摘要:

目的   比較研究炭疽病不同發(fā)病程度草莓根際微生物群落結(jié)構(gòu)并開展基因功能預測分析，為指導農(nóng)田管理及篩選草莓炭疽病拮抗菌提供理論依據(jù)和參考。

方法  采集設施栽培大棚中健康和不同發(fā)病程度的草莓植株根際土壤，利用Illumina Miseq測序比較分析不同發(fā)病程度對草莓根際細菌多樣性，根際細菌門、屬水平群落組成特征的影響。

結(jié)果  發(fā)病植株根際細菌多樣性顯著低于健康植株，其中輕度發(fā)病草莓根際細菌豐富度ACE降低了7.2%，而中度發(fā)病草莓根際細菌豐富度降低了11%。草莓根際土壤的優(yōu)勢菌群為變形菌門、放線菌門和擬桿菌門，不同發(fā)病程度的草莓根際土壤的細菌群落結(jié)構(gòu)發(fā)生了顯著改變，其中發(fā)病草莓顯著提高了根際土壤中芽孢菌屬和亞棲熱菌屬的相對豐度，而降低了與促生功能相關(guān)的黃桿菌屬和伯克氏菌屬的相對豐度。PICRUSt2分析表明，炭疽病顯著提高草莓根際細菌的細胞交流、細胞傳遞和分解，轉(zhuǎn)錄以及循環(huán)等功能，而降低了脂質(zhì)代謝和消化功能。

結(jié)論  本研究表明炭疽病顯著降低草莓根際細菌多樣性，并改變細菌群落結(jié)構(gòu)，降低了促進草莓營養(yǎng)吸收細菌種類的相對豐度，改變了根際細菌功能特征，研究結(jié)果為指導草莓生產(chǎn)管理和篩選抗病微生物提供參考。

Abstract:

Objective  The aim of this study is to compare the rhizosphere microbial community structure and function of strawberry among difference degrees of anthracnose, which is crucial for guiding farmland management and screening useful strawberry anthracnose antagonistic resource.

Method  The strawberry rhizosphere soil among difference degrees of anthracnose in the same field was collected in a greenhouse for studying the rhizosphere bacterial diversity and richness at phylum and genus levels by using Illumina sequencing and PICRUSt2.

Result  The results showed that anthracnose decreased rhizosphere bacterial diversity, with the mild degrees of anthracnose decreased 7.2% (ACE) and the middle decreased 11%. The phyla Proteobacteria, Actinobacteria and Bacteroidetes dominated in all strawberry rhizosphere soil. The different degrees of anthracnose changed rhizosphere soil bacterial composition, with an increase of Bacillus. spp and Meiothermus. Spp, while a decreased of Flavobacterium and Burkholderia. The PICRUSt2 analysis showed that the occurrence of anthracnose disease increased cell communication, transport and catabolism, transcription, but decreased lipid metabolism and digestive system.

Conclusion  The strawberry anthracnose change rhizosphere bacterial composition, functional traits and decrease diversity. In addition, the strawberry anthracnose also decreases the relative abundance of bacterial taxa and enhances plant nutrient absorption.

圖  1   健康及不同發(fā)病程度草莓根際細菌組成（門水平）

每組處理柱旁不同字母表示差異顯著（P < 0.05, Tukey’s test）

Figure  1.   Relative average abundances of bacteria in healthy and infected strawberries (at phylum level)

圖  2   健康及不同發(fā)病程度草莓根際細菌組成（屬水平）

每組處理柱上不同字母表示差異顯著（P < 0.05, Tukey’s test）

Figure  2.   Relative average abundances of bacteria in healthy and infected strawberry (at genus level)

圖  3   健康及不同發(fā)病程度草莓根際細菌PCoA分析（OTU水平）

Figure  3.   Results of PCoA analysis based on weighted Fast UniFrac distance at OTU level

圖  4   不同發(fā)病程度草莓根際細菌功能變化分析 (一級功能層)

Figure  4.   Changes of bacterial function profiles of different samples examined using changes of bacterial function profiles of different samples examined using PICRUSt2 （hierarchy level 1）

圖  5   不同發(fā)病程度草莓根際細菌功能變化分析 (二級功能層)

Figure  5.   Changes of bacterial function profiles of different samples examined using Changes of bacterial function profiles of different samples examined using PIRCUSt2 (Hierarchy level 2)

表  1   健康及不同發(fā)病程度草莓根際細菌多樣性

Table  1   Effects of healthy and different infected stage strawberry on rhizosphere bacterial α diversity

處理代號
CodeOTUsShannonACEChao1 T0 4194 ± 276 a 6.92 ± 0.07 a 5492 ± 238 a 5502 ± 191 a T1 3842 ± 352 ab 6.74 ± 0.10 b 5092 ± 346 b 5109 ± 325 b T2 3639 ± 289 b 6.74 ± 0.08 b 4890 ± 335 b 4915 ± 367 b 　　注：表中數(shù)據(jù)為平均值 ± 標準差，同列不同字母表示差異顯著（P < 0.05）。T0健康草莓根際土壤，T1初始發(fā)病狀態(tài)草莓根際土壤，T2發(fā)病中期草莓根際土壤?！　?

表  2   不同處理間細菌群落差異性分析

Table  2   The bacterial composition difference among different treatments under long-term fertilization

組別
Group差異性
Difference 系數(shù)
Coefficient (R)顯著性
Significance (P) T0 vs T1 vs T2 0.6393 0.001 T0 vs T1 0.5037 0.003 T0 vs T2 0.6198 0.003 T1 vs T2 0.5651 0.003 [1]

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