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Root tip detoxification and rapid propagation technology of strawberries

A strawberry and root tip technology, applied in plant regeneration, horticulture, horticultural methods, etc., can solve the problems of slow propagation speed of explant differentiation speed, detoxification test tube seedlings, low production of transplanted field, unstable quality of seedlings, etc. Seedling propagation speed and yield increase, pollution rate is low, and the effect of large-scale promotion of high yield

Inactive Publication Date: 2013-01-30
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Summary
  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, to detoxify the stem tip of strawberry, it is necessary to use surgical tools to continuously peel off more than ten layers of stem slices wrapped outside the stem (tip), which is difficult to operate and low in work efficiency. , but even experienced technicians are also difficult to completely strip and disinfect (tested by our multiple RT-PCR simultaneous rapid detection method, usually 60% (mixed) contamination rate, resulting in unstable quality of seedlings; and, using the above The method also has the problems of slow differentiation speed of explants and slow propagation speed of virus-free test-tube plantlets, weak growth and low production in transplanted fields.

Method used

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  • Root tip detoxification and rapid propagation technology of strawberries
  • Root tip detoxification and rapid propagation technology of strawberries
  • Root tip detoxification and rapid propagation technology of strawberries

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Example 1 Multiplex RT-PCR Simultaneous Rapid Detection of Virus Infection in Strawberry Tissue

[0068] Commissioned the synthesis of Strawberry virus-specific primer pairs with the following nucleotide sequences, and mixed them into two primers of 4 μM for SMoV, 5 μM for SMYEV, 9 μM for SVBV, and 9 μM for SCrV Mixed primer solution with two primer contents of 10 μM:

[0069] SMoV upstream primer P1: 5'-TAACGGTTCACGTCCTAGTCTGAC-3'

[0070] SMoV downstream primer P2: 5'-TCTTGGGCTTGGATCGTCACCTG-3'

[0071] SMYEV upstream primer P3: 5'-GTGTCCTCAATCCAGCCAG-3'

[0072] SMYEV downstream primer P4: 5'-CATGGCACTCATTGGACCTGGG-3'

[0073] SVBV upstream primer P5: 5'-GAATGGGACAATGAAATGAC-3'

[0074] SVBV downstream primer P6: 5'-GTGAGGAGAACTTAGGACA-3'

[0075] SCrV upstream primer P7: 5'-CCCTTCGCAGGCGCGCTAACA -3'

[0076] SCrV downstream primer P8: 5'-GGGGTCCAACTCATAAGCGAT -3'.

[0077] Take 1g of strawberry tissue, put it into 100mL containing 0.5%Triton X-100 and 0.4%Na ...

Embodiment 2

[0082] Example 2 Strawberry detoxification and tissue culture method 1

[0083] Prepare medium as follows:

[0084] 1) MS basic medium;

[0085] 2) Root tip induction medium: MS basic medium + NH 4 NO 3 400mg / L + KNO 3 500mg / L+KH 2 PO 4 50mg / L+BA 0.5mg / L+2,4-D 0.1mg / L+hydrolyzed milk protein 1 g / L+sucrose 20 g / L;

[0086] 3) Proliferation medium: MS basic medium + NH 4 NO 3 400mg / L + KNO 3 500 mg / L + KH 2PO 4 50 mg / L +BA 0.5 mg / L+ NAA 0.2mg / L+ hydrolyzed milk protein 1 g / L+ sucrose 20 g / L;

[0087] 4) Subculture growth medium: MS basic medium + NH 4 NO 3 400mg / L + KNO 3 500mg / L+KH 2 PO 4 50mg / 0.5L+BA 0.2mg / L + hydrolyzed milk protein 1 g / L+ sucrose 20 g / L;

[0088] 5) Rooting medium: MS basic medium + NH 4 NO 3 400mg / L+ KNO 3 500 mg / L + KH 2 PO 4 50 mg / L + NAA 0.5mg / L+ hydrolyzed milk protein 1 g / L+ sucrose 20 g / L;

[0089] Root tip detoxification and tissue culture steps are as follows:

[0090] 1) Material pretreatment: Put the strawberries in steril...

Embodiment 3

[0096] Example 3 Strawberry detoxification and tissue culture method 2

[0097] Prepare medium as follows:

[0098] 2) Root tip induction medium: MS basic medium + NH 4 NO 3 400mg / L + KNO 3 500mg / L+KH 2 PO 4 50mg / L+BA 0.8mg / L+2,4-D 0.2mg / L+hydrolyzed milk protein 1 g / L+sucrose 20 g / L;

[0099] 3) Proliferation medium: MS basic medium + NH 4 NO 3 400mg / L + KNO 3 500 mg / L + KH 2 PO 4 50 mg / L +BA 0.35 mg / L+NAA 0.35mg / L+hydrolyzed milk protein 1 g / L+sucrose 20 g / L;

[0100] 4) Subculture growth medium: MS basic medium + NH 4 NO 3 400mg / L + KNO 3 500mg / L+KH 2 PO 4 50mg / 0.5L+BA 0.35mg / L +hydrolyzed milk protein 1 g / L+sucrose 20 g / L;

[0101] 5) Rooting medium: MS basic medium + NH 4 NO 3 400mg / L+ KNO 3 500 mg / L + KH 2 PO 4 50 mg / L + NAA 0.3 mg / L+ hydrolyzed milk protein 1 g / L+ sucrose 20 g / L.

[0102] The root tip detoxification and tissue culture steps are the same as those in Example 2.

[0103]

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Abstract

The invention discloses a root tip detoxification and rapid propagation technology of strawberries. The method for detoxification and tissue culture of strawberries comprises the steps of: pre-processing the strawberries, to obtain disinfected root tips; carrying out induction culture on the root tips on a root tip inducing medium to obtain regenerated plant sprouts; carrying out multiplication culture on the sprouts into seedlings on a multiplication medium; culturing the seedlings into subculture seedlings on a subculture growth medium; culturing the subculture seedlings into test-tube seedlings on a rooting medium; and optionally detecting whether the test-tube seedlings are affected by virus. In addition, the invention discloses a multiple reverse transcription-polymerase chain reaction (RT-PCR) method for synchronous and rapid detection of strawberry virus, wherein a hybrid primer pair is adopted.

Description

technical field [0001] The invention belongs to the field of plant breeding. Specifically, the invention relates to a method for detoxification and tissue culture of strawberry root tips and a multiple RT-PCR synchronous rapid detection method that can be used to detect whether strawberries produced by the method are infected with viruses. [0002] Background technique [0003] my country is the country with the richest wild strawberry resources in the world. It has been using wild strawberries very early and has continued to this day. The cultivation of strawberry with large fruit in my country began in 1915, but it was not taken seriously in the past and developed slowly. Strawberry production has developed rapidly since the 1980s, and the current strawberry production area is about 70,000 hm 2 , The area ranks first in the world, and the main producing areas are distributed in Anhui, Liaoning, Hebei, Shandong, Jiangsu, Shanghai, Zhejiang, Sichuan, Xinjiang, Beijing and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 严成其陈剑平钱长根钱尖锐杨勇王栩鸣周洁张维林余初浪程晔王芳沈岚朱宏芬刘健张国芳黄坚
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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