Mutagenesis method of nicotiana tabacum L. pollen
A pollen and tobacco technology, applied in horticultural methods, botanical equipment and methods, applications, etc., can solve the problems of difficult absorption of mutagens, small variation range, and high difficulty in screening, so as to overcome obvious and recessive obstacles and mutations The effect of large range and saving breeding time
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Embodiment 1
[0021] Embodiment 1: Flue-cured tobacco material 1 pollen in vitro liquid culture mutagenesis, the operation steps are as follows:
[0022] (1) Sample collection: Take healthy age-appropriate flower buds on the main inflorescence of the flue-cured tobacco plant, with a length ranging from 2.0 to 2.2 cm, and quickly store them in an ice box at 4-7°C;
[0023] (2) Disinfection treatment: Take out the flower buds in the ice box in step (1), carefully peel off the anthers in the flower buds with tweezers on the ultra-clean workbench, pack the anthers with sterile gauze and immerse them in 70% ethanol for 15 seconds, Immerse in 10% saturated calcium hypochlorite solution for 8 minutes, rinse with sterile water 3 times, 5 minutes each time;
[0024] (3) Collect pollen: Put the anthers sterilized in step (2) into a large round-bottomed test tube, add 2ml of B5-13 liquid medium (the concentration of sucrose in B5 basic medium is 13%), and then add 1-2ml of mixed enzyme solution with ...
Embodiment 2
[0031] Embodiment 2, flue-cured tobacco material 2 pollen isolated liquid culture mutagenesis, its operation steps are as follows:
[0032] (1) Sample collection: Take healthy age-appropriate flower buds on the side inflorescences of flue-cured tobacco plants at the single-nucleus edge stage, with a length range of 2.1-2.3cm, and quickly store the collected flower buds in an ice box at 4-7°C;
[0033] (2) Disinfection treatment: Take out the flower buds in the ice box in step (1), carefully peel off the anthers in the flower buds on the ultra-clean workbench, put the anthers in sterile commercial stockings and immerse them in 75% ethanol for 10 seconds , and then immersed in 10% saturated calcium hypochlorite solution for 10 minutes, rinsed twice with sterile water, 5 minutes each time;
[0034] (3) Collect pollen: transfer the anthers sterilized in step (2) into a large round-bottomed test tube, add 3ml of B5-13 liquid medium (the sucrose concentration in B5 basic medium is 13%...
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