亚洲柑桔木虱若虫在寄主上的转移和扩散研究

黄龙病是世界柑桔产业的一种毁灭性病害,亚洲柑桔木虱Diaphorina citri Kuwayama为黄龙病亚洲种和美洲种的媒介昆虫,其成虫和高龄若虫都能传病。本文研究亚洲柑桔木虱若虫在感染黄龙病及健康寄主上的转移和扩散规律,发现3龄及3龄以上若虫在柑桔植株垂直和水平位置的嫩梢之间转移现象十分明显,但在黄龙病植株上转移速度较健康植株慢,且嫩梢间的转移速度受虫密度的影响,密度越高转移速度越快。在有选择的条件下,水平位置转移过程中若虫更趋向黄龙病植株。此外,在健康寄主上的木虱若虫有明显向植株下部转移的现象,向下部转移的若虫个体数显著高于黄龙病寄主。本文明确了木虱若虫的扩散规律和寄主感染黄龙病对其转移和扩散产生的影响,为柑桔木虱和黄龙病防治提供参考。     

亚洲柑桔木虱体色变化规律的研究

亚洲柑桔木虱Diaphorina citri Kuwayama为黄龙病的媒介昆虫。本文研究了其雌雄成虫在刚羽化、羽化后3 h、交配盛期和产卵盛期的腹部和前翅颜色变化规律,明确了每个时期的主要体色,同时探讨了不同季节、虫口密度和寄主条件对体色变化的影响。结果表明,雌雄木虱羽化后前翅颜色均由白色→黑色→红褐色渐变,木虱雌成虫腹部颜色随时间推移由黄绿色→灰白色→蓝色→橙色渐变,雄虫由黄绿色→灰黑色→橙色渐变。季节、寄主和虫口密度的变化对雌虫腹部颜色构成有一定的影响,但对雄虫基本上没有影响。不同条件下雌虫腹部的主要颜色为蓝色或橙色,雄虫均以橙色为主,其次为灰黑色,表明木虱成虫多处于交配期或产卵期。木虱5龄若虫的体色变化也较大。多数条件下木虱的性比接近于1∶1,仅春秋季雌性比例略低于雄性。通过观察木虱的体色变化规律可以初步判断其羽化情况和性别,估计羽化期、交配期和产卵盛期,作为木虱生物学特性研究和预测预报的参考。     

柑桔木虱及其防治

柑砧术虱是柑桔新梢期重要害虫,刺吸为害。福建福州在柑桔上一年发生8代,在月桔、九里香上一年约发生10代左右,世代重叠,以成虫在柑桔叶背群集越冬,无完全滞育。4—5月气温22．3℃世代历期42．5天．6-7月27．2℃为24．4天．8月28．1℃为23．2天。10-12月19.6℃为52．9天。成虫寿命长,约历1个月半,越冬代长达半年。田间种群消长与柑桔芽梢抽发期相对应．一年中虫口数量出现3个高峰期,第一峰期3-4月为柑桔春梢主要抽发期,第二峰期5-6月为夏梢主要抽发期,第三峰期8～9月为秋梢主要抽发期,其中以秋梢期虫口数量最大．秋芽受害最重,次为春梢期。本还对柑桔术虱天敌进行调查,并对柑桔术虱卵、若虫和成虫进行药剂防治试验。     

光、温湿度对柑桔木虱发育、繁殖与存活的影响

本研究结果表明,在光照强度为11000lx以下(6小时光照时为15000lx)、每天光照时间在18小时以内,光照强度越大,光照时间越长,柑桔木虱雌成虫的产卵前期越短,产卵量越大,死亡率越低。 温度为15—34℃,相对湿度为43—92％范围内,温湿度对柑桔木虱卵的孵化率影响较小。柑桔木虱若虫在高温(34℃)、高湿(85％,92％)下死亡率高;适温(20—30℃)、低湿(43—75％)下死亡率低。湿度对柑桔木虱卵及若虫发育历期影响不大。在温度为15—34℃范围内,温度与卵及若虫的发育历期呈抛物线关系。柑桔木虱卵及1—5龄若虫的发育起点温度分别是:9.41℃、8.30℃、9.72℃、8.92℃、9.61℃及9.07℃,有效积温分别是:60.03日度、39.78日度、26.82日度,33.23日度及74.49日度。     

中国部分地区柑桔木虱体内感染Wolbachia的检测及其系统发育分析

Wolbachia是一种广泛分布于节肢动物体内的共生细菌, 该共生菌经寄主卵的细胞质传播并参与多种寄主生殖活动调控, 对节肢动物的物种进化有着重要意义并可能应用于害虫的生物防治。本研究以柑桔黄龙病（Huanglongbing, HLB）的主要传播媒介——柑桔木虱Diaphorina citri为研究对象, 通过Wolbachia的16S rDNA、 细胞分裂基因（ftsZ）、 表面蛋白基因（wsp）的特异引物对来自于中国广西北海、 广西柳州、 广东深圳、 广东阳春、 福建福州5个地区的柑桔木虱进行PCR扩增, 并对扩增产物进行克隆测序, 与GenBank中相关序列进行比对分析, 建立了柑桔木虱共生Wolbachia的系统发育树。结果显示上述5个地区的柑桔木虱均存在Wolbachia感染, 通过Wolbachia的16S rDNA, ftsZ基因和wsp基因DNA序列的系统发育分析表明, 5个地区的柑桔木虱体内的Wolbachia均属于B组; 基于wsp基因的系统发育分析进一步表明5个地区的亚洲柑桔木虱体内的Wolbachia属于B组的Con亚组, 且不同地区柑桔木虱体内的Wolbachia的16S rDNA, ftsZ基因和wsp基因序列差异不明显。研究结果为认识柑桔黄龙病传播媒介昆虫的进化及其综合防治提供了重要的参考依据。     

亚洲柑橘木虱和柚喀木虱过冷却点和体液结冰点测定

亚洲柑橘木虱Diaphorina citri Kuwayama和柚喀木虱Cacopsylla citrisuga YangLi均为柑桔上的重要害虫,前者为黄龙病的主要媒介昆虫,后者也已证实为黄龙病菌的携带者。本文对这两种木虱各龄若虫和成虫的过冷却点和体液结冰点进行了测定,结果表明,柑橘木虱过冷却点和冰点均以1龄若虫最低,平均值分别为-26.32℃和-26.22℃;成虫的过冷却点和冰点最高,分别为-19.60℃和-18.71℃;其它虫态过冷却点和冰点由低到高依次为2龄若虫、4龄若虫、5龄若虫、3龄若虫。柚喀木虱过冷却点和冰点也均以1龄若虫最低,平均值分别为-25.30℃和-25.08℃;5龄若虫的过冷却点和冰点最高,平均分别为-19.50℃和-17.69℃;其它虫态过冷却点和冰点由低到高依次为2龄若虫、3龄若虫、4龄若虫、成虫。两种木虱的比较结果表明,各发育阶段的表现不一致,柚喀木虱的3龄若虫和成虫过冷却点显著低于柑桔木虱,而4龄、5龄若虫则显著高于柑桔木虱;3龄若虫体液结冰点也显著低于柑桔木虱,但1龄、4龄却显著高于柑桔木虱。柑桔木虱雌雄成虫的过冷却点和体液结冰点差异不显著。研究结果表明两种木虱的过冷却点、体液结冰点都较低,因而可能具有较强的耐寒能力。     

亚洲柑橘木虱内生殖系统形态变化规律研究

亚洲柑橘木虱Diaphorina citri Kuwayama是柑橘黄龙病的传播媒介。本文利用Ste REO Discovery V20体视显微镜对亚洲柑橘木虱成虫内生殖系统进行解剖观察,并探索了亚洲柑橘木虱雌雄成虫内生殖系统的形态变化规律。结果表明:雄虫内生殖系统由1对精巢、1对输精管、1个精泵、1个射精管、1对附腺和1个贮精囊组成。雌虫内生殖系统由1对卵巢、1对侧输卵管、1个中输卵管、2个附腺、1个黏腺和1个受精囊组成。交配前期和交配期的雄虫精巢饱满,精巢在交配后期明显萎缩。交配期和交配后期的雄虫贮精囊都明显大于交配前期的贮精囊。雌虫受精囊在交配前期、交配期和交配后期依次增大,交配前期的受精囊呈不饱满状态,交配期和交配后期受精囊呈饱满状态,内有白色精包。交配后期的雌虫卵巢内有大量成熟的橙黄色卵。     

烟粉虱对寄主的选择行为

烟粉虱Bemisia tabaci(Gennadius)成虫对寄主植物选择行为的实验结果表明,烟粉虱是通过植物广谱的绿色气体识别植物材料,并进一步受到绿色的诱导。通过烟粉虱在适宜与非适宜寄主植物上滞留时间的差异,可以看出烟粉虱是通过口针刺探的寄主评估过程逐渐实现寄主偏好性的,存在一个短的时滞。综合来看,烟粉虱的寄主选择是通过嗅觉、视觉和味觉共同参与的一个决策过程,以寄主植物的气味、颜色和质量为线索,逐步定位到适宜的寄主植物上,因此在寄主选择过程中存在很大的可塑性。     

柑橘木虱对12种寄主植物嫩梢及其挥发物成分的趋性反应

为了发现对柑橘木虱Diaphorina citri Kuwayama具有引诱活性的寄主植物嫩梢挥发物,采用Y型嗅觉仪在室内测定柑橘木虱对12种寄主植物嫩梢及其挥发物成分的趋性反应,采用固相微萃取技术(SPME)和气-质联用分析技术(GC-MS)分析鉴定嫩梢挥发物的共同成分.结果表明,柑橘木虱对九里香Murraya ex...     

寄主钾营养对烟粉虱发育、存活和寄主选择的影响

为了探讨寄主钾营养对烟粉虱发育、存活和寄主选择的影响,设置了K0（0 mg/L）、K30（30 mg/L）、K60（60 mg/L）、K120（120 mg/L）和K240（240 mg/L）5种不同钾浓度,研究了烟粉虱在不同处理黄瓜上的发育历期、存活率、体形以及成虫的寄主选择。结果表明：在温度（26±1）℃,相对湿度80%±5%的条件下,取食不同钾营养水平黄瓜的烟粉虱在发育历期、存活率、体形上均有差异,其中在K240处理黄瓜上烟粉虱若虫发育最慢,从卵到成虫的时间最长,为21.4天,而在K60处理黄瓜上最快,为18.3天。在不同钾浓度处理寄主上从卵到成虫存活率以K30处理黄瓜上最高（84.7%）,其次依次为K60（83.8%）、K120（76.2%）、K0（71.4%）和K240（64.8%）。在体形上以K30处理黄瓜上最大,K240处理黄瓜上最小。寄主选择性试验结果表明,在温室条件下,烟粉虱成虫喜欢在K60处理黄瓜上取食和产卵。这些结果提示在一定范围内可以通过调节寄主钾营养状况来调控烟粉虱种群。     

亚致死浓度吡虫啉和毒死蜱对柑橘木虱成虫组织蛋白酶基因表达的影响

昆虫组织蛋白酶属于多功能酶系,在昆虫生命活动中具有重要作用,但昆虫组织蛋白酶对化学农药的响应却鲜有报道。吡虫啉和毒死蜱是目前防治柑橘木虱Diaphorina citri常用的化学药剂。本研究旨在探究吡虫啉和毒死蜱亚致死浓度处理对柑橘木虱成虫组织蛋白酶基因表达的影响。本研究基于柑橘木虱的转录组数据库,同时结合NCBI数据库,通过Blast比对鉴定得到8个柑橘木虱组织蛋白酶基因,序列分析得出其均属于半胱氨酸蛋白酶家族且含有保守的组氨酸活性位点。研究采用浸叶法处理得到经吡虫啉和毒死蜱处理24 h对柑橘木虱成虫的致死中浓度LC50分别为97.88 mg/L和47.94 mg/L。采用荧光定量PCR（qPCR）分析这两种药剂亚致死浓度（LC20和LC50）胁迫下柑橘木虱成虫组织蛋白酶基因表达水平变化。结果表明吡虫啉胁迫显著下调柑橘木虱DcCath-B,DcCath-F,DcCath-L和DcCath-L1基因表达量;毒死蜱LC20浓度胁迫显著下调柑橘木虱DcCath-B,DcCath-L和DcCath-W基因表达量,LC50浓度胁迫下仅DcCath-B表达量显著下调,其他基因表达无明显差异。以上结果表明吡虫啉和毒死蜱亚致死浓度胁迫下,柑橘木虱成虫下调表达体内部分组织蛋白酶基因以响应此逆境。本研究为探索亚致死浓度吡虫啉和毒死蜱对柑橘木虱的毒理机制提供了一定的理论依据。     

植物挥发物对柑橘木虱趋避机制的研究进展

柑橘木虱Diaphorina citri为半翅目Hemiptera木虱科Psyllidae,主要危害芸香科植物,是柑橘黄龙病（Citrus Huanglongbing, HLB）的主要昆虫传播媒介。HLB在柑橘产业中破坏性极大,柑橘木虱的及时防控可有效降低HLB传播风险。目前,柑橘木虱对化学药剂已产生一定的抗药性,而柑橘木虱在植物挥发物作用下的行为反应可为其绿色防控提供一定的参考。本文就往年研究中不同植物挥发物对柑橘木虱的引诱、驱避作用以及柑橘植株、HLB、柑橘木虱三者间通过植株挥发物产生的互作进行论述,旨在为柑橘木虱的科学高效绿色防控提供参考。     

Secondary hosts of the Asian citrus psyllid,Diaphorina citri Kuwayama: Survivorship and preference

The Asian citrus psyllid, Diaphorina citri Kuwayama, is the most economically important and difficult to manage citrus pest in Florida due to its role as vector of huanglongbing or citrus greening disease. Spread of the disease is a function of dispersal patterns and thus influenced by availability of suitable hosts. Young leaves of citrus or related plants are required for reproduction, but in their absence, secondary hosts may provide needed resources to enhance survival. Therefore, survivorship on and preference for three abundant weed species was investigated. The suitability of potential secondary host plants Bidens alba (L.), DC, Ludwigia octovalvis (Jacqu.) P. H. Raven, and Eupatorium capillifolium (Lam.) Small was compared to a reproductive host, Murraya paniculata (L.) Jack, in no‐choice tests by assessing survivorship of D. citri adults confined to these plants in cages. Preference was evaluated by choice tests where D. citri adults were released into cages containing all three secondary hosts alone or with M. paniculata. Both B. alba and E. capillifolium increased D. citri survivorship by twofold compared to starvation conditions with only water available. Choice trials revealed no difference in initial selection between true and secondary hosts; however, the true host was favoured over time. This result suggested that hosts were selected initially by sight, and only later by taste and/or smell. While secondary hosts are unable to support reproduction or long‐term survival, these findings establish the ability of D. citri to use secondary hosts that are ubiquitous in Florida citrus groves as temporary reservoirs for food and moisture when ideal host conditions are scarce or absent.     

Biochemical analysis and pathogenicity of entomopathogenic fungi to Diaphorina citri Kuwayama (Hemiptera: Liviidae)

Asiatic citrus psylla, Diaphorina citri, Kuwayama (Hemiptera: Liviidae) is an economic pest of citrus groves and a vector of the bacterium, Candidatus Liberibacter spp., one of the causative agents of citrus greening. In order to estimate the infectivity of six different isolates of Beauveria bassiana, Metarhizium anisopliae and Isaria fumosorosea, fungal bioassay was performed on the adults of D. citri. Adults of D. citri were treated individually with 1 × 10⁵, 1 × 10⁶, 1 × 10⁷, 1 × 10⁸, 2 × 10⁸ spores/mL fungal concentrations by the immersion method. Subsequent to fungal bioassay, treated D. citri were used to determine the levels of esterase and glutathione S‐transferases (GST) enzymes over a period of 3–7 days. The mortality results suggest that I. fumosorosea isolates (If‐02) caused 82.2% mortality on the seventh day of treatment. However, B. bassiana isolate (Bb‐08) with lowest LC₅₀ (1.4 × 10⁷ spores/mL) proved to be highest potential isolate against D. citri. Biochemical determination of esterase and GST activity assay showed significant differences in activities after infection of fungi. Significantly high activity of esterase was observed by Bb‐01 (27.0 unit per mg protein) on the seventh day, while Ma‐11.1 and If‐2.3 (16.9 and 36.3 unit per mg protein) on the third day post treatment. However, maximum GST's activity was showen by isolates Bb‐08,Ma‐M₂ and If‐2.3 (37.6, 1.40 and 10.9 unit per mg protein) on the third day. The current investigation will help to explore the relations between the insect defense system and entomopathogenic fungi. Moreover, the determination of enzymatic activities will be useful for selecting the most pathogenic isolates.     

Twelve polymorphic microsatellite loci from the Asian citrus psyllid,Diaphorina citri Kuwayama,the vector for citrus greening disease,huanglongbing

Twelve polymorphic microsatellite markers were developed from microsatellite‐enriched DNA libraries and mined from an expressed sequence tags library of Diaphorina citri, the vector of the citrus greening disease (huanglongbing). Analysis of 288 individuals from Florida, Texas, and Brazil showed that allelic diversity ranged from three to eight alleles per locus and observed and expected heterozygosities ranged from 0.014 to 0.569 and from 0.052 to 0.653, respectively. These variable microsatellite loci can provide means for assessing overall genetic variation and migration patterns for this agriculturally important pest species. This information can be used to aid in developing successful management strategies.     

Huanglongbing pathogen Candidatus Liberibacter asiaticus exploits the energy metabolism and host defence responses of its vector Diaphorina citri

Asian citrus psyllid Diaphorina citri is the vector of the citrus Huanglongbing (HLB) associated bacterial agent ‘Candidatus Liberibacter asiaticus’ (CLas). The molecular interactions between CLas and D. citri remain unclear. In the present study, protein profiles of mitochondrial, microsomal and cytosolic fractions from uninfected and CLas‐infected adult D. citri are investigated using two‐dimensional gel electrophoresis. The comparative analysis reveals a total of 18, 24 and 20 protein spots that are unique or differentially expressed in mitochondrial, microsomal and cytosolic proteins fractions respectively. These proteins are successfully identified by mass spectrometry. Among the 62 identified proteins, 30 are up‐regulated, whereas 32 are down‐regulated. These proteins include important components in energy metabolism such as ATP synthase, ATPase, ATP/ADP carrier protein, etc.; host stress responses such as heat shock proteins; host detoxification processes (i.e., cytochrome P450 and glutathione S‐transferase); and the cytoskeleton (such as actin, tubulin, myosin and tropomyosin). These data suggest that, after CLas infection, several proteins of D. citri, especially energy metabolism and protein biosynthesis, are altered, and extensive host defence responses are induced. In conclusion, the present study reports proteomic information that is helpful in understanding the vector–pathogen relationship between CLas and D. citri, and could be used to identify potential targets for limiting the spread of CLas, as well as to provide new insights into HLB management.     

亚洲柑橘木虱与柑橘黄龙病菌互作的研究进展

亚洲柑橘木虱Diaphorina citri Kuwayama作为柑橘产业重要病害柑橘黄龙病的主要传播媒介,已经成为重点防治对象。该害虫与黄龙病之间的互作一直是相关研究的热点,本文就近年来该领域的研究进展做了一个总结,从亚洲柑橘木虱的获菌与传病机制、病原菌与柑橘木虱之间的互作以及病原菌感染寄主植物后对木虱的影响等方面进行了综述。期望为深入开展黄龙病相关研究、寻找防控新途径提供依据。