滴灌施用噻虫胺防治柑橘木虱研究

【目的】明确滴灌施药防治柑橘木虱Diaphorina citri Kuwayama的可行性,为田间控制柑橘黄龙病传播提供新的手段。【方法】通过室内模拟滴灌施用噻虫胺,测定了噻虫胺在土壤中的分布以及柑橘叶片中含量,研究了滴灌施药对柑橘木虱防治效果。通过田间试验,检测分析了柑橘叶片中噻虫胺含量。【结果】在室内条件下,一次滴灌施药对柑橘木虱的防治效果可长达4个月;噻虫胺在土壤中以滴头为起始点在土壤中呈半圆形迁移分布,当柑橘叶片中噻虫胺质量分数大于0.64 mg·kg–1时,对柑橘木虱的防治效果均高于80%。田间试验表明,在药后7～63 d柑橘上部叶片中噻虫胺的质量分数大于0.64 mg·kg–1。【结论】滴灌施用噻虫胺能有效防治柑橘木虱,是田间控制柑橘黄龙病传播的有效手段。     

四种铜制剂防治沃柑溃疡病的田间药效试验

试验通过测试美果铜、噻菌铜、加瑞农和可杀得叁仟四种铜制剂对沃柑溃疡病的防治田间效果,确定合理的使用浓度和技术,评价其对沃柑的安全性,为沃柑病虫害防控技术提供依据。结果表明：四种铜制剂均能有效防治沃柑溃疡病,其中美果铜对沃柑溃疡病的防治效果最显著,其平均药效高达95%,噻菌酮和加瑞农次之,可杀得叁仟效果相对最差。四个处理组对果实的防效分别为97.68%、94.32%、93.70%、93.03%,其中美果铜的防效与其他三组显著差异。同时,在新梢叶片转绿期、幼果直径1.5㎝开始喷药,下雨前后适时喷药,能有效防治溃疡病。     

Development of a yield monitoring system for citrus mechanical harvesting machines

A load cell based yield monitoring system was developed for the Oxbo citrus mechanical harvesting machines. The yield monitoring system consisted of a GPS receiver, a mass flow sensor and data processing and storage units. The mass flow sensor consisted of four load cells attached to a carbon-fiber plate which sensed the impact force created by the oranges hitting the plate. A mathematical model was developed to relate the impact force to fruit mass. Laboratory tests were conducted on a test rig that replicated the flow of oranges to measure the accuracy of the system under a controlled environment. The system performed very well under laboratory conditions (R ² = 0.99 and an average error of 3.3%). In addition, a field test was conducted in a citrus orchard in Florida to evaluate the performance of the system under field conditions. Of the 72 rows used in the field test, the first 10 rows were used to calibrate the computed weight. A correlation of R ² = 0.97 between the actual weight and the computed weight was found from the field data with an average error of 7.81%.     

柑橘黄龙病菌对沙田柚田间性状和果实品质的影响

【目的】明确柑橘黄龙病菌对沙田柚Citrus maxima (Burm.) Merr. cv. Shatian Yu田间性状和果实品质的影响。【方法】结合田间调查和室内试验系统研究沙田柚感染黄龙病菌后叶片症状、果实内外品质和风味感官品质的变化,及病菌浓度和树龄对病程发展的影响。【结果】沙田柚嫁接病芽后,黄龙病菌浓度保持在较低水平(Ct28)时,植株叶片的黄龙病症状表现不明显,老叶轻微斑驳黄化,新叶轻微均匀黄化,抽梢正常。与健康植株(Ct35)相比,低黄龙病菌浓度(28Ct32)的植株,叶片症状、果实产量和各品质指标均无显著变化;随着发病程度的加深,高黄龙病菌浓度(Ct26)的柚树叶片出现典型斑驳黄化症状,单株柚果总产量和总结果数显著降低,果实变小变轻,着色不均匀,可食率、出汁率、可溶性固形物含量、甜味度、果肉饱满度和综合风味显著下降,酸味度和异味度反而显著提高,失去食用价值。【结论】沙田柚黄龙病病程进展较慢,但随着发病程度加深,其经济价值受到严重影响。本研究可为评价柑橘黄龙病对沙田柚品质的影响提供科学依据,为解析柚类植物的黄龙病耐性机理提供理论支撑。     

猕猴桃细菌性溃疡病可视化LAMP检测方法的建立与应用

为了建立能够广泛应用于田间的猕猴桃细菌性溃疡病快速检测技术,参考国外经验,设计并优化LAMP反应的检测体系,并可将检测结果可视化;同时,测试评价优化后的检测体系的灵敏性及特异性;通过采集猕猴桃不同部位的组织样品,验证优化后的检测体系的田间实际应用效果。结果表明:该方法能够特异性地检测出Psa病原菌,检测灵敏度达到100 CFU/mL。经对田间样品检测测试,表明该方法田间应用效果好。     

柑桔黄龙病的直接荧光诊断法

试验证明,直接荧光诊断法是诊断柑桔黄龙病的一种快速、简便、准确的方法。 将健叶和黄龙病叶叶柄用保险刀片徒手切片,在透射式荧光显微镜下检查,可看到健叶叶柄的木质部导管细胞壁发黄色荧光,韧皮纤维细胞发绿色荧光,而病叶叶柄韧皮部中有1～多个鲜明的黄色或黄绿色荧光团块。这种特异性的荧光,在健叶柄韧皮部中是没有的,其它由病毒、类病毒、细菌、真菌等病原引起的病叶柄韧皮部中亦没有此种特异性荧光。     

柑橘木虱趋光行为及复眼结构分析

【目的】研究柑橘木虱Diaphorina citri Kuwayama对不同光波的趋光性,探索其复眼结构及在光适应和暗适应下的结构变化并阐明其趋光机制,为发展柑橘木虱非化学防控技术奠定基础。【方法】选用趋光反应装置测定柑橘木虱成虫对不同光波的趋光率,利用扫描电镜技术和组织切片方法对成虫复眼形态和结构进行观察。【结果】柑橘木虱成虫对不同波长光的趋光率不同,对紫外光(365～370 nm)的趋光率为66.62%～71.38%,蓝光(470～475 nm)为47.17%～50.88%,绿光(520～525 nm)为39.37%～44.26%,黄光(590～592 nm)为28.18%～31.32%,红光(620～630 nm)为14.68%～18.33%。成虫复眼呈半球形,位于头部两侧触角基部,表面光滑无辅助结构,属于典型的并列相眼。小眼由角膜、晶锥、视网膜细胞、基膜和色素细胞组成,位于复眼中心区域的小眼呈六边形,边缘区域的小眼呈近圆形。雌、雄成虫每个复眼的小眼数分别为225～254和238～252个。【结论】柑橘木虱成虫对紫外光、蓝光和绿光的趋光率显著高于白光,雌雄成虫在复眼结构和形态方面没有明显的差异。研究结果为阐明柑橘木虱趋光行为的机制提供了理论基础。     

超量表达CsGH3.6通过抑制生长素信号转导 增强柑橘溃疡病抗性

背景 由柑橘黄单胞杆菌柑橘亚种（Xanthomonas citri subsp. citri,Xcc）引起的柑橘溃疡病是柑橘生产上最具毁灭性的一种病害。植物生长素在调控柑橘溃疡病菌引起的寄主侵染部位脓疱形成中起重要作用。生长素早期响应基因GH3通过酰基化吲哚-3-乙酸（indole-3-acetic acid,IAA）调控植物激素动态平衡。前期研究发现柑橘CsGH3.6在调控生长素响应溃疡病侵染中起着重要作用。目的 通过对超量表达CsGH3.6转基因晚锦橙的抗病性、植株表型、细胞和激素变化进行分析,利用RNA-Seq解析CsGH3.6调控的信号通路,探明CsGH3.6调控激素动态平衡影响柑橘溃疡病抗性的内在机制。方法 利用针刺法对离体转基因叶片接种溃疡病菌Xcc,统计接种第10 天时病斑面积和病情指数,以野生型为对照,评价转基因植株的抗性水平;提取感病前后叶片内源激素,利用高效液相色谱技术(high performance liquid chromatography,HPLC )检测转基因植株中激素含量变化;温室中观察转基因植株表型变化;通过测量叶片纵径、横径和厚度分析转基因植株叶型变化特征;制备叶片表皮切片,显微观察表皮细胞和气孔,并统计转基因植株表皮细胞长度和气孔密度;采用RNA-Seq测序技术研究转基因植株转录组变化情况,并利用Nr、Nt、Pfam、COG、SwissProt和gene ontology (GO)数据库注释基因功能,进一步利用KEEG数据库和MapMan软件解析超量表达CsGH3.6影响的重要基因、功能和途径,阐明CsGH3.6调控柑橘溃疡病抗性的分子机制。结果 超量表达CsGH3.6显著增强转基因植株的溃疡病抗性;转基因植株分枝增多且下垂,叶片向上卷曲,变小,颜色浅;转基因植株气孔密度增加,表皮细胞变短;激素含量分析显示,转基因植株自由生长素（IAA）和茉莉酸（jasmonic acid,JA）含量显著降低,而水杨酸（salicylic acid,SA）含量显著增加;转录组测序分析表明,超量表达CsGH3.6显著抑制生长素信号转导相关基因表达,特别是所有注释的Aux/IAA家族基因均下调表达,相反,与生物胁迫相关基因的表达为上调,其中绝大部分基因为病程相关蛋白基因。结论 超量表达CsGH3.6通过酰基化自由IAA抑制生长素信号转导,调控JA和SA的动态平衡,改变细胞和植株的形态建成,从而增强柑橘对溃疡病的抗性。研究结果暗示调控激素动态平衡在柑橘抗病育种中具有潜在价值。     

转录因子CsWRKY61对柑橘溃疡病抗性的影响

【背景】柑橘溃疡病（citrus bacterial canker,CBC）是世界柑橘产业上危害最严重的病害之一,由柑橘黄单胞杆菌柑橘亚种（Xanthomonas citri subsp. citri,Xcc）引起,在国内外被列为检疫对象。由于柑橘分子病理研究相对滞后,导致可供利用的抗性基因资源相对匮乏。WRKY转录因子参与植物抵御生物和非生物胁迫反应,前期研究发现柑橘WRKY转录因子可能在调控寄主抗病反应中起着重要作用。【目的】通过对超量表达CsWRKY50、CsWRKY61和CsWRKY72转基因晚锦橙（Citrus sinensis）的溃疡病抗性进行评价,明确这些基因在柑橘响应溃疡病菌侵染中的生物学功能和抗病育种价值。进一步利用RNA-Seq解析CsWRKY61调控的信号通路。【方法】利用农杆菌介导法进行柑橘遗传转化,获得超量表达CsWRKY50、CsWRKY61和CsWRKY72的晚锦橙;利用实时荧光定量PCR（qRT-PCR）分析转基因植株中目的基因的表达水平以及拷贝数;以非转基因植株为对照,采用离体针刺接种评价转基因植株对溃疡病的抗性;通过比较超量表达CsWRKY61和野生型植株的转录组测序结果,探究CsWRKY61提高柑橘溃疡病抗性的内在机制。【结果】分别构建了CAMV 35S启动子控制CsWRKY50、CsWRKY61和CsWRKY72表达的植物表达载体,通过GUS组织化学染色和PCR鉴定分别获得了6、8和6株转基因晚锦橙。转基因植株中目的基因的表达量有不同程度的提高,大部分转基因植株中外源基因的拷贝数为1,只有超量表达CsWRKY61的转基因植株溃疡病抗性显著增强,其病斑面积明显小于野生型植株,而超量表达CsWRKY50和CsWRKY72的转基因植株抗病性与野生型相比无明显差异。转录组分析结果显示,超量表达CsWRKY61的转基因植株中生物胁迫相关途径（包括病原入侵的感知、活性氧爆发、转录因子、防御基因、激素、细胞壁和次生代谢等）和信号转导相关途径（主要是激酶受体）均被显著激活。【结论】超量表达CsWRKY61能够激活与生物胁迫和信号转导相关的途径,增强柑橘对溃疡病的抗性;CsWRKY61在柑橘抗病育种中存在潜在的应用价值。     

对福建省柑桔健康栽培示范园的初步评估

80年代，福建省在5个地区建立柑桔示范园（曾称为无病母本园）。按照健康栽培的观点严格实施，在隔离的地区建园，种植无病毒的苗木，对黄龙病、病毒病及其虫媒定期检测，及时铲除病株的防治虫媒，加强土壤、肥、水的管理措施。这样的柑桔健康栽培示范园，与对照柑桔园相比，前者的植株生长强壮而抽梢整齐，结果优质而高产，经济效益延长5a以上。在1982-1999年间，对示范园的观察与调研中发现，黄龙病和病毒病的发生，完全取决于能否阻止其虫媒的侵入。     

Distinguishing nitrogen deficiency and fungal infection of winter wheat by laser-induced fluorescence

One of the most important tasks in precision farming is the site-specific application of fertilisers and pesticides in heterogeneous large-area fields. For such site-specific crop management, effective remote sensing methods for the detection of crop diseases and nutrient deficiencies are required. The aim of the present work was to compare laser-induced fluorescence (LIF) parameters from nitrogen-deficient and pathogen (rust and mildew)-infected winter wheat (Triticum aestivum L.) plants and to assess the potential of LIF to detect and discriminate between these types of stress. Both long term nitrogen deficiency and pathogen infection resulted in an increase of the ratio of fluorescence at 686 and 740 nm (F686/F740) accompanied by a reduction of leaf chlorophyll content to approximately 35 μg cm⁻². A linear negative correlation between chlorophyll content and F686/F740 ratio (r² = 0.78) was found for leaves with chlorophyll content ranging between 17 and 52 μg cm⁻². Since chlorophyll breakdown appeared an unspecific symptom to both nitrogen deficiency and pathogen infection, it was not possible to discriminate between these types of stress only by means of the F686/F740 ratio. Specific for the pathogen-infected leaves was a large heterogeneity in the records of their spectral parameters caused by inhomogeneous, discrete lesions of fungi infection. Nitrogen-deficient plants with homogeneous reduction in chlorophyll content showed, in contrast, more uniform readings of the spectral parameters. Thus, mildew- and rust-infected plants, grown under sufficient nitrogen fertilisation could be distinguished from those grown under reduced nitrogen supply by the higher variance of their spectral readings. The simultaneous scanning multipoint mode measurements of LIF and laser light reflection characteristics with parallel estimation of their heterogeneity is proposed for the discrimination between nitrogen deficiency and pathogen infection under field conditions.     

糙米粉和柑橘皮粉的复配添加对牛肉饼品质的影响

通过单因素和复配试验,研究糙米粉和柑橘皮粉复配对牛肉饼品质的影响。将糙米粉（0%、1%、2%和3%）和柑橘皮粉（0%、1%、2%、3%和4%）分别添加到牛肉饼中,通过产品得率、颜色（L*、a*、b*）、质构（弹性、黏聚和回复性）等指标的测定,并结合感官评价,筛选出糙米粉和柑橘皮粉的适合添加量,分别为2%和3%,在此添加量下的牛肉饼与空白对照组相比均有最大的产品得率、最佳的质构特性以及最好的感官评分（P < 0.05）。再利用复配试验优化出糙米粉和柑橘皮粉的最适复合配比,结果表明糙米粉和柑橘皮粉的最适复配比例为：糙米粉1.5%,柑橘皮粉3.5%,在此配比下加工的牛肉饼具有良好的产品得率（92.73%）和感官评分。同时低场核磁弛豫时间（T₂）结果显示,此配比下的牛肉饼水分结合最牢固,保水性最好。     

转CiNPR4基因柑橘抗溃疡病的机制解析

【目的】病程相关基因非表达子1（non-expressor of pathogenesis-related gene 1,NPR1）是水杨酸（salicylic acid,SA）介导的系统获得性抗性信号转导途径中一个重要的转录因子,在调节植物的抗病方面发挥着关键性的作用。研究耐黄龙病的‘Jackson’葡萄柚（Citrus paradisi）中的NPR1-like基因CiNPR4对柑橘溃疡病的抗性,解析过表达CiNPR4的转基因晚锦橙（C. sinensis）抗柑橘溃疡病的机理。【方法】选取黄龙病抗性增强的CiNPR4转基因柑橘,采取完全成熟的叶片利用针刺法进行柑橘溃疡病的离体抗性评价分析;在此基础上,对溃疡病抗性增强的CiNPR4转基因植株利用针刺法离体接种溃疡病菌（Xanthomonas citri subsp. citri,Xcc）,统计接种Xcc后0、1、3、5、7和9 d的细菌数量;利用注射法对溃疡病抗性增强的CiNPR4转基因植株叶片接种Xcc,接种后0、3和5 d收集接种部位的叶片,测定叶片中SA和茉莉酸（jasmonic acid,JA）含量;同时,收集接种Xcc后0、3和5 d接种部位的叶片,利用实时荧光定量 PCR分析SA和JA分别介导的防御反应相关基因CsPR1和CsPDF1.2在Xcc诱导下表达水平的变化;根据CiNPR4蛋白与TGA转录因子相互作用网络预测CiNPR4互作蛋白为Ciclev10005080m和Ciclev10001081m,以甜橙为参考基因组,将这两个基因在https://www.citrusgenomedb.org/网站上进行blastx分析,预测CiNPR4在甜橙基因组中互作的候选蛋白,克隆CiNPR4和候选蛋白基因的cDNA,利用同源重组法分别构建诱饵载体和猎物载体,并将诱饵质粒和猎物质粒共转入酵母菌株Y2HGold中,进行点对点酵母双杂交分析。【结果】CiNPR4的超量表达减轻了转基因柑橘叶片上溃疡病的症状,柑橘溃疡病抗性增强的CiNPR4转基因植株叶片上Xcc的生长比较缓慢,在接种Xcc后9 d,转基因植株Xcc数量显著低于野生型（wild type,WT）晚锦橙;Xcc诱导0 d,溃疡病抗性增强的CiNPR4转基因植株含有与WT植株无差异的SA含量,随着Xcc诱导时间的延长,CiNPR4转基因植株中SA的含量显著增加;而Xcc处理0 d时,WT植株含有显著高于CiNPR4转基因植株的JA含量,随后,JA含量在CiNPR4转基因植株中逐步上升,Xcc处理5 d时,达到显著高于WT植株的水平;而WT植株在整个处理期间SA和JA的含量基本保持不变;溃疡病抗性增强的转基因植株受Xcc诱导3 d时CsPR1的表达水平迅速上升,达到与WT植株差异显著的水平,而CsPDF1.2的表达水平在Xcc诱导5 d时上升;WT植株受Xcc诱导后CsPR1的表达水平没有发生显著变化,而CsPDF1.2的表达水平在Xcc诱导5 d时上升,且显著高于CiNPR4转基因植株中CsPDF1.2的表达水平;酵母双杂交分析证实,CiNPR4与甜橙基因组中的CsTGA2转录因子互作。【结论】CiNPR4与CsTGA2转录因子互作,通过促进SA而抑制JA介导的防御反应,调控转基因柑橘对溃疡病的抗性。     

柠檬自交后代抗柑橘溃疡病的离体鉴定

为鉴定柠檬自交后代对柑橘溃疡病的抗性,以柠檬鲜叶为材料,通过注射法和针刺法接种不同浓度溃疡病菌菌液,对114株柠檬自交后代群体进行离体鉴定,并以感病材料冰糖橙、抗病材料枸橼C-05作为对照。结果表明:柠檬自交后代对溃疡病抗性存在个体间差异;其中,注射接种法中表现为抗10~4和10~5 CFU/m L病原菌的植株有24株,针刺接种法中表现为抗108 CFU/m L病原菌的植株有11株;其中,有6株柠檬自交后代在2种鉴定方法中均被认定为抗病株。综合2种鉴定方法,114株柠檬自交后代中有29株对溃疡病表现出抗10~4 CFU/m L病原菌。     

温度和多胺对柑橘溃疡病发生的影响及作用机制

【目的】溃疡病是严重危害柑橘的一种细菌性病害,通常在高温下容易发生。论文旨在阐明高温下柑橘易发生溃疡病的机制,揭示其代谢变化,为利用药剂防治溃疡病提供重要的理论指导。【方法】以感病的甜橙(Citrus sinensis)为研究对象,在21℃和30℃下预培养3 d,然后均接种同样浓度(108 cfu/m L)的柑橘溃疡病菌(Xanthomonas citri subsp.citri,Xcc)10μL,比较两组植株的发病率,采用半定量RT-PCR分析两种温度下4个抗病基因AOS、CHI、GPX和PR4A的表达量,利用HPLC测定预培养3 d后叶片内源多胺(腐胺、亚精胺和精胺)的含量。在此基础上,利用外源亚精胺(0.4 mmol·L-1)处理甜橙植株(以清水处理为对照),比较亚精胺和清水处理的植株接种溃疡病菌后的发病率和病情指数,分析亚精胺处理对内源多胺含量和抗病基因AOS、CHI、GPX和PR4A表达的影响。【结果】溃疡病菌接种后观察发现,21℃培养植株溃疡病的发病率在前期低于30℃培养的植株,至第10天时,两个处理组植株的发病率接近;同时HPLC测定发现,21℃培养植株叶片3种自由态多胺(腐胺、亚精胺和精胺)含量高于30℃培养植株;RT-PCR分析表明,CHI、GPX和PR4A这3个抗病基因的表达量在21℃培养植株中高于30℃培养植株,而AOS表达水平在两组材料中差异不明显。外源亚精胺处理显著增加了内源腐胺和亚精胺的含量,降低了所处理植株接种溃疡病菌后的发病率和病情指数,接种后14 d发病率比对照降低45%,病情指数比对照降低4.8,而由表型可见对照发病程度重于亚精胺处理材料。此外,亚精胺处理能够增强AOS、CHI、GPX和PR4A 4个抗病基因的表达量。【结论】高温下甜橙更易发生溃疡病的可能机制是高温抑制抗病基因的表达和多胺合成。外源多胺处理能够降低甜橙发生溃疡病,可能机制是多胺处理后增强了抗病基因的表达,诱发植株的抗病反应最终表现出抗病。因此,高温是影响溃疡病发生的一个关键环境因子,多胺有助于提高对柑橘溃疡病的抗性。     

柑橘黄龙病检测技术研究进展

冠以“柑橘（Ｃｉｔｒｕｓ ｒｅｔｉｃｕｌａｔａ Ｂｌａｎｃｏ．）癌症”之称的黄龙病暂被认为是由韧皮部难养细菌引起的对柑橘产业造成严重威胁的毁灭性病害,它给柑橘产业的健康发展造成了严重的威胁．鉴于柑橘黄龙病检测技术对柑橘黄龙病防治的重要性,文章对柑橘黄龙病及其病原、黄龙病的检测技术进行了综述,并对柑橘黄龙病的检测技术未来发展进行了展望.     

应用FBA染色法观察柑桔黄龙病病株中的病原难培养细菌

病株新梢主叶脉或嫩茎横切片用 FBA 染色工作液染色5～8min,在光学显微镜(油镜)下可见到其外向韧皮部筛管细胞中存在被染成红褐色的病原难培养细菌,菌体大小为0.35～0.65μm × 1.70～4.00μm,多为杆状,有些呈长亚铃形或长椭圆形。而健株、衰退病株、裂皮病株、溃疡病株,生理黄化株和田间天牛为害黄化株等的叶脉或嫩茎韧皮部筛管细胞中均未见到任何菌体存在。FBA 染色法快速、简便、准确,可作为诊断柑桔黄龙病的一种实用方法。本文还就超薄切片电镜观察黄龙病病原检出率低的原因进行了探讨。     

Detection of Huanglongbing (citrus greening) based on hyperspectral image analysis and PCR

Huanglongbing (HLB, citrus greening) is one of the most serious quarantine diseases of citrus worldwide. To monitor in real-time, recognize diseased trees, and efficiently prevent and control HLB disease in citrus, it is necessary to develop a rapid diagnostic method to detect HLB infected plants without symptoms. This study used Newhall navel orange plants as the research subject, and collected normal color leaf samples and chlorotic leaf samples from a healthy orchard and an HLB-infected orchard, respectively. First, hyperspectral data of the upper and lower leaf surfaces were obtained, and then the polymerase chain reaction (PCR) was used to detect the HLB bacterium in each leaf. The PCR test results showed that all samples from the healthy orchard were negative, and a portion of the samples from the infected orchard were positive. According to these results, the leaf samples from the orchards were divided into disease-free leaves and HLB-positive leaves, and the least squares support vector machine recognition model was established based on the leaf hyperspectral reflectance. The effect on the model of the spectra obtained from the upper and lower leaf surfaces was investigated and different pretreatment methods were compared and analyzed. It was observed that the HLB recognition rate values of the calibration and validation sets based on upper leaf surface spectra under 9-point smoothing pretreatment were 100% and 92.5%, respectively. The recognition rate values based on lower leaf surface spectra under the second-order derivative pretreatment were also 100% and 92.5%, respectively. Both upper and lower leaf surface spectra were available for recognition of HLB-infected leaves, and the HLB PCR-positive leaves could be distinguished from the healthy by the hyperspectral modeling analysis. The results of this study show that early and nondestructive detection of HLB-infected leaves without symptoms is possible, which provides a basis for the hyperspectral diagnosis of citrus with HLB.     

Feature extraction of hyperspectral images for detecting immature green citrus fruit

At an early immature growth stage of citrus, a hyperspectral camera of 369–1042 nm was employed to acquire 30 hyperspectral images in order to detect immature green fruit within citrus trees under natural illumination conditions. First, successive projections algorithm (SPA) were implemented to select 677, 804, 563, 962, and 405 nm wavebands and to construct multispectral images from the original hyperspectral images for further processing. Then, histogram threshold segmentation using NDVI of 804 and 677 nm was implemented to remove image backgrounds. Three slope parameters, calculated from the pairs 405 and 563 nm, 563 and 677 nm, and 804 and 962 nm were used to construct a classifier to identify the potential citrus fruit. Then, a marker-controlled watershed segmentation based on wavelet transform was applied to obtain potential fruit areas. Finally, a green fruit detection model was constructed according to Grey Level Co-occurrence Matrix (GLCM) texture features of the independent areas. Three supervised classifiers, logistic regression, random forest and support vector machine (SVM) were developed using texture features. The detection accuracies were 79%, 75%, and 86% for the logistic regression, random forest, and SVM models, respectively. The developed algorithm showed a great potential for identifying immature green citrus for an early yield estimation.