中国农业科学 ›› 2019, Vol. 52 ›› Issue (24): 4505-4517.doi: 10.3864/j.issn.0578-1752.2019.24.006
赵卫松,郭庆港,李社增,王亚娇,鹿秀云,王培培,苏振贺,张晓云,马平()
收稿日期:
2019-06-03
接受日期:
2019-06-28
出版日期:
2019-12-16
发布日期:
2020-01-15
通讯作者:
马平
作者简介:
赵卫松,Tel:0312-5927076;E-mail:zhaoweisong1985@163.com。
基金资助:
WeiSong ZHAO,QingGang GUO,SheZeng LI,YaJiao WANG,XiuYun LU,PeiPei WANG,ZhenHe SU,XiaoYun ZHANG,Ping MA()
Received:
2019-06-03
Accepted:
2019-06-28
Online:
2019-12-16
Published:
2020-01-15
Contact:
Ping MA
摘要:
【目的】研究土壤中添加西兰花残体(broccoli residues,BR)对棉花黄萎病及不同生育时期土壤细菌群落结构的影响,为棉花黄萎病的绿色生态防控和化学农药的减量施用提供新途径和思路。【方法】通过田间小区试验,分别以西兰花残体还田和不含有西兰花残体(CK)的耕层土壤中播种的棉花为试验对象,监测不同处理的棉花黄萎病的发生动态。采用实时荧光定量PCR(real-time PCR)和高通量测序(Illumina MiSeq)技术分别测定不同时期土壤中大丽轮枝菌(Verticillium dahliae)DNA拷贝数量和土壤细菌群落,进而分析西兰花残体还田对土壤中病原菌数量变化和土壤细菌群落的影响。采用主成分分析探索西兰花残体还田对不同生育时期土壤细菌群落在门和属水平的结构变化特征与规律。【结果】西兰花残体还田后棉花黄萎病的发病率和病情指数均呈现出下降趋势,且发病高峰时间推迟,在发病高峰期对黄萎病的防治效果达到70.77%,建立了整个生育期内病情指数-时间病程发展曲线,平均防治效果为57.21%。与空白对照相比,西兰花残体还田处理在棉花生长季和非生长季的4个阶段土壤中大丽轮枝菌DNA拷贝数量分别降低了10.96%、11.11%、25.95%和11.25%。高通量测序分析表明,不同生育时期西兰花残体还田处理显著提高了土壤细菌多样性,丰富度指数Chao1均显著上升,ACE指数在播种前期、花铃期和吐絮期显著上升。与空白对照相比,在不同生育时期内,西兰花残体还田后土壤中的放线菌门(Actinobacteria)菌群的相对丰度呈现显著上升。同时研究表明,西兰花残体还田后优势菌群组成在不同生育时期存在差别,在播种前期和苗期,厚壁菌门(Firmicutes)菌群上升为优势菌群;在蕾期和花铃期,蓝藻细菌门(Cyanobacteria)菌群成为优势菌群。主成分分析表明,西兰花残体还田改变了苗期、蕾期、花铃期和吐絮期的细菌群落结构。进一步分析表明,西兰花残体还田显著增加了链霉菌属(Streptomyces)和芽孢杆菌属(Bacillus)菌群的相对丰度。【结论】西兰花残体还田能够有效抑制棉花黄萎病的发生,降低土壤中大丽轮枝菌DNA拷贝的数量,改变土壤细菌群落结构,增加有益微生物的相对丰度,是一种有效控制棉花黄萎病的绿色措施。
赵卫松,郭庆港,李社增,王亚娇,鹿秀云,王培培,苏振贺,张晓云,马平. 西兰花残体还田对棉花黄萎病防治效果及其对不同生育时期土壤细菌群落的影响[J]. 中国农业科学, 2019, 52(24): 4505-4517.
WeiSong ZHAO,QingGang GUO,SheZeng LI,YaJiao WANG,XiuYun LU,PeiPei WANG,ZhenHe SU,XiaoYun ZHANG,Ping MA. Control Efficacy of Broccoli Residues on Cotton Verticillium Wilt and Its Effect on Soil Bacterial Community at Different Growth Stages[J]. Scientia Agricultura Sinica, 2019, 52(24): 4505-4517.
表1
西兰花残体还田对棉花不同生育时期细菌测序量和群落多样性的影响"
时期 Stage | 处理 Treatment | 测序 Reads | 丰富度Richness | 均匀度Evenness | ||
---|---|---|---|---|---|---|
Chao1指数 Chao1 index | ACE指数 ACE index | 辛普森指数 Simpson index | 香浓指数 Shannon index | |||
播种前期 Pre-sowing stage | 对照CK | 38251.50±293.50a | 2814.33±14.34a | 2813.33±69.14a | 0.9986±0.0002a | 10.67±0.13a |
西兰花残体BR | 39434.67±2787.82a | 3339.07±262.21b | 3419.18±147.49b | 0.9970±0.0007a | 10.18±0.10a | |
苗期 Seedling stage | 对照CK | 38800.00±206.00a | 2703.00±12.25a | 2669.00±41.67a | 0.9984±0.0001a | 10.56±0.08a |
西兰花残体BR | 42364.67±3689.72a | 2795.04±64.96b | 2691.80±68.21a | 0.9984±0.0000a | 10.56±0.06a | |
蕾期 Bud stage | 对照CK | 42593.50±495.50a | 3039.57±46.12a | 3143.00±80.87a | 0.9985±0.0002a | 10.67±0.21a |
西兰花残体BR | 41670.00±2906.67a | 3624.21±223.84b | 3173.46±99.20a | 0.9983±0.0000a | 10.16±0.44a | |
花铃期 Flowering and boll stage | 对照CK | 36694.00±35.00a | 2861.00±36.34a | 2861.00±31.38a | 0.9987±0.0000a | 10.71±0.25a |
西兰花残体BR | 41284.67±3638.72a | 3763.18±167.23b | 3946.36±97.67b | 0.9982±0.0002a | 10.49±0.11a | |
吐絮期 Boll-opening stage | 对照CK | 36369.50±75.50a | 2543.33±57.97a | 2558.00±60.21a | 0.9965±0.0006a | 10.33±0.33a |
西兰花残体BR | 34275.33±2063.41a | 2665.31±259.75b | 2813.93±279.95b | 0.9974±0.0001a | 10.12±0.34a |
图4
棉花不同生育时期土壤细菌群落结构的主成分分析 红色椭圆代表空白对照(1—15),绿色椭圆代表西兰花残体处理(16—30)The red ellipse represents the blank control (1-15), the green ellipse represents the broccoli residues treatment (16-30)。1—3、16—18:播种前期Pre-sowing stage;4—6、19—21:苗期Seedling stage;7—9、22—24:蕾期Bud stage;10—12、25—27:花铃期Flowering and boll stage;13—15、28—30:吐絮期Boll-opening stage。Proteobacteria:变形菌门;Actinobacteria:放线菌门;Acidobacteria:酸杆菌门;Gemmatimonadetes:芽单孢菌门;Chloroflexi:绿弯菌门;Planctomycetes:浮霉菌门;Nitrospirae:硝化螺旋菌门;Bacteroidetes: 拟杆菌门;Firmicutes:厚壁菌门;Cyanobacteria:蓝藻细菌门;Others:其他"
表2
属水平棉花不同生育时期细菌菌群相对丰度(10组中至少1组相对丰度>1%)"
属 Genus | 时期Stage | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
播种前期 Pre-sowing stage | 苗期 Seedling stage | 蕾期 Bud stage | 花铃期 Flowering and boll stage | 吐絮期 Boll-opening stage | ||||||
CKT0 | BRT0 | CKT1 | BRT1 | CKT2 | BRT2 | CKT3 | BRT3 | CKT4 | BRT4 | |
Haliangium | 1.93b | 0.96a | 2.16b | 1.58a | 0.75b | 0.54a | 1.21b | 1.07a | 1.48b | 0.87a |
鞘脂单胞菌属Sphingomonas | 2.18b | 1.57a | 2.73a | 2.66a | 2.13a | 2.32b | 2.51b | 2.33a | 1.24b | 1.16a |
Iamia | 1.28a | 1.16a | 0.40a | 0.65b | 0.51a | 0.90b | 0.82a | 1.01b | 0.63a | 1.66b |
芽单胞菌属Gemmatimonas | 0.98b | 0.34a | 1.08b | 0.88a | 1.49b | 0.81a | 2.27b | 1.01a | 0.57a | 1.12b |
硝化螺菌属Nitrospira | 0.81a | 1.03b | 1.34a | 1.32a | 0.70b | 0.51a | 1.07a | 0.93a | 1.03b | 0.61a |
红色杆菌属Rubrobacter | 0.72b | 0.55a | 1.11b | 0.82a | 1.20b | 0.68a | 0.93b | 0.75a | 0.85a | 0.84a |
Pontibacter | 0.77b | 0.22a | 1.36b | 0.61a | 0.96b | 0.56a | 0.96b | 0.47a | 0.20a | 0.13a |
游动放线菌属Actinoplanes | 0.40b | 0.28a | 0.48a | 0.55b | 0.67a | 1.74b | 1.81b | 0.50a | 0.76a | 0.76b |
Bryobacter | 0.55b | 0.45a | 0.98b | 0.73a | 0.76b | 0.50a | 1.20b | 0.61a | 0.53a | 0.58a |
气微菌属Aeromicrobium | 0.83b | 0.48a | 1.68b | 1.16a | 0.67a | 0.82b | 0.60b | 0.47a | 0.17a | 0.58b |
玫瑰弯菌属Roseiflexus | 0.29a | 0.43b | 0.53a | 0.65b | 0.63b | 0.30a | 1.13b | 0.67a | 0.61a | 0.87b |
Solirubrobacter | 0.46b | 0.35a | 0.60a | 0.62a | 0.50a | 0.54a | 1.01b | 0.70a | 0.54a | 0.53a |
类诺卡氏属Nocardioides | 0.53a | 0.69b | 0.42a | 0.40a | 0.56a | 0.68b | 0.84a | 1.03b | 0.34a | 0.63b |
溶杆菌属Lysobacter | 0.65a | 0.67a | 0.98b | 0.55a | 0.19a | 0.50b | 0.23a | 0.57b | 0.20a | 0.75b |
链霉菌属Streptomyces | 0.29a | 0.83b | 0.23a | 0.35b | 0.48a | 1.16b | 0.39a | 0.55b | 0.27a | 0.55b |
芽孢杆菌属Bacillus | 0.33a | 1.29b | 0.23a | 0.62b | 0.40a | 0.53b | 0.29a | 0.61b | 0.20a | 0.19a |
Nonomuraea | 0.26a | 2.58b | 0.01a | 0.07b | 0.14a | 0.30b | 0.06a | 0.07a | 0.07a | 0.74b |
其他Others | 86.75a | 86.10a | 83.66a | 85.77b | 87.25b | 86.61a | 82.67a | 86.65b | 90.64b | 87.43a |
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