Some Xylaria species on termite nests

Xylaria arenicola, X. brasiliensis, X. escharoidea, X. furcata, X. nigripes, X. piperiformis and X. rhizomorpha represent ancient names of fungi known to inhabit abandoned termite nests. We attempt to redescribe them and to reduce the confusion among them. Xylaria tanganyikaensis and X. readeri, species that might be associated with termite nests, are described. We describe a new variety, X. furcata var. hirsuta, and discuss an unnamed fungus that probably represents a new species. Photographs and a key are presented to aid the identification of these taxa.     

黑翅土白蚁与其它二种白蚁种群间的相容性

将黑翅土白蚁Odontotermes formosanus(Shiraki)与另外2个种群的白蚁按一定比例放置于同一培养皿中观察其斗争情况,研究黑翅土白蚁与台湾乳白蚁Coptotermes formosanus(Shiraki)和黑胸散白蚁Reticulitermes chinensisSnyder的相容性。结果表明,实验一开始,双方的工蚁和兵蚁都立即进入激烈的嘶咬斗争,在不同配比下,开始5min内的斗争次数均在40次左右,5h后开始5min内的斗争次数陡然下降,几乎不发生争斗。各种群白蚁在其数量占优势的情况下均表现出显著的斗争优势;而在双方数量相当时,黑翅土白蚁工蚁的斗争能力明显强于另外2个种群的工蚁。结果表明,自然界中不可能出现黑翅土白蚁与家白蚁和黑胸散白蚁群体的偶然性融合,黑翅土白蚁很可能在其群体建立的地区中占据优势。     

Distribution patterns of four Termitomyces species cultivated by a fungus-growing termite,Odontotermes formosanus,in Taiwan

Fungus-growing termites inoculate the obligate symbiotic fungus Termitomyces into the fungus comb in their colonies. In this study, Taiwanese Termitomyces species were determined by diagnostic PCR using the metagenome of the body of Odontotermes formosanus (the only host termite living in Taiwan) as a template. Phylogenetic analyses revealed that four different Termitomyces species are cultivated by O. formosanus in Taiwan. Three have previously been registered in the DNA database, but one was first recorded in Taiwan. Only Termitomyces sp. Type C was distributed in all areas investigated in Taiwan, whereas the other three species were distributed regionally. Field observations indicated that the flush period and the number of fruit bodies in each colony varied between species. The distribution patterns of Termitomyces spp. in Taiwan may be related to the Taiwanese climate and/or the fruiting pattern.     

黑翅土白蚁分飞蚁及蚁王、蚁后生殖系统结构观察

对黑翅土白蚁Odontotermes formosanus Shiraki雌雄分飞蚁及蚁后、蚁王的生殖系统进行解剖研究。结果表明,蚁后的卵巢管数量显著多于雌性分飞蚁的,蚁王的精巢管数量显著多于雄性分飞蚁的。蚁后的卵巢体积显著大于雌性分飞蚁的,卵巢也由分飞蚁时期的卵形发育到了蚁后时期的长形。蚁王的精巢体积显著大于雄性分飞蚁的,但在形状上没有明显的差异。     

黑翅土白蚁有翅成虫复眼的形态结构

采用组织切片法光镜下观察黑翅土白蚁Odontotermes formosanus(Shiraki)有翅成虫的复眼形态结构及光、暗适应条件下色素颗粒移动的规律。结果如下:(1)头正前方观,复眼外部形态略呈圆形。(2)有翅成虫复眼类型属于并列像眼,每只复眼约由360个小眼组成。(3)每个小眼是由1套屈光器(1个角膜和1个晶锥)、小网膜色素细胞、视杆和基细胞等几部分组成。小网膜色素细胞内均含有丰富的色素颗粒。(4)在光适应条件状态下,屈光器及视杆周围的色素颗粒主要分布在视杆部位的上侧,暗度适应条件状态时则较均匀地分布于视杆两侧上下;性别对色素颗粒分布无明显影响。     

黑翅土白蚁下唇腺解剖构造研究

本文利用显微解剖和测量,研究黑翅土白蚁Odontotermes formosanus下唇腺解剖构造及其在不同类型个体间职能的分化。黑翅土白蚁的下唇腺由左右对称的两部分组成,每侧构造由9～11个腺泡群、1个下唇腺囊及相关的导管组成。每侧的腺泡群导管与下唇腺囊导管首先汇合成分导管,两侧的分导管在舌基部后方汇合,形成一条主导管,主导管开口于舌基部下方。黑翅土白蚁下唇腺发达程度与个体的职能有关。有翅成虫下唇腺最发达,3龄无翅芽若虫下唇腺最小。巢内工蚁是口交哺物质的主要提供者,在下唇腺长度、腺泡直径及腺泡数量方面均极显著高于采食工蚁。兵蚁下唇腺及下唇腺囊转化为与防卫职能相关的器官。前兵蚁与兵蚁下唇腺囊均发达,老龄兵蚁下唇腺囊内充满黄色粘稠液体。冰冻保存后下唇腺明显缩小。冰冻保存后下唇腺腺泡直径的校正系数为1.19,腺泡群长度校正系数为1.14。     

黑翅土白蚁表皮碳氢化合物的品级特征

【目的】探索黑翅土白蚁Odontotermes formosanus(Shiraki)表皮碳氢化合物的品级特征。【方法】应用固相微萃取、气相色谱和气相色谱-质谱联用等技术,对不同品级、不同龄期个体表皮碳氢化合物进行检测。【结果】工蚁、兵蚁、幼蚁、长翅生殖蚁表皮碳氢化合物组分种类相同(32种),主要为正烷烃、支链烷烃和烯烃等。非生殖品级中组分含量变化特点表现在,工蚁(大工蚁和小工蚁):1-十九碳一烯显著低于幼蚁和兵蚁;大工蚁:三十一烷、三十四烷显著低于小工蚁和幼蚁;小工蚁(巢内工蚁):2,8,8-三甲基十烷显著高于、十六烷和十八烷低于其他非生殖品级;兵蚁:十四烷显著低于、2-me-1-C19:1高于其他非生殖品级,三十一烷、三十四烷显著低于小工蚁和幼蚁;1龄幼蚁:十六烷高于其他非生殖品级;2龄幼蚁:(E)-9-4-me C18高于其他非生殖品级;3龄幼蚁:1-十九碳一烯、四十四烷高于其他非生殖品级;此外,1龄、2龄、3龄幼蚁的3,3,6-三甲基十烷、2-me-(E)-7-C16:1显著高于、8-十七碳一烯显著低于其他非生殖品级。生殖品级与非生殖品级比较,十二烷、十四烷、十七烷、三十一烷、2,8,8-三甲基十烷、8-乙基十五烷、2,6-二甲基十七烷、8-十七碳一烯、1-十七碳一烯、1-十八碳一烯、9-十九碳一烯的含量差异显著。【结论】黑翅土白蚁不同品级个体表皮碳氢化合物种类相同,组分含量存在差异,可用于巢内品级间识别。不同龄幼蚁和不同发育阶段工蚁,表皮碳氢化合物存在特征性组分含量,表皮碳氢化合物分泌随个体发育而改变。     

Termite-regulated fungal monoculture in fungus combs of a macrotermitine termite Odontotermes formosanus

The mechanism of the exclusive growth of Termitomyces in fungus combs with fungi-growing termites, O. formosanus was examined using laboratory scale fungus combs. In the combs without the termites, vigorous growth of unidentified fungi was observed although no significant change was found in the case of the combs with termites. In addition, these results were reproducible even when incubated in a separated dish, suggesting that the physicochemical conditions were not the reason for the growth. With the molecular based analysis for the microbial communities in the combs, monoculture of the Termitomyces in the combs with termites was confirmed while the bacterial communities were independent either with or without termites. Possible mechanism of the exclusive growth of Termitomyces, such as the selective grazing of pathogenic fungi or contribution of antifungal activity giving actinomycetes were also discussed.     

黑翅土白蚁基因组DNA提取方法的优化

目的 为快速地提取到质量较好的黑翅土白蚁基因组DNA进行白蚁种群多样性的研究,对基因组DNA提取方法进行了比较与改进.方法 先初步采取CTAB法与蛋白酶K法对黑翅土白蚁基因组DNA的提取方法进行比较,再利用正交设计法对蛋白酶K法中裂解液、蛋白酶、RNA酶及作用时间4个因素进行优化.结果 蛋白酶K法获得的基因组DNA的质量与产量稍优于CTAB法;较佳的提取步骤组合为:裂解液150 μL,蛋白酶K 6μL,作用时间1h,RNA酶可不添加.结论 采用优化后的方法获得的基因组DNA为模板进行PCR扩增,得到了清晰、稳定的扩增谱带,完全可用于相关后续实验.     

氟虫腈、吡虫啉作为黑翅土白蚁诱杀药剂的效果

毒力测定结果表明,0.025~0.4μg/mL氟虫腈和吡虫啉分别在药后3 d和5 d对黑翅土白蚁Odontotermes formosanus表现出明显的毒杀效果,氟虫腈和吡虫啉药后1 d的LC50分别为药后5 d的509倍和63.8倍,2种药剂对黑翅土白蚁的毒杀效果均比较缓慢。毒性传递试验表明,0.5μg/g毒沙处理白蚁1 h后,氟虫腈和吡虫啉的致死毒性均可被传毒白蚁传递给受毒白蚁。驱避作用试验表明,50μg/mL氟虫腈对黑翅土白蚁无明显的驱避作用,而50μg/mL吡虫啉对黑翅土白蚁表现出了明显的驱避作用。可见,2种供试药剂中,氟虫腈是较理想的白蚁诱杀药剂。     

Purification of a laccase from fungus combs in the nest of Odontotermes formosanus

A new enzyme was isolated from the fungus combs in the nest of Odontotermes formosanus and identified as a laccase. The single laccase was purified with a purification factor of 16.83 by ammonium sulphate precipitation and anion exchange chromatography, to a specific activity of 211.11 U mg⁻¹. Its molecular mass was 65 kDa. The optimum pH value and temperature were 4.0 °C and 10 °C with ABTS as the substrate, respectively. The enzyme activity stabilized at temperatures between 10 °C and 30 °C and decreased rapidly when the temperature was above 30 °C. The V_max and K_m values were 3.62 μmol min⁻¹ mg⁻¹ and 119.52 μM, respectively. Ethanol concentration affected laccase activity, inhibiting 60% of enzyme activity at a concentration of 70%. Metal ions of Mg²⁺, Ba²⁺ and Fe²⁺ showed inhibition on enzyme activity of 17.2%, 5.3% and 9.4%, respectively, with the increase of metal ions concentration from 1 mM to 5 mM. Especially Fe²⁺ strongly inhibited enzyme activity up to 89% inhibition at a concentration of 1 mM.     

Levels of specificity of Xylaria species associated with fungus-growing termites: a phylogenetic approach

Fungus-growing termites live in obligate mutualistic symbiosis with species of the basidiomycete genus Termitomyces , which are cultivated on a substrate of dead plant material. When the termite colony dies, or when nest material is incubated without termites in the laboratory, fruiting bodies of the ascomycete genus Xylaria appear and rapidly cover the fungus garden. This raises the question whether certain Xylaria species are specialised in occupying termite nests or whether they are just occasional visitors. We tested Xylaria specificity at four levels: (1) fungus-growing termites, (2) termite genera, (3) termite species, and (4) colonies. In South Africa, 108 colonies of eight termite species from three termite genera were sampled for Xylaria . Xylaria was isolated from 69% of the sampled nests and from 57% of the incubated fungus comb samples, confirming high prevalence. Phylogenetic analysis of the ITS region revealed 16 operational taxonomic units of Xylaria , indicating high levels of Xylaria species richness. Not much of this variation was explained by termite genus, species, or colony; thus, at level 2–4 the specificity is low. Analysis of the large subunit rDNA region, showed that all termite-associated Xylaria belong to a single clade, together with only three of the 26 non-termite-associated strains. Termite-associated Xylaria thus show specificity for fungus-growing termites (level 1). We did not find evidence for geographic or temporal structuring in these Xylaria phylogenies. Based on our results, we conclude that termite-associated Xylaria are specific for fungus-growing termites, without having specificity for lower taxonomic levels.     

烯啶虫胺肠衣饵剂对黑翅土白蚁的防治效果

[目的]为了开发一种高效环保型白蚁饵剂.[方法]选定烯啶虫胺和溴虫腈对黑翅土白蚁Odontotermes formosanus进行室内毒力测定,根据室内毒力测定结果筛选毒力较高药剂进行驱避试验,并制备3种不同剂量的肠衣饵剂,开展室内白蚁毒杀效果试验和园林白蚁防治效果试验;选用合适剂量肠衣饵剂进行堤坝黑翅土白蚁诱杀试验,并用挖巢法检测肠衣饵剂对堤坝黑翅土白蚁的诱杀效果.[结果]1.6tg/mL烯啶虫胺处理黑翅土白蚁72 h后校正死亡率达100％,16 μtg/mL溴虫腈处理72 h后黑翅土白蚁校正死亡率也达到100％.室内毒力测定结果表明烯啶虫胺对黑翅土白蚁的LC5o值低于溴虫腈,100 μtg/mL烯啶虫胺处理8h后对黑翅土白蚁无显著驱避作用.3种剂量烯啶虫胺肠衣饵剂处理72 h后,黑翅土白蚁校正死亡率均在60％以上.将不同剂量的烯啶虫胺肠衣饵剂投放到园林中45 d后,肠衣饵剂基本被食空,施药点周围无白蚁及活动迹象.在福建省水库大坝周围投放60 μtg/g烯啶虫胺肠衣饵剂,3个月后肠衣饵剂未发霉且基本被食空,6个月后挖巢发现蚁道内无黑翅土白蚁活动,白蚁巢体出现死亡和坍塌情况.[结论]60 μg/g烯啶虫胺肠衣饵剂对园林和堤坝中的黑翅土白蚁具有良好的诱杀效果,是一种高效环保型白蚁肠衣饵剂.     

扩头蔡白蚁消化与生殖系统解剖及其与黑翅土白蚁的比较研究

扩头蔡白蚁Tsaitermes ampliceps(Wang&Li)和黑翅土白蚁Odotoermes formosanus(Shiraki)是分布广泛的食木性昆虫,群居生活,具有复杂的品级分化.本试验研究了低等白蚁扩头蔡白蚁不同品级的消化和生殖系统结构,并与高等白蚁黑翅土白蚁进行了比较.结果显示扩头蔡白蚁的消化、生殖系...     

Phylogenetic analysis of the gut bacterial microflora of the fungus-growing termite Odontotermes formosanus

We constructed a bacterial 16S rRNA gene clone library from the gut microbial community of O. formosanus and phylogenetically analyzed it in order to contribute to the evolutional study of digestive symbiosis and method development for termite control. After screening by restriction fragment length polymorphism (RFLP) analysis, 56 out of 280 clones with unique RFLP patterns were sequenced and phylogenetically analyzed. The representative phylotypes were affiliated to four phylogenetic groups, Firmicutes, the Bacteroidetes/Chlorobi group, Proteobacteria, and Actinobacteria of the domain Bacteira. No one clone affiliated with the phylum Spirochaetes was identified, in contrast to the case of wood-feeding termites. The phylogenetic analysis revealed that nearly half of the representative clones (25 phylotypes) formed monophyletic clusters with clones obtained from other termite species, especially with the sequences retrieved from fungus-growing termites. These results indicate that the presence of termite-specific bacterial lineages implies a coevolutional relationship of gut microbes and host termites.     

Phylogenetic Analysis of the Gut Bacterial Microflora of the Fungus-Growing Termite Odontotermes formosanus

We constructed a bacterial 16S rRNA gene clone library from the gut microbial community of O. formosanus and phylogenetically analyzed it in order to contribute to the evolutional study of digestive symbiosis and method development for termite control. After screening by restriction fragment length polymorphism (RFLP) analysis, 56 out of 280 clones with unique RFLP patterns were sequenced and phylogenetically analyzed. The representative phylotypes were affiliated to four phylogenetic groups, Firmicutes, the Bacteroidetes/Chlorobi group, Proteobacteria, and Actinobacteria of the domain Bacteira. No one clone affiliated with the phylum Spirochaetes was identified, in contrast to the case of wood-feeding termites. The phylogenetic analysis revealed that nearly half of the representative clones (25 phylotypes) formed monophyletic clusters with clones obtained from other termite species, especially with the sequences retrieved from fungus-growing termites. These results indicate that the presence of termite-specific bacterial lineages implies a coevolutional relationship of gut microbes and host termites.     

黑翅土白蚁的生物学特性及综合防治技术

从黑翅土白蚁Odontotermes formosanus(Shiraki)的形态特征、生物学特性、寄主与分布、综合防治技术和白蚁资源综合利用等5个方面着重论述近年来该虫的研究动态及其重要进展。     

Physicochemical conditions and metal ion profiles in the gut of the fungus-growing termite Odontotermes formosanus

The physicochemical conditions in an insect's gut microenvironment have been reported to play an important role in food processing and metabolisms. In this study, the profiles of oxygen, pH, redox potentials, and hydrogen in the isolated guts of the fungus-growing termite, Odontotermes formosanus Shiraki, were investigated with a microeletrode system. Compared with those in other termites, O. formosanus exhibited a relatively lower oxygen partial pressures in its gut system ranging from 0 to 8.6kPa. The pH profile in the different gut compartments was neutral (pH 6.1-7.4) except in the rectum region. The average redox potentials at the center of each gut region (except rectum) were high and ranged from approximately +70 to +310mV. Especially, as the central intermediate during lignocellulose degradation, hydrogen partial pressures in the hindgut paunch lumen were recorded as high as 10.4kPa. Furthermore, thirteen metal ion concentrations in the termite's gut system, nest symbiotic fungal combs, as well as the nest soil samples were evaluated with Inductively Coupled Plasma Mass Spectrometry (ICP-MS), which indicated that six metal ions (K, Mg, Mn, Ba, Se, and Mo) out of 13 ions recorded in the major digestive tract regions show some significant differences in their spatial distributions. A significant enrichment of some metal ions was also observed in the rectum, fungal combs, and the nest soil samples. The lower oxygen profiles, neutral pH, higher redox potentials, and higher hydrogen accumulation with the characterized spatial distributions for metal ions in the digestive tract of O. formosanus, highlighted the most important distinctiveness of the fungus-growing termites in its gut microenvironments, suggesting that the unique structure and functions of the intestinal ecosystem may present within its gut.     

密粘褶菌松木粉培养物对黑翅土白蚁的生物活性

【目的】研究密粘褶菌Gloeophyllum trabeum(Pers.:Fr.)Murr松木粉培养物对黑翅土白蚁Odontotermes formosanus(Shiraki)的生物活性。【方法】比较了密粘褶菌松木粉培养物不同溶剂提取液对采食工蚁的踪迹和引诱活性,研究不同浓度正己烷和乙酸乙酯提取液的引诱活性变化。【结果】正己烷提取液表现出高引诱活性和踪迹活性,乙酸乙酯、三氯甲烷、丙酮提取液具有高引诱活性和较低踪迹活性,甲醇、乙醇提取液表现出较高踪迹活性和较低引诱活性。正己烷提取液浓度与趋性反应率的回归方程为y=12.561ln(x)+50.11;乙酸乙酯提取液浓度与趋性反应率的回归方程为y=12.161ln(x)+51.411。【结论】密粘褶菌松木粉培养物对黑翅土白蚁采食工蚁具有明显的踪迹和引诱活性。正己烷和甲醇提取液的踪迹活性最强,乙酸乙酯和正己烷提取液的引诱活性最强。正己烷和乙酸乙酯提取液对采食工蚁的引诱活性具有很好的浓度相关性。     

Distribution pattern of Termitomyces types symbiotic with the fungus‐growing termite Odontotermes formosanus on Okinawa Island

Fungus‐growing termites (subfamily Macrotermitinae) cultivate the symbiotic basidiomycete fungus Termitomyces in their fungus comb to digest cellulosic materials and to supply nitrogen‐rich fungal diet. In Japan, the fungus‐growing termite Odontotermes formosanus is found on the Yaeyama Islands and Okinawa Island, Okinawa Prefecture. Odontotermes formosanus is thought to have been recently and artificially introduced to Okinawa Island as its distribution is discontinuous and restricted to small areas. Previous DNA analyses revealed that two types of Termitomyces, namely Termitomyces sp. Type A and Termitomyces sp. Type B, whose fruiting bodies correspond to Termitomyces microcarpus‐like pseudorhiza‐lacking small mushroom and Termitomyces intermedius, respectively, are cultivated by O. formosanus on the Yaeyama Islands. However, information about the Termitomyces types cultivated by O. formosanus on Okinawa Island is limited. To define the fungal types cultivated by O. formosanus on Okinawa Island, I developed a diagnostic polymerase chain reaction method using primer sets specific to the nuclear ribosomal DNA sequences consisting of the internal transcribed spacers (ITS1 and ITS2) and 5.8S rDNA of Termitomyces not using fungal mycelium, but using the termite gut metagenome including fungal DNA as a template. The results indicated that the same two Termitomyces types from Iriomote Island are cultivated by O. formosanus on Okinawa Island. The distribution pattern of Termitomyces types on Okinawa Island showed that Termitomyces sp. Type A is limited to the mountainous side of Sueyoshi Park, despite Termitomyces sp. Type B being widely distributed in the area in which O. formosanus is found. This finding implies that O. formosanus on Okinawa Island was recently introduced from Iriomote Island to Sueyoshi Park.