摘要
本研究旨在揭示基础日粮中不同浓度的卷丹百合珠芽对藏猪卵巢卵泡发育及相关LncRNA表达的影响。将同一批次健康的体重为(17.40±1.15)kg的9头雌性西藏藏猪,随机分为BW组(基础日粮)、BY组(基础日粮+1%百合珠芽)和BR组(基础日粮+2%百合珠芽),饲喂6周后,利用石蜡切片HE染色观察卵巢形态,测定卵巢器官指数和不同类型卵泡数量,并利用高通量测序技术对lncRNA进行转录组测序分析,最后对差异lncRNA进行荧光定量验证。结果显示:(1)卵巢表面与卵泡特征:BR组,藏猪卵巢表面出现大卵泡;卵巢指数,各组间无显著差异;卵泡比例,BW组原始卵泡比例最高,BY组二级卵泡比例最高,BR组成熟卵泡比例最高。(2)lncRNA筛选与表达:新预测lncRNA,筛选出5772个新预测的lncRNA;表达量分析,BY组平均表达量最高,BW组最低,3组表达趋势相似;组间相关性,BY组、BW组和BR组3个组间相关性较强。(3)差异表达分析:BW_vs_BY,筛选到2021个差异表达lncRNA基因(BY组相对BW组上调1197个,下调824个);BW_vs_BR,筛选到1308个差异表达lncRNA基因(BR组相对BW组上调829个,下调479个);BY_vs_BR,筛选到1330个差异表达lncRNA基因(BR组相对BY组上调646个,下调684个)。(4)lncRNA靶基因预测与功能分析:靶基因预测,预测得到CCNL2、MAP3K14、STUB1、IQANK1和PKP3;靶基因GO和KEGG富集分析,发现cAMP、MAPK、PI3K-Akt、Wnt等与卵巢发育信号相关的通路都被显著富集到,其中相关信号通路的基因有LOC110261342、MSTRG.26467、FXN、CLP1、Gem、PODNL1、TGFBR3L等。结果表明,在饲料中添加百合珠芽(特别是2%浓度)有助于促进藏猪卵巢成熟卵泡的发育,并显著调控lncRNA表达,参与卵巢发育的多条生物学通路,为进一步理解百合珠芽对藏猪生殖能力的提升提供了基础数据和理论支持。
This study aimed to reveal the effects of different concentrations of the bulbils of Lilium lancifolium in the basic diet on the development of ovarian follicles and the expression of related lncrnas in Xizang pigs.Xizang pigs were used as the research object,and the same batch of nine healthy Xizang pigs with a weight of(17.40±1.15)kg was randomly divided into three groups,of which the BW group was the control group,fed on the basal diet,the BY group and the BR group were the experimental groups,with 1%concentration lily bead buds and 2%concentration lily bead buds added to the basal diet,respectively,and after 6 weeks of experimental feeding,morphological observation,ovarian organ index measurement,and statistics of the number of different types of follicles in the ovaries were carried out to judge the ovarian development.The high-throughput omics transcriptome technology was used to sequence the ovarian LncRNA,and the differential LncRNA was verified in time quantitatively to verify the reliability of the transcriptome sequencing results.The results showed that Ovarian surface and follicle characteristics:In the BR group,large follicles appeared on the ovarian surface of Xizang pigs;there was no significant difference in the ovarian index among the groups.Follicle ratio analysis revealed that the BW group had the highest proportion of primordial follicles,the BY group had the highest proportion of secondary follicles,and the BR group had the highest proportion of mature follicles.LncRNA screening and expression revealed 5,772 new lncRNAs.Expression analysis revealed that the BY group displayed the highest average expression level,whereas the BW group had the lowest expression level,and the three groups had similar expression trends,with strong correlations among the BY,BW,and BR groups.Differential expression analysis revealed that the BW vs.BY comparison identified 2,021 differentially expressed lncRNA genes(1,197 upregulated and 824 downregulated in the BY group compared with the BW group);the BW vs.BR comparison identified 1,308 differentially expressed lncRNA genes(829 upregulated and 479 downregulated in the BR group compared with the BW group);the BY vs.BR comparison screened 1,330 differentially expressed lncRNA genes(646 upregulated and 684 downregulated in the BR group compared with the BY group);and LncRNA target gene prediction and functional analysis:Target gene prediction identified CCNL2,MAP3K14,STUB1,IQANK1,and PKP3.GO and KEGG enrichment analyses of target genes revealed significant enrichment of cAMP,MAPK,PI3KAkt,Wnt,and other ovarian development signaling pathways.The genes related to these pathways included LOC110261342,MSTRG.26467,FXN,CLP1,Gem,PODNL1,and TGFBR3L.The results showed that the addition of Lilium bulbiferous bulbs to the diet,especially at a 2%concentration,promoted the development of mature ovarian follicles in Xizang pigs and significantly regulated the expression of long noncoding RNAs(lncRNAs),which were involved in multiple biological pathways related to ovarian development.These results indicated that this study would provide fundamental data and theoretical support for further understanding the enhancement of Xizang pig reproductive capacity by Lilium bulbiferous bulbs.
作者
任子利
曹婷婷
阚勇
王雨佳
赵彦玲
REN Zili;CAO Tingting;KAN Yong;WANG Yujia;ZHAO Yanling(Animal Science College,Xizang Agriculture&Animal Husbandry University;Xizang Key Laboratory of Genetic Improvement and Reproduction Engineering of Xizang Pigs,Linzhi,Xizang 860000,China)
出处
《高原农业》
2025年第3期277-293,F0002,共18页
Journal of Plateau Agriculture
基金
国家自然科学基金资助项目[藏猪精子高原适应性的DNA甲基化机制(31760669)]
藏猪睾丸高原适应性的DNA甲基化机制(31660658)
西藏自治区科技厅重点研发计划项目[藏羚羊高原适应的特殊遗传机制研究(XZ202402ZY0025)]。
关键词
lncRNA
卷丹百合珠芽
西藏藏猪
靶基因
Long Noncoding RNA
Lilium bulbiferous bulbs
Xizang pigs
Target genes
作者简介
任子利(1969-),男,汉族,河南卫辉人,西藏农牧学院教授,博士,硕士生导师,研究方向:主要从事动物繁殖方面的研究。美国加州大学圣地亚哥分校访问学者。现任中国动物学会生殖生物学分会理事、中国畜牧兽医学会家畜生态学分会理事、西藏畜牧兽医学会理事,西藏自治区科协科技创新智库专家、林芝市标准化专家组专家,藏猪遗传改良与繁殖工程重点实验室管理团队成员。主要从事高原动物育种与繁殖、动物资源保护开发与利用的教学与科研工作。先后主持国家自然科学基金1项、西藏自治区科技厅项目4项、西藏自治区教育厅项目1项、微生物技术国家重点实验室(山东大学)开放课题1项,先后参加国家自然科学基金、科技部等科研项目10余项,获西藏自治区高等学校教学成果奖“三等奖”2项,以第一作者或通讯作者在《Proteome Science》、《Italian Journal of Animal Science》、《畜牧兽医学报》等期刊上发表论文40余篇,获发明专利1项,实用新型专利5项,主编教材、专著2部,参编教材、专著6部。
