Detection of a Latent Infectious Agent That Protects Against Infection by Chrysanthemum Chlorotic Mottle Viroid
Detection of a Latent Infectious Agent That Protects Against Infection by Chrysanthemum Chlorotic Mottle Viroid
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检测可防止菊花褪绿斑驳病毒感染的潜伏感染因子
DOI：
10.1094/phyto-65-1000
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发表时间：
1975
期刊：
影响因子：
3.2
通讯作者：
R. Horst
中科院分区：
文献类型：
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作者：
R. Horst
材料和方法：以金菊“深岭”为试材，以侵染CHCMV的深岭为接种源。机械接种的方法是：从表现出慢性症状(严重斑驳到完全黄化)的植物上取出新展开的幼叶，在预冷的砂浆中研磨叶片，加入一部分膨润土-硼酸盐缓冲液(0，1M HBO和0.1MNaBO调节到pH 8.0，与1%的膨润土溶液在Burrell腕上摇床上摇动30分钟，在4℃冷却)混合制成机械接种。所有的机械接种都按照Horst等人的描述进行(10)。组织植入接种如前所述由Dimock等人(5)使用14号和16号抽吸套管针(Popper&Sons，Inc.；300Benton Ave.，New Hyde Park，NY 11040)进行。接种后隔天记录出现症状的植株总数，连续21天。然后，将接种的植株修剪到离土壤表面约15厘米的地方，以诱导侧芽生长。最后的观察是在9天后进行的，数据被表示为感染性指数(L，3)；即30天后反应的植物数量的总和，但在这里报告的数据中没有使用稀释因子。感染力指数反映了在实验完成时做出反应的植物总数，以及出现症状的时间，环境生长室被用来保持恒定的温度和光照条件(23672lx)。温室试验的接种源植物和测试植物保持在大约24摄氏度。在冬季的几个月里，为了保持每天14个小时的至少10,760 lx的亮度，使用了辅助照明。植物受精了
MATERIALS AND METHODS.–Chrysanthernum morifolium “Deep Ridge'was used as a test plant, and ChCMV-infected Deep Ridge was used as a source of inoculum in all experiments. Inoculum for mechanical inoculations was prepared by removing young newly expanding leaves from plants exhibiting chronic symptoms (severe mottling to complete chlorosis) and grinding the leaves in a precooled mortar with one part (w/v) of bentonite-borate buffer (0, 1 M HBO, and 0.1 M Na BO adjusted to pH 8.0 and mixed 1: 1 with a 1% bentonite solution shaken on a Burrell Wrist-Action shaker for 30 minutes and chilled at 4C). All mechanical inoculations were made as described by Horst et al.(10). Tissue implantation inoculations were performed as previously described by Dimock et al.(5) using 14-and 16-gauge Aspiration Trocarsº (Popper & Sons, Inc.; 300 Benton Ave., New Hyde Park, NY 11040). The total number of plants that exhibited symptoms was recorded on alternate days for 21 days after inoculation. Inoculated plants were then trimmed to approximately 15 cm above the soil surface to induce new growth from lateral buds. Final observations were made 9 days later, and data were expressed as an infectivity index (l, 3); ie, a summation of the number of plants reacting after 30 days, but no dilution factor was used in the data reported here. The infectivity index reflects the total number of plants that had reacted at the completion of the experiment, and also the time at which symptoms appeared, Environmental growth chambers were used to maintain constant temperatures and light conditions (23,672 lx). Inoculum source plants and test plants for greenhouse experiments were maintained at approximately 24C. Supplemental lighting was used during the winter months to maintain a minimum of 10,760 lx for 14 hours of every day. Plants were fertilized