幼虫饲粮中添加叶酸对雌性意大利蜜蜂DNA甲基化及发育的影响  

Effect of adding folic acid to the larval diet of Apis mellifera ligustica on DNA methylation and female development

作　　者：景洪雨 姚玉风 韩凯 雷丽 孙桂云 王红芳[1] 胥保华[1] JING Hong-Yu;YAO Yu-Feng;HAN Kai;LEI Li;SUN Gui-Yun;WANG Hong-Fang;XU Bao-Hua(College of Animal Science and Technology,Shandong Agricultural University,Tai'an 271018,China)

机构地区：[1]山东农业大学动物科技学院,泰安271018

出　　处：《应用昆虫学报》2022年第3期587-598,共12页Chinese Journal of Applied Entomology

基　　金：泰山产业领军人才高效生态农业创新类项目(项目编号:LJNY202003);国家现代农业产业技术体系(CARS-44)。

摘　　要：【目的】本文旨在探究意大利蜜蜂Apis mellifera ligustica幼虫饲粮中添加叶酸对雌性蜜蜂DNA甲基化及发育的影响。【方法】从姊妹蜂群中选用2日龄意大利蜜蜂雌性幼虫,平均分为6组,任选一组以饲喂不添加叶酸的基础日粮为对照组(CK),其余5组为试验组,分别饲喂添加0.02%、0.04%、0.06%、0.08%和0.10%叶酸的基础日粮。在室内温度(34.5±0.5)℃,相对湿度为90%±5%的条件下,按照饲养流程饲养意大利蜜蜂至出房。取3-5日龄的幼虫,测定叶酸代谢指标及DNA甲基化相关指标,计算新蜂发育历期,称量新蜂重。【结果】(1)添加叶酸(FA)水平为0.04%的试验组,3、4和5日龄幼虫蜂体内叶酸(FA)和5-甲基四氢叶酸(5-MTHF)含量显著高于对照组(P<0.05),并且显著提高了幼虫时期蜂体的二氢叶酸还原酶(DHFR)基因、丝氨酸羟甲基转移酶(SHMT)基因和亚甲基四氢叶酸还原酶(MTHFR)基因的表达量(P<0.05),显著提高了MTHFR酶活性(P<0.05)。(2)与对照组相比,添加叶酸为0.04%的试验组显著提高了3日龄幼虫DNA甲基转移酶1a(Dnmt1a)基因的表达量(P<0.05),且显著提高了3日龄和4日龄幼虫的DNA甲基转移酶1(DNMT1)酶活性(P<0.05)。添加0.04%的叶酸试验组相较于对照组显著降低了3日龄幼虫DNA甲基转移酶3(DNMT3)的酶活性(P<0.05),显著降低了4日龄幼虫Dnmt3基因的表达量和DNMT3酶活性(P<0.05)。叶酸添加剂量为0.04%时,4日龄蜂体DNA甲基化水平显著降低(P<0.05)。(3)与对照组相比,添加0.04%的叶酸试验组的意大利蜜蜂平均发育历期(19.95 d)显著小于对照组(P<0.05),平均新蜂重(0.17 g)显著大于对照组(P<0.05)。【结论】在基础日粮中添加适量叶酸能够降低4日龄意大利蜜蜂幼虫的DNA甲基化水平,并能缩短发育历期和增加新蜂重。在饲粮中添加0.04%的叶酸有助于雌性意大利蜜蜂幼虫向蜂王方向发育。[Objectives]To investigate the effect of adding folic acid to the larval diet of Apis mellifera ligustica on DNA methylation and female development.[Methods]Two-day-old female A.m.ligustica larvae were randomly selected from sister colonies and divided into six equal-sized groups.The control group(CK)was fed a diet without additional folic acid,whereas the remaining five groups were fed the same diet with the addition of either 0.02%,0.04%,0.06%,0.08% or 0.10% folic acid.Larvae were reared at room temperature of(34.5±0.5)℃ at a relative humidity of 90%±5% until they emerged.Indices of folic acid metabolism and DNA methylation related indices were measured in 3-to 5-day-old larvae,and the developmental duration and weight of newly emerged bees was recorded.[Results](1)The 0.04%treatment group had significantly higher levels of folic acid(FA)and 5-methyltetrahydrofolate(5-MTHF)than the control group at 3,4 and 5 days of age(P<0.05).This group also had significantly higher expression of the dihydrofolate reductase(DHFR)gene,the serine hydroxymethyltransferase(SHMT)gene and the methylenetetrahydrofolate reductase(MTHFR)gene(P<0.05),and significantly higher MTHFR enzyme activity(P<0.05),in the larval period.(2)This treatment group also had significantly higher expression of the DNA methyltransferase 1a(Dnmt1a)gene in 3-day-old larvae(P<0.05),and significantly higher DNA methyltransferase 1(DNMT1)enzyme activity in 3-and 4-day-old larvae,than the control group(P<0.05).The 0.04% treatment group had significantly lower DNA methyltransferase 3(DNMT3)enzyme activity in 3-day-old larvae compared to the control group(P<0.05),and significantly lower Dnmt3 gene expression and DNMT3 enzyme activity in 4-day-old larvae(P<0.05).The level of DNA methylation in this treatment group was also significantly lower in 4-day-old bees(P<0.05).(3)The average developmental duration(19.95 d)of the 0.04%treatment group was significantly lower(P<0.05),and the average weight(0.17 g)of newly emerged bees was,significantly higher,than that of

关 键 词：意大利蜜蜂 叶酸 DNA甲基化 发育 

分 类 号：S894[农业科学—特种经济动物饲养]