兽类学报 ›› 2026, Vol. 46 ›› Issue (3): 337-348.DOI: 10.16829/j.slxb.151032
马英1, 魏有文1, 江峰2, 张爱萍1, 高红梅2, 李积德1, 王文龙1, 张鹏辉1, 李玲雯1, 谷昕玥1, 张同作2(
)
收稿日期:2024-11-28
接受日期:2025-02-24
出版日期:2026-05-30
发布日期:2026-06-04
通讯作者:
张同作
作者简介:马英(1973- ),女,硕士,主要从事鼠疫防控和动物遗传学研究;基金资助:
Ying MA1, Youwen WEI1, Feng JIANG2, Aiping ZHANG1, Hongmei GAO2, Jide LI1, Wenlong WANG1, Penghui ZHANG1, Lingwen LI1, Xinyue GU1, Tongzuo ZHANG2(
)
Received:2024-11-28
Accepted:2025-02-24
Online:2026-05-30
Published:2026-06-04
Contact:
Tongzuo ZHANG
摘要:
青海农业区主要集中在青海省东部地区,区内旱獭生活区域与人类活动范围有部分交叠。本研究通过宏病毒组测序技术对青海东部农业区6个地区的喜马拉雅旱獭(Marmota himalayana)肠道病毒组成、多样性及其宿主进行比对分析。研究结果可知,病毒以尾噬菌体门、菲克斯病毒门和单环编码病毒门为主,目水平上可分类病毒以微小噬菌体目和圆环样病毒目为主,科水平上可分类病毒以微小噬菌体科和维雅病毒科为主。近60%的病毒基因注释集中在蛋白质家族的遗传信息处理、信号和细胞过程以及复制与修复功能,宿主预测以肠杆菌科埃希氏菌属的大肠杆菌(Escherichia coli)和芽孢杆菌科芽孢杆菌属的侧孢短芽孢杆菌(Brevibacillus laterosporus)为主。本研究揭示了青海东部农业区喜马拉雅旱獭病毒组的多样性及地区差异,提示某些病毒科可能具有地域局限性和宿主特异性,为未来病毒监测和致病性研究提供了重要依据。
中图分类号:
马英, 魏有文, 江峰, 张爱萍, 高红梅, 李积德, 王文龙, 张鹏辉, 李玲雯, 谷昕玥, 张同作. 青海东部农业区喜马拉雅旱獭肠道病毒种群宏基因组学分析[J]. 兽类学报, 2026, 46(3): 337-348.
Ying MA, Youwen WEI, Feng JIANG, Aiping ZHANG, Hongmei GAO, Jide LI, Wenlong WANG, Penghui ZHANG, Lingwen LI, Xinyue GU, Tongzuo ZHANG. Metagenomic analysis of intestinal virome in Marmota himalayana in eastern Qinghai’s agricultural area[J]. ACTA THERIOLOGICA SINICA, 2026, 46(3): 337-348.
种群 Population | 采集地点 Region | 样本量(性别) Number (Gender) | 经度 Longitude | 纬度 Latitude | 海拔 Elevation/m |
|---|---|---|---|---|---|
| HDHZ | 海东市互助县 Huzhu County,Haidong City | 3(2♂ 1♀) | 102.2763°E | 36.7299°N | 2 983 |
| HDMH | 海东市民和县 Minhe County,Haidong City | 3(3♀) | 102.6256°E | 36.2035°N | 2 592 |
| HDPA | 海东市平安区 Pingan District,Haidong City | 3(1♂ 2♀) | 101.8478°E | 36.3422°N | 2 803 |
| XNHY | 西宁市湟源县 Huangyuan County,Xining City | 3(3♀) | 101.2291°E | 36.4249°N | 3 334 |
| XNHZ | 西宁市湟中县 Huangzhong County,Xining City | 3(1♂ 2♀) | 101.7416°E | 36.3320°N | 2 934 |
| XNDT | 西宁市大通县 Datong County,Xining City | 3(3♀) | 101.6220°E | 37.1058°N | 2 930 |
表1 本研究喜马拉雅旱獭采集样点信息
Table 1 Sampling information of Marmota himalayana used in the present study
种群 Population | 采集地点 Region | 样本量(性别) Number (Gender) | 经度 Longitude | 纬度 Latitude | 海拔 Elevation/m |
|---|---|---|---|---|---|
| HDHZ | 海东市互助县 Huzhu County,Haidong City | 3(2♂ 1♀) | 102.2763°E | 36.7299°N | 2 983 |
| HDMH | 海东市民和县 Minhe County,Haidong City | 3(3♀) | 102.6256°E | 36.2035°N | 2 592 |
| HDPA | 海东市平安区 Pingan District,Haidong City | 3(1♂ 2♀) | 101.8478°E | 36.3422°N | 2 803 |
| XNHY | 西宁市湟源县 Huangyuan County,Xining City | 3(3♀) | 101.2291°E | 36.4249°N | 3 334 |
| XNHZ | 西宁市湟中县 Huangzhong County,Xining City | 3(1♂ 2♀) | 101.7416°E | 36.3320°N | 2 934 |
| XNDT | 西宁市大通县 Datong County,Xining City | 3(3♀) | 101.6220°E | 37.1058°N | 2 930 |
样品 Samples | 原始数据 Raw data(G) | 原始reads数 Number of raw reads(PE) | 高质量数据 Clean data(G) | 高质量reads数 Number of clean reads(PE) | 高质量占比 Proportion of high quality/% |
|---|---|---|---|---|---|
| HDHZ | 12.5 | 41826533 | 8.9 | 29913139 | 71.52 |
| HDMH | 12.3 | 41085030 | 9.9 | 33035768 | 80.41 |
| HDPA | 11.4 | 37919759 | 8.1 | 27187912 | 71.70 |
| XNHY | 10.0 | 33475840 | 8.5 | 28559824 | 85.31 |
| XNHZ | 9.8 | 32780458 | 5.9 | 19772197 | 60.32 |
| XNDT | 11.3 | 37633105 | 9.1 | 30345487 | 80.64 |
表 2 宏病毒组测序reads数据汇总
Table 2 Summary of macro virus sequencing reads data
样品 Samples | 原始数据 Raw data(G) | 原始reads数 Number of raw reads(PE) | 高质量数据 Clean data(G) | 高质量reads数 Number of clean reads(PE) | 高质量占比 Proportion of high quality/% |
|---|---|---|---|---|---|
| HDHZ | 12.5 | 41826533 | 8.9 | 29913139 | 71.52 |
| HDMH | 12.3 | 41085030 | 9.9 | 33035768 | 80.41 |
| HDPA | 11.4 | 37919759 | 8.1 | 27187912 | 71.70 |
| XNHY | 10.0 | 33475840 | 8.5 | 28559824 | 85.31 |
| XNHZ | 9.8 | 32780458 | 5.9 | 19772197 | 60.32 |
| XNDT | 11.3 | 37633105 | 9.1 | 30345487 | 80.64 |
样品 Samples | 质控后的reads Clean reads | 比对到宿主基因组的read数 Number of reads mapped to the host genome | 最后剩余reads数 Remaining reads | 最后剩余reads数占比 Proportion of remaining reads/% |
|---|---|---|---|---|
| HDHZ | 29913139 | 487552 | 29425587 | 98.37 |
| HDMH | 33035768 | 484506 | 32551262 | 98.53 |
| HDPA | 27187912 | 1115333 | 26072579 | 95.90 |
| XNHY | 28559824 | 55236 | 28504588 | 99.81 |
| XNHZ | 19772197 | 56003 | 19716194 | 99.72 |
| XNDT | 30345487 | 124262 | 30221225 | 99.59 |
表3 去除宿主序列后reads数据统计
Table 3 Statistics of reads data after removing host sequences
样品 Samples | 质控后的reads Clean reads | 比对到宿主基因组的read数 Number of reads mapped to the host genome | 最后剩余reads数 Remaining reads | 最后剩余reads数占比 Proportion of remaining reads/% |
|---|---|---|---|---|
| HDHZ | 29913139 | 487552 | 29425587 | 98.37 |
| HDMH | 33035768 | 484506 | 32551262 | 98.53 |
| HDPA | 27187912 | 1115333 | 26072579 | 95.90 |
| XNHY | 28559824 | 55236 | 28504588 | 99.81 |
| XNHZ | 19772197 | 56003 | 19716194 | 99.72 |
| XNDT | 30345487 | 124262 | 30221225 | 99.59 |
样品 Samples | 组装获得的序列 碱基总数 Assembled base pairs | 组装contigs条数 Number of assembled contigs | contig长度最大值 Maximum contig length | contig长度最小值 Minimum contig length | N50值 Contig N50 | contigs GC平均含量 Average GC content of contigs |
|---|---|---|---|---|---|---|
| HDHZ | 138.51 | 150 585 | 132 879 | 300 | 1 009 | 42.17 |
| HDMH | 316.15 | 382 618 | 208 313 | 300 | 833 | 45.87 |
| HDPA | 84.25 | 92 097 | 232 212 | 300 | 984 | 43.12 |
| XNHY | 355.78 | 415 327 | 196 346 | 300 | 888 | 47.58 |
| XNHZ | 161.14 | 183 488 | 172 565 | 300 | 916 | 45.00 |
| XNDT | 212.15 | 234 266 | 304 333 | 300 | 976 | 41.98 |
表 4 去除宿主序列后的组装结果统计
Table 4 Statistics of assembly results after removing host sequences
样品 Samples | 组装获得的序列 碱基总数 Assembled base pairs | 组装contigs条数 Number of assembled contigs | contig长度最大值 Maximum contig length | contig长度最小值 Minimum contig length | N50值 Contig N50 | contigs GC平均含量 Average GC content of contigs |
|---|---|---|---|---|---|---|
| HDHZ | 138.51 | 150 585 | 132 879 | 300 | 1 009 | 42.17 |
| HDMH | 316.15 | 382 618 | 208 313 | 300 | 833 | 45.87 |
| HDPA | 84.25 | 92 097 | 232 212 | 300 | 984 | 43.12 |
| XNHY | 355.78 | 415 327 | 196 346 | 300 | 888 | 47.58 |
| XNHZ | 161.14 | 183 488 | 172 565 | 300 | 916 | 45.00 |
| XNDT | 212.15 | 234 266 | 304 333 | 300 | 976 | 41.98 |
基因名称 Name of gene | 样品Samples | |||||
|---|---|---|---|---|---|---|
| HDHZ | HDMH | HDPA | XNHY | XNHZ | XNDT | |
| SCD|contig_3238 | 0.53 | 0.02 | 0.86 | 0.03 | 0.07 | 0.01 |
| HDPA|contig_89993 | 0.42 | 0 | 0.63 | 0.01 | 0.03 | 0 |
| HNZK|contig_7503 | 0.02 | 0.16 | 33 327.41 | 0.03 | 0.07 | 0.06 |
| XNHY|contig_402915 | 293.08 | 1.95 | 0 | 319.12 | 39 730.32 | 46.19 |
| SMD|contig_147452 | 0 | 0.01 | 0 | 0 | 0 | 0 |
| HNZK|contig_65526 | 0 | 0 | 1.92 | 0.01 | 0.08 | 0.02 |
| HDPA|contig_81697 | 0 | 0.04 | 1.82 | 0 | 0.06 | 0 |
| SMD|contig_147636 | 0 | 1.62 | 0 | 172.60 | 34 127.26 | 0 |
| HDMH|contig_380818 | 0.02 | 1.97 | 0 | 176.59 | 33 716.08 | 0 |
| HNZK|contig_4700 | 0 | 0.06 | 21 873.11 | 0.31 | 0.13 | 12.75 |
表 5 前十位RPKM的contigs丰度统计
Table 5 Abundance statistics of the top 10 contigs by RPKM
基因名称 Name of gene | 样品Samples | |||||
|---|---|---|---|---|---|---|
| HDHZ | HDMH | HDPA | XNHY | XNHZ | XNDT | |
| SCD|contig_3238 | 0.53 | 0.02 | 0.86 | 0.03 | 0.07 | 0.01 |
| HDPA|contig_89993 | 0.42 | 0 | 0.63 | 0.01 | 0.03 | 0 |
| HNZK|contig_7503 | 0.02 | 0.16 | 33 327.41 | 0.03 | 0.07 | 0.06 |
| XNHY|contig_402915 | 293.08 | 1.95 | 0 | 319.12 | 39 730.32 | 46.19 |
| SMD|contig_147452 | 0 | 0.01 | 0 | 0 | 0 | 0 |
| HNZK|contig_65526 | 0 | 0 | 1.92 | 0.01 | 0.08 | 0.02 |
| HDPA|contig_81697 | 0 | 0.04 | 1.82 | 0 | 0.06 | 0 |
| SMD|contig_147636 | 0 | 1.62 | 0 | 172.60 | 34 127.26 | 0 |
| HDMH|contig_380818 | 0.02 | 1.97 | 0 | 176.59 | 33 716.08 | 0 |
| HNZK|contig_4700 | 0 | 0.06 | 21 873.11 | 0.31 | 0.13 | 12.75 |
图1 青海东部农业区喜马拉雅旱獭在门、目和科水平上的病毒组成相对丰度
Fig. 1 Relative abundance of virus composition at the phylum, order, and family levels of Marmota himalayana from the agricultural region of eastern in Qinghai Province
图2 青海东部农业区喜马拉雅旱獭肠道病毒Shannon指数. ns,无显著差异
Fig. 2 Shannon index for intestinal viruses of Marmota himalayana from the agricultural region of eastern in Qinghai Province. ns, no significant difference
图3 青海东部农业区喜马拉雅旱獭肠道病毒组间PCoA聚类分析. HD,海东市;XN,西宁市
Fig. 3 PCoA analysis among intestinal microbiomes of Marmota himalayana from the agricultural region of eastern in Qinghai Province. HD, Haidong City; XN, Xining City
图4 基于KEGG数据库的喜马拉雅旱獭肠道病毒在Level 1水平(A)和Level 2水平(B)上基因注释数目
Fig. 4 Gene annotation counts at Level 1 (A) and Level 2 (B) for intestinal viruses of Marmota himalayana on the KEGG database
图5 青海东部农业区喜马拉雅旱獭肠道病毒在科水平上的前10位宿主信息. A:互助;B:民和;C:平安;D:湟源;E:湟中;F:大通
Fig. 5 The top 10 host information at the family level for intestinal viruses in Marmota himalayana from the agricultural region of eastern in Qinghai Province.A: HDHZ; B: HDMH; C: HDPA; D: XNHY; E: XNHZ; F: XNDT
图6 青海东部农业区喜马拉雅旱獭肠道病毒在属水平上的前10位宿主信息. A:互助;B:民和;C:平安;D:湟源;E:湟中;F:大通
Fig. 6 The top 10 host information at the genus level for intestinal viruses in Marmota himalayana from the agricultural region of eastern in Qinghai Province. A: HDHZ; B: HDMH; C: HDPA; D: XNHY; E: XNHZ; F: XNDT
图7 青海东部农业区喜马拉雅旱獭肠道病毒在种水平上的前10位宿主信息. A:互助;B:民和;C:平安;D:湟源;E:湟中;F:大通
Fig. 7 The top 10 host information at the species level for intestinal viruses in Marmota himalayana from the agricultural region of eastern in Qinghai Province. A: HDHZ; B: HDMH; C: HDPA; D: XNHY; E: XNHZ; F: XNDT
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