US 12,010,963 B2
Medium and culture method for blueberry tissue culture
Yangyan Zhou, Rizhao (CN); Weijian Sun, Rizhao (CN); Wenxiu Chen, Rizhao (CN); Aowei Mo, Rizhao (CN); Peng Guo, Rizhao (CN); and Penghao Xu, Rizhao (CN)
Assigned to Shandong Dafengyuan Agriculture Co., Ltd., Rizhao (CN)
Filed by SHANDONG DAFENGYUAN AGRICULTURE CO., LTD., Rizhao (CN)
Filed on Apr. 14, 2022, as Appl. No. 17/720,652.
Claims priority of application No. 202110414403.0 (CN), filed on Apr. 16, 2021.
Prior Publication US 2022/0338435 A1, Oct. 27, 2022
Int. Cl. A01H 6/36 (2018.01); A01G 22/05 (2018.01); A01G 24/15 (2018.01); A01G 24/22 (2018.01); A01H 4/00 (2006.01); A01H 5/08 (2018.01); C12N 5/00 (2006.01); C12N 5/04 (2006.01)
CPC A01H 4/002 (2021.01) [A01G 22/05 (2018.02); A01G 24/15 (2018.02); A01G 24/22 (2018.02); A01H 4/001 (2013.01); A01H 5/08 (2013.01); A01H 6/368 (2018.05); C12N 5/0025 (2013.01); C12N 5/04 (2013.01); C12N 2500/05 (2013.01); C12N 2500/32 (2013.01); C12N 2500/34 (2013.01); C12N 2500/35 (2013.01); C12N 2500/38 (2013.01); C12N 2500/60 (2013.01); C12N 2501/30 (2013.01); C12N 2533/76 (2013.01)] 6 Claims
OG exemplary drawing
 
1. A blueberry tissue expansion propagation method, comprising:
inoculating cut, sterile blueberry leaf into a callus induction medium such that an abaxial surface of the cut, sterile blueberry leaf is adjacent to the callus induction medium, the callus induction medium consisting of
woody plant medium (WPM), 4-8 g/L agar, 20-30 g/L sucrose, 2.0-4.0 mg/L forchlorfenuron (CPPU) and 0.1-0.4 mg/L 2-isopentenyladenine (2-ip),
wherein the WPM consists of 400 mg/L Ammonium Nitrate, 370 mg/L Magnesium Sulfate Heptahydrate, 278 mg/L Calcium Nitrate Tetrahydrate, 200 mg/L Potassium Nitrate, 170 mg/L Potassium Dihydrogen Phosphate, 8.6 mg/L Zinc Sulfate Heptahydrate, 22.4 mg/L Manganese Sulfate Monohydrate, 0.25 mg/L Sodium Molybdate Dihydrate, 0.25 mg/L Copper Sulfate Pentahydrate, 73.4 mg/L Sodium Iron Ethylene Diamine Tetracetate, 100 mg/L Inositol, 2 mg/L Glycine, 0.5 mg/L Thiamine Hydrochloride, 0.5 mg/L Niacin and 0.1 mg/L Pyridoxine Hydrochloride,
the callus induction medium having a pH in a range of 5.0-5.5;
conducting an induction culture with the cut, sterile blueberry leaf in the callus induction medium for 12-22 days to form blueberry callus, during which the induction culture is maintained in a dark environment at a temperature of 22-26° C.;
inoculating callus lumps of the blueberry callus into a bud growth medium consisting of
the WPM, 4-8 g/L agar, 20-30 g/L sucrose, 1.0 mg/L CPPU and 0.4-0.6 mg/L 2-ip, with a pH falling in a range of 5.0-5.5,
conducting a bud growth culture with the inoculated callus lumps to form a blueberry plant growing adventitious buds;
inoculating the blueberry plant growing adventitious buds into a rooting medium and conducting a rooting culture with the inoculated blueberry plant growing adventitious buds to form a blueberry plant growing adventitious buds and roots, the rooting medium consisting of
1/2 Murashige and Skoog (MS) medium, 5-9 g/L agar, 15-25 g/L sucrose and 2.0-3.0 mg/L naphthyl acetic acid (NAA), the rooting medium having a pH in a range of 5.0-5.5,
wherein the 1/2 MS medium consists of 950 mg/L Potassium Nitrate, 166.1 mg/L Calcium Chloride, 825 mg/L Ammonium Nitrate, 90.35 mg/L Magnesium Sulfate, 85 mg/L Potassium Dihydrogen Phosphate, 0.025 mg/L Copper Sulphate, 6.2 mg/L Boric Acid, 16.9 mg/L Manganese Sulfate, 0.25 mg/L Sodium Molybdate, 8.6 mg/L Zinc Sulfate, 0.025 mg/L Cobalt Chloride, 0.83 mg/L Potassium Iodide, 27.8 mg/LFeSO4·7H2O, 37.26 mg/L Na2-EDTA·2H2O, 2.0 mg/L Glycine, 100 mg/L Inositol, 0.50 mg/L Niacin, 0.1 mg/L Thiamine Hydrochloride, and 0.5 mg/L Pyridoxine Hydrochloride.