Prevention technology for multiple blueberry viruses at early stage
A blueberry and virus technology, applied in the fields of plant regeneration, horticulture, horticultural methods, etc., can solve the problems of loss of early input of fruit farmers, missed optimal utilization period of land, etc., and achieves the effects of simple operation, low pollution rate, and various types.
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Embodiment 1
[0040] Example 1 Multiple simultaneous rapid RT-PCR detection of virus infection in blueberry tissue
[0041] Commission the synthesis of blueberry virus-specific primer pairs with the following nucleotide sequences, and mix them into two primers of BRRV with a content of 4 μM, BLScV with a content of 6 μM, BLShV with a content of 8 μM, and TRSV with a content of 8 μM Mixed primer solution with the contents of the two primers at 7 μM and the two primers at BLMV at 9 μM:
[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'-TCCGTCCAATCACGCGA...
Embodiment 2
[0057] Example 2 Blueberry detoxification and tissue culture method 1
[0058] Prepare the medium as follows:
[0059] 1) Root tip induction medium: MS basic medium + BA (6-bean aminopurine) 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;
[0060] 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;
[0061] 3) Rooting medium: MS basic medium + NAA 0.1 mg / L + hydrolyzed milk protein 1 g / L + sucrose 20 g / L.
[0062] Root tip detoxification and tissue culture steps are as follows:
[0063] 1) Material pretreatment: Put the blueberries into sterilized fine sand and heat-treat them at 37 ± 2°C for 5-8 weeks until the root system is pumped; take the 1-2cm long main root tip, and then pass through 70% (V / V) Alcohol disinfection for 30 seconds, shaking disinfection with...
Embodiment 3
[0067] Example 3 Blueberry detoxification and tissue culture method 2
[0068] Prepare the medium as follows:
[0069] 1) Root tip induction medium: MS basic medium + BA (6-bean aminopurine) 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;
[0070] 2) Proliferation medium: MS basic medium + BA 0.3 mg / L + ZT0.4 mg / L + NAA (naphthalene acetic acid) 0.07 mg / L + hydrolyzed milk protein 1 g / L + sucrose 20 g / L;
[0071] 3) Rooting medium: MS basic medium + NAA 0.2mg / L + hydrolyzed milk protein 1 g / L + sucrose 20 g / L.
[0072] The root tip detoxification and tissue culture steps are the same as those in Example 2.
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