Method for improving establishing high linolenic acid plant-Bonianghao
A technology of Artemisia somnifera and linolenic acid is applied in the field of improving and domesticating the cruciferous high-linolenic acid plant Artemisia somnifera, which can solve problems such as difficulty in cultivation of Artemisia somnifera, and achieve the effects of increasing the yield of linolenic acid and solving difficult artificial cultivation.
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Embodiment 1
[0035] The wild weed plant Artemisia persicae TX-1 (collected in the countryside of Taixing City, Jiangsu Province and named as TX-1) was selected. The protoplasts were extracted from the callus of Artemisia persicae that had been treated in the dark at 4°C for 24 hours, and the protoplasts were cultured in a shallow liquid layer using B5 medium. The callus was obtained and differentiated into shoots, and the protoplasts were established. body culture system. Low temperature and dark treatment is beneficial to obtain high-yield and high-quality protoplasts; 5% cellulase + 0.5% analyzase + 5mmol / L MES enzymolysis for 16h is the best enzymolysis treatment among the set treatments; Good protoplast culture effect; MS+2.0mg / L6-BA+0.2mg / L NAA+1.7mg / L AgNO 3 It is a suitable differentiation medium; MS+0.5mg / L IBA+0.5mg / L NAA is a suitable rooting medium.
Embodiment 2
[0037]The protoplasts were extracted from the leaves of Artemisia sobanifolia and the cotyledon NJ5424 of Brassica napus L. low erucic acid and low glucosinolate rapeseed line. The protoplasts were extracted by using PEG-high pH (10.5), high calcium and DMSO fusion method, and the liquid shallow layer was cultured statically, and obtained 19 regenerated plants. In the experiment, 4% cellulase + 0.5% isolated enzyme + 5mmol / L MES enzymatic hydrolysis for 10h and 1% cellulase + 0.2% isolated enzyme + 3mmol / L MES enzymatic hydrolysis for 14h respectively obtained high-yield Artemisia annua and Rapeseed Protoplast; 30%PEG+0.3mol / L glucose+50mmol / L CaCl 2 2H 2 O+15% DMSO obtained a fusion rate of 10%; the protoplast culture effect of the improved B5 medium is better; adding different fresh medium each time and adding MES has a good effect on preventing browning; suitable differentiation medium MS+4.0mg / L 6-BA+0.3mg / L NAA+5.0mg / L AgNO 3 ; Suitable rooting medium is MS+0.1mg / L IBA...
Embodiment 3
[0039] The regenerated plants obtained from the fusion of protoplasts of Brassica napus and Artemisia baicalensis were transplanted into the field, and their morphology, cytology and pollen viability were observed. The hybrid plant grows slowly, the plant height is between 95-155cm, the leaf shape is more like rapeseed, but the leaf margins are deeply cracked like Artemisia arborescens, and the flower buds are small and shrunk to purple like Artemisia annua; the flowering period is later, and the flowers are thinner Small, yellow, with 34.3% active pollen; late maturity, capable of fruiting, with an average of 6.5 seeds per silique; the maximum number of chromosomes is 66, indicating that the fusion is successful.
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