Artificial culture, breeding and preservation method of plant root-knot nematode

A technique for artificially cultivating and root-knot nematodes, applied in the field of laboratory reproduction and preservation away from host plants, can solve the problems of investing a lot of time, and achieve the effect of shortening time, strong operability, and fast propagation method

Inactive Publication Date: 2009-07-08
INST OF TROPICAL BIOSCI & BIOTECH CHINESE ACADEMY OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, in the related studies reported at home and abroad, the culture method gradually tends to leave the greenhouse and leave the soil and use the method of nutrient solution or culture medium in the labo

Method used

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  • Artificial culture, breeding and preservation method of plant root-knot nematode
  • Artificial culture, breeding and preservation method of plant root-knot nematode
  • Artificial culture, breeding and preservation method of plant root-knot nematode

Examples

Experimental program
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Effect test

Embodiment 1

[0019] Embodiment 1: the preparation of the medium for cultivating Fusarium oxysporum f.sp.cubense

[0020] Preparation of test tube slant medium: weigh 10 grams of glucose, 5 grams of peptone, KH 2 PO 4 1 g, MgSO 4 ·7H 2 O 0.5 g, dissolved in 1000 ml of water, add 15-18 g of agar, heat and boil until the agar melts, make up the volume to 1000 ml, after cooling properly, put it into test tubes, the filling volume is 1 / 5-1 of the test tube height / 6. Plug the test tube mouth with a test tube stopper, autoclave at 121°C for 25 minutes, take it out while it is hot, place it on an inclined plane until it cools down, and the height of the inclined plane should not exceed 1 / 3 of the height of the test tube.

[0021] Preparation of liquid seed medium: weigh 10 grams of glucose, 5 grams of peptone, KH 2 PO 4 1 g, MgSO 4 ·7H 2 O 0.5 g, dissolved in 1000 ml of water, then sub-packaged, the liquid volume is 20-30ml / 100ml triangular flask, plugged with a cotton plug, autoclaved a...

Embodiment 2

[0023] Embodiment 2: the preparation of Fusarium oxysporum f.sp.cubense spore liquid

[0024] The Fusarium oxysporum f.sp.cubense described in the present invention is a bacterial species preserved by the inclined-plane cold storage method in our laboratory.

[0025] a) Activation of Fusarium oxysporum f.sp.cubense strain

[0026] The method of activating the strains preserved on a slant: pick a strain with a diameter of about 8 mm from the strains preserved on a slant and inoculate it into the slant culture medium of a test tube for activation, cultivate it in an environment of 28 ± 2°C for 3-5 days and then use it for later use.

[0027] b) Cultivation of seed solution

[0028] From the Fusarium bacterium activated in the test tube, pick three bacterium pieces and inoculate them respectively in three 100ml triangular flasks with 25ml liquid seed culture medium, place them on a shaker, the rotating speed of the shaker is 180r / min, at 28 Continue to cultivate in the environm...

Embodiment 3

[0032] Embodiment 3: the preparation of double culture medium

[0033] After the prepared solid medium is hot or melted by heating, pour it into a 9cm sterile petri dish in a sterilized ultra-clean workbench. , too little will limit the living space of nematodes, and too much will not be conducive to the separation of nematodes. After the medium is cooled and solidified, add 0.1-0.2ml of Fusarium oxysporumf.sp.cubense spore liquid to each plate, spread evenly, seal with a parafilm, and incubate upside down in a constant temperature incubator at 28°C for 3-5 days, white bacteria After the silk grows all over the plate, it is the double medium.

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Abstract

The invention relates to a method for artificial culture and propagation of plant root-knot nematodes and for species preservation. The propagation and preservation of the root-knot nematodes in a lab depend on host plants all the time; the invention places a relatively small amount of root-knot nematode ovums or 2nd instar larvas in a particular dual culture medium rather than host roots and performs a upside-down culture under conditions of constant temperature and hermetic closure, the root-knot nematodes ingest, grow and propagate inside the culture medium and a large amount of nematomorphas can be obtained in a short term; additionally, a culture plate is timely transferred to be located under cold conditions, and dish change and subculture are implemented once within a half year so that the propagation and preservation processes of the root-knot nematodes are carried out without the host plants. The inventive advantages lie in: saving the culture program of the root-knot nematodes to host the plants without losing the infectivity of the root-knot nematodes on sensitive host plants; the employed materials have low costs and the experiment method is easy in operation; the invention provides a simple, convenient and fast propagation method for the species preservation of the root-knot nematodes, in particular for a large number of nematomorphas as required in the related researches of the root-knot nematodes.

Description

technical field [0001] The invention relates to a method for artificially cultivating and preserving species of root-knot nematodes of plants, in particular to a method for laboratory propagation and preservation of main root-knot nematodes separated from host plants suitable for plants. The invention belongs to the application field of plant root-knot nematode propagation and preservation. Background technique [0002] Root-knot nematode (Meloidogyne spp.) is a kind of plant root resident endoparasite widely distributed all over the world among nematodes. One of the genus of plant parasitic nematodes. There are more than 90 species of root-knot nematodes reported, among which the most widely distributed are M.incognita, M.arenaria, and M.javanica. and four dominant populations such as root-knot nematode (M.hapla). Root-knot nematodes have a wide range of parasitism, and can infect more than 3,000 species of plants, belonging to 114 families, including monocotyledonous pl...

Claims

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Application Information

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IPC IPC(8): A01K67/033A23K1/18C12N1/14C12R1/77
Inventor 鲍时翔曾庆飞方哲黄惠琴朱军吴晓鹏孙前光
Owner INST OF TROPICAL BIOSCI & BIOTECH CHINESE ACADEMY OF TROPICAL AGRI SCI
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