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为有效筛选烟草抗赤星病细胞突变体,本研究建立了一种双层培养基筛选系统。即在下层培养基中接种烟草赤星病(Alternaria alternata),在上层培养基中接种烤烟品种 NC89和净叶黄的花药(NC89高感赤星病,净叶黄高抗赤星病)。在26±1℃培养40天后发现,NC89的花药出苗率和花药平均出苗数与对照(不接种赤星病的培养基)相比,分别由30%和53颗降到2%和12颗,而净叶黄的花药出苗率和平均出苗数基本没有变化。对筛选出来的 NC89花培突变体进行了田间赤星病抗性鉴定,其中80%对赤星病抗性较 NC89有所提高,甚至部分接近高抗。这说明该双层培养基系统可以有效地对抗烟草赤星病的细胞突变体进行筛选。本系统尤其适应于(1)需长期培养;(2)毒素不易制备或易失活的病原菌的抗性筛选。
In order to effectively screen the tobacco plants against the mutant of brown spot disease, a double-layer medium screening system was established in this study. That is, Alternaria alternata was inoculated into the lower culture medium, and the upper culture medium was used to inoculate the flue-cured tobacco varieties NC89 and the net leaf-yellow anthers (NC89 highly susceptible to the disease, net leaf yellow high against brown spots disease). After cultured for 40 days at 26 ± 1 ℃, the emergence rate of anther and the average number of anther emergence of NC89 decreased from 30% and 53 to 2% and 12, respectively, compared with the control (medium not inoculated with brown spot) Net leaf yellow anther emergence rate and the average number of emergence did not change. The NC89 flower cultivars mutants were identified as brown spot disease resistance in the field, of which 80% of resistant to brown spot disease than NC89 increased, and some close to high resistance. This indicates that the two-layer culture system can effectively screen for the mutant of tobacco brown spot disease. The system is especially adapted to (1) require long-term culture; (2) toxins are not easy to prepare or easy to inactivate pathogenic bacteria screening.