Role of c-Jun NH_(2)-terminal kinase-mediated mitogen-activated protein kinase pathway in response to pesticides in Apis cerana cerana

The mitogen-activated protein kinase(MAPK)cascade pathway plays an important role in regulating stress responses.The function of the c-Jun NH_(2)-terminal kinase(JNK),a component of the MAPK cascade pathway,in Apis cerana cerana(Acc)remains unclear.Here,JNK was isolated and identified from Acc.Bioinformatics analyses revealed there is a typical serine/threonine protein kinase catalytic domain in the AccJNK protein.An expression profile analysis showed that AccJNK was significantly induced by pesticide treatments.To further explore the functional mechanisms of AccJNK,a yeast 2-hybrid screen was performed,activator protein-1(AP-1)was screened as the interaction partner of AccJNK,and the interaction relationship was further verified by pull-down assay.Quantitative real-time polymerase chain reaction showed the expression pattern of AccAP-I was similar to that of AccJNK.After a knockdown of AccJNK or AccAP-I by RNA interference,the survival rate of Acc after pesticide treatments increased.Additionally,the expression levels of antioxidant-related genes and the activities of antioxidant enzymes increased,suggesting that the knockdown of AccJNK or AccAP-I increased the antioxidant capacity of bees.Our study revealed that the JNK-mediated MAPK pathway responds to pesticide stress by altering the antioxidant capacity of Acc.

Expression of a bee venom phospholipase A_2 from Apis cerana cerana in the baculovirus-insect cell

Bee venom phospholipase A2(BvPLA2) is a lipolytic enzyme that catalyzes the hydrolysis of the sn-2 acyl bond of glycerophospholipids to liberate free fatty acids and lysophospholipids.In this work,a new BvPLA2(AccPLA2) gene from the Chinese honeybee(Apis cerana cerana) venom glands was inserted into bacmid to construct a recombinant transfer vector.Tn-5B-4(Tn) cells were transfected with the recombinant bacmid DNA for expression.Sodium dodecylsulfate-polyacrylamide gel electrophoresis(SDS-PAGE) analysis revealed a double band with molecular weights of 16 and 18 kDa.Products of hexahistidine AccPLA2 fusion protein accumulated up to 5.32% of the total cellular proteins.The AccPLA2 fusion protein was cross reactive with the anti-AmPLA2(BvPLA2 of the European honeybee,Apis mellifera) polyclonal serum.The reaction resulted in a double glycosylation band,which agrees with the band generated by the native AmPLA2 in Western blot analysis.The PLA2 activity of the total extracted cellular protein in the hydrolyzing egg yolk is about 3.16 μmol/(min·mg).In summary,the recombinant AccPLA2 protein,a native BvPLA2-like structure with corresponding biological activities,can be glycosylated in Tn cells.These findings provided fundamental knowledge for potential genetic engineering to produce AccPLA2 in the pharmaceutical industry.

Defining honeybee subspecies in an evolutionary context warrants strategized conservation

Despite the urgent need for conservation consideration,strategic action plans for the preservation of the Asian honeybee,Apis cerana Fabricius,1793,remain lacking.Both the convergent and divergent adaptations of this widespread insect have led to confusing phenotypical traits and inconsistent infraspecific taxonomy.Unclear subspecies boundaries pose a significant challenge to honeybee conservation efforts,as it is difficult to effectively prioritize conservation targets without a clear understanding of subspecies identities.Here,we investigated genome variations in 362 worker bees representing almost all populations of mainland A.cerana to understand how evolution has shaped its population structure.Whole-genome single nucleotide polymorphisms(SNPs)based on nuclear sequences revealed eight putative subspecies,with all seven peripheral subspecies exhibiting mutually exclusive monophyly and distinct genetic divergence from the widespread central subspecies.Our results demonstrated that most classic morphological traits,including body size,were related to the climatic variables of the local habitats and did not reflect the true evolutionary history of the organism.Thus,such morphological traits were not suitable for subspecific delineation.Conversely,wing vein characters showed relative independence to the environment and supported the subspecies boundaries inferred from nuclear genomes.Mitochondrial phylogeny further indicated that the present subspecies structure was a result of multiple waves of population divergence from a common ancestor.Based on our findings,we propose that criteria for subspecies delineation should be based on evolutionary independence,trait distinction,and geographic isolation.We formally defined and described eight subspecies of mainland A.cerana.Elucidation of the evolutionary history and subspecies boundaries enables a customized conservation strategy for both widespread and endemic honeybee conservation units,guiding colony introduction and breeding.

中华蜜蜂养殖业在我国的发展方向和解决方案

分析中华蜜蜂养殖规模小和多点性特点,提出解决我国中华蜜蜂养殖业的方案。建议采用活节叠加式仿生蜂箱,使养蜂技术标准化和简单化、门槛降低,可扩大养殖的规模;使用网络技术、大数据分析技术实现养殖的精准管理;最终在国家政策的引领下,实现中华蜜蜂产业集团化。

多花黄精花粉萌发与传粉生物学特性

以同质园移栽的多花黄精Polygonatum cyrtonema Hua为研究对象,对其开花物候、访花昆虫种类、访花行为及传粉过程进行观察,并采用单因素和正交设计研究不同培养基(蔗糖、H_(3)BO_(3)、CaCl_(2))对多花黄精离体花粉萌发的影响,比较I_(2)-KI染色法、MTT染色法与TTC染色法测定花粉活力的差异,检测其花粉活力与柱头可授性,并统计其自然结实率。结果表明:(1)多花黄精花期为4月末到6月初,盛花期在5月中旬,单株、花序和单花水平的花期依次为8~13、3~7、1~3 d,开花振幅为多峰曲线式样,开花同步性较高,花期同步指数为0.81±0.12;(2)离体花粉萌发的最适条件是150.00 g/L蔗糖+200.00 mg/L H_(3)BO_(3)+250.00 mg/L CaCl_(2);(3)TTC染色法是快速且适宜的测定多花黄精离体花粉活力的方法;(4)花粉在开花前1~2 d开始有活力,在开花后2~3 d花粉活力最高,此后随着花药散粉活力逐渐下降,花枯萎后花粉活力丧失;(5)柱头在开花前1~2 d至开花后4~5 d均有可授性,其中开花后2~3 d柱头可授性最高;(6)多花黄精的传粉昆虫为中华蜜蜂Apis cerana、黑带食蚜蝇Episyrphus balteatus和熊蜂Bombus latreille,中华蜜蜂是其主要的传粉昆虫;(7)多花黄精自然结实率为46.35%。

重庆彭水县中华蜜蜂形态指标测定及比较研究

采集重庆彭水县中华蜜蜂(Apis cerana cerana)主要栖息区内18个乡镇的蜜蜂样本,对工蜂吻长、前翅长等6项主要形态指标进行测定,并将彭水县中华蜜蜂形态指标数据与湖北、江西等17地已报道的中华蜜蜂形态指标进行比较。结果显示,彭水县中华蜜蜂的翅长、翅宽、T3+T4背板长平均值处于中上水平,暗示其采集能力和花蜜储存能力有一定优势。试验结果可为当地中华蜜蜂遗传资源调查与保护提供形态学数据参考。