葡萄黑痘病病原菌生长特性研究  

Growth characteristics of pathogen causing grape black pox

作　　者：林玲[1] 陈立[1,2] 李玮[1] 谢蜀豫 文仁德[1] 陈国品[1] 李洪艳[1] 韩佳宇[1] 盘丰平[1] 白先进[3] 曹慕明[1] 

机构地区：[1]广西农业科学院葡萄与葡萄酒研究所,南宁530007 [2]广西大学农学院,南宁530005 [3]广西农业科学院,南宁530007

出　　处：《南方农业学报》2016年第4期571-575,共5页Journal of Southern Agriculture

基　　金：国家自然科学基金项目(31501716);广西自然科学基金项目(2014GXNSFBA118069)

摘　　要：【目的】探索葡萄黑痘病病原菌在不同培养条件下的生长情况,优化其培养条件,以缩短培养该菌纯培养物的周期,提高培养效率。【方法】通过常规培养手段,从患黑痘病黑巴拉多病叶上分离葡萄黑痘病病原菌,PCR扩增该菌ITS片段,经测序后在NCBI比较该片段序列。用同一规格培养皿培养病原菌,每隔7 d观察并统计该病原菌在4种不同培养基组分、5个不同温度梯度、4个不同蔗糖浓度梯度、4种光照时间、5个不同p H条件下共计21 d的生长情况。【结果】分离到病原菌并扩增获得其ITS片段,经比对,该片段序列与NCBI已公布葡萄痂囊腔菌序列AY826763的相似度达99.9%,证实所分离病原菌为葡萄痂囊腔菌(Elsinoe ampelina)。葡萄痂囊腔菌的生长最适培养基为PSA,其次是YEB;最适培养温度为25℃、最适蔗糖浓度为4.5 g/L、最适p H为8,在最适培养温度、蔗糖浓度和p H下病原菌的生长速度分别为0.07、0.03和0.01 cm/d;光照时间对葡萄痂囊腔菌生长影响不明显。【结论】葡萄痂圆孢菌生长受环境条件影响,通过培养条件优化能显著加快其生长速度,提高培养效率。[Objective]The present experiment was conducted to study growth situation of pathogen causing grape black pox under different culture conditions, and optimize culture conditions, in order to shorten culture cycle of pathogen pure cultures and improve culture efficiency. [Method ]The strain was isolated from leaves of grape variety Heibaladuo suffering from grape black pox by conventional culture method. The ITS sequence of isolated strain was amplified by using PCR technique, then sequenced and blasted on NCBL The isolated strain was cultured in the petri dishes with same spec- ification, and its growth situation was observed every 7 days under different culture conditions including 4 kinds of media with different compositions, 5 temperature gradients, 4 sucrose concentration gradients, 4 illumination time gradients and 5 pH gradients, totaling 21 days. [Result]The ITS sequencing result of isolated strain showed that, it shared 99.9% similarity with E. ampelina＇s sequence AY826763,. therefore, the isolated strain was identified as Elsinoe ampelina （de Bary） Shear causing grape black pox. Furthermore, the optimized culture conditions showed that, the optimum medium for pathogen growth was PSA, then YEB; the optimum temperature was 25℃, the optimum sucrose concentration was 4.5 g/ L, the optimum pH was 8. Under the above optimum culture conditions, the growth rates of E. ampelina were 0.07, 0.03 and 0.01 cm/d, the illumination time had little effect on E. ampelina growth. [Conclusion]The culture conditions have a certain effect on E. ampelina growth, so its growth rate can be improved significantly by optimizing culture conditions.

关 键 词：葡萄黑痘病 葡萄痂囊腔菌 培养条件 生长特性 

分 类 号：S436.8[农业科学—农业昆虫与害虫防治]