Method for cultivating meloidogyne hapla by adopting peanut rooting

A technology for root-knot nematodes and peanut roots, which is applied in animal husbandry and other directions to achieve the effects of high reproduction rate, simple operation, and beneficial to infection and reproduction

Active Publication Date: 2015-11-04
SHANDONG PEANUT RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Using the hairy root system to detect plant resistance to nematodes has been used in crops such as sugar beet, soybean, potato, tomato and sweet potato, but no one has tried to use the peanut hairy root system to cultivate root-knot nematode.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] a Preparation of peanut hairy roots

[0033] Select the seeds of Huayu No. 45 with large and plump seeds, wash them with tap water for 20 minutes, then soak them in alcohol with a mass percentage concentration of 75% for 1 minute under sterile conditions, and then soak them in 0.1% mercury liter for 15 minutes to carry out surface disinfection. Finally, wash with sterile water 3 times, 3min each time. After disinfection, use a sterile knife to peel off the seed coat on sterile filter paper, and inoculate it on the culture medium.

[0034] Under sterile conditions, use an inoculation loop to pick up the Agrobacterium rhizogenes A4 strain frozen in a low-temperature refrigerator at -20°C, inoculate it on a solid plate of LB+50mg / L Kan, and then culture it at a constant temperature of 28°C Cultivate in the dark in the box. After a single colony grows, pick a single colony and transfer it to 10ml LB liquid medium (with 50mg / L Kan added), and culture and recover in the dark...

Embodiment 2

[0040] a Preparation of peanut hairy roots

[0041] Select the large and plump Huayu No. 45 seeds, wash them with tap water for 30 minutes, then soak them in alcohol with a mass percentage concentration of 75% for 0.5 minutes under sterile conditions, and then soak them in 0.1% mercury liter for 15 minutes to carry out surface disinfection. , and finally washed 4 times with sterile water, 3 min each time. After disinfection, the seed coat was peeled off on sterile filter paper with a sterile knife, and inoculated on hormone-free MSB5 medium for culture. MSB5 medium is MS inorganic salt + B5 organic component (referred to as MSB5).

[0042] Under sterile conditions, use an inoculation loop to pick up the Agrobacterium rhizogenes A4 strain frozen in a low-temperature refrigerator at -20°C, inoculate it on a solid plate of LB+50mg / L Kan, and then culture it at a constant temperature of 28°C Cultivate in the dark in the box. After a single colony grows, pick a single colony and ...

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PUM

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Abstract

The invention discloses a method for cultivating meloidogyne hapla by adopting peanut rooting. The method comprises the following steps that number 45 seeds are selected, flowered and seeded on an MSB5 culture medium for cultivation; agrobacterium rhizogenes strains A4 are taken for cultivation and activation to achieve a logarithmic growth period; the agrobacterium rhizogenes strains A4 are adopted for infecting the sterile culture 6d flowered number 45 peanut cotyledon for inducing hairy roots; peanut roots with multiple insect galls are selected to be immersed into a NaCLO solution and broken to form a mixed solution; the mixed solution is poured onto combination sieves, wherein the combination sieves are of the types of 40 meshes, 200 meshes, 325 meshes and 500 meshes in sequence from top to bottom, and sterile water is used for spraying and washing, so that the meloidogyne hapla is concentrated to one side; then, a small amount of sterile water is used for washing oversize products from the back faces of the sieves, and liquid is collected into a container to obtain meloidogyne hapla suspension; the meloidogyne hapla suspension is evenly inoculated on a peanut hairy root culture dish, and cultivation is performed to obtain a large number of eggs and second-stage larvae. The method solves the problem that a large amount of purified meloidogyne hapla cannot be obtained easily, and the reproduction rate is high.

Description

technical field [0001] The invention relates to a method for cultivating root-knot nematode, in particular to a method for cultivating root-knot nematode northern by using peanut hairy roots. Background technique [0002] Peanut is one of the most widely planted economic crops in the world, and it is the second largest edible vegetable oil and protein resource in my country. In northern my country, root-knot nematodes are mainly harmful to peanut production, especially Shandong, a major peanut province, has the largest incidence area and the most serious losses. The damaged peanuts generally have a reduction in yield of about 20%-30%, and in severe cases it can reach 70%-80%, or even no harvest (Xu Xiujuan, 2009). Peanut nematode disease is an important disease in peanut production. So far, the occurrence and damage of nematode disease cannot be well controlled. [0003] After the root-knot nematode infects peanuts, it stimulates the production of giant cells, which leads ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/033
CPCA01K67/033
Inventor 任艳袁美石延茂尹亮李双铃崔潇李磊
Owner SHANDONG PEANUT RES INST
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