Prevention technology for multiple blueberry viruses at early stage

A blueberry and virus technology, applied in the direction of microbial determination/inspection, plant regeneration, biochemical equipment and methods, etc., can solve the problems of loss of early input of fruit farmers, missed optimal utilization period of land, etc., and achieves easy operation and low pollution rate. , the effect of obvious planting advantages

Inactive Publication Date: 2015-05-06
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The harm of these viruses is extremely harmful, not only depleting the early investment of fruit farmers, but also making the land miss the best use period, and the loss is irreparable

Method used

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  • Prevention technology for multiple blueberry viruses at early stage
  • Prevention technology for multiple blueberry viruses at early stage
  • Prevention technology for multiple blueberry viruses at early stage

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Multiple simultaneous rapid RT-PCR detection of virus infection in blueberry tissue

[0041] Entrusted to synthesize the blueberry virus-specific primer pairs with the nucleotide sequence as follows, and mix them into two BRRV primers with a content of 4μM, BLScV two primers with a content of 6μM, BLShV two primers with a content of 8μM, TRSV A mixed primer solution with two primers content of 7μM and BLMV two primers content of 9μM:

[0042] BRRV upstream primer P1: 5‘-GTAGTATTTAATTATATAGTTG-3’

[0043] BRRV downstream primer P2: 5‘-GGTATATATTCGAATTTTGG-3’

[0044] BLScV upstream primer P3: 5‘-CAGTTATGCCTCCGAAAG-3’

[0045] BLScV downstream primer P4: 5‘-CCCGCATTTCGATGATTGCG-3’

[0046] BLShV upstream primer P5: 5‘-TTTATTCAATTTCGAAGCGAA-’

[0047] BLShV downstream primer P6: 5‘-TTCGCTTCGAAATTGAATAAA-’

[0048] TRSV upstream primer P7: 5‘-GACGAAGTTATCAATA -3’

[0049] TRSV downstream primer P8: 5‘-TCCGTCCAATCACGCGAATA -3’

[0050] BLMV upstream primer P9: 5‘-TGAC...

Embodiment 2

[0056] Example 2 Blueberry detoxification and tissue culture method 1

[0057] Prepare the following medium:

[0058] 1) Root tip induction medium: MS basic medium + BA (6-benaminopurine) 0.1mg / L+ ZT (Zeatin, zeatin) 0.3 mg / L+2,4-D (2,4-dichlorobenzene) Oxyacetic acid) 0.1mg / L + hydrolyzed milk protein 1 g / L + sucrose 20 g / L;

[0059] 2) Proliferation medium: MS basic medium + BA 0.2 mg / L+ ZT0.3 mg / L+ NAA (naphthalene acetic acid) 0.05 mg / L+ hydrolyzed milk protein 1 g / L+ sucrose 20 g / L;

[0060] 3) Rooting medium: MS basic medium + NAA 0.1 mg / L + hydrolyzed milk protein 1 g / L + sucrose 20 g / L.

[0061] The steps of root tip detoxification and tissue culture are as follows:

[0062] 1) Material pretreatment: Put blueberries in sterilized fine sand at 37 soil 2℃ for heat treatment for 5-8 weeks, until the roots are taken out; take the 1-2cm long taproot tips, and then pass 70% (V / V) Sterilize with alcohol for 30 seconds, disinfect with 0.1% mercuric chloride plus a drop of Tween...

Embodiment 3

[0066] Example 3 Blueberry detoxification and tissue culture method 2

[0067] Prepare the following medium:

[0068] 1) Root tip induction medium: MS basic medium + BA (6-benaminopurine) 0.2mg / L+ ZT0.4mg / L + 2,4-D (2,4-dichlorophenoxyacetic acid) + 0.2mg / L+hydrolyzed milk protein 1 g / L+sucrose 20 g / L;

[0069] 2) Proliferation medium: MS basic medium + BA 0.3 mg / L + ZT 0.4 mg / L + NAA (naphthalene acetic acid) 0.07 mg / L + hydrolyzed milk protein 1 g / L + sucrose 20 g / L;

[0070] 3) Rooting medium: MS basic medium + NAA 0.2mg / L + hydrolyzed milk protein 1 g / L + sucrose 20 g / L.

[0071] The steps of root tip detoxification and tissue culture were the same as those in Example 2.

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Abstract

The invention provides a method for cultivating blueberries, which is used for preventing or reducing viral pollution during the process of cultivating blueberries. The method comprises the following steps: performing pretreatment on the blueberries so as to obtain disinfectant root tips; culturing the root tips into regeneration plant sprouts on a culture medium; culturing the sprouts into plantlets on the culture medium; enabling the plantlets to grow into test-tube plantlets on the culture medium; grouping and sampling randomly, and detecting whether samples are infected viruses, especially specific viruses or not. In addition, the invention further provides a method for detecting blueberry viruses synchronously and quickly through multiple RT-PCR, and a mixed prime pair is adopted.

Description

technical field [0001] The present invention belongs to the field of plant breeding. Specifically, the present invention relates to a method for cultivating blueberries, or a method for preventing or reducing (multiple) virus contamination in blueberry cultivation, and also relates to the synchronous rapid detection of multiple (species) viruses matched with it technology. Background technique [0002] Blueberry is a shrub of the Rhododendronaceae Vaccinium genus. Its fruit is a berry with small particles but high anthocyanin content. This is a completely different plant from Rosaceae herbs such as strawberries. [0003] The history of artificial cultivation of blueberries is relatively short, and there are relatively few studies on diseases and insect pests, especially viral diseases. Fruit growers lack knowledge and experience in the prevention and control of viral diseases. At present, the rooting and survival of blueberry seedlings are usually used to evaluate whether th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68A01H4/00
Inventor 严成其陈剑平钱长根钱尖锐朱宏芬毛东海王芳沈岚刘健张国芳黄坚杨勇王栩鸣余初浪周洁张维林羊健程晔
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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