伴生细菌在入侵种桉树枝瘿姬小蜂克服桉树抗性中的作用

【目的】桉树枝瘿姬小蜂是我国近年来发现的一种主要危害桉属树种的外来有害生物。本研究旨在通过探究中国桉树枝瘿姬小蜂主要伴生细菌在桉树枝瘿姬小蜂成功定殖中的作用。【方法】测定不同抗性品系桉树的次生代谢物质黄酮和单宁的含量以及易感品系桉树在枝瘿姬小蜂危害前后的含量变化。通过体外抑菌和化学物质降解实验,探究桉树枝瘿姬小蜂主要伴生细菌对抗虫性物质黄酮和单宁的耐受性及降解能力。【结果】抗性品系桉树的黄酮和单宁的含量明显高于易感品系,易感品系在桉树枝瘿姬小蜂危害后黄酮和单宁的含量显著提高;高浓度的黄酮和单宁会抑制桉树枝瘿姬小蜂伴生细菌的生长,在中低浓度黄酮和单宁的条件下,主要伴生细菌能够适应,并继续繁殖;桉树枝瘿姬小蜂伴生细菌具有一定的降解黄酮和单宁的能力,其中细菌Staphylococcus cohnii降解黄酮的能力比Pseudomonas geniculate稍强,而Bacillus wiedannii、Serratia macescens对单宁具有较强的降解能力。【结论】桉树枝瘿姬小蜂侵染桉树后,可以诱导桉树产生抗性,桉树产生大量的次生代谢物质来抵御桉树枝瘿姬小蜂的危害,而桉树枝瘿姬小蜂部分伴生细菌可降解桉树次生代谢物质来帮助小蜂克服植物抗性完成定殖。     

寄主植物对桉树枝瘿姬小蜂生物学特性的影响

桉树枝瘿姬小蜂Leptocybe invasa Fisher&La Salle是桉属(Eucalyptus)植物的重要枝叶害虫。自2007年在我国发现以来,对广西、海南及广东地区的桉树种植已造成严重危害。本文在野外调查的基础上,研究了海南和广东地区共5种桉树上桉树枝瘿姬小蜂的虫体大小、寿命及怀卵量。结果表明,桉树品种能够显著影响桉树枝瘿姬小蜂的虫体大小;用40%蜂蜜水进行补充营养时,不同寄主植物上桉树枝瘿姬小蜂雌成虫的寿命差异显著,而桉树枝瘿姬小蜂雄虫在不同寄主间无显著性差异;不同寄主植物上桉树枝瘿姬小蜂雌成虫的怀卵量不同,并且随着日龄的延长,其怀卵量呈现上升的趋势。     

2种不同寄主对桉树枝瘿姬小蜂体内细菌多样性的影响

[目的]桉树枝瘿姬小蜂是一种林业的重要入侵害虫,为了探明桉树枝瘿姬小蜂的寄主适应机制,研究2种不同寄主植物对桉树枝瘿姬小蜂体内细菌群落组成及多样性的影响.[方法]以分别取食巨园桉DH 201-2和窿缘桉的桉树枝瘿姬小蜂雌性成虫为材料,采用Illumina HiSeq高通量测序技术对桉树枝瘿姬小蜂体内细菌16S rDNA...     

桉树不同无性系叶片内含物变化与枝瘿姬小蜂抗性的关系

通过对24个桉树无性系在桉树枝瘿姬小蜂（Leptocybe invasa Fisher& La Salle）生长周期连续5个月危害情况及其营养物质含量的变化进行研究,根据受害等级将24个桉树无性系分为高感、中抗和高抗三种类型。三种抗性类型在未受危害前,可溶性蛋白含量和可溶性糖含量均无显著差异;在受到危害后,高感、中抗无性系的可溶性蛋白含量、可溶性糖含量均出现一定增长。与对照相比,高感无性系内含物含量随桉树枝瘿姬小蜂危害加剧显著增加。内含物含量增长排序均表现为：高感类型>中抗类型>高抗类型,说明桉树叶片内含物含量的增加是桉树对病虫害防御的重要生理反应。     

桉树枝瘿姬小蜂幼虫林间化学防治试验

选用70%吡虫啉水分散粒剂、40%毒死蜱乳油、25%噻虫嗪水分散粒剂以及这3种农药的混配剂,分别稀释成5、10、20倍液,采用注干法对桉树枝瘿姬小蜂幼虫进行林间药效试验。试验表明,施药30 d后,相同浓度的不同药剂对桉树枝瘿姬小蜂幼虫的校正死亡率则存在极显著差异。混配剂5倍液的防治效果最好,在30 d内对桉树枝瘿姬小蜂幼虫的校正死亡率为81.63%。施药后5～10 d,桉树枝瘿姬小蜂幼虫的死亡率增长速度最快;施药10 d后,死亡率的增长速度明显变缓。同一种药剂的浓度越高,对桉树枝瘿姬小蜂幼虫的致死时间越短。     

桉树枝瘿姬小蜂产卵行为研究

桉树枝瘿姬小蜂Leptocybe invasa Fisher&La Salle喜好在桉属(Eucalyptus)植物的新生部位产卵。本研究以尾赤桉为寄主研究其产卵模式发现,当天羽化且没有产卵经验的雌成虫在经过寄主识别(116.87 s)和寄主评估(208.27 s)后即进行产卵循环(平均5.87次),每个产卵循环产2粒卵,且搜索行为的发生一般伴随产卵位置的变换。在被观察的15头雌成虫中,平均每头产卵13.60粒,每次产卵持续17.31 s。在产卵过程中,其刺探行为所占总时间的比例和发生频次最大,而静息行为最小。在不同寄主植物间,桉树枝瘿姬小蜂喜好在尾赤桉的叶柄和叶脉处产卵,且2个部位间无显著性差异,而在尾叶桉、巨尾桉、隆缘桉和小果灰桉上,其喜好在叶柄处产卵且显著大于其它2个部位。     

生态因子对赣南桉树枝瘿姬小蜂雄成虫比率的影响

桉树枝瘿姬小蜂Leptocybe invasa具有偏雌性的特点,了解桉树枝瘿姬小蜂的性比规律,有利于监测和掌控该蜂的种群动态,便于及时有效防控该蜂.2017年至2019年,采用在桉树Eucalyptus林中悬挂黄色粘虫板的方法,监测、观察桉树枝瘿姬小蜂成虫的发生动态,通过分析该蜂雄成虫比率及其影响因子,以弄清生态因子对...     

桉树枝瘿姬小蜂连续世代种群生命表

桉树枝瘿姬小蜂在广州地区一年发生4-5代,且世代重叠,其越冬期为11月中旬到翌年3月底,越冬虫态为大龄幼虫和蛹.在广州自然条件下,通过人工接种和自然接种相结合的方法,桉树枝瘿姬小蜂在2010年3月至2011年3月间共完成4个世代.利用生命表的方法分析桉树枝瘿姬小蜂连续世代可知,第1代的净增值率R0最大,而第2代的最小;第3代的内禀增长率rm和周限增长率λ最大,其世代平均历期T最小;第4代的rm和λ最小,其T最大.这说明第3代桉树枝瘿姬小蜂的种群增长能力最大,对桉树的危害最强.该代害虫的发生时间为7-9月,因此在防治上要注意该阶段的防控和监测.     

桉树枝瘿姬小蜂危害对桉树次生代谢产物含量的影响

通过对24个桉树品系遭受桉树枝瘿姬小蜂危害前后次生物质含量单宁和类黄酮含量动态变化研究,结果表明桉树品系遭受枝瘿姬小蜂危害后,叶片内类黄酮含量明显上升,高感品系类黄酮含量和增长变幅明显低于其他抗性类型的品系;而桉树叶片内单宁含量的动态变化规律表现为：高抗品系单宁含量明显高于高感品系,但随着进入小蜂危害期,其叶片内单宁含量出现一定程度下降,随后升高。次生物质的动态变化在不同抗性品系间差异显著,表明桉树叶片内单宁和类黄酮含量与抗枝瘿姬小蜂有显著关系,可作为检测桉树对枝瘿姬小蜂抗性的指标。     

桉树枝瘿姬小蜂胚胎发育及胚后发育

【目的】对桉属树木的重要害虫桉树枝瘿姬小蜂Leptocybe invasa胚胎发育及胚后发育生物学特征进行详细调查,以期深入了解该害虫的生物学特征及发生为害规律。【方法】对桉树枝瘿姬小蜂雌性生殖系统、卵、虫瘿进行解剖,观察其胚胎发育和胚后发育过程,测量其各发育阶段的虫瘿、虫室和幼体的体积及薄壁细胞层的厚度。【结果】桉树枝瘿姬小蜂完成胚胎发育约需138 h,由3个阶段组成：产卵后0-24 h为营养物质聚集期;36-84 h为胚胎期;96-138 h为孵化期。桉树枝瘿姬小蜂在虫瘿内完成发育,其发育过程与虫瘿发育过程保持一致。虫瘿结构由3个基本层组成,由外至内分别为表皮层、中间层和薄壁细胞层。桉树枝瘿姬小蜂幼虫体积增长速率随发育时间呈现单峰型,在30 d时增长速率达到最大;虫瘿体积的增长速率呈抛物线状,并且虫瘿内薄壁细胞层厚度随发育时间也呈现抛物线状,其前期增长较快,后期下降变缓。【结论】桉树枝瘿姬小蜂发育至25 d时处于取食量较小的低龄阶段,而虫瘿内薄壁细胞层(食物)厚度已达到最大,为幼虫体积快速增大阶段作好准备。     

桉树枝瘿姬小蜂虫瘿解剖特征与寄主叶片生理指标的变化

采用人工接蜂、桉树组织石蜡切片和生化组分测定等方法研究了桉树受桉树枝瘿姬小蜂诱导后的生理生化响应机制.结果表明:DH201-2叶柄角质层厚度和油囊数量均较GL-UG9的厚、多,且差异均达到极显著水平(P角质层厚度=0.00001 ＜0.01,P油囊数量=0.00209＜0.01),但是GL-UG9叶柄表皮细胞厚度和维管束数量均较DH201-2厚、多,差异分别达到显著水平(P=0.015215＜0.05)和极显著水平(P=0.002375＜0.01),DH201-2茎皮层薄壁细胞厚度、油囊直径、维管束数量较GL-UG9厚、多,差异分别为显著(P皮层薄壁细胞厚度=0.04071 ＜0.05)、极显著(P油囊直径=000016＜0.05)、极显著(P维管束数量=0.00000＜0.01).而GLUG9茎角质层和表皮细胞厚度均较DH201-2厚,差异达到了极显著水平(P角质层厚度=0.00167,P表皮细胞厚度=0.00000＜0.01);桉树枝瘿姬小蜂虫瘿为组织瘿,由内到外可以分为:营养组织层、薄/厚壁组织层、维管束层、皮层、表皮等结构;接种桉树枝瘿姬小蜂成虫后,桉树叶片可溶性总糖、游离氨基酸、叶绿素含量均升高,差异达到了极显著水平(P＜0.01),但是蛋白质、pH值、类黄酮、总酚的变化没有达到显著水平(P＞0.05);吲哚乙酸氧化酶、过氧化物、过氧化氢酶活力均有不同程度的升高.研究显示桉树枝瘿姬小蜂入侵诱导了桉树一系列的生理生化变化,实验结果为抗桉树枝瘿姬小蜂桉树无性系的选育技术体系构建提供了理论基础.     

Assessment performance of Eucalyptus clones attacked by the recent invasion of Leptocybe invasa (Hymenoptera: Eulophidae): Implications to invasion pest management

Eucalyptus gall wasp, Leptocybe invasa is one of the most important invasive pests of Eucalyptus (Myrtaceae) and has been reported in new cultivation areas worldwide. This pest causes severe damage to susceptible plants of the genus Eucalyptus, where it induces galls in the midribs and petioles of young leaves and the internodes of branch and apices. In this work, we evaluated the response of Eucalyptus clones (E. urophylla (Urophylla) and two clone hybrids, one of the E. urophylla × E. camaldulensis (Urocam) and other of E. urophylla × E. grandis (Urograndis) crossings.) submitted to L. invasa collected from areas where L. invasa was recently reported. Our results show that the eucalypt materials cultivated in the areas where L. invasa were reported sensitive to L. invasa attacks. However, the wasp causes fewer galls in the Urocam clone. Thus, the Urocam clone can be preferred for use to reduce population growth of L. invasa in Brazilian savannah areas.     

桉树杂交种对桉树枝瘿姬小蜂的抗性变异分析

植物种间杂交是一种普遍自然现象,杂交往往造成植物表型及生理变异,从而改变杂种抗虫性。与亲本种相比,杂种抗虫性可能增强或减弱,也有可能处于与亲本相似水平。初生、次生代谢物的质变与量变是引起杂种抗虫性变异的重要原因。近年来,桉树杂交育种已在世界范围内广泛应用并取得了显著成效,桉树杂交种间抗虫性表现参差不齐,因此,桉树是研究杂交种抗虫性变异机制的理想材料。以2个桉树杂交种巨细桉DH201-2、巨尾桉G9及桉树重要害虫桉树枝瘿姬小蜂为研究对象,比较了2个杂交种与其纯亲本种[(巨桉×细叶桉),(巨桉×尾叶桉)]间的抗虫性差异;同时,综合比较了品系间叶片性状(叶片厚度、含水率、比叶面积)、初生化合物(C、N、可溶性糖、可溶性蛋白)及次生化合物(总酚、单宁)差异,以研究桉树杂交种抗虫性变异的理化机制。结果表明:DH201-2感染桉树枝瘿姬小蜂的虫瘿数目显著高于其双亲本种,而G9上虫瘿数目显著低于其双亲本种。DH201-2与G9的叶片厚度与巨桉相近,而显著薄于另一亲本种。DH201-2叶片含水率显著高于细叶桉、与巨桉相近;G9叶片含水率则显著低于其双亲本种。相似的是,DH201-2和G9的比叶面积均显著高于其双亲本种。初生化合物方面,DH201-2叶片可溶性糖和可溶性蛋白含量均显著高于其亲本种,N含量则仅高于细叶桉;而G9叶片可溶性蛋白含量虽高于其双亲本种,可溶性糖含量则无显著差异,N含量显著低于其双亲本种。次生化合物方面,DH201-2叶片总酚和单宁含量显著低于其双亲本种,而G9则显著高于其双亲本种。因此,与其亲本种相比,DH201-2感虫性增加,而G9抗虫性增加;与桉树枝瘿姬小蜂发育相关的营养指标(如含水率、可溶性糖、N含量)及次生防御物质(如总酚、缩合单宁)在桉树杂交种组织内的含量差异影响了桉树杂交种对桉树枝瘿姬小蜂的抗性。在全球推行桉树杂交育种且桉树害虫数量逐年增加的大背景下,应加强对桉树杂交种抗虫性机制研究,为选育高抗品系及桉树产业可持续发展提供理论指导。     

桉树及其杂交种叶片形态的遗传变异特征

为了解桉树重要性状在杂交种亲本、子代间的遗传变异规律,以尾叶桉(Eucalyptus urophylla)为母本、粗皮桉(E. pellita)和细叶桉(E. tereticornis)为父本、8个全同胞杂交子代为材料,对亲本和子代的叶片形态性状进行了分析。结果表明,亲缘关系近的尾叶桉和粗皮桉的叶片性状大多无显著差异;但他们与亲缘关系远的细叶桉间的差异显著;尾粗桉与父母本间的性状无显著差异或仅与母本差异显著,属偏父本型;尾细桉与双亲间的多数性状都差异显著,且多偏向于母本。在树种内的家系间或个体间,叶片数量性状差异显著,而质量性状仅叶面革质厚度、叶面朝向差异显著;杂交种家系部分性状有超亲现象,且尾细桉家系超亲频率显著高于尾粗桉。叶长、叶长宽比、形状因子、叶色、叶面革质是区别叶片形态特征的决定性状。     

内生真菌对羽茅生长及光合特性的影响

通过温室栽培实验,以感染两种内生真菌(Neotyphodium sibiricum和Neotyphodium gansuence)和未感染内生真菌的羽茅(Achnatherum sibiricum)为实验材料,分析感染不同种内生真菌对宿主植物的生长及光合特性的影响。结果表明,感染两种内生真菌羽茅的株高和CO₂补偿点显著低于未染菌的羽茅,而染菌羽茅的蒸腾速率和气孔导度显著高于未染菌羽茅,但对于感染不同种内生真菌的羽茅,无论是分蘖数与生物量的积累还是光合生理值之间均无显著差异。     

巨桉根系分解对菊苣生长及光合特性的影响

巨桉是我国退耕还林过程中采用的重要速生树种之一,被广泛用于人工造林。采用盆栽试验,研究了巨桉根系分解初期对菊苣幼苗生长和光合生理特性的影响。试验设置A1(50 g/盆)、A2(100 g/盆)和对照(CK)3个根系水平,将各处理的根系分别与10 kg土壤混合后装盆,播种菊苣。待A2处理植株的第3片真叶完全展开后测定菊苣光合生理指标及相关生长指标。结果表明:在巨桉根系分解初期,明显抑制了菊苣高生长、根生长、生物量积累、叶面积扩展及光合色素的合成,且随着根系添加量的增加抑制作用加大;菊苣叶片胞间CO2浓度(Ci)增加,而净光合速率(Pn)、气孔导度(Gs)和蒸腾速率(Tr)均显著低于对照;随土壤中根系含量的增加,除CO2补偿点(CCP)呈增加趋势外,其他光响应和CO2响应的特征参数均呈明显的下降趋势,并与对照差异显著;各生长指标除与胞间CO2浓度呈现出负相关外,与其它光合特征参数、光合色素以及相应的响应曲线参数之间均呈现出显著或极显著的正相关关系;通过GC-MS检测表明,巨桉根系中含有2,6-二叔丁基对甲酚、N-甲基苯乙胺等多种具有化感潜力化学物质,在其分解过程中,这些化感物质逐步释放并作用于受体植物,抑制其光合色素合成和光合作用,降低其环境适应能力,从而抑制菊苣的生长。     

Inhibitory effects of leachate from Eupatorium adenophorum on germination and growth of Amaranthus retroflexus and Chenopodium glaucum

Amaranthus retroflexus L. and Chenopodium glaucum L. are two widely distributed destructive weeds. Their strong adaptability and massive seed production make them the hardest weeds to deal with. This present study intended to investigate the effect of leachate from Eupatorium adenophorum on the growth of these weeds and explore the potential to develop an environmental friendly strategy to use the leachate to control the weeds. Seeds of A. retroflexus L. and C. glaucum L. were soaked in solutions containing 0%, 0.6%, 1.25%, 2.5%, and 5% leachate from E. adenophorum leaves. A. retroflexus and C. glaucum seedlings grown in pots were sprayed with leachate solutions in the same concentration range. The effects of these leachate solutions on membrane permeability and germination of seeds, and growth and physiological characteristics of the seedlings were investigated. The highest concentration of leachate (5%) caused significant damage to the cell membrane of seeds of both weed species, whereas lower concentrations (0.6%) promoted repair of the membrane system, as reflected by higher and lower than control in relative conductivity (RC), respectively. Different concentrations of leachate showed distinct allelopathic inhibitory effects on the two weed species; lower concentrations showed weak inhibitory or even positive effects, whereas higher concentrations showed stronger inhibitory effects. Higher concentrations of leachate (2.5% and 5%) delayed germination and significantly decreased the emergence rate of the seeds, survival rate, and dry matter accumulation of the seedlings. When treated by 5% leachate, the emergence date of A. retroflexus was delayed by 3.6 d, emergence rate of the seeds and survival rate was 69.1% and 70.6% of the control, respectively, seedling dry matter was 48.6% less than the control; In the case of C. glaucum, the emergence date was delayed by 2.7 d, emergence rate of the seeds and survival rate was 45.1% and 58.6% of the control, respectively, seedling dry matter was 44.7% less than the control. There were significant interactions among the different concentrations of leachate and the length of treatment period with respect to activities of antioxidant enzymes, malondialdehyde (MDA) contents, and chlorophyll contents. Seedlings treated with 0.6%, 1.25%, or 2.5% leachate solution for 24–72 h showed increased superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) activities. When seedlings were treated with leachate solutions for 96 h, antioxidant enzyme activities and chlorophyll content decreased in A. retroflexus, but only CAT activity decreased in C. glaucum. When seedlings of the two weed species were treated with 5% leachate solution, CAT activity and chlorophyll content decreased and MDA content gradually increased with longer treatment times (from 24 to 96 h). The two weed species showed different allelopathic responses to E. adenophorum; A. retroflexus was more sensitive than C. glaucum. Based on the investigation, it could be speculated that the delayed germination and low germination rate of the weeds after treatment by leachate could be due to the fact that leachate damaged the membrane system of the seeds. By delaying germination, lowering the germination rate of the weeds and inhibiting seedling growth, leachate from E. adenophorum could provide an effective way of controlling the weeds.     

Axenization and optimization of in vitro growth of clonal cultures of Tetratrichomonas gallinarum and Trichomonas gallinae

A rapid and simple procedure was established to obtain clonal axenic cultures of Tetratrichomonas gallinarum and Trichomonas gallinae and to optimize their in vitro growth conditions. Medium 199 was used for axenization of two genetically different clones of T. gallinarum and T. gallinae. Six different media were used to optimize the growth behaviour of axenically grown parasites: Medium 199, TYM, TYI-S-33, Hollander fluid (HF), Trichomonas vaginalis (TV) and modified TV media. The highest cell yields for both axenic clones of T. gallinarum were obtained in modified TV medium without antibiotics. The maximum numbers of trophozoites of T. gallinae were obtained in an optimized HF medium. This study demonstrated that axenic cultures for T. gallinarum and T. gallinae could be obtained avoiding the migration technique through a V-tube. Following axenization and optimization, both clones of T. gallinarum and T. gallinae could be propagated both aerobically and anaerobically.