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取花蕾长3.0~5.5mm的花序,7%(W/V)饱和次氯酸钙溶液(加一滴吐温20)表面消毒15min,无菌水洗3次。然后轻压花蕾,过滤,收集滤液,离心后,将小孢子悬浮于经过滤灭菌的改良的1/2NLN培养基中。石蜡膜封口,置温度梯度培养箱作32.5℃、1d的高温热激诱导处理,后置25℃下继续培养。本试验对13份绿菜花供试材料的游离小孢子培养能力进行初步研究,其中8种基因型(占全部材料的61.5%)有胚胎发生,并获得5种基因型的再生株,共207株。基因型对绿菜花游离小孢子培养的影响很大,基因型“巴绿青花菜”出胚率最高,达37.08胚/蕾。同时,对培养基中加入活性炭悬浮液和更换培养基的作用进行了研究。
Take flower bud length 3.0 ~ 5.5mm inflorescence, 7% (W / V) saturated calcium hypochlorite solution (plus a drop Tween 20) surface disinfection 15min, sterile washed 3 times. The buds were then lightly pressed, filtered and the filtrate was collected. After centrifugation, the microspores were suspended in filter-sterilized modified 1/2 NTL medium. Parafilm seal, temperature gradient incubator for 32.5 ℃, 1d heat shock induction treatment, followed by 25 ℃ continue to train. In this study, 13 microspore culture ability of greencrops were studied. Among them, 8 genotypes (61.5% of all materials) had embryogenesis and regenerated 5 genotypes 207 strains. The genotype had a great influence on the culture of green microspores from microspores. The genotype “Pak Green Broccoli” had the highest rate of embryogenesis at 37.08 embryo / bud. At the same time, the effect of adding activated carbon suspension and medium change was studied.