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苹果盘二孢Marssonina coronaria引起的褐斑病是造成我国苹果树早期落叶的主要原因,由于该病菌分离培养困难,阻碍了对其生物学特性的研究,进而影响了对其防治机理和流行规律的研究。本研究应用4种培养基质,探索了3种方法对苹果盘二孢的分离效果。结果表明,3种方法均可分离到病原菌,但组织块分离法和分生孢子团分离法成功率仅有10%左右,而单孢分离法污染少,成功率高达到90%以上,明显优于其他两种方法。不同培养基上菌落形态、大小和产孢情况差异也很大,培养1个月(25℃)后PDA上菌落黑褐色隆起,表面蚯蚓粪状,无气生菌丝,无子实体和基内菌丝;10%V8培养基上菌落中央隆起,黑褐色,表面生少量气生菌丝,边缘放射状,基内菌丝深褐色,有子实体;苹果叶片葡萄糖琼脂培养基(LDA)上菌落平坦,黄褐色,表面生茂密的金黄色气生菌丝,基内菌丝深褐色,有子实体;苹果叶片煎汁葡萄糖琼脂培养基(LEDA)上菌落有明显的不规则隆起,黄褐色至黑褐色,表面生少许气生菌丝,菌落生长缓慢,无基内菌丝,分生孢子盘菌落表面生,菌落直径仅2mm左右,而在其他培养基上的菌落直径可达6-8mm,说明培养基质、分离方法均对苹果盘二孢的分离培养和生长发育有明显的影响。
Apple brown spot caused by Marssonina coronaria is the main cause of early deciduous apple tree in our country, because of the difficulty of isolation and culture of the bacterium, hindering the study of its biological characteristics, thus affecting its prevention and control of the law of the law the study. In this study, four culture media were used to explore the separation effects of three methods on the conidia of apple. The results showed that all the three methods could isolate the pathogenic bacteria, but the success rate of tissue block separation and conidia separation was only about 10%, while that of single spore separation was less, the success rate was more than 90%, which was obviously superior In the other two methods. The morphology, size and sporulation of colonies on different media were also different. After cultured for 1 month (25 ℃), the colonies on PDA were brownish-brown, vermicompost-like, apathetic, Mycelium; 10% V8 medium colonies central uplift, dark brown, a small amount of aerial surface of raw air mycelium, the radial edge of the base mycelium dark brown, fruiting bodies; Apple leaf glucose agar medium (LDA) on the colonies flat , Yellowish brown, the surface of the lush golden aerial mycelium, mycelium dark brown, fruiting bodies; apple leaf decoction dextrose agar medium (LEDA) on the colonies have obvious irregular uplift, brown to black Brown, the surface of a small amount of aerial mycelium, colonies grow slowly, no base mycelium, colonies of conidia colony surface, the colony diameter of only about 2mm, while the colonies on other media up to 6-8mm diameter, indicating Culture medium and isolation methods all had obvious effects on the isolation, culture and growth of the conidia of apple.