利用激光光源的荧光成像系统高通量计数大豆孢囊线虫褐化孢囊  

A high-throughput method for counting brown cysts of Heterodera glycines using a Fluorescence-based imaging system with laser excitation

作　　者：张凤 油清波 李泽宇[3] 陈井生[3] 代文君 程怡慧 周新安[1] 郭葳[1] 矫永庆[1] ZHANG Feng;YOU Qing-bo;LI Ze-yu;CHEN Jing-sheng;DAI Wen-jun;CHENG Yi-hui;ZHOU Xin-an;GUO Wei;JIAO Yong-qing(Oil Crops Research Institute,Chinese Academy of Agriculture Sciences,Key Laboratory of Biology and Genetic Improvement of Oil Crops,Ministry of Agriculture,Wuhan 430062,China;Graduate School of Chinese Academy of Agricultural Sciences,Beijng 100081,China;Daqing Branch,Chinese Academy of Sciences of Heilongjiang Province,Daqing 163316,China)

机构地区：[1]中国农业科学院油料作物研究所/农业部油料作物生物学与遗传育种重点实验室,湖北武汉430062 [2]中国农业科学院研究生院,北京100081 [3]黑龙江省农业科学院大庆分院,黑龙江大庆163316

出　　处：《中国油料作物学报》2018年第3期420-425,共6页Chinese Journal of Oil Crop Sciences

基　　金：国家自然科学基金(31501334;31522042和31371654);国家重点研发计划(2016YFD0100603-5);湖北省自然科学基金(2016CFA049);中国农业科学院基本科研业务费(Y2017JC14);科技创新工程

摘　　要：高效准确的孢囊计数方法对大豆孢囊线虫研究具有重要意义。为改善普通的荧光成像系统激发光能量偏低,褐化孢囊被激发的荧光减弱,难以清晰成像且计数效果欠佳的问题,本研究改用激光为激发光源,在470nm的激发波长和535nm的发射波长组合或532nm激发波长和620nm发射波长组合条件下,可得到褐化孢囊对比度高且清晰的图片,配合使用自动计数软件,可以准确高效地进行孢囊计数。A high-throughput and accurate counting of cysts is an important prerequisite in the study of soybean cyst nematode（ SCN,Heterodera glycines Ichinohe）. The fluorescence imaging technology has been used to count the cysts using halogen or Xenon as the excitation light source. However,because of the low energy of excitation light source,this technology only has good counting results on fresh white cysts rather than old brown cysts with weak fluorescence. In this study,we used laser as the excitation light source and explored two pairs of combinations of excitation and emission wavelengths,which were 470 nm and 535 nm,and 532 nm and 620 nm,respectively. The results showed that,under these conditions,the brown cysts could also be obtained with high contrast and clear pictures. Combined with the use of automatic counting software,cyst-counting could be conducted in an accurate and high-throughput way.

关 键 词：大豆 大豆孢囊线虫 荧光成像系统 激光 

分 类 号：S432.6[农业科学—植物病理学] S565.1[农业科学—农业昆虫与害虫防治]