浙江农业学报 ›› 2026, Vol. 38 ›› Issue (8): 1648-1659.DOI: 10.3969/j.issn.1004-1524.20250418
张思怡1,2(
), 陆丽2, 陈昭良2, 吕卫光2,3,4, 张翰林2,3,4, 李双喜2,3,4, 张停林5, 孙丽娟2, 白娜玲2,3,4,*(
)
收稿日期:2025-06-02
出版日期:2026-08-25
发布日期:2026-09-14
作者简介:张思怡,主要从事微生物资源化利用与机制研究。E-mail:18235645196@163.com
通讯作者:
*白娜玲,E-mail: bainaling@saas.sh.cn
基金资助:
ZHANG Siyi1,2(
), LU Li2, CHEN Zhaoliang2, LYU Weiguang2,3,4, ZHANG Hanlin2,3,4, LI Shuangxi2,3,4, ZHANG Tinglin5, SUN Lijuan2, BAI Naling2,3,4,*(
)
Received:2025-06-02
Published:2026-08-25
Online:2026-09-14
摘要:
本研究从宁夏某枸杞种植基地采集盐碱地土壤样本,通过高通量测序分析土壤细菌群落特征,利用稀释涂布平板法在中盐碱度(NaCl质量分数8%、pH值8.5)的培养条件下从土壤中分离纯化耐/嗜盐碱细菌,鉴定其系统分类学地位,分析菌株的耐/嗜盐碱特性,并对其中效果最优菌的生物学特性、降盐降碱等功能进行研究。结果显示,样本盐碱土中门水平上的优势细菌为放线菌门(Actinobacteriota)、变形菌门(Proteobacteria)、绿弯菌门(Chloroflexi)、芽单胞菌门(Gemmatimonadota)。在中盐碱度条件下,从样本土壤中筛选到12株耐/嗜盐碱菌株,其中,Bacillus haynesii nx-11菌株可在高盐(NaCl质量分数8%~18%)或高碱(pH值9.0~12.0)条件下生长;Halobacillus andaensis nx-9菌株盐碱耐受性最强,不仅可在高盐或高碱条件下生长,在兼具高盐碱度(NaCl质量分数8%~16%、pH值8.5~10.0)的条件下生长良好,其降碱率为12.17%~18.25%,降盐率为4.88%~12.02%。本研究结果丰富了对宁夏地区盐碱土壤微生物特征的认知,可为生物治理盐碱土提供微生物资源。
中图分类号:
张思怡, 陆丽, 陈昭良, 吕卫光, 张翰林, 李双喜, 张停林, 孙丽娟, 白娜玲. 宁夏枸杞盐碱地土壤细菌群落特征及可培养耐/嗜盐碱菌筛选[J]. 浙江农业学报, 2026, 38(8): 1648-1659.
ZHANG Siyi, LU Li, CHEN Zhaoliang, LYU Weiguang, ZHANG Hanlin, LI Shuangxi, ZHANG Tinglin, SUN Lijuan, BAI Naling. Characteristics of bacterial communities in saline-alkali soil of Lycium barbarum fields in Ningxia of China and screening of culturable halotolerant/haloalkaliphilic bacteria[J]. Acta Agriculturae Zhejiangensis, 2026, 38(8): 1648-1659.
图1 相对丰度排名前10的门水平优势物种(a)和排名前20的属水平优势物种(b) Actinobacteriota,放线菌门;Proteobacteria,变形菌门;Chloroflexi,绿弯菌门;Gemmatimonadota,芽单胞菌门;Bacteroidota,拟杆菌门;Acidobacteriota,酸杆菌门;Bacillota,芽孢杆菌门;Myxococcota,黏球菌门;Planctomycetota,浮霉菌门;Verrucomicrobiota,疣微菌门;Others,其他。Unclassified_f_Euzebyaceae,未分类尤泽比氏菌科;Unclassified_f_Nocardioidaceae,未分类类诺卡氏菌科;Unclassified_c_Acidimicrobiia,未分类酸微菌纲;Nocardioides,类诺卡氏菌属;Unclassified_c_Alphaproteobacteria,未分类α-变形菌纲;Unclassified_Gemmatimonadaceae,未分类芽单胞菌科。
Fig.1 The top 10 dominant species at phylum level (a) and top 20 dominant species at genus level (b) in terms of relative abundance
图2 Bugbase(a)和PICRUSt2(b)功能预测分析 Stress_Tolerant,胁迫耐受;Forms_Biofilms,生物膜形成;Gram_Negative,革兰氏阴性;Aerobic,好氧;Gram_Positive,革兰氏阳性;Potentially_Pathogenic,潜在致病;Anaerobic,厌氧;Facultatively_Anaerobic,兼性厌氧;Contains_Mobile_Elements,携带移动遗传元件。Biosynthesis,生物合成;Degradation/Utilization/Assimilation,降解/利用/同化;Generation of precursor metabolite and energy,前体代谢物与能量生成;Superpathways,超级通路;Macromolecule modification,大分子修饰。
Fig.2 Functional prediction analysis by Bugbase (a) and PICRUSt2 (b)
| 菌株 Strain | 不同NaCl质量分数下菌液的D600值 D600 values of bacterial suspensions under different mass fractions of NaCl | ||||||
|---|---|---|---|---|---|---|---|
| 8% | 10% | 12% | 14% | 16% | 18% | 20% | |
| nx-1 | 1.046 | 0.723 | 0.728 | 0.654 | 0.518 | — | — |
| nx-2 | 0.812 | 0.604 | — | — | — | — | — |
| nx-3 | 0.752 | 0.842 | 0.532 | 0.233 | — | — | — |
| nx-4 | 0.501 | — | — | — | — | — | — |
| nx-5 | 0.334 | 0.719 | 0.574 | 0.364 | — | — | — |
| nx-6 | 0.872 | 0.765 | 0.835 | 0.505 | — | — | — |
| nx-7 | 0.791 | 0.729 | 0.641 | 0.512 | — | — | — |
| nx-8 | 0.808 | 0.753 | 0.587 | 0.412 | — | — | — |
| nx-9 | 1.068 | 1.037 | 1.083 | 0.872 | 0.524 | 0.378 | — |
| nx-10 | 1.079 | 1.192 | 0.848 | 0.500 | — | — | — |
| nx-11 | 1.054 | 0.747 | 0.641 | 0.659 | 0.524 | 0.215 | — |
| nx-12 | 0.823 | 0.581 | 0.771 | 0.616 | 0.663 | — | — |
表1 不同菌株的耐盐能力
Table1 Salt tolerance capacity of different strains
| 菌株 Strain | 不同NaCl质量分数下菌液的D600值 D600 values of bacterial suspensions under different mass fractions of NaCl | ||||||
|---|---|---|---|---|---|---|---|
| 8% | 10% | 12% | 14% | 16% | 18% | 20% | |
| nx-1 | 1.046 | 0.723 | 0.728 | 0.654 | 0.518 | — | — |
| nx-2 | 0.812 | 0.604 | — | — | — | — | — |
| nx-3 | 0.752 | 0.842 | 0.532 | 0.233 | — | — | — |
| nx-4 | 0.501 | — | — | — | — | — | — |
| nx-5 | 0.334 | 0.719 | 0.574 | 0.364 | — | — | — |
| nx-6 | 0.872 | 0.765 | 0.835 | 0.505 | — | — | — |
| nx-7 | 0.791 | 0.729 | 0.641 | 0.512 | — | — | — |
| nx-8 | 0.808 | 0.753 | 0.587 | 0.412 | — | — | — |
| nx-9 | 1.068 | 1.037 | 1.083 | 0.872 | 0.524 | 0.378 | — |
| nx-10 | 1.079 | 1.192 | 0.848 | 0.500 | — | — | — |
| nx-11 | 1.054 | 0.747 | 0.641 | 0.659 | 0.524 | 0.215 | — |
| nx-12 | 0.823 | 0.581 | 0.771 | 0.616 | 0.663 | — | — |
| 菌株 Strain | 不同pH值条件下菌液的D600值 D600 values of bacterial suspensions under different pH value | |||||
|---|---|---|---|---|---|---|
| 7.0 | 9.0 | 10.0 | 11.0 | 12.0 | 13.0 | |
| nx-1 | 0.627 | 0.749 | 0.935 | 0.978 | 0.553 | — |
| nx-2 | 0.701 | 0.679 | 0.824 | 1.023 | 0.907 | — |
| nx-3 | — | 0.789 | 0.815 | 0.765 | 0.323 | — |
| nx-4 | 0.501 | 0.245 | — | — | — | — |
| nx-5 | — | 0.652 | 0.822 | 0.881 | 0.312 | — |
| nx-6 | 0.612 | 0.744 | 1.021 | 0.915 | 0.739 | — |
| nx-7 | 0.703 | 0.779 | 0.801 | 0.448 | — | — |
| nx-8 | 0.673 | 0.546 | 0.531 | 0.537 | — | — |
| nx-9 | — | 0.929 | 0.789 | 0.683 | 0.657 | — |
| nx-10 | 0.738 | 0.816 | 1.034 | 0.976 | 0.904 | — |
| nx-11 | 0.766 | 0.785 | 1.116 | 1.037 | 0.908 | — |
| nx-12 | 0.516 | 0.608 | 0.804 | — | — | — |
表2 不同菌株的耐碱能力
Table 2 Alkali tolerance capacity of different strains
| 菌株 Strain | 不同pH值条件下菌液的D600值 D600 values of bacterial suspensions under different pH value | |||||
|---|---|---|---|---|---|---|
| 7.0 | 9.0 | 10.0 | 11.0 | 12.0 | 13.0 | |
| nx-1 | 0.627 | 0.749 | 0.935 | 0.978 | 0.553 | — |
| nx-2 | 0.701 | 0.679 | 0.824 | 1.023 | 0.907 | — |
| nx-3 | — | 0.789 | 0.815 | 0.765 | 0.323 | — |
| nx-4 | 0.501 | 0.245 | — | — | — | — |
| nx-5 | — | 0.652 | 0.822 | 0.881 | 0.312 | — |
| nx-6 | 0.612 | 0.744 | 1.021 | 0.915 | 0.739 | — |
| nx-7 | 0.703 | 0.779 | 0.801 | 0.448 | — | — |
| nx-8 | 0.673 | 0.546 | 0.531 | 0.537 | — | — |
| nx-9 | — | 0.929 | 0.789 | 0.683 | 0.657 | — |
| nx-10 | 0.738 | 0.816 | 1.034 | 0.976 | 0.904 | — |
| nx-11 | 0.766 | 0.785 | 1.116 | 1.037 | 0.908 | — |
| nx-12 | 0.516 | 0.608 | 0.804 | — | — | — |
| 菌株 Strain | 不同条件下菌液的D600值D600 values of bacterial suspensions under different conditions | |||||||
|---|---|---|---|---|---|---|---|---|
| 8%+8.5 | 8%+9.5 | 10%+8.5 | 10%+9.5 | 14%+9.0 | 14%+10.0 | 16%+9.0 | 16%+10.0 | |
| nx-1 | 0.589 | 0.956 | 0.833 | 0.674 | — | — | — | — |
| nx-2 | 0.791 | 0.848 | 0.912 | 0.876 | — | — | — | — |
| nx-3 | 0.639 | 0.871 | 0.536 | 0.805 | — | — | — | — |
| nx-4 | 0.227 | 0.541 | 0.369 | — | — | — | — | — |
| nx-5 | 0.696 | 0.927 | 0.701 | 0.848 | — | — | — | — |
| nx-6 | 0.766 | 0.804 | 0.523 | — | — | — | — | — |
| nx-7 | 0.703 | 0.812 | 0.659 | 0.663 | — | — | — | — |
| nx-8 | 0.645 | 0.859 | 0.812 | 0.708 | — | — | — | — |
| nx-9 | 1.085 | 0.947 | 1.143 | 0.806 | 0.541 | 0.607 | 0.235 | 0.209 |
| nx-10 | 0.717 | 0.831 | 0.824 | 0.809 | — | — | — | — |
| nx-11 | 0.589 | 1.006 | 0.823 | 0.605 | — | — | — | — |
| nx-12 | 0.518 | 0.391 | 0.216 | — | — | — | — | — |
表3 不同菌株的耐盐碱能力
Table 3 Salt and alkali tolerance capacity of different strains
| 菌株 Strain | 不同条件下菌液的D600值D600 values of bacterial suspensions under different conditions | |||||||
|---|---|---|---|---|---|---|---|---|
| 8%+8.5 | 8%+9.5 | 10%+8.5 | 10%+9.5 | 14%+9.0 | 14%+10.0 | 16%+9.0 | 16%+10.0 | |
| nx-1 | 0.589 | 0.956 | 0.833 | 0.674 | — | — | — | — |
| nx-2 | 0.791 | 0.848 | 0.912 | 0.876 | — | — | — | — |
| nx-3 | 0.639 | 0.871 | 0.536 | 0.805 | — | — | — | — |
| nx-4 | 0.227 | 0.541 | 0.369 | — | — | — | — | — |
| nx-5 | 0.696 | 0.927 | 0.701 | 0.848 | — | — | — | — |
| nx-6 | 0.766 | 0.804 | 0.523 | — | — | — | — | — |
| nx-7 | 0.703 | 0.812 | 0.659 | 0.663 | — | — | — | — |
| nx-8 | 0.645 | 0.859 | 0.812 | 0.708 | — | — | — | — |
| nx-9 | 1.085 | 0.947 | 1.143 | 0.806 | 0.541 | 0.607 | 0.235 | 0.209 |
| nx-10 | 0.717 | 0.831 | 0.824 | 0.809 | — | — | — | — |
| nx-11 | 0.589 | 1.006 | 0.823 | 0.605 | — | — | — | — |
| nx-12 | 0.518 | 0.391 | 0.216 | — | — | — | — | — |
图3 菌株nx-9在中盐碱度(NaCl质量分数8%、pH值8.5)和高盐碱度(NaCl质量分数16%、pH值10.0)条件下的细胞形态 a、b分别为中、高盐碱度条件下的冷冻场发射扫描电子显微镜(Cryo-FE-SEM)观测结果;c、d分别为中、高盐碱度条件下的场发射扫描电子显微镜(FE-SEM)观测结果。
Fig.3 Cell morphology of strain nx-9 under medium (NaCl mass fraction of 8%, pH value of 8.5) and high (NaCl mass fraction of 16%, pH value of 10.0) saline-alkaline conditions Panels a and b represent cryo-field-emission scanning electron microscopy (Cryo-FE-SEM) images under moderate and high saline-alkaline condition, respectively. Panels c and d show field-emission scanning electron microscopy (FE-SEM) images under moderate and high saline-alkaline conditions, respectively.
| 酶Enzyme | 结果Results | 酶Enzyme | 结果Results |
|---|---|---|---|
| 碱性磷酸酶Alkaline phosphatase | + | 酸性磷酸酶Acid phosphatase | - |
| 酯酶(C4)Esterase (C4) | weak | 萘酚-AS-BI-磷酸水解酶Naphthol-AS-BI-phosphohydrolase | weak |
| 类脂酯酶(C8)Lipase (C8) | weak | α-半乳糖苷酶α-Galactosidase | - |
| 类脂酯酶(C14)Lipase (C14) | - | β-半乳糖苷酶β-Galactosidase | + |
| 亮氨酸芳胺酶Leucine arylamidase | - | β-葡萄糖醛酸苷酶β-Glucuronidase | - |
| 缬氨酸芳胺酶Valine arylamidase | - | α-葡萄糖苷酶α-Glucosidase | + |
| 胱氨酸芳胺酶Cystine arylamidase | - | β-葡萄糖苷酶β-Glucosidase | - |
| 胰蛋白酶Trypsin | - | N-乙酰-葡萄糖胺酶N-Acetylglucosaminidase | - |
| α-胰凝乳蛋白酶α-Chymotrypsin | - | α-岩藻糖苷酶α-Fucosidase | - |
表4 菌株nx-9的酶活特性
Table 4 Enzymatic characteristics of strain nx-9
| 酶Enzyme | 结果Results | 酶Enzyme | 结果Results |
|---|---|---|---|
| 碱性磷酸酶Alkaline phosphatase | + | 酸性磷酸酶Acid phosphatase | - |
| 酯酶(C4)Esterase (C4) | weak | 萘酚-AS-BI-磷酸水解酶Naphthol-AS-BI-phosphohydrolase | weak |
| 类脂酯酶(C8)Lipase (C8) | weak | α-半乳糖苷酶α-Galactosidase | - |
| 类脂酯酶(C14)Lipase (C14) | - | β-半乳糖苷酶β-Galactosidase | + |
| 亮氨酸芳胺酶Leucine arylamidase | - | β-葡萄糖醛酸苷酶β-Glucuronidase | - |
| 缬氨酸芳胺酶Valine arylamidase | - | α-葡萄糖苷酶α-Glucosidase | + |
| 胱氨酸芳胺酶Cystine arylamidase | - | β-葡萄糖苷酶β-Glucosidase | - |
| 胰蛋白酶Trypsin | - | N-乙酰-葡萄糖胺酶N-Acetylglucosaminidase | - |
| α-胰凝乳蛋白酶α-Chymotrypsin | - | α-岩藻糖苷酶α-Fucosidase | - |
| 底物 Substrate | 结果 Results | 底物 Substrate | 结果 Results | 底物 Substrate | 结果 Results |
|---|---|---|---|---|---|
| 甘油Glycerol | weak | 甘露醇Mannitol | weak | D-蜜三糖D-Melitriose | + |
| 赤藓醇Erythritol | - | 山梨醇Sorbitol | - | 棉子糖Raffinose | + |
| D-阿拉伯糖D-Arabinose | - | α-甲基-D-甘露糖苷α-Methyl-D-mannoside | - | 糖原Glycogen | + |
| L-阿拉伯糖L-Arabinose | - | α-甲基-D-葡萄糖苷α-Methyl-D-glucoside | - | 木糖醇Xylitol | - |
| 核糖Ribose | - | N-乙酰葡糖胺N-Acetylglucosamine | + | β-龙胆二糖β-Gentiobiose | - |
| D-木糖D-Xylose | weak | 苦杏仁苷Amygdalin | - | D-松二糖D-Panose | - |
| L-木糖L-Xylose | - | 熊果苷Arbutin | - | D-来苏糖D-Lyxose | - |
| 核糖醇Adonitol | - | 七叶苷Esculin | + | D-塔格糖D-Tagatose | - |
| β-甲基-木糖苷β-Methyl-xyloside | - | 水杨苷Salicin | - | D-岩藻糖D-Fucose | - |
| 半乳糖Galactose | + | 纤维二糖Cellobiose | - | L-岩藻糖L-Fucose | - |
| D-葡萄糖D-Glucose | + | 麦芽糖Maltose | + | D-阿拉伯醇D-Arabitol | - |
| D-果糖D-Fructose | + | 乳糖Lactose | - | L-阿拉伯醇L-Arabitol | - |
| D-甘露糖D-Mannose | + | 蜜二糖Melibiose | + | 葡萄糖酸Gluconic acid | - |
| L-山梨糖L-Sorbose | - | 蔗糖Sucrose | + | 2-酮基-葡萄糖酸2-Ketogluconic acid | - |
| 鼠李糖Rhamnose | - | 海藻糖Trehalose | + | 5-酮基-葡萄糖酸5-Ketogluconic acid | - |
| 半乳糖醇Galactitol | - | 菊糖Inulin | + | ||
| 肌醇Myo-inositol | - | 松三糖Melezitose | + |
表5 菌株nx-9的底物利用特性
Table 5 Substrate utilization characteristics of strain nx-9
| 底物 Substrate | 结果 Results | 底物 Substrate | 结果 Results | 底物 Substrate | 结果 Results |
|---|---|---|---|---|---|
| 甘油Glycerol | weak | 甘露醇Mannitol | weak | D-蜜三糖D-Melitriose | + |
| 赤藓醇Erythritol | - | 山梨醇Sorbitol | - | 棉子糖Raffinose | + |
| D-阿拉伯糖D-Arabinose | - | α-甲基-D-甘露糖苷α-Methyl-D-mannoside | - | 糖原Glycogen | + |
| L-阿拉伯糖L-Arabinose | - | α-甲基-D-葡萄糖苷α-Methyl-D-glucoside | - | 木糖醇Xylitol | - |
| 核糖Ribose | - | N-乙酰葡糖胺N-Acetylglucosamine | + | β-龙胆二糖β-Gentiobiose | - |
| D-木糖D-Xylose | weak | 苦杏仁苷Amygdalin | - | D-松二糖D-Panose | - |
| L-木糖L-Xylose | - | 熊果苷Arbutin | - | D-来苏糖D-Lyxose | - |
| 核糖醇Adonitol | - | 七叶苷Esculin | + | D-塔格糖D-Tagatose | - |
| β-甲基-木糖苷β-Methyl-xyloside | - | 水杨苷Salicin | - | D-岩藻糖D-Fucose | - |
| 半乳糖Galactose | + | 纤维二糖Cellobiose | - | L-岩藻糖L-Fucose | - |
| D-葡萄糖D-Glucose | + | 麦芽糖Maltose | + | D-阿拉伯醇D-Arabitol | - |
| D-果糖D-Fructose | + | 乳糖Lactose | - | L-阿拉伯醇L-Arabitol | - |
| D-甘露糖D-Mannose | + | 蜜二糖Melibiose | + | 葡萄糖酸Gluconic acid | - |
| L-山梨糖L-Sorbose | - | 蔗糖Sucrose | + | 2-酮基-葡萄糖酸2-Ketogluconic acid | - |
| 鼠李糖Rhamnose | - | 海藻糖Trehalose | + | 5-酮基-葡萄糖酸5-Ketogluconic acid | - |
| 半乳糖醇Galactitol | - | 菊糖Inulin | + | ||
| 肌醇Myo-inositol | - | 松三糖Melezitose | + |
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