首页|不同日龄内蒙古白绒山羊瘤胃及粪便真菌多样性差异研究

不同日龄内蒙古白绒山羊瘤胃及粪便真菌多样性差异研究

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本试验旨在采用高通量测序技术,测定内蒙古白绒山羊不同日龄时瘤胃及粪便真菌多样性的定植规律,为后续绒山羊早期营养调控相关研究提供数据依据.选取按照场区正常生产管理并饲养的10、30、50、150日龄体重分别为(4.99±0.13)、(7.71 士0.52)、(13.83±0.18)、(20.60±0.81)kg的内蒙古白绒山羊母羔各3只(每只为1个重复,共12只),进行屠宰、取瘤胃液(R组)及粪便(F组).采用特异性引物对ITS1F/ITS2R扩增瘤胃和粪便真菌ITS区,用Illumina Miseq PE300平台进行测序.结果表明:1)本试验24个样共获得1 636 055条优化序列(Clean tags),平均每个样含有(68 168±24 317)条,每条序列平均长度为(244±28)bp.2)随日龄的增加,瘤胃和粪便真菌总OTUs以及与后一个日龄共享的OTUs开始时稍有降低,但随后逐渐增加.10、30、50、150日龄时瘤胃和粪便共享的真菌OTUs分别占同日龄检测到的瘤胃和粪便OTU总数的34.22%、20.87%、27.63%和20.53%.3)随日龄的增加,瘤胃真菌群落Shannon、Simpson、Ace、Chao指数差异不显著(P>0.05),而50、150日龄时粪便中真菌Shannon和Simpson指数分别显著高于和低于10日龄组(P<0.05).50日龄时瘤胃中真菌Shannon和Simpson指数分别显著高于和低于粪便组(P<0.05).4)50日龄和150日龄瘤胃真菌菌群β多样性极显著聚为一类(P<0.01),并与10日龄和30日龄瘤胃真菌菌群β多样性极显著分开(P<0.01).50日龄和150日龄粪便真菌群落 β多样性分别极显著独立分布且与10日龄和30日龄粪便真菌群落β多样性分开(P<0.01).另外,150日龄瘤胃及粪便组内真菌群落β多样性相似度大于其他日龄组.5)当日龄为10、30、50、150d时,在门水平上瘤胃及粪便真菌组成相似,均为子囊菌门(Ascomycota)、未分类的真菌(Unclassified_k_fungi)、新美鞭菌门(Neocallimas-tigomycota)、担子菌门(Basidiomycota)和被孢霉门(Mortierellomycota),其中子囊菌门占绝对优势.而在属水平上瘤胃及粪便真菌组成稍有差异.瘤胃中丰度最高的物种在10、50日龄时均为未分类的真菌,在30、150日龄时分别为荚孢腔菌属(Sporormiella)和酵母菌属(Saccharomyces).粪便中丰度最高的物种10、50日龄时分别为未分类的真菌和新美鞭菌属(Neocallimastix),30、150日龄时分别为Cleistothelebolus和枝孢菌层(Cladosporium).6)在门水平上的进一步LefSe分析发现,新美鞭菌门无论是在150日龄内蒙古白绒山羊瘤胃中还是在50日龄内蒙古白绒山羊粪便中均是显著富集的物种(P<0.05);比较同日龄瘤胃和粪便真菌群落时,只有在150日龄瘤胃和粪便中分别显著富集了未分类的真菌和子囊菌门.7)基于FUNGuild功能预测,瘤胃真菌在羔羊10、50日龄时分别以未分类营养型(Unclassified)和病理营养型(Pathotroph)为主,30、150日龄时均以腐生营养型(Saprotroph)为主.粪便真菌在10、150日龄时均以病理-腐生-共生营养型(Pathotroph-Saprotroph-Symbiotroph)为主,30、50日龄时均以腐生营养型为主.由此可见,随着内蒙古白绒山羊羔羊的发育,瘤胃真菌群落逐渐有利于饲料中纤维的降解.幼龄时期瘤胃和粪便中还存在很多丰度较高且未分类的物种,其功能还未明确,因此还需要更深一步的研究.
Study on the Differences of Fungal Diversity in Rumen and Feces of Inner Mongolia Cashmere Goats at Different Ages
The purpose of this experiment was to determine the colonization of fungal diversity in rumen and feces of Inner Mongolia white cashmere goats at different ages by high-throughput se-quencing technology,and to provide data basis for subsequent research on early nutritional regu-lation of cashmere goats.Three female lambs of Inner Mongolia white cashmere goats of each days of age(10,30,50 and 150)with body weight of(4.99±0.13),(7.71±0.52),(13.83±0.18)and(20.60±0.81)kg respectively were selected according to the normal production man-agement and feeding in the farm area,each lamb was a replicate,all together 12 lambs.They were slaughtered and rumen fluid(group R)and feces(group F)were taken.The ITS region of rumen and fecal fungi was amplified by specific primers ITS1F/ITS2R,and sequenced by Illumi-na Miseq PE300 platform.The results showed that:1)A total of 1 636 055 clean tags were ob-tained from 24 samples in this experiment,with an average of(68 168±24 317)clean tags per sample and an average length of(244±28)bp per sequence.2)With the increase of age,the to-tal OTU of rumen and fecal fungi and the OTU shared with the next day-of-age decreased slightly at first,but then increased gradually.The fungal OTUs shared by rumen and feces at 10,30,50 and 150 days of age accounted for 34.22%,20.87%,27.63%and 20.53%of the total number of OTUs,respectively.3)There was no significant difference in Shannon,Simpson,Ace and Chao indexes of rumen fungal community with the increase of age(P>0.05),but the Shannon and Simpson indexes of fecal fungi at 50 and 150 days of age were significantly higher and lower than those at 10 days of age,respectively(P<0.05).At 50 days of age,the Shannon and Simpson index of fungi in rumen were significantly higher and lower than those in fecal group(P<0.05).4)The β diversity of rumen fungi at 50 days of age and 150 days of age was significantly clustered into one group(P<0.01),and was significantly separated from the β diversity of rumen fungi at 10 and 30 days of age(P<0.01).The β diversity of fecal fungi of 50-day-old and 150-day-old were significantly independent distribution and were seperated from 10-day-old and 30-day-old(P<0.01).In addition,the similarity of fungi β diversity in rumen and feces groups at 150 days of age was higher than that in other age groups.5)At 10,30,50 and 150 days of age,the compo-sition of rumen and fecal fungi was similar at the phylum level,which were Ascomycota,Unclas-sified_k_fungi,Neocallimastigomycota,Basidiomycota and Mortierellomycota,among which As-comycota was dominant.At the genus level,the composition of rumen and fecal fungi was slight-ly different.The species with the highest abundance in rumen were Unclassified_k_fungi at 10 and 50 days of age,Sporormiella and Saccharomyces at 30 and 150 days of age,respectively.The species with the highest abundance in feces were Unclassified_k_fungi and Neocallimastix at 10 and 50 days of age,respectively,and Cleistothelebolus and Cladosporium at 50 and 150 days of age,respectively.6)Further LefSe analysis at the phylum level showed that Neocallimastigo-mycota was significantly enriched in the rumen aged 150 days and in feces aged 50 days(P<0.05).When comparing the rumen and fecal fungal communities at the same age,Unclassified_k_fungi and Ascomycota were significantly enriched only in the rumen and feces at 150 days of age.7)Based on FUNGuild function prediction,rumen fungi were dominated by Unclassified and Pathotroph at 10 and 50 days of age,respectively,and Saprotroph was dominant at 30 and 150 days of age.Fecal fungi were mainly Pathotroph-Saprotroph-Symbiotroph at 10 and 150 days of age,and Saprotroph at 30 and 50 days of age.It can be concluded that with the development of Inner Mongolia Cashmere Goat lambs,the rumen fungal community is gradually conducive to the degradation of fiber in the feed,and the digestion ability is significantly enhanced.There are still many high-abundance and unclassified species in the rumen and feces of young age,and their functions are not yet clear,so further research is needed.

high-throughput sequencing technologyInner Mongolia white cashmere goatdiffer-ent days of agerumenfecesfungal diversity

娜梅拉、李科南、杜海东、郭文亮、娜仁花

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内蒙古农业大学动物科学学院,呼和浩特 010018

高通量测序技术 内蒙古白绒山羊 不同日龄 瘤胃 粪便 真菌多样性

内蒙古自治区自然科学基金

2021MS03009

2024

畜牧兽医学报
中国畜牧兽医学会

畜牧兽医学报

CSTPCD北大核心
影响因子:0.729
ISSN:0366-6964
年,卷(期):2024.55(8)
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