中国农业科技导报 ›› 2022, Vol. 24 ›› Issue (6): 58-69.DOI: 10.13304/j.nykjdb.2021.0692
王莉莉1,5(), 殷丛培2(
), 李峰3, 杨志敏3, 刘芳明1, 林柏松1, 刘晓静1, 刘海军4, 孙靖4, 单东东4, 崔江慧2,5(
), 张振清4,5(
)
收稿日期:
2021-08-12
接受日期:
2021-11-15
出版日期:
2022-06-15
发布日期:
2022-06-21
通讯作者:
崔江慧,张振清
作者简介:
王莉莉 E-mail:546156736@qq.com基金资助:
Lili WANG1,5(), Congpei YIN2(
), Feng LI3, Zhimin YANG3, Fangming LIU1, Baisong LIN1, Xiaojing LIU1, Haijun LIU4, Jing SUN4, Dongdong SHAN4, Jianghui CUI2,5(
), Zhenqing ZHANG4,5(
)
Received:
2021-08-12
Accepted:
2021-11-15
Online:
2022-06-15
Published:
2022-06-21
Contact:
Jianghui CUI,Zhenqing ZHANG
摘要:
为研究马铃薯根际土壤细菌的群落结构及其对干旱胁迫的响应,采用盆栽试验,以开花期马铃薯根际土壤为研究对象,提取其总DNA,通过Illumina Miseq对细菌 V3~V4区进行高通量测序,对干旱处理下马铃薯根际微生物的群落结构进行分析。结果表明,共鉴定有效序列1 263 889条,6 785个OTUs归属33个门、94个纲、267个目、462个科、919个属、1 930个种。与对照土壤相比,种植马铃薯后根际土壤中的变形菌门(Proteobacteria)、放线菌门(Actinobacteria)、酸杆菌门(Acidobacteria)、绿弯菌门(Chloroflexi)、拟杆菌门(Bacteroidetes)、厚壁菌门(Firmicutes)和芽单胞菌门(Gemmatimonadetes)为优势菌门,其中放线菌门(Proteobacteria)的相对丰度显著增加。干旱处理提高了马铃薯根际土壤中变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)、芽单胞菌门(Gemmatimonadetes)和假单胞菌属(Pseudomonas)的相对丰度。PCoA(principal coordinates analysis)分析显示,种植马铃薯和干旱胁迫是造成土壤细菌群落结构变异的重要因素。PICRUSt(phylogenetic investigation of communities by reconstruction of unobserved states)分析表明,干旱处理后根际土壤细菌的变化与马铃薯植株的生理代谢、信号转导、防御机制和基本生命活性等密切相关,对植物的生存和耐旱性具有重要意义,为通过改良土壤微生物环境来提高植物胁迫耐受性提供了重要参考。
中图分类号:
王莉莉, 殷丛培, 李峰, 杨志敏, 刘芳明, 林柏松, 刘晓静, 刘海军, 孙靖, 单东东, 崔江慧, 张振清. 马铃薯根际土壤细菌群落结构及其对干旱胁迫的响应[J]. 中国农业科技导报, 2022, 24(6): 58-69.
Lili WANG, Congpei YIN, Feng LI, Zhimin YANG, Fangming LIU, Baisong LIN, Xiaojing LIU, Haijun LIU, Jing SUN, Dongdong SHAN, Jianghui CUI, Zhenqing ZHANG. Microbial Community Structure of Potato Rhizosphere Soil and Its Response to Drought Stress[J]. Journal of Agricultural Science and Technology, 2022, 24(6): 58-69.
品种Variety | 处理Treatment | 净光合速率Pn/(µmol·m-2·s-1) | 蒸腾速率 Tr/(mmol·m-2·s-1) | 气孔导度 Gs/(mol·m-2·s-1) | 胞间CO2浓度 Ci/(µmol·mol-1) |
---|---|---|---|---|---|
冀张薯8号 Jizhangshu 8 | T0 | 22.00±2.12 a | 2.35±0.33 a | 13.83±1.10 a | 170.33±10.66 a |
T1 | 24.80±9.04 a | 2.27±0.21 a | 10.33±0.74 b | 178.53±16.03 a | |
T2 | 17.44±3.10 ab | 2.01±0.21 ab | 8.50±0.89 bc | 182.67±9.18 a | |
T3 | 13.91±1.08 b | 1.79±0.05 ab | 6.00±0.47 c | 195.41±9.50 a | |
夏波蒂 Shepody | T0 | 28.01±2.22 a | 2.79±0.16 a | 13.76±1.35 a | 137.42±11.03 b |
T1 | 18.63±3.66 a | 2.17±0.45 b | 11.67±1.06 ab | 159.00±3.65 ab | |
T2 | 14.73±2.00 b | 1.58±0.32 b | 8.87±1.51 bc | 173.14±7.11 a | |
T3 | 8.97±1.82 c | 1.03±0.11 c | 4.80±1.08 c | 180.00±13.12 a |
表1 不同处理下马铃薯叶片的光合特性
Table 1 Photosynthetic characteristics of potato leaves under different treatments
品种Variety | 处理Treatment | 净光合速率Pn/(µmol·m-2·s-1) | 蒸腾速率 Tr/(mmol·m-2·s-1) | 气孔导度 Gs/(mol·m-2·s-1) | 胞间CO2浓度 Ci/(µmol·mol-1) |
---|---|---|---|---|---|
冀张薯8号 Jizhangshu 8 | T0 | 22.00±2.12 a | 2.35±0.33 a | 13.83±1.10 a | 170.33±10.66 a |
T1 | 24.80±9.04 a | 2.27±0.21 a | 10.33±0.74 b | 178.53±16.03 a | |
T2 | 17.44±3.10 ab | 2.01±0.21 ab | 8.50±0.89 bc | 182.67±9.18 a | |
T3 | 13.91±1.08 b | 1.79±0.05 ab | 6.00±0.47 c | 195.41±9.50 a | |
夏波蒂 Shepody | T0 | 28.01±2.22 a | 2.79±0.16 a | 13.76±1.35 a | 137.42±11.03 b |
T1 | 18.63±3.66 a | 2.17±0.45 b | 11.67±1.06 ab | 159.00±3.65 ab | |
T2 | 14.73±2.00 b | 1.58±0.32 b | 8.87±1.51 bc | 173.14±7.11 a | |
T3 | 8.97±1.82 c | 1.03±0.11 c | 4.80±1.08 c | 180.00±13.12 a |
品种Variety | 处理 Treatment | 超氧化物歧化酶活性 SOD activity/(U·min-1) | 过氧化物酶活性 POD activity /(U·min-1) | 过氧化氢酶活性 CAT activity /(U·min-1) |
---|---|---|---|---|
冀张薯8号 Jizhangshu 8 | T0 | 118.04±8.77 b | 73.50±2.45 c | 5.38±0.20 b |
T1 | 123.74±6.62 b | 93.76±4.71 b | 6.70±0.61 ab | |
T2 | 145.98±7.02 b | 98.35±5.03 b | 8.01±0.30 a | |
T3 | 240.01±9.22 a | 119.98±6.73 a | 5.30±0.40 b | |
夏波蒂 Shepody | T0 | 44.98±5.66 c | 74.03±2.07 c | 4.66±0.31 a |
T1 | 58.06±4.08 b | 94.03±3.99 b | 4.76±0.58 a | |
T2 | 67.02±5.31 ab | 97.77±3.22 b | 4.72±0.57 a | |
T3 | 87.96±6.33 a | 118.03±4.14 a | 3.13±0.71 b |
表 2 干旱胁迫下马铃薯叶片保护酶的活性
Table 2 Enzyme activities of potato leaves under different treatments
品种Variety | 处理 Treatment | 超氧化物歧化酶活性 SOD activity/(U·min-1) | 过氧化物酶活性 POD activity /(U·min-1) | 过氧化氢酶活性 CAT activity /(U·min-1) |
---|---|---|---|---|
冀张薯8号 Jizhangshu 8 | T0 | 118.04±8.77 b | 73.50±2.45 c | 5.38±0.20 b |
T1 | 123.74±6.62 b | 93.76±4.71 b | 6.70±0.61 ab | |
T2 | 145.98±7.02 b | 98.35±5.03 b | 8.01±0.30 a | |
T3 | 240.01±9.22 a | 119.98±6.73 a | 5.30±0.40 b | |
夏波蒂 Shepody | T0 | 44.98±5.66 c | 74.03±2.07 c | 4.66±0.31 a |
T1 | 58.06±4.08 b | 94.03±3.99 b | 4.76±0.58 a | |
T2 | 67.02±5.31 ab | 97.77±3.22 b | 4.72±0.57 a | |
T3 | 87.96±6.33 a | 118.03±4.14 a | 3.13±0.71 b |
处理 Treatment | 有效序列数 Sequence number | 碱基数 Base number | 平均长度 Mean length/bp | 最短序列长度 Min. length/bp | 最长序列长度 Max. length/bp |
---|---|---|---|---|---|
对照土CK soil | 173 462 | 72 743 138 | 419.36 | 257 | 480 |
S-T0 | 150 075 | 62 852 665 | 418.84 | 276 | 472 |
S-T1 | 125 549 | 52 301 567 | 416.60 | 289 | 455 |
S-T2 | 159 305 | 66 521 294 | 417.57 | 243 | 476 |
S-T3 | 178 086 | 74 359 015 | 417.53 | 271 | 471 |
JZS-T0 | 171 403 | 71 496 233 | 417.13 | 214 | 468 |
JZS-T1 | 157 105 | 65 548 212 | 417.20 | 271 | 474 |
JZS-T2 | 161 144 | 67 378 509 | 418.09 | 240 | 504 |
JZS-T3 | 161 222 | 67 470 566 | 418.48 | 235 | 490 |
表3 不同处理下根际细菌群落的测序质量
Table 3 Sequencing quantity of rhizosphere bacteria under different treatments
处理 Treatment | 有效序列数 Sequence number | 碱基数 Base number | 平均长度 Mean length/bp | 最短序列长度 Min. length/bp | 最长序列长度 Max. length/bp |
---|---|---|---|---|---|
对照土CK soil | 173 462 | 72 743 138 | 419.36 | 257 | 480 |
S-T0 | 150 075 | 62 852 665 | 418.84 | 276 | 472 |
S-T1 | 125 549 | 52 301 567 | 416.60 | 289 | 455 |
S-T2 | 159 305 | 66 521 294 | 417.57 | 243 | 476 |
S-T3 | 178 086 | 74 359 015 | 417.53 | 271 | 471 |
JZS-T0 | 171 403 | 71 496 233 | 417.13 | 214 | 468 |
JZS-T1 | 157 105 | 65 548 212 | 417.20 | 271 | 474 |
JZS-T2 | 161 144 | 67 378 509 | 418.09 | 240 | 504 |
JZS-T3 | 161 222 | 67 470 566 | 418.48 | 235 | 490 |
处理 Treatment | 测序深度 Sequencing depth coverage | 丰度指数 Richness index | 多样性指数 Diversity index | 均匀度指数 Evenness index | ||||
---|---|---|---|---|---|---|---|---|
Sobs | Chao | ACE | Shannon | Simpson | Shannon | Simpson | ||
对照土CK soil | 0.956 3 | 2 871.00±46.89 a | 4 019.46±124.69 a | 4 226.19±454.79 a | 6.30±0.04 e | 0.007±0.001 b | 0.791±0.003 e | 0.049±0.005 cd |
S-T0 | 0.963 4 | 3 034.00±126.57 a | 4 300.03±193.64 a | 4 285.13±178.17 a | 6.64±0.04 b | 0.004±0.001 bcd | 0.828±0.007 b | 0.076±0.009 b |
S-T1 | 0.962 7 | 2 914.33±332.18 a | 4 168.09±393.31 a | 4 306.80±129.02 a | 6.75±0.03 a | 0.003±0.000 d | 0.846±0.008 a | 0.117±0.016 a |
S-T2 | 0.969 5 | 2 891.67±143.07 a | 4 154.81±341.77 a | 4 412.31±699.81 a | 6.49±0.04 cd | 0.004±0.000 bcd | 0.813±0.001 bcd | 0.078±0.002 b |
S-T3 | 0.972 9 | 3 093.67±146.73 a | 4 318.33±190.86 a | 4 331.86±175.96 a | 6.53±0.02 cd | 0.005±0.000 bcd | 0.813±0.002 cd | 0.066±0.001 bc |
JZS-T0 | 0.967 6 | 3 123.00±187.28 a | 4 409.18±296.88 a | 4 391.73±254.61 a | 6.46±0.12 d | 0.010±0.004 a | 0.802±0.015 de | 0.036±0.018 d |
JZS-T1 | 0.970 9 | 2 999.00±184.52 a | 4 260.66±277.70 a | 4 209.69±243.27 a | 6.58±0.03 bc | 0.004±0.000 cd | 0.822±0.003 bc | 0.085±0.010 b |
JZS-T2 | 0.970 0 | 3 011.67±127.44 a | 4 273.25±239.49 a | 4 240.67±228.24 a | 6.52±0.06 cd | 0.005±0.001 bcd | 0.814±0.009 bcd | 0.075±0.015 b |
JZS-T3 | 0.969 6 | 3 015.67±100.95 a | 4 256.61±110.02 a | 4 253.03±136.65 a | 6.45±0.07 d | 0.006±0.001 bc | 0.804±0.010 de | 0.054±0.009 cd |
表4 各处理根际土壤样本 Alpha 多样性指数
Table 4 Alpha diversity index of rhizosphere soil samples in each treatment
处理 Treatment | 测序深度 Sequencing depth coverage | 丰度指数 Richness index | 多样性指数 Diversity index | 均匀度指数 Evenness index | ||||
---|---|---|---|---|---|---|---|---|
Sobs | Chao | ACE | Shannon | Simpson | Shannon | Simpson | ||
对照土CK soil | 0.956 3 | 2 871.00±46.89 a | 4 019.46±124.69 a | 4 226.19±454.79 a | 6.30±0.04 e | 0.007±0.001 b | 0.791±0.003 e | 0.049±0.005 cd |
S-T0 | 0.963 4 | 3 034.00±126.57 a | 4 300.03±193.64 a | 4 285.13±178.17 a | 6.64±0.04 b | 0.004±0.001 bcd | 0.828±0.007 b | 0.076±0.009 b |
S-T1 | 0.962 7 | 2 914.33±332.18 a | 4 168.09±393.31 a | 4 306.80±129.02 a | 6.75±0.03 a | 0.003±0.000 d | 0.846±0.008 a | 0.117±0.016 a |
S-T2 | 0.969 5 | 2 891.67±143.07 a | 4 154.81±341.77 a | 4 412.31±699.81 a | 6.49±0.04 cd | 0.004±0.000 bcd | 0.813±0.001 bcd | 0.078±0.002 b |
S-T3 | 0.972 9 | 3 093.67±146.73 a | 4 318.33±190.86 a | 4 331.86±175.96 a | 6.53±0.02 cd | 0.005±0.000 bcd | 0.813±0.002 cd | 0.066±0.001 bc |
JZS-T0 | 0.967 6 | 3 123.00±187.28 a | 4 409.18±296.88 a | 4 391.73±254.61 a | 6.46±0.12 d | 0.010±0.004 a | 0.802±0.015 de | 0.036±0.018 d |
JZS-T1 | 0.970 9 | 2 999.00±184.52 a | 4 260.66±277.70 a | 4 209.69±243.27 a | 6.58±0.03 bc | 0.004±0.000 cd | 0.822±0.003 bc | 0.085±0.010 b |
JZS-T2 | 0.970 0 | 3 011.67±127.44 a | 4 273.25±239.49 a | 4 240.67±228.24 a | 6.52±0.06 cd | 0.005±0.001 bcd | 0.814±0.009 bcd | 0.075±0.015 b |
JZS-T3 | 0.969 6 | 3 015.67±100.95 a | 4 256.61±110.02 a | 4 253.03±136.65 a | 6.45±0.07 d | 0.006±0.001 bc | 0.804±0.010 de | 0.054±0.009 cd |
图1 3种分组方式的组内和组间差异ANOSIM分析A:品种;B:空白对照土与马铃薯根际土;C:干旱胁迫
Fig. 1 ANOSIM analysis of differences between groups and groups in 3 grouping methodsA: variety;B: CK soil and planted soil;C: drought stress
分组Group | R值R value | P值P value |
---|---|---|
A | 0.416 7 | 0.001 |
B | 0.774 4 | 0.001 |
C | 0.907 2 | 0.001 |
表5 相似性分析
Table 5 Analysis of similarties
分组Group | R值R value | P值P value |
---|---|---|
A | 0.416 7 | 0.001 |
B | 0.774 4 | 0.001 |
C | 0.907 2 | 0.001 |
图2 种植马铃薯后土壤细菌多样性A:OTUs分析;B:门水平优势细菌的相对丰度;C:组间差异;D:PCoA分析。*、**、***分别表示在P<0.05、P<0.01和P<0.001水平差异显著。
Fig. 2 Effects of planted potato on bacterial diversityA:OTUs analysis;B:Relative abundance of dominant bacteria at phylum level;C:Differences between CK and rhizosphere soils;D:PCoA analysis.*,** and *** indicate significant differences at P<0.05,P<0.01 and P<0.001 levels,respectively.
图3 干旱胁迫下马铃薯根际细菌群落结构A:OTUs 分析;B:门水平;C:属水平
Fig. 3 Bacterial community abundance in potato rhizosphere under drought stressA: OTUs analysis; B: Phylum level; C: Genus level in potato rhizosphere
图4 不同干旱胁迫下马铃薯根际菌群组间差异注:A—门水平;B—属水平;C:门水平PCoA分析;*、**、***分别表示在P<0.05、P<0.01和P<0.001水平差异显著。
Fig. 4 Difference and distribution of rhizosphere soils under drought stressNote:A—Phylum level; B—Genus level; C—PCoA analysis on Phylum level. *, ** and *** indicate significant differences at P<0.05, P<0.01 and P<0.001 levels, respectively.
图5 根际微生物的系统发生树及功能预测A:系统进化分析;B:根际微生物的功能预测
Fig. 5 Taxonomic analysis through phylogenetic tree and microbial functional featuresA: Phylogenetic analysis; B: Functional features of rhizosphere microbe
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