中国熊蜂多样性、人工利用与保护策略

熊蜂是众多野生植物和农作物的重要传粉者, 在维持自然和农业生态系统平衡中发挥着十分重要的作用。中国地形多样、植被丰富, 是全球熊蜂多样性的热点地区, 但过去缺乏系统调查研究, 中国熊蜂多样性现状及其利用特性不详。本文介绍了近20年在中国熊蜂资源系统调查及其人工利用方面取得的主要研究结果: (1) 2002-2017年采集熊蜂标本5万余号, 先后揭示了一系列疑难类群的分类地位, 确认中国已知熊蜂125种, 占全球熊蜂已知物种总数的50%, 表明中国是全球熊蜂物种资源最丰富的国家; (2)首次报道了中国熊蜂物种名录, 其中22种为中国特有种。我国青藏高原东部向黄土高原、秦岭山地和四川盆地过渡的地带, 是全球熊蜂多样性分布中心; (3) 1998-2017年, 明亮熊蜂(Bombus lucorum)、密林熊蜂(B. patagiatus)、红光熊蜂(B. ignitus)、火红熊蜂(B. pyrosoma)、重黄熊蜂(B. picipes)和兰州熊蜂(B. lantschouensis) 6种本土熊蜂先后被驯养成功, 其中密林熊蜂和兰州熊蜂的繁殖性状优良, 已应用于我国设施作物传粉服务。同时, 我们从栖息地、食物资源、外来物种、农药使用等方面提出了保护中国熊蜂资源的策略, 以期为野生传粉昆虫特别是熊蜂资源的保护与可持续利用提供参考资料。     

A new electrophoretic approach to biochemical systematics of bees

SDS electrophoresis of soluble proteins from 49 species of bees from 8 families produce a conservative character set. Differences between species appear to be actual differences in polypeptides and their concentrations rather than spurious differences. The data set based on the absorption values of bands proves to be more useful than does a data set based on the molecular weights of bands present or one based on the presence and absence of bands. Numerical analysis of a matrix of distance coefficients based on absorption values of bands shows much overlap between families of bees. Also, some taxa of questionable affinities when compared morphologically can be shown to be biochemically quite similar to other members of their traditional morphological families. Other taxa, which appear very similar morphologically, are highly differentiated biochemically. Examples are presented to compare and contrast the data from the SDS-separated character set to predictions based on the theories of regulatory and constant rate protein evolution. The results of the study largely agreed with predictions based on constant rate protein evolution. Exceptions may be understood in the framework of regulatory evolution. Consequences of discordance between morphological and biochemical phenetics are discussed as they affect systematic classifications.     

A survey of alkaloids in the genera Harpalyce and Brongniartia (Fabaceae—Brongniartieae)

The presence of alkaloids in six species of Brongniartia and three species of Harpalyce is reported. This survey revealed remarkable qualitative differences in the alkaloid profiles of these two genera. B. discolor, B. lupinoides, B. sousae and B. intermedia showed a typical -pyridone pattern, with cytisine, anagyrine and baptifoline as major alkaloids. In leaves of the first three species ormosanine-type alkaloids occurred additionally. B. flava and B. vazquezii are devoid of -pyridones, but accumulate lupanine, hydroxylated lupanines and ester alkaloids. All three species of Harpalyce were similar in accumulating -pyridones, but H. formosa differed from H. brasiliana and H. pringlei in the presence of epilupinine. In general the alkaloid profiles of Brongniartia and Harpalyce show similarities to those of the Australian genera Hovea, Lamprolobium, Plagiocarpus and Templetonia and support therefore the actual concept of the enlarged tribe Brongniartieae.     

武夷山自然保护区土壤可培养芽胞杆菌 的物种多样性及分布

为了解武夷山自然保护区土壤中可培养芽胞杆菌的分布状况, 2012年6月从该保护区的黄岗山顶部、中部、底部和桐木关、挂墩、大竹岚等6个地点采集土样75份。用80℃水浴加热、稀释平板法进行芽胞杆菌的分离, 并根据16S rRNA基因序列分析对菌株进行初步鉴定。从土样中分离出芽胞杆菌418株, 鉴定为8个属42个种, 其中Bacillus属的种数最多, 有20种, Paenibacillus属和Lysinibacillus属分别有8种和7种。不同地点分离到的芽胞杆菌在种类、数量上存在差异: 从大竹岚土壤中分离到的芽胞杆菌种类最多, 从黄岗山中部和底部分离到的种类数则较少; 挂墩、大竹岚土壤中芽胞杆菌的数量较大, 达3.6×10⁶ cfu/g以上, 而黄岗山顶部和中部土壤中的数量则少于4.9×10⁵ cfu/g。Bacillus cereus、B. mycoides、B. thuringiensis和Lysinibacillus xylanilyticus等4个种在6个地点的土样中均有分离到, 其中B. thuringiensis、L. xylanilyticus是该保护区土壤中的优势种。桐木关土壤中芽胞杆菌的种类多样性和均匀度指数都比其他5个地点的高, 而挂墩土壤中芽胞杆菌的Shannon-Wiener多样性、均匀度和优势度指数都最低。B. mycoides和B. thuringiensis的数量与海拔显著相关, 相关系数分别为0.852和-0.834, B. cereus、B. mycoides、B. thuringiensis的分离频度与海拔的相关性极显著, 相关系数分别为0.960、0.952和-0.931。研究结果表明, 武夷山自然保护区土壤中可培养芽胞杆菌的种类丰富、数量较大, 具有较高的多样性。     

Responses of four Brassica species to sodium chloride

Responses in sand culture of four Brassica species, B. campestris, B. carinata, B. juncea and B. napus to NaCl were assessed for growth and yield. B. napus produced significantly greater fresh and dry biomass than the other three species in NaCl treatments and also a higher seed yield in both absolute and relative terms when grown at 125 mol/m³ NaCl. In contrast, B. campestris had the lowest biomass production, seed yield and oil content. B. campestris did not differ significantly from B. juncea and B. carinata in biomass production for growth, but B. carinata had a significantly higher seed yield (1.60 g/plant) than that of B. juncea and B. campestris (0.69 and 0.61 g/plant, respectively). The higher seed yielding species, B. napus (1.74 g/plant) and B. carinata (1.60 g/plant) accumulated lower amounts of Na⁺ and Cl⁻ in their tissues and had significantly higher K selectivity (S_K,Na) in their shoots than did those of B. campestris and B. juncea, B. napus and B. carinata also had higher leaf succulence than that of B. campestris. In view of the results presented here it can be concluded that B. napus and B. carinata are relatively tolerant to NaCl whereas B. campestris and B. juncea are relatively sensitive to NaCl.     

Schistosoma bovis: Variability of cercarial production as related to the snail hosts: Bulinus truncatus, B. wrighti and Planorbarius metidjensis

Development of Schistosoma bovis from Spain in different species or genus of intermediate hosts (Bulinus truncatus, B. wrighti and Planorbarius metidjensis) modifies cercarial productivity and its dynamics. From B. truncatus to B. wrighti and to P. metidjensis, cercarial productivity decreases while the length of the production period is increased. Variations in the dynamics are less obvious between the two species of Bulinus than between Bulinus spp. and P. metidjensis. In the latter the emission pattern is characterized by a 45–48-day production rhythm. These differences are explained in terms of larval demographic strategies and biotic capacities of the hosts. The validity of employing cercarial production as an indicator of host-parasite compatibility is discussed.     

Microhabitat selection in intertidal gobiid fishes: Species- and size-associated variation

Patterns of space use and the individual-based behaviour of microhabitat selection were investigated in three intertidal gobiid fishes, Bathygobius fuscus, Chaenogobius annularis and C. gulosus, from Kyushu, southern Japan. While the three species tended to occupy slightly different types of tidepool, their patterns of distribution largely overlapped in the field. Laboratory experiments involving choice of shelter (i.e. underneath a stone plate) and four different substrate types were conducted to examine size- and time-related variation in habitat selection. The shelter area was preferred by small- and large-sized C. gulosus (day and night), large C. annularis (day and night) and small C. annularis (daytime only), while no preference was evident in small B. fuscus (day and night) and small C. annularis (night). Patterns of substrate choice also differed among species, size groups and between day and night. Size differences in substrate use were evident in B. fuscus and C. gulosus but not in C. annularis, while diel differences were shown by all species groups except large B. fuscus. The gravel and sand substrates tended to be used more frequently than the bare rock substrate, but the strength of preference of a particular substrate type varied among individuals/species. Our results demonstrate that habitat selection by the three gobiid species is variable depending on species, body size and time of day, which must ultimately bear upon mitigating intra-/interspecific interactions in tidepool environments.     

Development of a rapid and sensitive immunoassay for detection and subsequent recovery of Bacillus anthracis spores in environmental samples

Bacillus anthracis is considered a major threat as an agent of bioterrorism. B. anthracis spores are readily dispersed as aerosols, are very persistent, and are resistant to normal disinfection treatments. Immunoassays have been developed to rapidly detect B. anthracis spores at high concentrations. However, detection of B. anthracis spores at lower concentrations is problematic due to the fact that closely related Bacillus species (e.g., B. thuringiensis) can cross-react with anti-B. anthracis antibodies, resulting in false positive detections. Subsequent polymerase chain reaction (PCR) analysis is required to differentiate virulent strains. We report here on a protocol for the rapid, sensitive detection of B. anthracis spore using the Integrating Waveguide Biosensor followed by a method for the rapid release and germination of immunocaptured spores. A detection limit of ca. 10³ spores was achieved by incubating spores simultaneously with capture and detection antibodies (“liquid-phase” assay) prior to capture on capillary tubes/waveguides. Subsequent incubation with BHI broth directly in capillary tubes allowed for rapid germination, outgrowth, and release of spores, resulting in vegetative cells for PCR analysis.     

Triassic ginkgoalean wood from the South Rewa Gondwana Basin, India

A new species of Baieroxylon, viz. B. cicatricum, is described from the Tiki Formation, Middle Gondwana, Upper Triassic of the South Rewa Gondwana Basin, Madhya Pradesh, India. The outer surface of the wood is marked with closely spaced “eye-shaped scars” and exhibits small, distinct central protuberances. These may probably represent vascular traces. The present described wood differs from all the known species of the genus by its characteristics stem scars. Dadoxylon graminovillae Zimmermann has been revised and a new combination is proposed, viz. Baieroxylon graminovillae (Zimmermann) Prasad et Lele comb. nov.     

岩白菜属植物规模化繁殖及遗传稳定性

根据市场需求和野生资源现存状况, 筛选厚叶岩白菜(Bergenia crassifolia)、秦岭岩白菜(B. scopulosa)和岩白菜(B. purpurascens)进行规模化繁殖, 并利用ISSR分子标记对组培苗进行遗传稳定性分析。以顶芽为外植体, 筛选出MS+0.5 mg·L^-1 6-BA+0.01 mg·L^-1 NAA+2.0 mg·L^-1 V_C为最佳增殖培养基, 3种岩白菜属植物增殖系数分别为3.10、2.50和2.10; 在1/2MS+1.0 mg·L^-1 IBA+2.0 mg·L^-1 V_C培养基上, 3种岩白菜属植物生根率分别为85%、80%和75%; 在腐殖土:黄沙:珍珠岩=2:1:1 (v/v/v)的混合基质中, 移栽成活率分别为90%、85%和80%。规模化繁殖厚叶岩白菜20万株, 秦岭岩白菜2万株, 岩白菜1万株, 目前还在持续生产中。ISSR分子标记结果表明, 岩白菜后代遗传变异较大, 秦岭岩白菜后代遗传变异较小, 3个种在继代至第20代时出现了遗传变异; 岩白菜和秦岭岩白菜的平均遗传变异率随继代次数的增加而增加, 厚叶岩白菜的平均遗传变异率随继代次数的增加呈现不规律变化。     

The chemistry of the abdominal gland secretion of six species of the rove beetle genus Bledius

The chemistry of the abdominal defensive secretions of six species of the rove beetle genus Bledius was examined. In all species the secretion contains the solid toxin p-toluquinone and its precursor p-toluhydroquinone dissolved in different solvents. In B. furcatus, B. tricornis and B. dissimilis these solvents are mainly alkenes and lactones, especially 1-undecene and γ-dodecalactone. In addition to alkenes and lactones the secretion contains decanoic acid and 3-methyl-2-butenoic acid in B. opacus and B. subterraneus. In B. arenarius the quinones are dissolved in octanoic acid and octyloctanoate, exclusively. In B. arenarius and B. opacus the secretion's content of p-toluquinone is reduced in comparison with the other species. Application experiments with the natural predators revealed that in B. arenarius this quinone reduction does not involve a reduction of irritation efficiency. This is probably due to the irriation properties of the B. arenarius solvent octanoic acid. The results are discussed with respect to the evolution of the different secretion types within the genus Bledius and within the subfamily Oxytelinae.     

Overexpression of serine alkaline protease encoding gene in Bacillus species: performance analyses

Bacillus species carrying subC gene encoding serine alkaline protease (SAP) enzyme were developed in order to increase the yield and selectivity in the bioprocess for SAP production. For this aim, subC gene was cloned into pHV1431 Escherichia coli–Bacillus shuttle vector, and transferred into nine host Bacillus species, i.e. B. alvei, B. amyloliquefaciens, B. badius, B. cereus, B. coagulans, B. firmus, B. licheniformis, B. sphaericus and B. subtilis. The influence of the host Bacillus species on SAP production on a defined medium with glucose was investigated in bioreactor systems. For each of the recombinant (r-) Bacillus species, effects of initial glucose concentration on cell growth and SAP production were investigated; and, physiological differences and similarities between the wild-type and r-Bacillus species are discussed. The highest biomass concentration was obtained with r-B. coagulans as 3.8 kg m⁻³ at the initial glucose concentration of C_Go=20 kg m⁻³ and the highest volumetric SAP activity was obtained with r-B. amyloliquefaciens as 1650 U cm⁻³ at C_Go=20 kg m⁻³. Overall SAP activity per amount of substrate consumed was the highest for r-B. sphaericus (137 U g⁻¹ cm⁻³) and r-B. licheniformis (130 U g⁻¹ cm⁻³). Among the r-Bacillus species the highest activity increase compared to the wild types was obtained with r-B. sphaericus while the lowest increase was obtained with r-B. amyloliquefaciens and r-B. licheniformis due to high SAP production potential of the wild-type strains. During storage of the host microorganisms, r-B. alvei and r-B. amyloliquefaciens were not able to bear the recombinant plasmid, probably, due to the restriction enzymes synthesized. Due to the highest stable volumetric activities r-B. licheniformis (950 U cm⁻³) and r-B. sphaericus (820 U cm⁻³) appear to be the favorable hosts for the production of SAP. All the r-Bacillus species excreted organic acids oxaloacetic and succinic acids, but, none excreted the amino acid valine. The variations in by-product distributions with each recombinant organism were also discussed.     

Expression of human dihydrofolate reductase cDNA and its induction by chloramphenicol in Bacillus subtilis

A bifunctional plasmid (pMP358) able to replicate and to express cloned human dihydrofolate reductase cDNA (cDHFR) in both Escherichia coli and Bacillus subtilis was constructed. The expression of cDHFR in B. subtilis was the result of a deletion that placed the cDNA fragment under the control of the chloramphenicol acetyltransferase (CAT) gene promoter of Staphylococcus aureus plasmid pC194. By sequence analysis of plasmid pMP358, we observed a gene fusion occurring between the cDHFR and the 32nd codon of the CAT gene. We report that such a “hybrid” gene is able to direct the synthesis of a 25-kDal “hybrid” protein, which was found to be inducible by supplementing B. subtilis cells with sublethal doses of chloramphenicol.     

A biochemical approach toward the systematics of the Leptothorax “muscorum”Group in North America (Hymenoptera: Formicidae)

Enzyme patterns have been used to distinguish among a number of morphologically very similar ant species belonging to the Leptothorax “muscoru” group in North America. Not counting the already described species L. retractus, L. sphagnicolus and L. crassipilis, the complex apparently consists of at least three or four more different taxa.     

中国东部和南部小檗属植物之研究

我国小檗属植物种类十分丰富,据文献记载共有200余种,主要分布于西部和南部地区。东部和南部种类虽然较少,但迄今尚未见有较全面的整理和报导,故名称亦混乱。作者在编写《广东植物志》的过程中,对这一地区(包括广西、广东、湖南、江西、安徽、江苏、浙江、福建)的小檗属进行了初步的研究,发现了一些新分类群和分布新纪录,归并了B.chekiangensis Ahrendt和B.pingjiangensis Q.L.Chen et B.M.Yang,已知这一地区共有14种,2亚种和1变种。现将结果整理成文,内容包括已知种类的检索表、产地、分布以及对部分种类的评注和讨论。文中所引证的标本,凡未加标本室缩写代号者,均存华南植物研究所标本室。     

Berberine contents and alkaloid profile of Berberis species from different altitudes

Roots, stem and leaves of eight species of Berberis collected from different altitudes in Garhwal, Himilaya, were analysed for berberine. Lowland species and types were found to have considerably higher berberine contents in all plant parts than sub-alpine and alpine species. In all, 12 or more alkaloids were separable in these species. Based on the similarity coefficient of leaf alkaloids, B. nepalensis was found to be identical to B. asiatica and B. vulgaris was identical to B. lycium. It is suggested that the concentration of berberine is probably a result of adaptive response to altitudinal gradient in addition to it being a genetic character of different species of Berberis.     

五个三年桐品种过氧化物酶同功酶的酶谱分析

五个三年桐品种的过氧化物酶同功酶经聚丙烯酰胺凝胶电泳后,分离出了清晰的酶带,根据酶带的强弱、位置,它可划为三个区段,品种间的强带Rf值极为接近,表明它们之间的紧密亲缘关系,无疑可归于同一个种,但它们各区段中的弱带数及其Rf值又各不相同,表明了它们在性状上的差异,可划分成不同的品种。千年桐较之五个三年桐品种,不仅酶带少,无明显的强带,而且不呈现类似三年桐酶带的区段,无可非议的可成为另一个种。     

白及根部内生真菌多样性研究

白及是地生兰科多年生草本植物,是常见的中草药之一。白及根是白及植株吸收养分和水分的主要场所,开展其根部内生真菌的群落组成及多样性分析,对了解云南原生境白及与根部内生真菌间的共生关系具有重要意义。本研究采用形态学和ITS rDNA分子系统发育分析相结合的方法进行菌株鉴定。结果表明,3个采样点的白及根部分离到可培养内生真菌32株,包括9个目、13个科、16个属。其中木霉属Trichoderma、镰刀菌属Fusarium、拟盘多毛孢属Pestalotiopsis为优势类群,分别占总物种数的21.88%、18.75%、9.38%;中碳垫菌属Nemania、丛赤壳属Nectria、炭角菌属Xylaria、紫霉属Purpureocillium、刺盘孢属Colletotrichum、黑孢子菌属Nigrospora、Biscogniauxia、腐质霉属Humicola、脉孢菌属Neurospora、拟茎点霉属Phomopsis、植物腐霉Phytopythium、毛霉属Mucor、伞状霉属Umbelopsis为常见内生真菌类群。α多样性指数分析表明：各采样点的白及根部内生真菌物种多样性最高的是昆明市水源保护区采样点（D=0.799,H’=1.698）,保山市龙陵县采样点物种多样性最低（D=0.787,H’=1.580）。β多样性分析采样点之间的Jaccard（0.912-0.993）、Bray-Curtis（0.838-0.986）和UniFrac（加权0.618-0.631;不加权0.770-0.799）距离,结果表明3个采样点之间的内生真菌群落组成存在差异。3个采样点的不同地理环境和土壤类型给白及提供了不同的生境条件,是影响白及根部内生真菌物种丰度和群落组成差异的主要因素之一。     

Intra- and intraspecific variation of Juniperus virginiana and J. scopulorum seedling based on volatile oil composition

Comparisons of volatile oil constituents were made among samples of juvenile foliage collected from 78 Juniperus virginiana and 28 J. scopulorum seedling sources growing in a “common garden” environment. A canonical variate analysis, a principal coordinates analysis and hybrid distance diagrams of 30 chemical characters indicate both taxa are good species and that they exhibit clinical patterns in the Great Plains. In addition, a possible evoluitonary link between present-day J. virginiana populations in southern Texas and ancient J. scopulorum populations is indicated.     

三叶鬼针草与不同本地植物竞争对土壤 微生物和土壤养分的影响

入侵植物三叶鬼针草(Bidens pilosa)对我国农牧业生产造成了重大的损失。本文主要研究三叶鬼针草入侵与不同本地植物竞争对土壤微生物群落结构和土壤养分的影响。利用磷脂脂肪酸方法(phospholipid fatty acids, PLFAs)测定土壤微生物群落组成, 同时测定土壤养分和酶活性, 并利用Canoco4.5软件分析了土壤微生物、土壤养分和土壤酶活性的相关性。结果表明: (1)三叶鬼针草对革兰氏阳性菌、革兰氏阴性菌、丛枝菌根真菌等土壤微生物具有较强的聚集能力, 且其根际土壤聚集的微生物类群与本地植物种类密切相关。(2)三叶鬼针草入侵显著增加了入侵地土壤的有机碳含量, 降低了铵态氮的含量; 土壤中的速效钾、速效磷和硝态氮的含量则与本地植物种类密切相关。(3)相关性分析表明, 16:00和16:1 ω5c对铵态氮的含量影响较大, 而三叶鬼针草入侵地16:00和16:1 ω5c的含量显著高于裸土对照, 进而推测这一状况导致了铵态氮含量的降低。(4) 15:1 anteiso A和18:1 ω5c与速效钾的含量呈显著正相关, 而其含量在狗尾草(Setaria viridis)中显著高于其他处理, 三叶鬼针草与狗尾草混种处理中土壤中速效钾的含量高于其他处理。以上结果说明, 三叶鬼针草通过改变土壤微生物群落结构影响了土壤酶活性和土壤养分, 且这种改变与入侵地本地植物种类有关。