摘要
以易感染枣疯病的阜平大枣为试材，对影响枣疯病病原荧光显微观察的主要因素进行了系统研究。结果表明：苯胺蓝（ａｎｉｌｉｎｅ ｂｌｕｅ）、 ＤＡＰＩ（４’，６－ｄｉａｍｉｄｉｎｏ－２－ｐｈｅｎｙｌｉｎｄｏｌｅ· ２ＨＣｓｄ１）和阿的平（ｑｕｉｎａｃｒｉｎｅ）３种荧光染料均可用于枣疯病病原鉴定，其中以 ＤＡＰＩ效果最佳。 ＤＡＰＩ用于定性分析的最适浓度为 ０．３μｇ·ｍＬ－１，用于定量分析的最适浓度为１．０μｇ·ｍＬ－１；对于尚未木质化或木质化程度很低的幼嫩组织切片应至少染色２０ｍｉｎ，对于高度木质化程度的组织切片应至少染色５０ｍｉｎ；带病原试材在４℃冰箱保存２０～３０ｄ，在５％戊二醛中固定４５～６０ｄ对观察结果无明显影响。以ＤＡＰＩ为染料对枣头、枣吊、叶柄、叶主脉、花梗、枝、树干皮、枣股、托叶刺、根、幼果以及接近成熟果实的组织切片进行了荧光显微观察，初步选定枝条作为最佳取材部位。
The main factors which affected the observation of phytoplasma infections in Chinese jujube trees were studied in a systematic way with fluorescence microscope. Fuping Dazao, an important commercial variety which is prone to be infected by phytoplasma was chosen as materials. The results showed that aniline blue, DAPI and quinacrine were all effective in determining phytoplasma infections. Among the three dye liquors mentioned above, DAPI was the most effective. The most fittable concentration of DAPI for qualitative analysis was 0.3 μg·mL-1 and 1.0 μg·mL-1 for quantitative analysis. The sections of young tissues should be dyed at least for twenty minutes and at least fifty minutes for old ones. The shoots infected by phytoplasma could be stored in refrigerator (4℃) for twenty to thirty days with no obvious difference. They could also be fixed in 5% glutaraldehyde in 0.1M phosphate buffer(pH 7.0) in refrigerator(4 ℃) for forty-five to sixty days. The sections of growing shoot, bearing shoot, petiole, main leaf vein, pedicel, shoot, bark, condensed shoot with bearing shoot, spine, root, young fruit, mature-green fruit stained by DAPI were observed under fluorescence microscope. And the shoot of jujube trees was chosen as the most fittable samples.
出处
《河北农业大学学报》
CAS
CSCD
北大核心
1999年第4期46-49,共4页
Journal of Hebei Agricultural University
基金
国家自然科学基金!39670520
河北农业大学9816项目资助
关键词
荧光显微技术
枣疯病
类菌原体
病原鉴定
Fluorescence microscopy
Witches' -broom disease
Phytoplasma (Mycoplasma like organisms)
Diagnosis
