture bottle does not need to be replaced, and the operation is simple; compared with traditional root-damaged inoculation of field plants or greenhouse plants, the method has the advantages that the processes of seedling hardening and plant culture are reduced, the influence of the environment temperature and humidity on the identification result is avoided, and the identification result is accurate and reliable; and the tissue culture seedlings are subjected to disease resistance screening in the indoor tissue culture bottle, the conditions are easy to control, the identification period is greatly shortened, the speed is high, and a large amount of manpower, material resources and space can be saved.

Description

technical field [0001] The invention relates to a method for identifying plant disease resistance, in particular to a method for identifying the tropical type resistance of different banana varieties to Fusarium wilt No. 4 physiological race by using tissue culture seedlings, and belongs to the fields of agricultural production and plant protection. Background technique [0002] Banana is one of very important economic crops and food crops in tropical and subtropical countries. However, the development of the global banana industry is being seriously threatened by Fusarium wilt. Banana Fusarium wilt is a fungal disease of vascular necrosis caused by Fusarium oxysporumf.sp.cubense infection. control measures. Especially race 4 tropical type (T R4 ) harm is particularly serious, a large number of banana plantations due to T R4 And was abandoned. The asexual reproduction and single genetic basis of bananas have increased the difficulty in the application of traditional met...

Claims

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Application Information

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IPC IPC(8): C12Q1/18
CPCC12Q1/18G01N2333/37Y02P60/21
Inventor 刘立娜郑泗军李成云胡奈杨宝明黄玉玲李永平曾莉何平
Owner INST OF AGRI ENVIRONMENT & RESOURCES YUNNAN ACAD OF AGRI SCI
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