枯草芽孢杆菌YX-1对铝胁迫的响应及人参幼苗生长的影响

doi:10.16420/j.issn.0513-353x.2024-0964

摘要/Abstract

摘要： 从林下人参根际土壤分离得到一株枯草芽孢杆菌（Bacillus subtilis YX-1，Bs），通过测定其耐铝性、铝胁迫下生物膜形成、胞外多糖（exopolysaccharides，EPS）含量和相关基因表达量以及接种后人参根系形态、生理指标的变化，探究菌株YX-1对铝胁迫的响应及其对人参生长的影响。结果表明，随着铝浓度增加，菌株YX-1的生长逐渐受到抑制。随着铝处理时间的延长，菌种YX-1逐渐形成生物膜，且胞外多糖含量也逐渐增加；铝处理下菌株YX-1中与EPS合成和分泌相关的基因epsA、epsB、wzx和wzy的表达量呈先升高后降低的趋势，且在12 h表达量最高。与单独铝处理（100 mmol · L^-1Al）相比，先接种Bs再添加100 mmol · L^-1Al处理的人参叶片叶绿素含量显著增加，地上部和根系的干质量分别增加28.33%和11.65%，叶片和根系磷含量分别增加13.89%和16.24%，而铝含量分别降低32.11%和11.90%。表明B. subtilis YX-1通过合成和分泌胞外多糖促进生物膜的形成，可有效缓解铝胁迫对人参生长的影响。

关键词: 人参, 铝胁迫, 枯草芽孢杆菌YX-1, 生物膜, 胞外多糖

Abstract:

A strain of Bacillus subtilis YX-1 was isolated from the rhizosphere soil of understory Panax ginseng. The response of strain YX-1 to Al stress，the biofilm formation，extracellular polysaccharide content，the expression of related genes and ginseng root morphology were investigated，which explores the response of strain YX-1 to Al stress and its effects on ginseng growth. The results showed that with the increase of Al concentration，the growth ability of strain YX-1 was significantly inhibited. With the extension of Al treatment time，strain YX-1 gradually formed a biofilm，and the content of extracellular polysaccharides（EPS）also significantly increased. Under Al treatment，the expression levels of epsA，epsB，wzx and wzy in strain YX-1，which were responsible for EPS synthesis and secretion，initially increased and then decreased，with the highest expression observed at 12 h. Compared to Al treatment alone（100 mmol · L^-1），ginseng inoculated Bs and the addition of 100 mmol · L^-1 Al significantly increased the chlorophyll content of leaves，the dry weight of aboveground part and roots increased by 28.33% and 11.65%，respectively. Phosphorus content in leaves and roots increased by 13.89% and 16.24%，respectively，while the Al content in leaves and roots decreased by 32.11% and 11.90%，respectively. This study demonstrates that B. subtilis YX-1 promotes biofilm formation through EPS synthesis and secretion，mitigating Al stress on ginseng growth.

Key words: Panax ginseng, aluminum stress, Bacillus subtilis YX-1, biofilm, exopolysaccharides

梁池嘉, 丁冠中, 周帆, 穆朋, 张悦, 郑毅东, 王海涛, 王岩, 刘宁. 枯草芽孢杆菌YX-1对铝胁迫的响应及人参幼苗生长的影响[J]. 园艺学报, 2025, 52(9): 2507-2518.

LIANG Chijia, DING Guanzhong, ZHOU Fan, MU Peng, ZHANG Yue, ZHENG Yidong, WANG Haitao, WANG Yan, LIU Ning. Response of Bacillus subtilis YX-1 to Aluminum Stress and Its Effects on the Growth of Panax ginseng[J]. Acta Horticulturae Sinica, 2025, 52(9): 2507-2518.

图/表 8

表1 本研究中所用的qRT-PCR引物序列

Table 1 qRT-PCR primer sequence in this study

基因Gene	引物序列（5′-3′）Primer sequence	参考文献Reference
rpsJ	F：GAAACGGCAAAACGTTCTGG R：GTGTTGGGTTCACAATGCG	Jordan et al.,2006
epsA	F：GCTGAGATTGCGAACACGTT R：GACCATATTTCGCAGCCTGG	Belda et al.,2013
epsB	F：CCGGCAGAACTGTTGTCTTC R：TTCCGCTTAAAACGACGAGC	Belda et al.,2013
wzx	F：CCGCAAGGAGGTGT R：ATGATGAGCGACTGG	Johnson et al.,2015
wzy	F：TCCCACTCCGACTC R：CCAATGCCAATGAA	Johnson et al.,2015

图1 Bacillus subtilis YX-1的系统发育进化树（A）及耐铝性比较（B）进化树分支点上的数字代表置信值。不同小写字母表示处理间差异显著（P < 0.05）。下同

Fig. 1 Phylogenetic tree（A）of Bacillus subtilis YX-1 and comparsion on Al tolerance（B） The number on the branch point represents the confidence value. Different lowercase letters indicate significant differences among the treatments（P < 0.05）.The same below

图2 铝胁迫下不同时间点Bacillus subtilis YX-1生物膜形成的荧光成像图

Fig. 2 Fluorescent imaging of Bacillus subtilis YX-1 biofilm formation at different time points under Al stress

图3 铝胁迫下不同时间Bacillus subtilis YX-1胞外多糖含量

Fig. 3 The EPS content of Bacillus subtilis YX-1 at different time points under Al stress * P < 0.05

图4 铝胁迫对Bacillus subtilis YX-1生物膜形成相关基因表达量的影响 ns表示差异不显著，* P < 0.05，** P < 0.01

Fig. 4 Effects of Al stress on the expression of biofilm-related genes in Bacillus subtilis YX-1 ns indicates no significant difference，* P < 0.05，** P < 0.01

图5 铝胁迫下Bacillus subtilis YX-1对人参表型的影响

Fig. 5 Effects of Al stress on Panax ginseng phenotype with or without Bacillus subtilis YX-1

图6 铝胁迫下Bacillus subtilis YX-1对人参生长的影响

Fig. 6 Effects of Al stress on Panax ginseng growth with or without Bacillus subtilis YX-1

表2 铝胁迫下Bacillus subtilis YX-1对人参根际土壤理化性质的影响

Table 2 Effects of Al stress on the physical and chemical properties of rhizosphere soil of Panax ginseng with or without Bacillus subtilis YX-1

处理 Treatment	土壤pH Soil pH	土壤有机质含量/（g • kg^-1） Soil organic matter content	速效磷含量/（g • kg^-1） Available P content	活性铝含量/（mg • kg^-1） Active Al content
H₂O（对照 Control）	5.27 ± 0.06 b	1.26 ± 0.02 a	10.76 ± 2.52 ab	477.74 ± 4.98 c
Bs	5.50 ± 0.06 a	1.31 ± 0.02 a	14.75 ± 1.61 a	446.10 ± 8.64 d
100 mmol · L^-1Al	4.48 ± 0.03 d	1.26 ± 0.02 a	8.34 ± 1.70 b	719.68 ± 16.60 a
Bs + 100 mmol · L^-1Al	4.91 ± 0.03 c	1.26 ± 0.03 a	9.16 ± 1.71 b	675.05 ± 16.58 b

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