特种经济动物营养与饲料 SPECIAL ECONOMIC ANIMAL NUTRITION AND FEED

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

  • 荀利杰 ,
  • 黄新球 ,
  • 李琼艳 ,
  • 杨爽 ,
  • 王艳辉
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  • 云南省农业科学院蚕桑蜜蜂研究所, 蒙自 661101
荀利杰(1984-),男,河北邢台人,助理研究员,硕士,研究方向为蜜蜂分子生物学。E-mail:xunlijiecom@163.com

收稿日期: 2019-07-18

  网络出版日期: 2020-02-21

基金资助

国家蜂产业技术体系专项(CARS-44-SYZ16);云南省科技厅重点新产品开发计划项目(2015BB010)

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

  • XUN Lijie ,
  • HUANG Xinqiu ,
  • LI Qiongyan ,
  • YANG Shuang ,
  • WANG Yanhui
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  • Institute of Sericulture and Apiculture, Yunnan Academy of Agricultural Sciences, Mengzi 661101, China

Received date: 2019-07-18

  Online published: 2020-02-21

摘要

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。

本文引用格式

荀利杰 , 黄新球 , 李琼艳 , 杨爽 , 王艳辉 . 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[J]. 动物营养学报, 2020 , 32(2) : 856 -869 . DOI: 10.3969/j.issn.1006-267x.2020.02.041

Abstract

This experiment was conducted to study the effects of different bee pollens on the expression of major 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 Hymenodictyon flaccidum pollen, and those in control group were not fed pollen, but collected pollen (mainly Bidens 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 expression levels of mrjp1 to mrjp9 and yellow-e3, and the yield and quality, the contents and composition of water-soluble proteins and amino acids of royal jelly from each group were measured. The results showed as follows: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 control group, respectively. Compared with the control group, feeding Brassica napus pollen and Rosa ragosa pollen 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). However, 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 function 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.

参考文献

[1] NOZAKI R,TAMURA S,ITO A,et al.A rapid method to isolate soluble royal jelly proteins[J].Food Chemistry,2012,134(4):2332-2337.  
[2] YOU Z Y,QIAN Q J,WANG Y R,et al.Transgenic silkworms secrete the recombinant glycosylated MRJP1 protein of Chinese honeybee,Apis cerana cerana[J].Transgenic Research,2017,26(5):653-663.  
[3] TAKENAKA T,ECHIGO T.Chemical composition of royal jelly[J].Honeybee Sci,1982,3(2):69-74.
[4] ALBERT S,BHATTACHARYA D,KLAUDINY J,et al.The family of major royal jelly proteins and its evolution[J].Journal of Molecular Evolution,1999,49(2):290-297.  
[5] CHEN C,CHEN S Y.Changes in protein components and storage stability of royal jelly under various conditions[J].Food Chemistry,1995,54(2):195-200.  
[6] LI J K,WANG T,ZHANG Z H,et al.Proteomic analysis of royal jelly from three strains of western honeybees (Apis mellifera)[J].Journal of Agricultural and Food Chemistry,2007,55(21):8411-8422.  
[7] TAKENAKA T,ECHIGO T.Proteins and peptides in royal jelly[J].Journal of the agricultural Chemical Society of Japan,1983,57(12):1203-1209.  
[8] SCHMITZOVÁ J,KLAUDINY J,ALBERT Š,et al.A family of major royal jelly proteins of the honeybee Apis mellifera L.[J].Cellular and Molecular Life Sciences CMLS,1998,54(9):1020-1030.  
[9] BUTTSTEDT A,MORITZ R F,ERLER S.Origin and function of the major royal jelly proteins of the honeybee (Apis mellifera) as members of the yellow gene family[J].Biological Reviews,2014,89(2):255-269.  
[10] DRAPEAU M D,ALBERT S,KUCHARSKI R,et al.Evolution of the yellow/major royal jelly protein family and the emergence of social behavior in honey bees[J].Genome Research,2006,16(11):1385-1394.  
[11] KAMAKURA M.Royalactin induces queen differentiation in honeybees[J].Nature,2011,473(7348):478-483.  
[12] THOMPSON G J,KUCHARSKI R,MALESZKA R,et al.Towards a molecular definition of worker sterility:differential gene expression and reproductive plasticity in honey bees[J].Insect Molecular Biology,2006,15(5):537-644.  
[13] CHEN X,HU Y,ZHENG H Q,et al.Transcriptome comparison between honey bee queen- and worker-destined larvae[J].Insect Biochemistry and Molecular Biology,2012,42(9):665-673.  
[14] 李晓燕,杨文静,王琦,等.西方蜜蜂(Apis mellifera L.)体内王浆蛋白MRJP6基因的表达[J].基因组学与应用生物学,2016,35(6):1415-1420.
[15] BUTTSTEDT A,MORITZ R F A,ERLER S.More than royal food-major royal jelly protein genes in sexuals and workers of the honeybee Apis mellifera[J].Frontiers in Zoology,2013,10:72.
[16] BLANK S,BANTLEON F I,MCINTYRE M,et al.The major royal jelly proteins 8 and 9(Api m 11) are glycosylated components of Apis mellifera venom with allergenic potential beyond carbohydrate-based reactivity[J].Cliznical & Experimental Allergy,2012,42(6):976-985.  
[17] ALBERT Š,KLAUDINY J.The MRJP/YELLOW protein family of Apis mellifera:identification of new members in the EST library[J].Journal of Insect Physiology,2004,50(1):51-59.  
[18] SHEN L R,XING L P,YANG Y X,et al.Sequence analysis of functional apisimin-2 cDNA from royal jelly of Chinese honeybee and its expression in Escherichia coli[J].Asia Pacific Journal of Clinical Nutrition,2007,16(Suppl.1):222-226.
[19] 刘建涛,赵利,苏伟,等.蜂花粉生物活性物质的研究进展[J].食品科学,2006,27(12):909-912.
[20] LINSKENS H F,JORDE W.Pollen as food and medicine:a review[J].Economic Botany,1997,51(1):78-86.  
[21] ALMEIDA-MURADIAN L B,PAMPLONA L C,COIMBRA S,et al.Chemical composition and botanical evaluation of dried bee pollen pellets[J].Journal of Food Composition and Analysis,2005,18(1):105-111.  
[22] EL-WAHAB T E A,GOMAA A M.Application of yeast culture (Candida tropicalis) as pollen substitute in feeding honey bee colonies (Apis mellifera L.) in egypt[J].Journal of Applied Sciences Research,2005,1(5):386-390.
[23] HUANG Z Y.Pollen nutrition affects honey bee stress resistance[J].Terrestrial Ecology,2012,5(2):1-15.
[24] 陈盛禄.中国蜜蜂学[M].北京:中国农业出版社,2001:209-214.
[25] SHUMKOVA R,ZHELYAZKOVA I.Stimulating feeding and development of hypopharyngeal and thoracic glands of honeybees (Apis mellifera L.)[J].Agricultural Science and Technology,2015,7(3):309-312.
[26] CRAILSHEIM K.Interadult feeding of jelly in honeybee (Apis mellifera L.) colonies[J].Journal of Comparative Physiology B,1991,161(1):55-60.  
[27] STANDIFER L N.Honey bee nutrition and supplemental feeding[J].Beekeeping in the United States,Agriculture Handbook,1980,335:39-45.
[28] CRAILSHEIM K.The protein balance of the honey bee worker[J].Apidologie,1990,21(5):417-429.  
[29] 吴银松.云南蜜蜂科学饲养[M].昆明:云南科技出版社,2019:47.
[30] 王改英.意大利蜜蜂产浆期代用花粉中适宜蛋白质水平的研究[D].硕士学位论文.泰安:山东农业大学,2012:41.
[31] 李晓燕,梁庆环,杨文静,等.王浆主蛋白MRJP7基因在意大利蜜蜂体内的表达[J].环境昆虫学报,2016,38(2):371-377.
[32] LIU F,LI W F,ZHANG S G,et al.High-abundance mRNAs in Apis mellifera:comparison between nurses and foragers[J].Journal of Insect Physiology,2011,57(2):274-279.  
[33] ALBERT Š,KLAUDINY J.MRJP9,an ancient protein of the honeybee MRJP family with non-nutritional function[J].Journal of Apicultural Research,2007,46(2):99-104.
[34] PEIREN N,DE GRAAF D C,VANROBAEYS F,et al.Proteomic analysis of the honey bee worker venom gland focusing on the mechanisms of protection against tissue damage[J].Toxicon,2008,52(1):72-83.  
[35] 丁桂玲.蜜蜂的守卫行为[J].中国蜂业,2012,63(31):49,53.
[36] KUMOVA U,KORKMAZ A,BERKIN Ö,et al.An investigation about the effects of various factors on royal jelly production in different honeybee (Apis mellifera L.) genotypes[J].Mellifera,2005,5(9):56-64.
[37] SAHINLER N,KAFTANOGLU O.The effects of season and honey bee (Apis mellifera) genotype on grafting rates and royal jelly production[J].Turkish Journal of Veterinary and Animal Sciences,2005,29(2):499-503.
[38] DE GROOT A P.Protein and amino acid requirements of the honeybee (Apis mellifica L.)[J].Physiologia Comparata et Oecologia,1953,3:197-285.
[39] PERNAL S F,CURRIE R W.Pollen quality of fresh and 1-year-old single pollen diets for worker honey bees (Apis mellifera L.)[J].Apidologie,2000,31(3):387-409.  
[40] STANDIFER L N.A comparison of the protein quality of pollens for growth-stimulation of the hypopharyngeal glands and longevity of honey bees,Apis mellifera L.(Hymenoptera:apidae)[J].Insectes Sociaux,1967,14(4):415-425.  
[41] MAURIZIO A.Pollenernährung und lebensvorgänge bei der honigbiene (Apis mellifica L.)[J].Landwirtschaftliches Jahrbuch der Schweiz,1954,68:115-182.
[42] 程艳华,刘亚男,胡福良,等.蛋白质营养水平对蜂王浆产量和成分的影响[J].蜜蜂杂志,2008,28(4):7-9.
[43] ROULSTON T A H,CANE J H.The effect of pollen protein concentration on body size in the sweat bee Lasioglossum zephyrum (Hymenoptera:Apiformes)[J].Evolutionary Ecology,2002,16(1):49-65.  
[44] 胡福良,李英华,朱威.影响蜂王浆质量的因素——兼析春季油菜王浆质优的原因[J].蜜蜂杂志,2004(6):28-29.
[45] CRICK F H.On protein synthesis[J].Symposia of the Society for Experimental Biology,1958,12:138-163.
[46] CRICK F H.Central dogma of molecular biology[J].Nature,1970,227(5258):561-563.  
[47] LIU Y,BEYER A,AEBERSOLD R.On the dependency of cellular protein levels on mRNA abundance[J].Cell,2016,165(3):535-550.  
[48] BIESSMANN H.Molecular analysis of the yellow gene (y) region of Drosophila melanogaster[J].Proceedings of the National Academy of Sciences of the United States of America,1985,82(21):7369-7373.  
[49] BREHME K S.The effect of adult body color mutations upon the larva of drosophila melanogaster[J].Proceedings of the National Academy of Sciences of the United States of America,1941,27(6):254-261.  
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