基于衍射重构技术的作物真菌病害孢子微型检测装置
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摘要
为了解决现有孢子检测系统体积大、成本高等问题,提出了一种基于衍射重构技术的作物真菌病害孢子检测方法。根据惠更斯-菲涅尔原理、角谱理论,利用衍射成像复合重构计算设计了一种包含富集、进样机构的真菌病害孢子检测系统。该系统可以定时完成富集、进样、拍摄、重构和检测等操作,并通过重构算法实现对真菌病害孢子原像的重建,根据重构后的图像提取面积(Area)、细度(Thinness ratio)两个重要形态学特征,对稻瘟病孢子进行检测识别。实验结果表明,所设计装置对稻瘟病孢子的检测结果与人工显微镜识别结果高度线性相关,决定系数为0. 99,平均检测误差为5. 91%,具有较好的准确性。
Collecting airborne spores of rice blast in rice fields using spore trap devices has currently become an important approach for devising strategies early and effectively controlling rice blast. In order to solve the problems of large volume and high cost of existing spore detection system,a spore detection method for crop fungal diseases was proposed based on diffraction reconstruction technology. Based on Huygens-Fresnel principle and angular spectrum theory,a spore detection system for fungal diseases,including enrichment and sampling mechanism,was designed by using diffraction imaging complex reconstruction calculation. The system can complete a series of operations such as enrichment,sampling,shooting,reconstructing and detection,and reconstruct the original spore image of fungal diseases by reconstruction algorithm. According to the morphological characteristics of reconstructed images,two important parameters,area and thinness ratio,were extracted to detect and identify spores. Rice blast spores were selected as the research object to carry out detection and verification experiments. The experimental results showed that the system could capture the micro-images of diffractions of rice blast spores,with 2 592 pixels × 1 944 pixels resolution. The experiments validated that the correlation coefficient between the detection results of rice blast spores and the identification results of artificial microscope can reach 0. 99,while the average detection error rate was 5. 91%, which had good accuracy. The research provided a design scheme for the research and development of low-cost early warning equipment for crop fungal diseases.
Collecting airborne spores of rice blast in rice fields using spore trap devices has currently become an important approach for devising strategies early and effectively controlling rice blast. In order to solve the problems of large volume and high cost of existing spore detection system,a spore detection method for crop fungal diseases was proposed based on diffraction reconstruction technology. Based on Huygens-Fresnel principle and angular spectrum theory,a spore detection system for fungal diseases,including enrichment and sampling mechanism,was designed by using diffraction imaging complex reconstruction calculation. The system can complete a series of operations such as enrichment,sampling,shooting,reconstructing and detection,and reconstruct the original spore image of fungal diseases by reconstruction algorithm. According to the morphological characteristics of reconstructed images,two important parameters,area and thinness ratio,were extracted to detect and identify spores. Rice blast spores were selected as the research object to carry out detection and verification experiments. The experimental results showed that the system could capture the micro-images of diffractions of rice blast spores,with 2 592 pixels × 1 944 pixels resolution. The experiments validated that the correlation coefficient between the detection results of rice blast spores and the identification results of artificial microscope can reach 0. 99,while the average detection error rate was 5. 91%, which had good accuracy. The research provided a design scheme for the research and development of low-cost early warning equipment for crop fungal diseases.
引文
[1]王文斌,张荣胜,罗楚平,等.中国主要稻区稻曲病菌的生物学特性及群体遗传多样性[J].中国农业科学,2014,47(14):2762-2773.WANG Wenbin,ZHANG Rongsheng,LUO Chuping,et al.Biological characteristics and genetic diversity of Ustilaginoidea virens from rice regions in China[J].Scientia Agricultura Sinica,2014,47(14):2762-2773.(in Chinese)
[2]俞咪娜,陈志谊,于俊杰,等.来源于同一穗不同稻曲球的稻曲病的致病性及遗传多样性[J].植物病理学报,2013,43(6):561-573.YU Mina,CHEN Zhiyi,YU Junjie,et al.Genetic diversity and pathogenicity of Ustilaginoidea virens isolated from different rice false smut balls of a diseased spike[J].Acta Phytopathologica Sinica,2013,43(6):561-573.(in Chinese)
[3]BLOCK E K,LACHER T E,BREWER L W,et al.Population responses of Peromyscus resident in Iowa cornfields treated with the organophosphorus pesticide COUNTER[J].Ecotoxicology,1999,8(3):189-200.
[4]傅泽田,祁力钧.国内外农药使用状况及解决农药超量使用问题的途径[J].农业工程学报,1998,14(2):7-12.FU Zetian,QI Lijun.Over-use of pesticide and approaches to reduce pesticide dosage[J].Transactions of the CSAE,1998,14(2):7-12.(in Chinese)
[5]SUGENG A J,BEAMER P I,LUTZ E A,et al.Hazard-ranking of agricultural pesticides for chronic health effects in Yuma County,Arizona[J].Science of the Total Environment,2013,463(5):35-41.
[6]WAHID F A,WICKLIFFE J,WILSON M,et al.Presence of pesticide residues on produce cultivated in Suriname[J].Environmental Monitoring and Assessment,2017,189(6):303.
[7]杨燕,何勇.基于高光谱图像的稻瘟病抗氧化酶值早期预测[J].农业工程学报,2013,29(20):135-141.YANG Yan,HE Yong.Early prediction of antioxidant enzyme value of rice blast based on hyper-spectral image[J].Transactions of the CSAE,2013,29(20):135-141.(in Chinese)
[8]ZHANG Y,YANG J,LU Y,et al.A competitive direct enzyme-linked immunosorbent assay for the rapid detection of deoxynivalenol:development and application in agricultural products and feedstuff[J].Food and Agricultural Immunology,2017,28(3):516-527.
[9]YANG F,WANG G P,XU W X,et al.A rapid silica spin column-based method of RNA extraction from fruit trees for RT-PCRdetection of viruses[J].Journal of Virological Methods,2017,247:61-67.
[10]王军军,谢海芬,周嘉,等.微悬臂梁在化学及生物传感器中的应用进展[J].微电子学,2004,34(5):493-496.WANG Junjun,XIE Haifen,ZHOU Jia,et al.Development of micro cantilever-based chemical and biological sensors[J].Microelectronics,2004,34(5):493-496.(in Chinese)
[11]NUGAEVA N,GFELLER K Y,BACKMANN N,et al.Micromechanical cantilever array sensors for selective fungal immobilization and fast growth detection[J].Biosensors&Bioelectronics,2005,21(6):849-856.
[12]李鹏,李昕欣,王跃林.用于化学气体检测的压阻检测式二氧化硅微悬臂梁传感器[J].传感技术学报,2007,20(10):2174-2177.LI Peng,LI Xinxin,WANG Yuelin.Piezeoresistive silicon dioxide microcantilever sensor for chemical gas detection[J].Journal of Transduction Technology,2007,20(10):2174-2177.(in Chinese)
[13]李仪芳,杨冠玲,李丰果,等.大气PM2.5显微图像的处理与分析[J].光电工程,2005,32(11):54-58.LI Yifang,YANG Guanling,LI Fengguo,et al.Processing and analysis for micrograph of PM2.5 in atmosphere[J].OptoElectronic Engineering,2005,32(11):54-58.(in Chinese)
[14]李小龙,马占鸿,孙振宇,等.基于图像处理的小麦条锈病菌夏孢子模拟捕捉的自动计数[J].农业工程学报,2013,29(2):199-206.LI Xiaolong,MA Zhanhong,SUN Zhenyu,et al.Automatic counting for trapped urediospores of Puccinia striiformis f.sp.tritici based on image processing[J].Transactions of the CASE,2013,29(2):199-206.(in Chinese)
[15]齐龙,蒋郁,李泽华,等.基于显微图像处理的稻瘟病菌孢子自动检测与计数方法[J].农业工程学报,2015,31(12):186-193.QI Long,JIANG Yu,LI Zehua,et al.Automatic detection and counting method for spores of rice blast based on micro image processing[J].Transactions of the CASE,2015,31(12):186-193.(in Chinese)
[16]姜玉英,罗金燕,罗德平,等.远程控制病菌孢子捕捉仪对小麦气传病害的监测效果[J].植物保护,2015,41(6):163-168.JIANG Yuying,LUO Jinyan,LUO Deping,et al.Monitoring effect of remote-controlled spore trap on wheat aero-borne diseases[J].Plant Protection,2015,41(6):163-168.(in Chinese)
[17]雷雨,姚志凤,何东健.小麦条锈病菌夏孢子显微图像远程采集系统设计与试验[J/OL].农业机械学报,2018,49(11):39-47.LEI Yu,YAO Zhifeng,HE Dongjian.Design and experiment of micro-image remote acquisition system of uredinispores of Puccinia striiformis f.sp.tritici[J/OL].Transactions of the Chinese Society for Agricultural Machinery,2018,49(11):39-47.http:∥www.j-csam.org/jcsam/ch/reader/view_abstract.aspx?file_no=20181105&flag=1&journal_id=jcsam.DOI:10.6041/j.issn.1000-1298.2018.11.005.(in Chinese)
[18]MELO C,LOPES J G,ANDRADE A O,et al.Semi-automated counting of arbuscular mycorrhizal fungi spores using artificial neural network[J].IEEE Latin America Transactions,2017,15(8):1566-1573.
[19]ROY M,JIN G,SEO D.A simple and low-cost device performing blood cell counting based on lens-free shadow imaging technique[J].Sensors&Actuators B Chemical,2014,201(4):715-723.
[20]SEO D,ROY M,KIM J.High-throughput and real-time microalgae monitoring platform using lens-free shadow imaging system(LSIS)[C]∥Sensors,IEEE,2016:1-3.
[21]ZHANG X,KHIMJI I,GURKAN U A.Lensless imaging for simultanrous microfluidic sperm monitoring and sorting[J].Lab on a Chip,2011,11(15):2535-2540.
[22]LI G X,ZHANG R B,YANG N,et al.An approach for cell viability online detection based on the characteristics of lensfree cell diffraction fingerprint[J].Biosensors and Bioelectronics,2018,107:163-169.
[23]THOMAS B P,RAJENDREN K V,PILLAI S A.Wholefield NDT of porous materials using digital holography[C]∥Proc.National Seminar on Non-Destructive Evaluation.Hyderabad:Indian Society for Non-Destructive Testing,2006:229-234.
[24]LEE Y H,KASSAM S A.Generalized median filtering and related nonlinear filtering techniques[J].Acoustics Speech&Signal Processing IEEE Transactions on,1985,33(3):672-683.
[25]MUDANYALI O,TSENG D,OH C.Compact,Light-weight and cost-effective microscope based on lensless incoherent holography for telemedicine applications[J].Lab on a Chip,2010,10(11):321-328.
[26]许燎原,赵丽稳,胡宇峰,等.稻瘟病菌孢子q PCR方法的建立及用于监测气传菌源的研究[J].浙江农业学报,2016,28(8):1368-1373.XU Liaoyuan,ZHAO Liwen,HU Yufeng,et al.Development of a q PCR detection method for monitoring conidial density of rice blast fungus in the air[J].Acta Agriculturae Zhejiangensis,2016,28(8):1368-1373.(in Chinese)
[27]陈浩,康晓慧,张梅,等.稻瘟病菌空中孢子量与气象因子的关系[J].湖北农业科学,2009,48(8):1869-1871.CHEN Hao,KANG Xiaohui,ZHANG Mei,et al.Relationship between spatial spore amount of Pyricularia oryzae cavara and meteorological factors[J].Hubei Agricultural Science,2009,48(8):1869-1871.(in Chinese)
[2]俞咪娜,陈志谊,于俊杰,等.来源于同一穗不同稻曲球的稻曲病的致病性及遗传多样性[J].植物病理学报,2013,43(6):561-573.YU Mina,CHEN Zhiyi,YU Junjie,et al.Genetic diversity and pathogenicity of Ustilaginoidea virens isolated from different rice false smut balls of a diseased spike[J].Acta Phytopathologica Sinica,2013,43(6):561-573.(in Chinese)
[3]BLOCK E K,LACHER T E,BREWER L W,et al.Population responses of Peromyscus resident in Iowa cornfields treated with the organophosphorus pesticide COUNTER[J].Ecotoxicology,1999,8(3):189-200.
[4]傅泽田,祁力钧.国内外农药使用状况及解决农药超量使用问题的途径[J].农业工程学报,1998,14(2):7-12.FU Zetian,QI Lijun.Over-use of pesticide and approaches to reduce pesticide dosage[J].Transactions of the CSAE,1998,14(2):7-12.(in Chinese)
[5]SUGENG A J,BEAMER P I,LUTZ E A,et al.Hazard-ranking of agricultural pesticides for chronic health effects in Yuma County,Arizona[J].Science of the Total Environment,2013,463(5):35-41.
[6]WAHID F A,WICKLIFFE J,WILSON M,et al.Presence of pesticide residues on produce cultivated in Suriname[J].Environmental Monitoring and Assessment,2017,189(6):303.
[7]杨燕,何勇.基于高光谱图像的稻瘟病抗氧化酶值早期预测[J].农业工程学报,2013,29(20):135-141.YANG Yan,HE Yong.Early prediction of antioxidant enzyme value of rice blast based on hyper-spectral image[J].Transactions of the CSAE,2013,29(20):135-141.(in Chinese)
[8]ZHANG Y,YANG J,LU Y,et al.A competitive direct enzyme-linked immunosorbent assay for the rapid detection of deoxynivalenol:development and application in agricultural products and feedstuff[J].Food and Agricultural Immunology,2017,28(3):516-527.
[9]YANG F,WANG G P,XU W X,et al.A rapid silica spin column-based method of RNA extraction from fruit trees for RT-PCRdetection of viruses[J].Journal of Virological Methods,2017,247:61-67.
[10]王军军,谢海芬,周嘉,等.微悬臂梁在化学及生物传感器中的应用进展[J].微电子学,2004,34(5):493-496.WANG Junjun,XIE Haifen,ZHOU Jia,et al.Development of micro cantilever-based chemical and biological sensors[J].Microelectronics,2004,34(5):493-496.(in Chinese)
[11]NUGAEVA N,GFELLER K Y,BACKMANN N,et al.Micromechanical cantilever array sensors for selective fungal immobilization and fast growth detection[J].Biosensors&Bioelectronics,2005,21(6):849-856.
[12]李鹏,李昕欣,王跃林.用于化学气体检测的压阻检测式二氧化硅微悬臂梁传感器[J].传感技术学报,2007,20(10):2174-2177.LI Peng,LI Xinxin,WANG Yuelin.Piezeoresistive silicon dioxide microcantilever sensor for chemical gas detection[J].Journal of Transduction Technology,2007,20(10):2174-2177.(in Chinese)
[13]李仪芳,杨冠玲,李丰果,等.大气PM2.5显微图像的处理与分析[J].光电工程,2005,32(11):54-58.LI Yifang,YANG Guanling,LI Fengguo,et al.Processing and analysis for micrograph of PM2.5 in atmosphere[J].OptoElectronic Engineering,2005,32(11):54-58.(in Chinese)
[14]李小龙,马占鸿,孙振宇,等.基于图像处理的小麦条锈病菌夏孢子模拟捕捉的自动计数[J].农业工程学报,2013,29(2):199-206.LI Xiaolong,MA Zhanhong,SUN Zhenyu,et al.Automatic counting for trapped urediospores of Puccinia striiformis f.sp.tritici based on image processing[J].Transactions of the CASE,2013,29(2):199-206.(in Chinese)
[15]齐龙,蒋郁,李泽华,等.基于显微图像处理的稻瘟病菌孢子自动检测与计数方法[J].农业工程学报,2015,31(12):186-193.QI Long,JIANG Yu,LI Zehua,et al.Automatic detection and counting method for spores of rice blast based on micro image processing[J].Transactions of the CASE,2015,31(12):186-193.(in Chinese)
[16]姜玉英,罗金燕,罗德平,等.远程控制病菌孢子捕捉仪对小麦气传病害的监测效果[J].植物保护,2015,41(6):163-168.JIANG Yuying,LUO Jinyan,LUO Deping,et al.Monitoring effect of remote-controlled spore trap on wheat aero-borne diseases[J].Plant Protection,2015,41(6):163-168.(in Chinese)
[17]雷雨,姚志凤,何东健.小麦条锈病菌夏孢子显微图像远程采集系统设计与试验[J/OL].农业机械学报,2018,49(11):39-47.LEI Yu,YAO Zhifeng,HE Dongjian.Design and experiment of micro-image remote acquisition system of uredinispores of Puccinia striiformis f.sp.tritici[J/OL].Transactions of the Chinese Society for Agricultural Machinery,2018,49(11):39-47.http:∥www.j-csam.org/jcsam/ch/reader/view_abstract.aspx?file_no=20181105&flag=1&journal_id=jcsam.DOI:10.6041/j.issn.1000-1298.2018.11.005.(in Chinese)
[18]MELO C,LOPES J G,ANDRADE A O,et al.Semi-automated counting of arbuscular mycorrhizal fungi spores using artificial neural network[J].IEEE Latin America Transactions,2017,15(8):1566-1573.
[19]ROY M,JIN G,SEO D.A simple and low-cost device performing blood cell counting based on lens-free shadow imaging technique[J].Sensors&Actuators B Chemical,2014,201(4):715-723.
[20]SEO D,ROY M,KIM J.High-throughput and real-time microalgae monitoring platform using lens-free shadow imaging system(LSIS)[C]∥Sensors,IEEE,2016:1-3.
[21]ZHANG X,KHIMJI I,GURKAN U A.Lensless imaging for simultanrous microfluidic sperm monitoring and sorting[J].Lab on a Chip,2011,11(15):2535-2540.
[22]LI G X,ZHANG R B,YANG N,et al.An approach for cell viability online detection based on the characteristics of lensfree cell diffraction fingerprint[J].Biosensors and Bioelectronics,2018,107:163-169.
[23]THOMAS B P,RAJENDREN K V,PILLAI S A.Wholefield NDT of porous materials using digital holography[C]∥Proc.National Seminar on Non-Destructive Evaluation.Hyderabad:Indian Society for Non-Destructive Testing,2006:229-234.
[24]LEE Y H,KASSAM S A.Generalized median filtering and related nonlinear filtering techniques[J].Acoustics Speech&Signal Processing IEEE Transactions on,1985,33(3):672-683.
[25]MUDANYALI O,TSENG D,OH C.Compact,Light-weight and cost-effective microscope based on lensless incoherent holography for telemedicine applications[J].Lab on a Chip,2010,10(11):321-328.
[26]许燎原,赵丽稳,胡宇峰,等.稻瘟病菌孢子q PCR方法的建立及用于监测气传菌源的研究[J].浙江农业学报,2016,28(8):1368-1373.XU Liaoyuan,ZHAO Liwen,HU Yufeng,et al.Development of a q PCR detection method for monitoring conidial density of rice blast fungus in the air[J].Acta Agriculturae Zhejiangensis,2016,28(8):1368-1373.(in Chinese)
[27]陈浩,康晓慧,张梅,等.稻瘟病菌空中孢子量与气象因子的关系[J].湖北农业科学,2009,48(8):1869-1871.CHEN Hao,KANG Xiaohui,ZHANG Mei,et al.Relationship between spatial spore amount of Pyricularia oryzae cavara and meteorological factors[J].Hubei Agricultural Science,2009,48(8):1869-1871.(in Chinese)