Fusarium species stored on silica gel and soil for ten years
Fusarium species stored on silica gel and soil for ten years
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镰刀菌在硅胶和土壤中保存十年
DOI：
10.1080/00275514.1993.12026240
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发表时间：
1993
期刊：
影响因子：
2.8
通讯作者：
T. Kommedahl
中科院分区：
文献类型：
--
作者：
C. Windels;P. Burnes;T. Kommedahl
有几种技术可用于长期保存镰刀菌物种，包括冷冻干燥(Fisher等人，1982)、液氮(Booth，1971)、矿物油(Lima，1991)、无菌土壤(Booth，1971；Tousourn and Nelson，1976)和硅胶(Windels等人，1988)。虽然这些方法在保存镰刀菌物种方面是有效的，但每一种方法都有局限性。冷冻干燥和液氮确保了长期的活力和最小的培养差异，但设备昂贵，培养物不能回到储藏中重复使用。矿物油使用起来很乱。储存在土壤中有突变的风险，这可能导致形态特征的丧失，有时还会导致致病性(Tousourn和Nelson，1976)。无水硅胶已被用作保存细菌(Sleesman和Leben，1978；特罗洛普，1975)、几种真菌(Perkins，1962；特罗洛普，1975)和藻类(Grivell和Jackson，1969)的便捷方法。在一项为期5年的研究中，17种镰刀菌中有15种在硅胶和无菌土壤上存活得相当好(Windels等人，1988年)。本文报道了同一种培养物保存10年后的存活率。用单孢子或菌丝顶端方法纯化镰刀菌培养物(Nelson等人，1983)，并将其储存在硅胶和无菌土壤中，温度为4.5？0.5℃(Windels等人，1988)。在458个培养物中，有18种(Nelson et al.，1983)：尖孢镰刀菌、燕麦镰刀菌、镰刀镰刀菌、二分体镰刀菌、木霉、禾谷镰刀菌、侧枝镰刀菌、尖孢镰刀菌、串珠镰刀菌、尖孢镰刀菌、Poae镰刀菌、增殖镰刀菌、松花镰刀菌、半技形镰刀菌、茄尼镰刀菌、孢囊藻、亚粘性镰刀菌和三切镰刀菌。在保存10年后，通过在Difco马铃薯-葡萄糖琼脂(PDA)上撒一些颗粒来测试每个培养物的活力。如果第一次尝试不成功，就重复这个过程，直到出现一个可行的菌落，或者直到储存的材料耗尽。培养在3个40W荧光灯管和1个黑色灯管下培养，F40BLB系列(通用电气，克利夫兰，俄亥俄州)，每天12h，连续培养7d。大多数镰刀菌的存活。在土壤和硅胶上5年后(分别为94%和92%)令人满意；在土壤上保持活力，但在硅胶上10年后下降(分别为92%和83%)(表I)。除燕麦、茄尼和三裂镰刀菌外，所有菌种在土壤中保存10年后的存活率均为90%。在硅胶上保存10年，除燕麦F.、F.culmorum、F.dimerum、F.graminearum、F.lateritium、F.poae、F.sambuum、F.Semitecum、F.solani和F.subherinans外，其余种的存活率均为90%。贮藏10年后，4种金针菇(F.merismoides)、尖孢镰刀菌(F.oxporum)、立枯丝核菌(F.solani)和三裂镰刀菌(F.tricincum)在硅胶上的存活率高于土壤，而在土壤中的存活率与土壤相当。其余11种植物在土壤上的存活率高于在硅胶上的存活率。10年后，禾谷镰刀菌在土壤中的存活率为100%。保存的禾谷镰刀菌培养物
Several techniques are available for long-term preservation of Fusarium species including lyophilization (Fisher et al., 1982), liquid nitrogen (Booth, 1971), mineral oil (Lima, 1991), sterile soil (Booth, 1971; Toussoun and Nelson, 1976), and silica gel (Windels et al., 1988). While these methods are effective in preserving Fusarium species, each has limitations. Lyophilization and liquid nitrogen ensure long-term viability with minimal cultural variation, but the equipment is expensive and cultures cannot be returned to storage for reuse. Mineral oil is messy to use. Storage in soil carries a risk of mutation which can result in loss of morphological characters and, sometimes, pathogenicity (Toussoun and Nelson, 1976). Anhydrous silica gel has been used as a convenient method for preservation of bacteria (Sleesman and Leben, 1978; Trollope, 1975), several genera of fungi (Perkins, 1962; Trollope, 1975), and algae (Grivell and Jackson, 1969). In a 5-yr study, 15 of 17 species of Fusarium survived comparably well on silica gel and sterile soil (Windels et al., 1988). This article reports on the viability of the same culture collection after storage for 10 yr. Fusarium cultures were purified by the singlespore or hyphal-tip method (Nelson et al., 1983) and stored on both silica gel and sterile soil at 4.5 ? 0.5 C (Windels et al., 1988). Of 458 cultures, 18 species (Nelson et al., 1983) were represented: F. acuminatum, F. avenaceum, F. culmorum, F. dimerum, F. equiseti, F. graminearum, F. lateritium, F. merismoides, F. moniliforme, F. oxysporum, F. poae, F. proliferatum, F. sambucinum, F. semitectum, F. solani, F. sporotrichioides, F. subglutinans, and F. tricinctum. After storage for 10 yr, each culture was tested for viability by sprinkling a few particles onto Difco potato-dextrose agar (PDA). If the first attempt was unsuccessful, the procedure was repeated until a viable colony appeared or until the stored material was depleted. Cultures were incubated under three fluorescent 40 W tubes supplemented with one black tube, F40 BLB series (General Electric, Cleveland, Ohio), for 12 h per day for 7 days. Survival of most Fusarium spp. was satisfactory on soil and silica gel after 5 yr (94 and 92%, respectively); viability was maintained on soil but declined on silica gel after 10 yr (92 and 83%, respectively) (TABLE I). Viability of cultures stored in soil for 10 yr was > 90% for all species except F. avenaceum, F. solani, and F. tricinctum. Viability of cultures stored on silica gel for 10 yr was > 90% for all species except F. avenaceum, F. culmorum, F. dimerum, F. graminearum, F. lateritium, F. poae, F. sambucinum, F. semitectum, F. solani, and F. subglutinans. After 10 yr of storage, survival was greater on silica gel than on soil for four species (F. merismoides, F. oxysporum, F. solani, and F. tricinctum) and equal to soil for three species (F. moniliforme, F. proliferatum, F. sporotrichioides). Survival was greater on soil than on silica gel for the other 11 species. Survival of F. graminearum on soil was 100% after 10 yr. Cultures of F. graminearum stored