两种穿山甲外形量衡度的测定及比较

There are 7 species of pangolin in the world. This paper deah with the measurement of some morphology indices in Manis petadactyla and Mania javanica. Measured variables included the body weight, the total body length, the length of head and body, the length of tail,the length of protruding rim of external ear, the length of hind feet, the length of middle claws of fore feet, the length of middle claws of hind feet, the number of rows of scales round mid-body, the number of single flank scales of edge tail. All variables measured were compared respectively between this 2 species. The results indicated that: ( 1 ) there were no signifieant differences in body weight and length of hind feet between this 2 species, but differences in the other variables were extremely significant. (21 The ratio of the length of middle claws of hind feet and fore feet in Mania pentadactyla was less than 1/2 ( n = 48 ), while Mania jaw, nica was more than 1/2 ( n = 15 ) ; the length of protruding rim of external ear in Mania pentadactyla was over 10 mm { n = 38 ), whereas Manis javanica was under 10 mm { n = 13 ) ; the number of single flank scales of edge tail in Mania pentadactyla was not beyond 21 (n = 65), and Mania javanica exeeeded 21 (n = 14) . So the ratio of the length of middle claws of hind feet and fore feet, the length of protruding rim of external ear, arid the ntunber of single flank scales of edge tail should be regard as distinctive features and taxonomic indices between this 2 species. (3) The relationships between body weight ( Y) and the total body length, the length of head and body, the length of tail {X) were positive correlation, and satisfied relation equation Y= aX^b.     

意大利工蜂咽下腺结构观察

通过研究意大利蜜蜂Apis mellifera工蜂咽下腺的结构及形态,为蜜蜂形态学和蜜蜂生理学以及王浆分泌生理提供新知识采用光学显微镜、扫描电镜、HE染色石蜡切片和透射电镜,从形态学和细胞学水平对意大利工蜂咽下腺的结构和功能做了详尽探究和分析.意大利工蜂咽下腺由多个卵圆形腺泡组成,每个腺泡的直径约为120×180 μm,腺泡之间有微气管相连,腺泡外侧有神经细胞,神经细胞的胞体直径约为12μm.每个腺泡是由多细胞构成,最外层有0.1 ～0.2 μm的结缔组织膜,内部有壁细胞、酶原细胞和王浆分泌细胞,其中王浆分泌细胞分布较多,细胞直径约为16 ～ 18 μm,细胞核直径约为6μm. 由咽下腺的形态结构和细胞类型推断,意大利工蜂咽下腺主要分泌王浆,也具有分泌消化酶的功能,同时了解到分泌消化酶和王浆的途径.     

Spatial distribution of Vespa velutina individuals hunting at domestic honeybee hives： heterogeneity at a local scale

Since its recent introduction into Europe, the yellow-legged hornet, Vespa velutina, has become a major predator of the domestic honeybee, Apis mellifera, but little is known about its hunting behavior. We studied V. velutina hunting behavior by a capture- mark-recapture procedure in an experimental apiary. A total of 360 hornets were captured and tagged, and we determined： （i） the number of hornets visiting the apiary and the changes in time, （ii） the average number of individual visits per half-day and the time elapsed between consecutive recaptures, and （iii） the individual and global distribution of the hornets in the apiary. More than 50% of the marked hornets were recaptured at least once, this increased to 74% in considering the first marked individuals. We estimated 350 hornets visiting the patch daily with at least 1 visit per half-day. The number of marked hornets decreased over time while the number of unmarked ones increased, suggesting a turnover of individuals. The reduction of the delay between consecutive visits indicates that hornets became more efficient over time. Most of the hornets （88%） were recaptured in front of different hives but, overall, the global distribution was aggregative. Hornets were mainly recaptured in front of 1 hive which was neither the smallest nor the biggest colony, suggesting that the major cue used by hornets is not the amount of food. We hypothesize that the defensive behavior of the honeybee colony could explain our results which may be promising to further studies.     

西方蜜蜂四个亚种酯酶同工酶型和苹果酸脱氢酶Ⅱ同工酶基因型的遗传差异

The esterase (EST) and the malate dehydrogenase Ⅱ (MDHⅡ) isozymes in A. mellifera carpatica (C), A. mellifera ssp.(D), “Zhejiang Agricultural University No.1” A. mellifera ligustica (Ea) and A. mellifera carnica (K), and the genotypes of MDHⅡ isozymes in four subspecies of A. mellifera were analysed by IEF PAGE. The results indicate that the four subspecies of A. mellifera showed the same EST zymogram, while A. cerana showed a different EST zymogram, which suggests that the genotypes of EST isozymes in A. mellifera are different from those in A. cerana. The genotypes of the MDHⅡ isozymes in four subspecies of A. mellifera were aa, ab, ac, bb, bc and cc. The C and Ea subspecies exhibited high homozyosity, while D and K displayed high heterozyosity. The allele b appeared to be the highest frequency in C, while the allele c had the highest frequency in Ea. The frequencies of the alleles a, b and c in sub species D were very similar. The alleles a and c were common, while allele b was rare in sub species K. There were highly significant differences in genotype frequencies, allele frequencies, homozyosity and heterozyosity among these four subspecies.     

Comparative susceptibility and immune responses of Asian and European honey bees to the American foulbrood pathogen,Paenibacillus larvae

American foulbrood (AFB) disease is caused by Paenibacillus larvae. Currently, this pathogen is widespread in the European honey bee— Apis mellifera. However, little is known about infectivity and pathogenicity of P. lan'ae in the Asiatic cavity-nesting honey bees, Apis cerana. Moreover, comparative knowledge of P. larvae infectivity and pathogenicity between both honey bee species is scarce. In this study, we examined susceptibility, larval mortality, survival rate and expression of genes encoding antimicrobial peptides (AMPs) including defensin, apidaecin, abaecin, and hymenoptaecin in A. mellifera and A. cerana when infected with P. larvae. Our results showed similar effects of P. larvae on the survival rate and patterns of AMP gene expression in both honey bee species when bee larvae are infected with spores at the median lethal concentration (LC5 0 ) for A. mellifera. All AMPs of infected bee larvae showed significant upregulation compared with noninfected bee larvae in both honey bee species. However, larvae of A. cerana were more susceptible than A. mellifera when the same larval ages and spore concentration of P. larvae were used. It also appears that A. cerana showed higher levels of AMP expression than A. mellifera. This research provides the first evidence of survival rate, LC50 and immune response profiles of Asian honey bees, A. cerana, when infected by P. larvae in comparison with the European honey bee, A. mellifera.     

本期重点推介

正蜜蜂Apis spp.是具有级型分化的社会性昆虫。揭示其级型分化及调控机理,对认识社会性昆虫的演化形成机制、不同级型的发育和维持机理以及对其更好地加以应用均有重要的参考价值。已有研究表明,蜂王浆的主要蛋白成分———王浆主蛋白(major royal jelly proteins,MRJPs)在蜜蜂的级型分化中具有重要的功能。mrjp8是mrjps家族中较晚发现的一个成员。为了进一步明确mrjp8基因表达与蜜蜂级型分化的关系,福建农林大学蜂学学院李江红等对西方蜜蜂Apis mellifera不同发育龄期工蜂体内,以及成年工蜂、新出房蜂王和雄蜂不同组织中的mrjp8表达水平进行了检测,发现     

Daily number of bee louse （Braula coeca） in honey bee （Apis mellifera carnica and A. m. syriaca） colonies maintained under semi-arid conditions

Experimental work was conducted at two apiaries located in Irbid district and in Shuna North, Jordan, during the years 2004-2006. The aims of these investigations were to estimate the seasonal changes in the infestation rates of the bee louse （Braula sp.） and to develop an easy and rapid method of estimating the infestation rate on workers with bee Braula. Two major honey bee subspecies are reared in Jordan; Apis mellifera carnica and Apis mellifera syriaca were used in this study. The results showed that the infestation rate began to increase rapidly in May, reaching the season＇s maximum rate of 16.2%, 15.8% and 17.4% forA. m. carnica and 22.6%, 23.9% and 22.9% forA. m. syriaca in December of 2004, 2005 and 2006, respectively. The maximum adult numbers of bees were found in April and June, whereas the minimum for the year was in January in both honey bee subspecies colonies during the study period. The actual population of the bee louse could be estimated by counting the daily dropped lice and multiplying by a factor of 158. This factor is valid for the experimental colonies of both subspecies kept for 3 years under semi-arid Mediterranean conditions.     

Isolation and characterization of glacier VMY22, a novel lytic cold-active bacteriophage of Bacillus cereus

As a unique ecological system with low temperature and low nutrient levels, glaciers are considered a "living fossil" for the research of evolution. In this work, a lytic cold-active bacteriophage designated VMY22 against Bacillus cereus MYB41-22 was isolated from Mingyong Glacier in China, and its characteristics were studied. Electron microscopy revealed that VMY22 has an icosahedral head(59.2 nm in length, 31.9 nm in width) and a tail(43.2 nm in length). Bacteriophage VMY22 was classified as a Podoviridae with an approximate genome size of 18 to 20 kb. A one-step growth curve revealed that the latent and the burst periods were 70 and 70 min, respectively, with an average burst size of 78 bacteriophage particles per infected cell. The pH and thermal stability of bacteriophage VMY22 were also investigated. The maximum stability of the bacteriophage was observed to be at pH 8.0 and it was comparatively stable at p H 5.0–9.0. As VMY22 is a cold-active bacteriophage with low production temperature, its characterization and the relationship between MYB41-22 and Bacillus cereus bacteriophage deserve further study.     

Age, growth and reproduction of Sarcocheilichthys nigripinnis from the Qingyi Stream in the Huangshan Mountains

Identifying the life-history strategies of fish and their associations with the surrounding environment is the basic foundation in the conservation and sustainable utilization of fish species.We examined the age,growth,and reproduction of Sarcocheilichthys nigripinnis using 352 specimens collected monthly from May 2009 to April 2010 in the Qingyi Stream.We found the sex ratio of this study population was 0.58:1(female:male),significantly different from expected 1:1.Females and males both comprised four age groups.The annuli on the scales were formed during February and March.No obvious between-sex difference was observed in length-weight and length-scale-radius relationships.The total length in back-calculation significantly increased with age for both sexes,but did not differ significantly at each age between the two sexes.An inflection point was observed in the growth curves given by the von Bertalanffy growth function for total weight.At this inflection point,fish were 3.95 years.Both sexes reach their 50% sex maturity at age 2,when females and males were 94.7 mm and 103.0 mm total length.The temporal pattern of the gonado-somatic index corresponded to a spawning period that occurred from April through July.The non-synchronicity of egg diameter in each mature ovary during the breeding period suggested these fish may be batch spawners.The absolute fecundity increased significantly with total length and weight,whereas no significant correlation was observed between the relative fecundity and body size.     

王浆主蛋白MRJP7基因在意大利蜜蜂体内的表达

王浆蛋白是蜂王浆生物功能的物质基础,是由王浆蛋白基因家族(mrjps)编码合成的。但部分家族成员如MRJP7在王浆中的含量极少甚至检测不到。基因功能与其在生物体内的时空表达特性相关,为探究mrjp7的生物学功能,本研究利用荧光定量PCR技术对mrjp7在不同发育时期的工蜂和成年工蜂、雄蜂和蜂王的不同组织部位的表达进行定量检测。结果显示mrjp7在成年雄蜂体内的表达水平最低,成年蜂王次之,且在它们的各不同组织部位之间的表达量差异较小。该基因在工蜂幼虫和蛹期的表达同样较低,但在羽化后9日龄前后的哺育蜂王浆腺和头部特异性高表达,这与哺育蜂分泌蜂王浆哺育幼虫和蜂王的功能是相适应的,该结果在转录水平上证实了mrjp7的营养功能,为进一步的研究和应用打下了理论基础。     

Transcriptome differences in the hypopharyngeal gland between Western Honeybees (Apis mellifera) and Eastern Honeybees (Apis cerana)

Background

Apis mellifera and Apis cerana are two sibling species of Apidae. Apis cerana is adept at collecting sporadic nectar in mountain and forest region and exhibits stiffer hardiness and acarid resistance as a result of natural selection, whereas Apis mellifera has the advantage of producing royal jelly. To identify differentially expressed genes (DEGs) that affect the development of hypopharyngeal gland (HG) and/or the secretion of royal jelly between these two honeybee species, we performed a digital gene expression (DGE) analysis of the HGs of these two species at three developmental stages (newly emerged worker, nurse and forager).

Results

Twelve DGE-tag libraries were constructed and sequenced using the total RNA extracted from the HGs of newly emerged workers, nurses, and foragers of Apis mellifera and Apis cerana. Finally, a total of 1482 genes in Apis mellifera and 1313 in Apis cerana were found to exhibit an expression difference among the three developmental stages. A total of 1417 DEGs were identified between these two species. Of these, 623, 1072, and 462 genes showed an expression difference at the newly emerged worker, nurse, and forager stages, respectively. The nurse stage exhibited the highest number of DEGs between these two species and most of these were found to be up-regulated in Apis mellifera. These results suggest that the higher yield of royal jelly in Apis mellifera may be due to the higher expression level of these DEGs.

Conclusions

In this study, we investigated the DEGs between the HGs of two sibling honeybee species (Apis mellifera and Apis cerana). Our results indicated that the gene expression difference was associated with the difference in the royal jelly yield between these two species. These results provide an important clue for clarifying the mechanisms underlying hypopharyngeal gland development and the production of royal jelly.

Electronic supplementary material

The online version of this article (doi:10.1186/1471-2164-15-744) contains supplementary material, which is available to authorized users.     

Proteome comparison of hypopharyngeal gland development between Italian and royal jelly producing worker honeybees (Apis mellifera L.)

The hypopharyngeal gland (HG) of the honeybee (Apis mellifera L.) produces royal jelly (RJ) that is essential to feed and raise broods and queens. A strain of bees (high royal jelly producing bee, RJb) has been selected for its high RJ production, but the mechanisms of its higher yield are not understood. In this study, we compared HG acini size, RJ production, and protein differential expressions between the RJb and nonselected honeybee (Italian bee, ITb) using proteomics in combination with an electron microscopy, Western blot, and quantitative real-time PCR (qRT-PCR). Generally, the HG of both bees showed age-dependent changes in acini sizes and protein expression as worker behaviors changed from brood nursing to nectar ripening, foraging, and storage activities. The electron microscopic analysis revealed that the HG acini diameter of the RJb strain was large and produced 5 times more RJ than the ITb, demonstrating a positive correlation between the yield and HG acini size. In addition, the proteomic analysis showed that RJb significantly upregulated a large group of proteins involved in carbohydrate metabolism and energy production, those involved in protein biosynthesis, development, amino acid metabolism, nucleotide and fatty acid, transporter, protein folding, cytoskeleton, and antioxidation, which coincides with the fact that the HGs of the RJb strain produce more RJ than the ITb strain that is owing to selection pressure. We also observed age-dependent major royal jelly proteins (MRJPs) changing both in form and expressional intensity concurrent with task-switching. In addition to MRJPs, the RJb overexpressed proteins such as enolase and transitional endoplasmic reticulum ATPase, protein biosynthesis, and development proteins compared to the ITb strain to support its large HG growth and RJ secretion. Because of selection pressure, RJb pursued a different strategy of increased RJ production by involving additional proteins compared to its original counterpart ITb. To our knowledge, this morphological and proteomic comparison study on the HG of the two strains of worker honeybees associated with their age-dependent division of labor is the first of its kind. The study provided not only the quantity and quality differences in the HG from the RJb and the ITb, but also addressed the cellular and behavioral biology development question of how the RJb strain can produce RJ more efficiently than its wild type strain (ITb).     

意大利蜂营养腺的形态结构研究

研究了意大利蜂分泌蜂王浆的主要腺体──营养腺的形态结构及腺体随工蜂日龄增加而变化的规律。结果表明,工蜂羽化后,在各个日龄段随其职能分工的不同,营养腺形态结构也有相应的变化。从羽化出房的幼峰到哺育蜂到采集蜂,其营养腺要经历发育─→饱满发达─→衰退萎缩的过程。分泌细胞中的粗面内质网变化更为明显。     

幼虫信息素中三种酯类对中华蜜蜂和意大利蜜蜂工蜂哺育和封盖行为以及蜂王发育影响

为研究蜜蜂幼虫信息素中3种酯类成分（甲基棕榈酸酯、乙基棕榈酸酯和乙基油酸酯）对中华蜜蜂Apis cerana cerana和意大利蜜蜂Apis mellifera ligustica工蜂哺育行为、封盖行为以及蜂王发育影响, 在人造蜂蜡王台中加入1%和0.1%（w/w）的3种酯类作为实验组, 以不添加酯类（0%）的为对照组, 移入1日龄工蜂幼虫, 测定王台接受率、单个王台中幼虫和王浆重量; 另将分别添加1%和0.1%（w/w）3种酯类的石蜡假幼虫放入工蜂巢房中, 同样设对照组, 测定假幼虫的封盖率; 在新鲜王浆中以1%和0.1%（w/w）分别加入3种酯类作为实验组, 以不添加酯类（0%）的作为对照组, 再分别在1日龄、2日龄和3日龄幼虫王台中加入0.01 mL含有酯类的蜂王浆, 并测定蜂王初生重和卵巢管数量。结果表明: 0.1%甲基棕榈酸酯可以显著提高中蜂和意蜂幼虫重量; 意蜂的甲基棕榈酸酯和乙基油酸酯两个实验组(1.0%, 0.1%)假幼虫封盖率都极显著高于对照组; 乙基油酸酯两个实验组(1.0%, 0.1%)都显著降低了中蜂和意蜂蜂王初生重和卵巢管数量。这说明不同蜜蜂幼虫信息素具有不同的生物学效应。     

Conservation of nutrients in larval tissue by cannibalizing honey bees

ABSTRACT. ¹⁴C-phenylalanine appeared in royal jelly secreted by worker honey bees ( Apis mellifera , L.) directly after eating ¹⁴-phenyl-alanine labelled larvae. The amount of labelled phenylalanine in royal jelly was associated directly with the volume of jelly produced. Loss of radio-label from the workers, in the form of O₂¹⁴C, increased when royal jelly secretion ended. These patterns show that conservation of nutrients in cannibalized tissue is enhanced when the cannibal bees are producing royal jelly. Specific activity of radiolabel in worker bee hypopharyngeal glands was higher than in thoracic muscle, showing selective movement within the bees of a nutrient acquired by cannibalism.     

蜜蜂体内王浆主蛋白基因mrjp9的转录表达研究

蜜蜂体内有9种王浆蛋白基因(major royal jelly protein,MRJPs1～9),其中MRJPs1～5在蜂王浆中含量较高,是蜂王浆生物学功能的基础。MRJPs6～9在王浆中没有或含量极少,且功能未知。为研究非王浆蛋白组分的MRJP9的生物学功能,本研究用RT-PCR的方法对意大利蜜蜂Apis mellifera ligustica Spinola不同组织,不同部位,不同级型样本中mrjp9的转录水平进行检测和定量。结果发现mrjp9在蜜蜂的幼虫、蛹和成年蜜蜂的各组织部位均广泛转录表达,但其在幼虫、蛹和刚出房的成年蜜蜂体内表达水平较低,而在成年采集蜂体内表达水平则较高,其表达与蜜蜂的发育时期有关。通过对在成年蜜蜂体内各组织部位的表达水平进行检测的结果显示该基因主要在蜜蜂的头、胸和王浆腺等组织部位的表达较高,其他组织部位表达较少。此外,该基因也在雄蜂和蜂王体内广泛表达,不受蜜蜂性别和级型的影响。这些结果说明mrjp9是一与蜜蜂发育有关的基因,可能与蜜蜂的行为发育和分工调控有关。     

RNAi沉默王浆主蛋白1(Mrjp1)基因对意大利蜜蜂工蜂学习和记忆的影响

【目的】前期研究发现经吡虫啉处理的意大利蜜蜂Apis mellifera ligustica(简称“意蜂”)工蜂学习能力下降,转录组学分析表明王浆主蛋白1(major royal jelly protein 1, MRJP1)基因在吡虫啉处理的蜜蜂脑中显著下调,MRJP1可能参与调控蜜蜂学习能力。本研究旨在采用RNA干扰(RNAinference, RNAi)技术将Mrjp1特异性沉默,验证MRJP1在意蜂工蜂嗅觉学习中的关键作用。【方法】通过克隆技术获得Mrjp1基因cDNA 序列,经测序验证后,设计引物,合成用于RNAi干扰Mrjp1基因表达的dsRNA。注射dsMrjp1的意蜂工蜂作为处理组(dsMrjp1注射组),注射dsEGFP的意蜂工蜂作为对照组(dsEGFP注射组),随后通过伸吻反应(proboscis extension response, PER)实验比较两组的嗅觉学习与记忆能力差异。最后采用实时荧光定量PCR(quantitative real-time PCR, qRT-PCR)检测注射dsMrjp1后意大利蜜蜂工蜂脑中Mrjp1的相对表达量。【结果】dsMrjp1注射组与dsEGFP注射组意蜂工蜂学习能力差异显著,dsMrjp1注射组意蜂工蜂的学习能力显著降低。学习后2 h,两组意蜂工蜂的记忆力无显著差异。qRT-PCR结果显示Mrjp1的表达水平在dsMrjp1注射组意蜂工蜂脑中显著低于dsEGFP注射组,表明学习能力降低的处理组意蜂脑内对应的Mrjp1表达水平也降低。【结论】通过RNAi抑制意蜂工蜂Mrjp1基因的表达后,其嗅觉学习能力受到显著性抑制,但记忆力未受到显著影响,提示Mrjp1可能是调控意蜂学习的重要基因之一。本研究结果有助于后续进一步研究蜜蜂嗅觉学习相关的分子机制。     

人工注射Dnmt3 siRNA对意大利蜜蜂雌蜂发育的影响

为了研究人工注射DNA甲基化转移酶3 (DNA methyltransferase3, Dnmt3) siRNA对意大利蜜蜂Apis mellifera ligustica雌蜂的Dnmt3酶活性、 Dnmt3 mRNA表达量、 dynactin p62基因甲基化水平以及蜜蜂形态指标之间的影响和关系, 分别给3组3日龄的意大利蜜蜂幼虫人工注射50 ng Ringer, uth siRNA和Dnmt3 siRNA溶液。在幼虫的发育过程中, 测定每组幼虫头部的Dnmt3酶活性、 Dnmt3 mRNA表达量、 dynactin p62基因甲基化水平以及刚羽化雌性蜜蜂的初生重、 体长、 前翅长、 前翅宽、 吻长、 第3背板长等形态指标。结果表明: 人工注射Dnmt3 siRNA不仅可以显著降低意大利蜜蜂幼虫头部Dnmt3酶活性、 Dnmt3 mRNA表达量和dynactin p62基因整体甲基化水平, 同时显著提高了刚羽化雌性蜜蜂的初生重、 体长、 第3背板长。结果说明Dnmt3的改变可以影响雌性意蜂幼虫的发育。     

Functional flexibility of the honey bee hypopharyngeal gland in a dequeened colony

The role of the worker honey bee Apis mellifera L. changes depending on age after eclosion (age polyethism): young workers (nurse bees) take care of their brood by synthesizing and secreting brood food (royal jelly), while older workers (foragers) forage for nectar and process it into honey. Previously, we showed that the major proteins synthesized in the hypopharyngeal gland of the worker change from brood food proteins to alpha-glucosidase at the single secretory cell level in parallel with this age polyethism [Kubo et al., J. Biochem. 119, 291-295 (1996); Ohashi et al., Eur. J. Biochem. 249, 797-802 (1997)]. Here, we examined whether the function of the hypopharyngeal gland has flexibility depending on the colony conditions, by creating a dequeened colony in which the older workers were compelled to feed the drone larvae. It was found that most of the older workers in the dequeened colony synthesized brood food proteins as did nurse bees. Furthermore, the percentage of workers that synthesized brood food proteins was maintained at 80-90% of the total workers for at least two months, as in a normal colony. These results indicate that the function of the hypopharyngeal gland cells of the worker has flexibility and can, if necessary, be maintained as that of the nurse bee, depending on the condition of the colony.