人工饲粮对幼年工蜂生长发育及生理代谢的影响

Effects of Artificial Diets on Growth and Development and Physiological Metabolism of Young Worker Bees

本试验利用液相色谱-质谱联用技术(LC-MS)对幼年工蜂进行代谢组学分析,旨在探究不同人工饲粮对幼年工蜂生长发育及生理代谢的影响,为蜂王邮寄的人工饲粮配制提供一定理论依据。试验选取1日龄的意大利蜜蜂(Apis mellifera ligustica)300只,分为5组,分别饲喂5种人工饲粮,每组30只工蜂。对照组饲粮:蔗糖粉∶蜂蜜=3∶1;试验组A饲粮:蔗糖粉∶蜂粮∶蜂蜜=6∶5∶1;试验组B饲粮:蔗糖粉∶蜂粮∶蜂蜜=6∶3∶1;试验组C饲粮:蔗糖粉∶花粉∶蜂蜜=6∶5∶1;试验组D饲粮:蔗糖粉∶花粉∶蜂蜜=6∶3∶1。用5种人工饲粮进行室内喂养,记录蜜蜂7 d内的死亡数量;解剖试验组A和对照组第1、3、5、7天的工蜂咽下腺,测定其平均重量、颜色和饱满度;采用LC-MS方法检测试验组A和对照组蜜蜂饲喂7 d后的代谢物差异,对检测数据进行模式识别分析,筛选并鉴别差异代谢物。结果表明:1)对照组和试验组A蜜蜂第1~5天时均没有出现死亡,在第6和7天时试验组A蜜蜂死亡数量均显著低于对照组(P<0.05)。试验组B、C、D的蜜蜂在6 d内全部死亡。2)试验组A第7天时的蜜蜂咽下腺平均重量显著高于对照组(P<0.05),咽下腺颜色为全部乳白色,且咽下腺小体饱满。3)采用主成分分析(PCA)、偏最小二乘法判别方法(PLS-DA)和正交偏最小二乘方-判别分析(OPLS-DA)分析代谢组数据,结果显示试验组A和对照组明显分离,共鉴定出23个差异代谢物,包括氨基酸、脂类、糖类等,其中有10种代谢物上调,13种代谢物下调。差异代谢物通路分析发现,差异极显著的通路有牛磺酸和亚牛磺酸生物合成通路、赖氨酸降解通路、戊糖磷酸盐代谢通路、戊糖和葡萄糖醛酸相互转化通路(P<0.01),差异显著的通路有氨基酸的生物合成通路,色氨酸代谢通路,碳代谢通路,丁酸代谢通路,丙氨酸、天冬氨酸和谷氨酸代谢通路,丙酮酸代谢通路(P<0.05)。由此可见,试验组A的饲粮更适合蜜蜂工蜂的生长发育和生存,在邮寄环境中可代替常用的炼糖饲粮。

To study the effects of different artificial diets on growth and development and physiological metabolism of young worker bees,the metabonomics analysis was conducted for worker bees based on liquid chromatography-mass spectrometry(LC-MS),and to provide some theoretical basis for the preparation of artificial diet of mailed queen bee.A total of 300 one-day old Apis mellifera ligustica was selected and divided to 5 groups with 30 worker bees in each group,and fed with Five kinds of artificial diets.Control group diet:sucrose powder∶honey=3∶1;experimental group A diet∶sucrose powder∶bee feed∶honey=6∶5∶1;experimental group B diet:sucrose powder∶bee feed∶honey=6∶3∶1;experimental group C diet:sucrose powder∶pollen∶honey=6∶5∶1;experimental group D diet:sucrose powder∶pollen∶honey=6∶3∶1.Five kinds of artificial diet were used for indoor feeding,recorded the death number of honeybees within 7 days.The hypopharyngeal glands of worker bees on days 1,3,5 and 7 were dissected and measured the average weighing,color and plumpness.The metabolite differences of honeybees after feeding 7 days between experimental group A and control group were detected by LC-MS method,the detected data were carry out the pattern recognition analysis,screened and identified the different metabolites.The results showed as follows:1)the honeybees in control group and experimental group A were no death during days 1 to 5,the death number of honeybees on days 6 and 7 of experimental group A was significantly lower than that of the control group(P<0.05).The honeybees in experimental groups B,C and D were all died within 6 days.2)The mean weight of hypopharyngeal gland of honeybees on day 7 of experimental group A was significantly higher than that of the control group(P<0.05),the hypopharyngeal gland colour were all milky white,and hypopharyngeal gland corpuscle was plump.3)The metabolomic data were analyzed by principal component analysis(PCA),partial leastsquares discriminant analysis(PLS-DA)and orthogonal partial leastsquares-discriminant analysis(OPLS-DA),the results showed that the experimental group A and the control group were significantly separated,the 23 different metabolites were identified,included amino acid,lipid and saccharides,among then,10 metabolites were up-regulated and 13 metabolites were down-regulated.The differential metabolite pathways analysis showed that the pathways with significant differences were taurine and hypotaurine metabolism pathway,alysine degradation pathway,pentose phosphate metabolism pathway and pentose and glucuronate interconversions pathway(P<0.01),and the pathways with significant differences were biosynthesis of amino acids pathway,tryptophan metabolism pathway,carbon metabolism pathway,butanoate metabolism pathway,alanine,aspartate and glutamate metabolism pathway and pyruvate metabolism pathway(P<0.05).In conclusion,the diet of experimental group A is more suitable for the growth,development and survival of worker bees,it can replace the common sugar refining diet in mailing environment.