不同蜂花粉对西方蜜蜂蜂王浆主蛋白基因表达及蜂王浆产量、质量和组成的影响

Effects of Different Bee Pollens on Expression of Major Royal Jelly Protein Genes and Yield,Quality and Composition of Royal Jelly of Apis mellifera

本试验旨在研究不同蜂花粉对西方蜜蜂(Apis mellifera)蜂王浆主蛋白基因(mrjps)表达及蜂王浆产量、质量和组成的影响。选取蜂王、群势基本一致的意大利蜜蜂9群,随机分为3个组,每组3群蜜蜂:油菜花粉组,饲喂油菜花粉;土连翘花粉组,饲喂土连翘花粉;对照组,不饲喂花粉,自然采粉(主要为鬼针草花粉)。试验周期为2个月,试验时间为2017年9月中旬至11月中旬。利用凯氏定氮法和水解法分别检测油菜花粉、土连翘花粉和鬼针草花粉粗蛋白质含量和氨基酸组成与含量;每组选3只9日龄工蜂进行mrjp1~mrjp9和黄体基因e3(yellow⁃e3)相对表达量的检测;检测各组蜂王浆产量、质量及水溶性蛋白和氨基酸组成与含量。结果显示:3种蜂花粉粗蛋白质含量、总氨基酸含量均为油菜花粉>土连翘花粉>鬼针草花粉,且组间差异显著(P<0.05),3种蜂花粉单一氨基酸(组氨酸除外)含量也表现为油菜花粉>土连翘花粉>鬼针草花粉,且组间差异显著(P<0.05)。在9日龄工蜂头部,mrjp1、mrjp2、mrjp3、mrjp4、mrjp5和mrjp7相对高表达,mrjp6、mrjp8和mrjp9相对低表达,油菜花粉组、土连翘花粉组和对照组mrjp1、mrjp2、mrjp3、mrjp4、mrjp5和mrjp7的相对表达量之和分别占mrjps总相对表达量的99.94%、99.94%和98.60%。与对照组相比,饲喂油菜花粉和土连翘花粉显著提高了mrjp1、mrjp2、mrjp3、mrjp4、mrjp5、mrjp7和yellow⁃e3的相对表达量(P<0.05),且油菜花粉组上述基因的相对表达量显著高于土连翘花粉组(P<0.05);饲喂油菜花粉和土连翘花粉显著降低了mrjp6、mrjp8、mrjp9的相对表达量(P<0.05)。油菜花粉组蜂王浆产量最高,其次为土连翘花粉组,对照组最低,各组之间差异显著(P<0.05)。油菜花粉组和土连翘花粉组蜂王浆的质量与对照组相比无显著差异(P>0.05)。3组蜜蜂所产蜂王浆水溶性蛋白和氨基酸组成基本一致。综上所述,不同粗蛋白质含量的蜂花粉对西方蜜蜂mrjps基因的表达及蜂王浆产量的影响不同,对蜂王浆质量和组成的影响不大。3组蜂群相比,采食油菜花粉组具有营养功能的mrjps基因的相对表达量和蜂王浆产量最高。由此可见,对于西方蜜蜂繁育或蜂王浆生产,油菜花粉可作为一种优质的营养饲料。

This experiment was conducted to study the effects of different bee pollens on the expression of ma⁃jor royal jelly protein genes(mrjps)and the yield,quality and composition of royal jelly of Apis mellifera.Nine colonies of Italian honeybee with approximately the same queen and colony potential were selected and randomly divided into three groups,and each group had three colonies of bees.Bees in Brassica napus pollen group were fed with Brassica napus pollen,bees in Hymenodictyon flaccidum pollen group were fed with Hym⁃enodictyon flaccidum pollen,and those in control group were not fed pollen,but collected pollen(mainly Bi⁃dens pilosa pollen)naturally.The experiment period was 2 months,from mid⁃September to mid⁃November,2017.The content of crude protein and the contents and composition of amino acids in Brassica napus pollen,Hymenodictyon flaccidum pollen and Bidens pilosa pollen were determined by Kjeldahl method and hydrolysis method,respectively.Three 9⁃day⁃old worker bees were selected from each group to determine the relative ex⁃pression levels of mrjp1 to mrjp9 and yellow⁃e3,and the yield and quality,the contents and composition of wa⁃ter⁃soluble proteins and amino acids of royal jelly from each group were measured.The results showed as fol⁃lows:the contents of crude protein and total amino acids of the three bee pollons were Brassica napus pollon>Hymenodictyon flaccidum pollon>Bidens pilosa pollon,and differences were significant among groups(P<0.05);all the contents of single amino acids(except histidine)of the three bee pollons were Brassica napus pollon>Hymenodictyon flaccidum pollon>Bidens pilosa pollon,and differences were significant among groups(P<0.05).In the head of 9⁃day⁃old bees,the mrjp1,mrjp2,mrjp3,mrjp4,mrjp5 and mrjp7 had relatively high expression,while the mrjp6,mrjp8 and mrjp9 had relatively low expression.The total relative expression level of the mrjp1,mrjp2,mrjp3,mrjp4,mrjp5 and mrjp7 accounted for 99.94%,99.94%and 98.60%of the total relative expression level of mrjps in Brassica napus pollen group,Rosa ragosa pollen group and con⁃trol group,respectively.Compared with the control group,feeding Brassica napus pollen and Rosa ragosa pol⁃len significantly increased the relative expression levels of mrjp1,mrjp2,mrjp3,mrjp4,mrjp5,mrjp7 and yellow⁃e3(P<0.05),but significantly decreased the relative expression levels of mrjp6,mrjp8 and mrjp9(P<0.05).The royal jelly yield of Brassica napus pollen group was the highest,the Rosa ragosa pollen group was higher and the control group was the lowest,and differences were significant among groups(P<0.05).How⁃ever,there was no significant difference in the quality of royal jelly produced by bees from of Brassica napus pollen group and Rosa ragosa pollen group compared with the control group(P>0.05),and the composition of water⁃soluble protein and amino acids in three kinds of royal jelly were basically the same.In conclusion,the effects of bee pollen with different crude protein contents on the expression of mrjps and the yield of royal jelly are different,but the effects of the three groups of bee pollens on the quality and composition of royal jelly are not significant.Comparison of three groups of bee colonies,the expression of mrjps with nutritional func⁃tion and the yield of royal jelly in the Brassica napu pollen group were the highest.Collectively,for the Apis mellifera breeding or royal jelly production,the Brassica napu pollen can be used as a high⁃quality nutritional feed.