不同温度组合热处理治疗柑橘黄龙病的田间效果分析

[目的]研究不同温度组合热处理对柑橘黄龙病的控制效果,为柑橘黄龙病防治提供新思路.[方法]以12株感染柑橘黄龙病的纽荷尔脐橙为材料,设4个处理(处理1:对照,不作任何处理;处理2:套膜,薄膜中间不打孔;处理3:薄膜中间打10个半径为1.5 cm的孔洞;处理4:薄膜中间打10个半径为3.0 cm的孔洞),每处理4株病树,分别于2017年8月6、12、19、25日和9月2日进行5次热处理试验,热处理时间为温度较高的14:00~15:00,每隔5 min记录一次温度.处理前每组采集样品作为参照组,5次热处理后再次采集样品,并通过实时荧光定量PCR(qPCR)定量分析5次热处理后病菌含量相对其参照组的倍数,分析柑橘黄龙病菌在经过热处理后的变化趋势.[结果]处理2和处理3的平均温度均超过38.0℃,最高温度分别为49.3和42.3℃;5次热处理后病树体内病菌含量显著下降(P<0.05,下同);病情指数明显降低,病情明显好转;价值果产量下降较缓,平均损失率分别为17.71%和14.52%;但处理2的温度过高,在治疗的同时易对树体造成伤害,树冠部分出现萎蔫、焦黄等现象.处理4的最高温度仅为37.3℃,未达到治疗柑橘黄龙病的有效温度,其病情指数上升,病情加重,病菌含量也有所上升,价值果平均损失率为41.18%.对照组病情持续加重,树势明显衰退,病菌含量显著升高;价值果产量下降最明显,平均损失率为72.22%.[结论]经过5次热处理后,处理3的柑橘黄龙病树病情指数显著降低,症状明显减轻,树体病菌含量明显下降,树势明显好转,对柑橘黄龙病的治疗效果最佳.在发病严重的柑橘园可通过该方式延长果树的挂果周期,从而减少经济损失.     

云南柑橘黄龙病病原检测及其 １６ＳｒＤＮＡ序列分析

利用柑橘黄龙病菌的特异引物,对采自云南红河州建水县和个旧市、玉溪市华宁县、大理州宾川县、昭通市盐津县、德宏州瑞丽市以及保山市隆阳区等地共 １５１个疑似柑橘黄龙病样品进行了黄龙病菌的检测,其中建水县有 ５个样品检测结果为阳性,检出率为 ３３３％;个旧市、华宁县和宾川县各有 １个样品为阳性,检出率分别为 ８.３％,５.５％,６.７％;而昭通、德宏和保山的样品均为阴性。表明黄龙病在云南的主要柑橘产区建水、个旧、华宁和宾川均有不同程度的发生,其中建水县发生率较高,生产上应尽快引起重视。对所获得的黄龙病菌 １６ＳｒＤＮＡ进行了克隆和序列测定,发现 ８个云南柑橘黄龙病菌 １６ＳｒＤＮＡ的序列一致性均为１００％,与柑橘黄龙病菌亚洲种 （ＣａｎｄｉｄａｔｕｓＬｉｂｅｒｉｂａｃｔｅｒａｓｉａｔｉｃｕｓ）相应序列的一致性为 ９８.７％ ～１００％,与柑橘黄龙病菌非洲种 （ＣａｎｄｉｄａｔｕｓＬｉｂｅｒｉｂａｃｔｅｒａｆｒｉｃａｎｕｓ）的一致性为 ９７.５％ ～９８.７％,而与柑橘黄龙病菌美洲种（ＣａｎｄｉｄａｔｕｓＬｉｂｅｒｉｂａｃｔｅｒａｍｅｒｉｃａｎｕｓ）的一致性均为 ９６.５％。表明云南主要柑橘产区发生的黄龙病菌均属于柑橘黄龙病菌亚洲种。     

湿度对田间自然热罩防治柑橘黄龙病的影响

比较加湿处理后热罩和自然条件下热罩对黄龙病的防治效果,结果表明自然条件处理后30 d叶片平均病菌浓度下降了85％以上,而加湿条件下叶片平均病菌浓度仅下降3.7％,与自然条件处理间差异达到显著水平;自然条件热罩处理后90 d叶片平均病菌浓度下降了97％以上,而加湿处理病树叶片平均病菌浓度下降了38％,与自然条件处理间的差异达到极显著水平。对照组叶片平均病菌浓度处理后30 d和90 d均有略微增加;加湿未热罩处理叶片平均病菌浓度在处理后30 d和90 d分别增加了4.5倍和8.7倍,但与对照组无显著差异。本研究结果表明加湿热罩处理病树显著减弱了其杀灭黄龙病菌效果,同时表明增加土壤湿度对黄龙病菌繁殖可能具有一定的促进作用。     

柑橘黄龙病菌与柑橘根系关系研究进展

由柑橘黄龙病菌(Candidatus Liberibacter spp.)引起的柑橘黄龙病是一种极具破坏性的柑橘病害,严重影响柑橘产业的可持续发展。目前关于柑橘黄龙病的研究多以柑橘叶片为研究对象,而柑橘黄龙病是一种系统性病害,其症状在柑橘叶片、果实和根系上均有表现。近年来,越来越多的研究从根系的营养、基因和蛋白表达、根际微生物等角度研究柑橘黄龙病菌对柑橘根系的影响,因此就柑橘黄龙病菌与柑橘根系关系的最新研究结果进行综述,以期为系统开展柑橘黄龙病菌与柑橘根系互作关系的研究提供有益的借鉴和参考。     

柑橘黄龙病病情消长动态与立体式防效探讨

通过对涌泉镇2004-2012年全镇柑橘黄龙病全面普查,结果表明,无核蜜橘柑橘黄龙病病菌感染至显症潜伏期可达1年以上,其年度扩散消长呈单峰型曲线变化规律。坚持每年"一挖两治"(挖除病树阻断菌源,加强种苗接穗管治及柑橘木虱防治阻断菌链)和"三防五关"(春季清园和严防从病区采入接穗高接换种,夏季进行夏梢柑橘木虱防治,秋冬季进行秋梢柑橘木虱防治和彻底查挖病树,狠抓种苗检疫关、治虫防病关、疫情普查关、病树挖除关和健身控病关)立体式防控,其防控效果可达87%~93%,从而保障柑橘产业健康持续发展。     

庆元甜橘柚黄龙病菌的Real-time PCR检测与分析

为了解庆元县甜橘柚柑橘黄龙病的发生分布情况,为该病害的有效防控提供依据,通过Real-time PCR方法检测了庆元不同地区甜橘柚样品中黄龙病菌的存在情况。结果显示:在所检测的69个样品中,有13个样品表现为柑橘黄龙病阳性,阳性率达18.8%;其中松源街道和屏都街道样品的黄龙病菌检出率最高,可达33.3%。此检测结果表明,柑橘黄龙病已在庆元甜橘柚果园中普遍存在,应加强对该病害的防控。     

台州柑橘黄龙病亚洲种及亚洲柑橘木虱防控研究

文章主要回顾阐述2002—2017 年台州对柑橘黄龙病亚洲种和亚洲柑橘木虱防控研究结果。指出黄龙病在果园空间分布入侵初期呈均匀分布,后呈聚集分布,亚洲柑橘木虱在果园内也呈聚集分布状态;认为黄龙病对柑橘产量与品质影响是随着病级的上升增加。得出黄龙病疫情自然入侵扩散流行呈周期性变化规律,建立积年发病率(PI)与入侵年数(N)模型I=84.6740/[1+EXP(3.540-0.5838X)](n=14,r=0.9946**);检测发现黄龙病树体内和亚洲柑橘木虱带菌量在11 月和12 月最高,亚洲柑橘木虱带菌率同样以12 月最高。提出防治亚洲柑橘木虱除抓好春夏秋三梢期外,冬季和初春清园也是一个重要环节。台州探索实践的“九坚持”和“三防五关”防控模式对黄龙病防控和柑橘产业的可持续发展起到了积极作用,可在相似柑橘生态区推广应用。     

一种快速鉴定柑橘黄龙病耐性种质材料的方法

【目的】研发一种快速鉴定柑橘黄龙病耐性种质材料的方法,为加快柑橘黄龙病耐性育种进程和提高育种效率打下基础。【方法】以收集的36份疑似对柑橘黄龙病具有耐性的柑橘种质为材料,采用直接高接于感染黄龙病菌柑橘树上的方法(高接染毒鉴定法),通过田间症状观察结合定量PCR检测对试验材料的耐病性进行鉴定评价。【结果】春季高接供试材料1个月后接芽萌发,3个月后(2018年6月5日)首次在KH-14上出现典型的叶片斑驳型黄化症状,4个月后(2018年7月5日)观察有23份种质材料的叶片出现斑驳型黄化症状,另外11份种质材料未表现症状;6个月后(2018年9月4日)观察发现KH-18、KH-12、KHY-4、KHY-5和KHY-6等5份种质材料的生长虽然较正常,但叶片已出现黄化症状,只有KH-21的枝梢生长良好,无黄梢和斑驳型黄化叶,初步判定为对黄龙病具有耐性的种质材料。对6份种质材料的定性PCR检测结果表明,KH-21为阴性,其他5份材料为阳性;实时荧光定量PCR检测结果,KH-21的平均黄龙病菌含量为1870.0个细胞/μg DNA,对照材料平均黄龙病菌含量为372285.5个细胞/μg DNA,表明KH-21对柑橘黄龙病具有耐性。【结论】高接染毒鉴定法鉴定周期在6个月左右,具有通量大、时间短、效率高、成本低及结果准确等优点,能达到快速鉴定柑橘品种对柑橘黄龙病耐性的目的,可在柑橘黄龙病耐性鉴定中推广使用。     

江华县柑橘黄龙病发生特点及防控对策

2008年以来,江华瑶族自治县大力发展柑橘产业,现已形成3 334 hm2的规模,但柑橘黄龙病却成为柑橘进一步发展的制约因素。对近年来江华瑶族自治县柑橘黄龙病发生情况、症状特点及其发生的影响因素进行了认真分析,并提出了详细的防控对策,即加强检疫,培育无病苗木,挖除病树,减少病菌来源,防治柑橘木虱,消灭传播媒介,加强果园管理,培育健壮树势。     

柑橘黄龙病菌基因组学的研究进展

柑橘黄龙病(Citrus Huanglongbing,HLB)是柑橘生产上最具毁灭性的病害,严重威胁着世界柑橘产业的发展。该病害由难培养细菌——候选韧皮部杆菌(Candidatus Liberibacter spp.)所引起,通过带病苗木和媒介昆虫进行传播蔓延。近年来,随着DNA测序技术和生物信息学的发展,基因组学被广泛应用于柑橘黄龙病的研究,并为克服黄龙病菌难培养所造成的研究瓶颈提供了新的方法和思路。柑橘黄龙病菌基因组学的研究,不仅有助于探究黄龙病菌与寄主植物的互作关系,而且可为抗病栽培和病害早期检测提供重要的理论基础。本文结合当前柑橘黄龙病的研究概况、病原全基因组的测序策略及基因组的特征描述,重点综述基因组学在黄龙病致病机理、病原遗传多样性与分子检测等方面的研究进展。     

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.     

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。     

Development of a sensitive and reliable droplet digital PCR assay for the detection of ‘Candidatus Liberibacter asiaticus'

Citrus Huanglongbing(HLB, yellow shoot disease) is one of the most serious citrus diseases worldwide. To better improve the detection sensitivity, a droplet digital PCR(ddPCR) assay was developed for the rapid detection of ‘Candidatus Liberibacter asiaticus'(Las), the putative causal agent of HLB. The detection of sensitivity comparison using positive plasmid indicated that dd PCR was superior to quantitative PCR(qPCR) for detecting and quantifying Las at low concentrations. The Las detection of 40 field samples also showed that six of 13 asymptomatic samples(46.15%) with high Ct value(35) were positive by dd PCR. This methodology showed great potential for early HLB infection diagnosis.     

柚类果树叶片斑驳病的病原鉴定与检测

柚类是柑桔中的一类重要品种,在我国有广泛的分布、近年来,在福建等产区发现柚类病树日趋２增多、基病状为叶片斑驳、小果、植株矮化、暂称为叶片斑驳病。本有通过组织嫁接从病柚树传染到柚健苗和芦柑健苗。在电下观察,其病原功体的形态和细胞壁构造与柑桔黄龙病的病原十分要似。应用柑桔黄龙病病原ＤＮＡ的特异性引物进行ＰＣＲ试验产生的ＰＣＲ产物,其分子量与柑桔黄龙病病原ＤＮＡＰＣＲ产物的基本相同,为５６３ｂｐ;以及应     

Spectral difference analysis and airborne imaging classification for citrus greening infected trees

Citrus greening, also called Huanglongbing (HLB), became a devastating disease spread through citrus groves in Florida, since it was first found in 2005. Multispectral (MS) and hyperspectral (HS) airborne images of citrus groves in Florida were acquired to detect citrus greening infected trees in 2007 and 2010. Ground truthing including field and indoor spectral measurement, infection status along with GPS coordinates was conducted for both healthy and infected trees. Ground spectral measurements showed that healthy canopy had higher reflectance in the visible range, and lower reflectance in the near-infrared (NIR) range than HLB infected canopy. Red edge position (REP) also showed notable difference between healthy and HLB canopy. But the difference in the NIR range and REP were comparably more sensitive to the environment or the background noise. Accuracy for separating HLB and healthy samples reached more than 90% when a simple REP threshold method was implemented in the ground reflectance datasets, regardless of field or indoor measurement; but it did not work well with the HS images because of its low spatial resolution. Support vector machine (SVM) was able to provide a fast, easy and adoptable way to build a mask for tree canopy. High positioning error of the ground truth in the 2007 HS image led to validation accuracy of less than 50% for most of classification methods. In the 2010 image from Southern Gardens (SG) grove, with better ground truth records, higher classification accuracies (about 90% in training sets, more than 60% in validation sets for most of the methods) were achieved. Disease density maps were also generated from the classification results of each method; most of them were able to identify the severely infected areas. Simpler classification methods such as minimum distance (MinDist) and Mahalanobis distance (MahaDist) showed more stable and balanced detection accuracy between the training and validation sets in the 2010 images. Their similar infection trend with ground scouted maps showed a promising future to manage HLB disease with airborne spectral imaging.     

梅州市柑橘黄龙病的调查与综合防控措施

柑橘黄龙病(cirtus Huanglongbing,HLB)是细菌性病害,本文作者通过聚合酶链式反应(PCR)对梅州地区柑橘黄龙病发生情况进行调查,对梅州地区一共761个疑似黄龙病样本进行检测分析,并在此基础上提出梅州市柑橘黄龙病的综合防治策略,为梅州的柑橘产业发展提供参考。     

浙江省亚洲柑橘木虱黄龙病病原分布与消长规律

亚洲柑橘木虱Diaphorina citri Kuwayama是黄龙病病原Candidatus Liberibacter asiaticus(CLas)的主要传播媒介.为更好地探清亚洲柑橘木虱上的CLas阳性分布特性及其动态变化,提出适宜治虫防病的窗口期,特针对浙江省柑橘果园的分布特点,于2002—2019年采取定园定...     

柑橘黄龙病对砂糖橘果实品质的影响

为了综合评价黄龙病（Huanglongbing,HLB）对砂糖橘果实品质的影响,文章以健康砂糖橘植株和具有HLB症状植株上采的果实为试验材料,对两类果实的外观、内在、感官、果皮及果肉着色等品质指标进行量化比较。结果表明,与健康砂糖橘相比,感染HLB的果实单果重明显变小变轻,果形发生显著变化,可溶性固形物、维生素C含量、固酸比、可食率和出汁率显著减小,而可滴定酸含量及果实硬度明显增加;两类果实在果皮及果肉着色上差异显著。通过人为感官评价,感HLB果实的综合风味明显下降,果实萎缩,部分变僵硬,果实有异味,食用价值变低。HLB对砂糖橘的大小、果形、果实硬度、果皮果肉色泽亮度、感官及内质存在显著影响,造成砂糖橘品质严重下降,应高度重视砂糖橘感染黄龙病的防控。     

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.