兽类学报 ›› 2026, Vol. 46 ›› Issue (5): 744-758.DOI: 10.16829/j.slxb.151162
王斌鹏1(
), 张丹辉2, 杨志梅3, 潘月婷3, 罗晓斌2, 莫然1, 昝林森3, 王晓宇2(
), 梅楚刚1(
)
出版日期:2026-09-30
发布日期:2026-08-12
通讯作者:
王晓宇,梅楚刚
作者简介:王斌鹏(2000- ),男,硕士,主要从事微生物功能基因组学研究. E‐mail:wangbp0127@163.com
基金资助:
Binpeng WANG1(
), Danhui ZHANG2, Zhimei YANG3, Yueting PAN3, Xiaobin LUO2, Ran MO1, Linsen ZAN3, Xiaoyu WANG2(
), Chugang MEI1(
)
Online:2026-09-30
Published:2026-08-12
Contact:
Xiaoyu WANG, Chugang MEI
摘要:
本研究旨在通过宏基因组技术探究秦岭圈养大熊猫(Ailuropoda melanoleuca)幼年期与成年期肠道菌群结构和功能差异,为阐明秦岭圈养大熊猫的肠道菌群对不同阶段饮食的适应性响应,开发针对提高秦岭圈养大熊猫的促消化益生菌产品和改善其营养健康提供重要的理论依据。本研究采集秦岭大熊猫研究中心饲养的3只幼年(2 ~ 4月龄)和3只成年(62 ~ 63月龄)大熊猫粪便样本,通过宏基因组测序技术,对粪便细菌进行物种注释、多样性分析和功能注释与分析。测序共获得424 716 776条有效序列,构建非冗余基因后共获得318 605个基因。α多样性表明,与幼年组相比,成年组Chao1和Ace指数极显著提高(P < 0.01),而Shannon和Simpson指数极显著降低(P < 0.01);β多样性分析表明,两组样本肠道菌群结构差异明显;物种注释结果表明,幼年组以乳杆菌属(Lactobacillus)为主,成年组则以埃希氏菌属(Escherichia)和链球菌属(Streptococcus)为主,并在成年组中发现地衣芽孢杆菌(Bacillus licheniformis)、半乳糖地质杆菌(Geobacillus galactosidasius)和粪堆梭菌(Thermoclostridium stercorarium)等具有木质纤维素降解潜力的低丰度细菌。功能注释结果显示,幼年组GT4、GH43、GH73、碳水化合物代谢、氨基酸代谢、翻译、脂质代谢、核糖体结构和生物合成等通路富集程度显著高于成年组(P < 0.05)。本研究基于宏基因组学解析了秦岭圈养大熊猫幼年期与成年期肠道菌群特征与差异,挖掘到与木质纤维素分解相关的菌群,为后续促消化菌剂产品的研发,及助力大熊猫精准营养管理和肠道健康提供重要依据。
中图分类号:
王斌鹏, 张丹辉, 杨志梅, 潘月婷, 罗晓斌, 莫然, 昝林森, 王晓宇, 梅楚刚. 秦岭圈养大熊猫幼年期与成年期肠道菌群结构及功能差异解析[J]. 兽类学报, 2026, 46(5): 744-758.
Binpeng WANG, Danhui ZHANG, Zhimei YANG, Yueting PAN, Xiaobin LUO, Ran MO, Linsen ZAN, Xiaoyu WANG, Chugang MEI. Analysis of structural and functional differences in the gut microbiota between juvenile and adult captive giant pandas in the Qinling region[J]. ACTA THERIOLOGICA SINICA, 2026, 46(5): 744-758.
| 组名 Group | 编号 Number | 年龄 Age | 日粮组成 Diet composition | 出生地 Birthplace |
|---|---|---|---|---|
幼年组 Juvenile | A1 | 2月龄 2 months old | 母乳,每天3次,每次100 g Breast milk, 3 times/day, 100 g/time | 秦岭大熊猫研究中心 (陕西省珍稀野生动物救护基地) Research Center for the Qinling Giant Panda (Shaanxi Rare Wildlife Rescue Base) |
| A2 | 3月龄 3 months old | 母乳,每天3次,每次130 g Breast milk, 3 times/day, 130 g/time | ||
| A3 | 4月龄 4 months old | 母乳,每天2次,每次220 g Breast milk, 2 times/day,220 g/time | ||
成年组 Adult | B1 | 62月龄 62 months old | 竹子30 kg,苹果0.25 kg,胡萝卜1 kg, 南瓜1 kg,窝窝头300 g Bamboo 30 kg, apple 0.25 kg, carrot 1 kg, pumpkin 1 kg, steamed corn bun 300 g | |
| B2 | 63月龄 63 months old | |||
| B3 | 63月龄 63 months old |
表1 实验大熊猫基本信息表
Table 1 The basic information of the tested pandas
| 组名 Group | 编号 Number | 年龄 Age | 日粮组成 Diet composition | 出生地 Birthplace |
|---|---|---|---|---|
幼年组 Juvenile | A1 | 2月龄 2 months old | 母乳,每天3次,每次100 g Breast milk, 3 times/day, 100 g/time | 秦岭大熊猫研究中心 (陕西省珍稀野生动物救护基地) Research Center for the Qinling Giant Panda (Shaanxi Rare Wildlife Rescue Base) |
| A2 | 3月龄 3 months old | 母乳,每天3次,每次130 g Breast milk, 3 times/day, 130 g/time | ||
| A3 | 4月龄 4 months old | 母乳,每天2次,每次220 g Breast milk, 2 times/day,220 g/time | ||
成年组 Adult | B1 | 62月龄 62 months old | 竹子30 kg,苹果0.25 kg,胡萝卜1 kg, 南瓜1 kg,窝窝头300 g Bamboo 30 kg, apple 0.25 kg, carrot 1 kg, pumpkin 1 kg, steamed corn bun 300 g | |
| B2 | 63月龄 63 months old | |||
| B3 | 63月龄 63 months old |
| 样品 Sample | 高质量reads数 Clean reads | 有效碱基总数 Clean data/bp | 质控后碱基GC含量 GC/% | Q20/% | Q30/% |
|---|---|---|---|---|---|
| A1 | 85 996 110 | 12 891 207 670 | 49.17 | 97.36 | 92.62 |
| A2 | 80 685 546 | 12 096 032 915 | 53.26 | 97.77 | 93.60 |
| A3 | 65 251 992 | 9 780 335 742 | 43.53 | 97.13 | 92.08 |
| B1 | 62 875 460 | 9 425 792 981 | 45.35 | 97.64 | 93.12 |
| B2 | 66 260 324 | 9 930 960 623 | 45.48 | 97.43 | 92.69 |
| B3 | 63 647 344 | 9 542 239 488 | 45.28 | 97.66 | 93.19 |
表2 样本测序信息
Table 2 Sample sequencing information
| 样品 Sample | 高质量reads数 Clean reads | 有效碱基总数 Clean data/bp | 质控后碱基GC含量 GC/% | Q20/% | Q30/% |
|---|---|---|---|---|---|
| A1 | 85 996 110 | 12 891 207 670 | 49.17 | 97.36 | 92.62 |
| A2 | 80 685 546 | 12 096 032 915 | 53.26 | 97.77 | 93.60 |
| A3 | 65 251 992 | 9 780 335 742 | 43.53 | 97.13 | 92.08 |
| B1 | 62 875 460 | 9 425 792 981 | 45.35 | 97.64 | 93.12 |
| B2 | 66 260 324 | 9 930 960 623 | 45.48 | 97.43 | 92.69 |
| B3 | 63 647 344 | 9 542 239 488 | 45.28 | 97.66 | 93.19 |
样品 Sample | Contigs总数 Contigs No. | Contigs总长 Contigs total length/bp | Contigs平均长度 Contigs average length/bp | Contigs长度最大值 Contigs maximum contigs length/bp | Contigs长度最小值 Contigs minimum contigs length/bp | N50值 N50/bp | N90值 N90/bp |
|---|---|---|---|---|---|---|---|
| A1 | 11 001 | 49 624 552 | 4 510.91 | 830 781 | 500 | 25 851 | 1 381 |
| A2 | 20 059 | 75 770 877 | 3 777.40 | 544 716 | 500 | 11 720 | 1 170 |
| A3 | 22 253 | 52 059 617 | 2 339.44 | 830 985 | 500 | 11 969 | 686 |
| B1 | 35 335 | 59 385 152 | 1 680.63 | 92 515 | 500 | 2 988 | 621 |
| B2 | 53 247 | 72 411 477 | 1 359.92 | 140 979 | 500 | 1 840 | 577 |
| B3 | 43 970 | 64 211 550 | 1 460.35 | 116 002 | 500 | 2 207 | 591 |
表3 组装结果统计表
Table 3 Statistical table of assembly results
样品 Sample | Contigs总数 Contigs No. | Contigs总长 Contigs total length/bp | Contigs平均长度 Contigs average length/bp | Contigs长度最大值 Contigs maximum contigs length/bp | Contigs长度最小值 Contigs minimum contigs length/bp | N50值 N50/bp | N90值 N90/bp |
|---|---|---|---|---|---|---|---|
| A1 | 11 001 | 49 624 552 | 4 510.91 | 830 781 | 500 | 25 851 | 1 381 |
| A2 | 20 059 | 75 770 877 | 3 777.40 | 544 716 | 500 | 11 720 | 1 170 |
| A3 | 22 253 | 52 059 617 | 2 339.44 | 830 985 | 500 | 11 969 | 686 |
| B1 | 35 335 | 59 385 152 | 1 680.63 | 92 515 | 500 | 2 988 | 621 |
| B2 | 53 247 | 72 411 477 | 1 359.92 | 140 979 | 500 | 1 840 | 577 |
| B3 | 43 970 | 64 211 550 | 1 460.35 | 116 002 | 500 | 2 207 | 591 |
组名 Group | Chao1指数 Chao1 index | Ace指数 Ace index | Shannon指数 Shannon index | Simpson指数 Simpson index |
|---|---|---|---|---|
| A | 102.083 | 102.706 | 2.411 | 0.719 |
| B | 270.833 | 271.204 | 1.369 | 0.385 |
| P值 P-value | < 0.01 | < 0.01 | < 0.01 | < 0.01 |
表4 粪便微生物α多样性分析结果
Table 4 α diversity analysis fecal microbial community
组名 Group | Chao1指数 Chao1 index | Ace指数 Ace index | Shannon指数 Shannon index | Simpson指数 Simpson index |
|---|---|---|---|---|
| A | 102.083 | 102.706 | 2.411 | 0.719 |
| B | 270.833 | 271.204 | 1.369 | 0.385 |
| P值 P-value | < 0.01 | < 0.01 | < 0.01 | < 0.01 |
图1 基于种水平的肠道菌群β多样性分析. a:PCA分析;b:PCoA分析;c:PERMANOVA分析;d:ANOSIM分析
Fig.1 Beta diversity analysis of gut microbiota at the species level. a: PCA; b: PCoA; c: PERMANOVA; d: ANOSIM
图2 幼年与成年大熊猫肠道微生物组成. a:门水平韦恩图;b:属水平韦恩图;c:种水平韦恩图;d:门水平相对丰度;e:属水平相对丰度;f:种水平相对丰度
Fig. 2 Gut microbial composition of juvenile and adult giant pandas. a: Venn diagram at the phylum level; b: Venn diagram at the genus level; c: Venn diagram at the species level; d: Relative abundance at the phylum level; e: Relative abundance at the genus level; f: Relative abundance at the species level
图3 幼年与成年大熊猫肠道微生物相对丰度差异分析. a:门水平;b:属水平;c:种水平
Fig. 3 Differences in gut microbial relative abundance between juvenile and adult giant pandas at the phylum, genus, and species levels. a: Phylum level; b: Genus level; c: Species level
图4 CAZy数据库注释结果. a:Level 1水平上的CAZy数据库注释结果;b:Level 2水平上的CAZy数据库注释结果;c:LEfSe分析结果;d:Level 2水平上的功能差异分析结果
Fig. 4 Annotation results of the CAZy database. a: Annotation results at Level 1 of the CAZy database; b: Annotation results at Level 2 of the CAZy database; c: LEfSe analysis results; d: Differential functional analysis results at Level 2
图5 KEGG数据库注释结果. a:Level 1水平上的注释结果;b:Level 2水平上的注释结果;c:Level 1水平上的功能差异分析结果;d:Level 2水平上的功能差异分析结果
Fig. 5 Annotation results of the KEGG database. a: Level 1 annotation; b: Level 2 annotation; c: Differential analysis at Level 1; d: Differential analysis at Level 2
图6 EggNOG数据库注释结果.a:EggNOG数据库第一层级功能注释结果;b:EggNOG数据库第一层级功能差异分析结果
Fig. 6 Annotation results of the EggNOG database. a: Functional annotation results at Level 1; b: Differential functional analysis results at Level 1
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