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果树学报 Journal of Fruit Science ISSN 1009-9980,CN 41-1308/S 果树学报网络首发论文 题目 江苏省不同区域草莓连作土壤养分及微生物区系分析 作者 王廷峰，赵密珍，关玲，赵帆，于红梅，庞夫花 DOI 10.13925/ki.gsxb.20180273 收稿日期 2018-07-17 网络首发日期 2018-11-19 引用格式 王廷峰，赵密珍，关玲，赵帆，于红梅，庞夫花．江苏省不同区域草莓连作土壤养分及微生物区系分析[J/OL]．果树学报. https//doi.org/10.13925/ki.gsxb.20180273 网络首发在编辑部工作流程中，稿件从录用到出版要经历录用定稿、排版定稿、整期汇编定稿等阶段。录用定稿指内容已经确定，且通过同行评议、主编终审同意刊用的稿件。排版定稿指录用定稿按照期刊特定版式（包括网络呈现版式）排版后的稿件，可暂不确定出版年、卷、期和页码。整期汇编定稿指出版年、卷、期、页码均已确定的印刷或数字出版的整期汇编稿件。录用定稿网络首发稿件内容必须符合出版管理条例和期刊出版管理规定的有关规定；学术研究成果具有创新性、科学性和先进性，符合编辑部对刊文的录用要求，不存在学术不端行为及其他侵权行为；稿件内容应基本符合国家有关书刊编辑、出版的技术标准，正确使用和统一规范语言文字、符号、数字、外文字母、法定计量单位及地图标注等。为确保录用定稿网络首发的严肃性，录用定稿一经发布，不得修改论文题目、作者、机构名称和学术内容，只可基于编辑规范进行少量文字的修改。 出版确认纸质期刊编辑部通过与中国学术期刊（光盘版）电子杂志社有限公司签约，在中国学术期刊（网络版）出版传播平台上创办与纸质期刊内容一致的网络版，以单篇或整期出版形式，在印刷出版之前刊发论文的录用定稿、排版定稿、整期汇编定稿。因为中国学术期刊（网络版）是国家新闻出版广电总局批准的网络连续型出版物（ISSN 2096-4188，CN 11-6037/Z），所以签约期刊的网络版上网络首发论文视为正式出版。 DOI10.13925/ki.gsxb.20180273 江苏省不同区域草莓连作土壤养分及微生物区系分析1王廷峰1,2，赵密珍1*，关 玲1，赵 帆1，于红梅1，庞夫花11江苏省农业科学院果树研究所 /江苏省高效园艺作物遗传改良重点实验室，南京 210014； 2南京农业大学园艺学院，南京 210095） 摘要 【 目的 】探究不同区域草莓连作障碍的发生原因。 【 方法 】用典型调查法调查并统计江苏省邳州港上镇、东海石榴镇和溧水白马镇草莓连作土壤的养分含量及微生物区系，并采集当地连作土与非连作土盆栽草莓，观察植株茎粗、鲜质量、株高、根长和根质量的生长状况。 【 结果 】港上镇和石榴镇的草莓连作土壤有机质和碱解氮含量缺乏；港上镇 010 cm 土层的土壤有机质，有效钙、镁、铁和硼含量比 1020 cm 土层低 10.5054.65，根际土壤养分流失严重；港上镇土壤有效锰含量缺乏，白马镇的土壤硼含量很缺乏；港上镇和白马镇的草莓连作土壤已发生轻度盐渍化，石榴镇土壤已发生中度盐渍化；各地连作土壤微生物均以细菌为主，真菌的数量最低；连作会给植株茎粗、鲜质量、株高、根质量和根长带来显著的负影响。 【 结论 】草莓连作会使土壤理化性质改变、养分失衡以及微生物群落结构失衡，影响植株生长。生产上应尽可能采取措施避免连作，并及时对连作土壤进行修复改良。 关键词 草莓；连作；土壤微生物；土壤养分 中图分类号 S668.4 文献标志码 A 文章编号 1009-9980201901-0001-08 Analysis of nutrients content and microbial flora in the continuous cropping soil of strawberry in different regions of Jiangsu province WANG Tingfeng1,2, ZHAO Mizhen1*, GUAN Ling1, ZHAO Fan1, YU Hongmei1, PANG Fuhua1 1Institute of Pomology, Jiangsu Academy of Agricultural Sciences/Jiangsu Key Laboratory of Horticultural Crops Genetic Improvement, Nanjing 210014, China; 2College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China Abstract 【 Objective】 In recent years, different levels of continuous cropping obstacles have occurred in the strawberry production of China. Continuous cropping could reduce the content of soil organic matter, and the gap in the abundance of trace elements could be widened. The proportion of bacteria, fungi, and actinomycetes in the soil could become unbalanced. As a result, strawberry plants will be susceptible to pests and diseases, making huge losses for strawberry producers. In our study, the strawberry continuous cropping soil was collected from Gangshang Town, Shiliu Town, and Baima Town, which are the main strawberry producing areas in Jiangsu Province. By investigating the nutrient contents and microflora status of continuous cropping soils in these area among different soil layers, we plan to analyze the effect of continuous cropping on strawberry plants, and to discover the mechanism and corresponding improvement s of continuous cropping obstacles, in order to provide a theoretical basis for overcoming continuous cropping obstacles in strawberry. 【 】 Soil samples were collected from strawberry greenhouses in Gangshang Town, Shiliu Town, and Baima Town in Jiangsu Province at the end 收稿日期 2018-07-17 接受日期 2018-10-10 1基金项目江苏省现代农业研究开发示范类项目（ BE2016369） ；江苏省三新工程（编号 SXGC[2017]261） 作者介绍王廷峰，男，在读硕士研究生，研究方向果树生理栽培， E-mail *通信作者 Author for correspondence. E-mail 网络首发时间2018-11-19 170035网络首发地址http// May 2016, and these areas have been continuously cropping strawberries for more than seven years. The greenhouses at different locations in the strawberry base in each region were used as three replicate experiments, and the soil of 0 to 10 cm, 10 to 20 cm, and 20 to 30 cm were collected using a five-point sampling . The soil samples were divided into two equal parts, one of which was used to detect soil microorganisms, and the other was used to determine soil physical and chemical properties. On September 5th of the same year, 20 strawberry plants were planted in continuous cropping soil in each area in a potted manner, and the non-continuous cropping soil in the same area was used as an experimental control. On the 57th day after planting November 1st, the status of plant growth was investigated. 【 Result】 The organic matter content g·kg-1 of surface soil 0 to 20 cm in Gangshang Town, Shiliu Town, and Baima Town was 18.40±5.42, 18.26±1.28, and 28.63±8.04, respectively, and the content of organic matter in Gangshang Town and Shiliu Town is lacking. As the depth of soil increases, the soil organic matter content in Shiliu Town and Baima Town showed a decreasing trend, while the content of soil organic matter in Gangshang Town increased first and then decreased. The content of available nitrogen mg·kg-1 in the surface of Gangshang Town, Shiliu Town, and Baima Town was 75.37±1.24, 81.25±3.09, and 113.75±14.21, respectively, the content of available nitrogen in Gangshang Town and Shiliu Town is relatively lacking, and the content of available nitrogen in Baima Town is moderate. The available soil nitrogen in the three regions was mainly concentrated in the 0 to 20 cm soil layer, and the available nitrogen content in the deep soil layer decreased by 33-45. The available manganese content mg·kg-1 in the surface of Gangshang Town, Shiliu Town, and Baima Town was 3.76±0.56, 44.15±3.20, and 13.06±0.85, respectively, and the abundances were respectively lacking, abundant, and moderate. The boron concentration mg·kg-1 in continuous cropping soil in Gangshang Town, Shiliu Town, and Baima Town was 0.63±0.27, 0.51±0.03 and 0.25±0.03, respectively. The soil boron content was moderate in Gangshang Town and Shiliu Town, but lacking in Baima Town. The content of available potassium, available phosphorus, available magnesium, available iron and available zinc are very high in the three regions. According to the measured soil electrical conductivity EC, the continuous cropping soil in Gangshang Town and Baima Town had mild salinization, and the soil in Shiliu Town had moderate salinization. According to the measured soil pH, the soil in Gangshang Town and Shiliu Town belonged to the neutral soil, while Baima Town soil belongs to the slightly alkaline soil. As the depth of soil increases, the abundance of soil actinomycetes in Gangshang Town, Shiliu Town and Baima Town showed a decreasing trend; the abundance of soil bacteria in Gangshang Town and Shiliu Town tends to decrease as the soil depth deepens, while that in Baima Town increases; interestingly, with the deepening of the soil, the abundance of fungi in Gangshang Town first decreased and then increased, the Shiliu Town showed a decreasing trend, while Baima Town showed an increasing trend. Continuous cropping will have a significant adverse effect on plant stem diameter, fresh plant weight, plant height, root weight and root length. However, by some means, this situation can be effectively improved. 【 Conclusion】 Strawberry Continuous cropping will result in the loss of soil nutrients and the imbalance of microbial community structure, which will have a significant impact on production. As far as possible, continuous cropping should be avoided in the production, and we should improve the continuous cropping soil in time. Key words Strawberry; Continuous cropping; Soil microorganism; Soil nutrients 草莓（ Fragaria ananassa Duch.）属蔷薇科（ Rosaceae）多年生草本植物，其果实酸甜可口、营养丰富，深受人们喜爱。江苏省是我国草莓的主产区之一，其栽培面积和产量均居国内前列。近年来，由于可利用土地紧缺、园艺设施搬动成本高，江苏各地的草莓生产均发生了不同程度的连作障碍[1]。研究表明，随着连作年限的增加，土壤有机质含量逐渐下降，微量元素丰缺差距逐渐扩大，土壤中的细菌、真菌和放线菌比例逐渐失衡[2-3]。土壤养分的缺失和根际微生物群落结构的破坏使草莓植株易受病虫害侵染，植株长势和抗病力逐年变弱，甚至出现整株死亡的现象，使草莓生产损失巨大[4-5]。连作障碍已成为阻滞农业可持续发展的主要因素之一[6]。 土壤有机质主要来源于土壤中的动物、植物和微生物残体，是植物摄取养分的主要来源之一，有机质含量高低可以决定土壤的理化性质，在一定范围内与土壤肥力呈正相关关系[7]。土壤微生物也是植株正常生长不可或缺的环境组份，在土壤养分循环中起到关键作用，微生物群落结构中有益菌和有害菌的比例影响着植株根系吸收养分和抵御疾病的能力[8-9]。笔者采集江苏草莓主产区的邳州港上镇、东海石榴镇和溧水白马镇的草莓连作土壤，分析各地区不同深度连作土的养分含量和微生物区系状况，并结合土壤理化性质分析连作土对草莓植株的影响，以明确草莓连作障碍的形成机理和改善措施，为克服江苏乃至华东地区草莓连作障碍提供理论依据。 1 材料和方法 1.1 土壤材料 土壤样品于 2016 年 5 月底采于江苏省邳州港上镇、东海石榴镇和溧水白马镇的草莓连作大棚，其中港上镇连作 7 a（年） ，前茬品种为‘甜查理’ ，栽前土壤进行太阳能闷棚，石榴镇连作 9 a，前茬品种为‘红颊’ ，栽前土壤进行太阳能闷棚，白马镇连作 8 a，前茬品种为‘宁玉’ ，栽前土壤进行有机质 石灰氮结合太阳能闷棚。设置各地区 3 个不同位置的草莓连作大棚为实验重复，每个连作棚按照“ S”型均匀布置 5 个点采集土样，每个取样点按照土层深度 010 cm、 1020 cm 和 2030 cm 采集土壤。将各地区不同土层土分别混合用于当年 9 月初的盆栽试验，剩余土壤分成两份，其中一份用以分析土壤微生物区系（ 4 ℃ 保存） ，另一份自然风干后研磨，过 0.8 mm 尼龙筛，取 100 g 筛后样品再次研磨后过 0.16 mm 尼龙筛，收集筛后样品用以测定土壤理化性质。取各点相邻的露地小麦田土壤为盆栽试验的对照 CK。 1.2 盆栽试验 同年 9 月 5 日，各处理定植草莓品种‘宁玉’ 20 株（ 1 株 /盆） ，在定植后的第 57 天（ 11 月 1 日） ，调查植株茎粗、鲜质量、株高、根长和根质量等生长指标。 1.3 测定方法 采用常规方法测定土壤养分含量[10]、 pH 值和 EC 值[11]；使用常规方法培养并计数土壤微生物数量[12]。 1.4 土壤 pH 值、EC 值及养分分级标准 土壤 pH 值分级标准参考美国土壤调查手册[13]，土壤 EC 值的分级标准参考张鹤航等[14]的研究结果，土壤有机质、大量和微量元素的养分分级指标参考中国第二次土壤普查推荐的养分分级标准[15]，其中有效钙和有效镁含量分级指标参考章永松等[16]的研究结果，各养分分级标准见表 1。 1.5 数据处理 使用 SPSS 17.0 软件对相关数据进行多重比较（ LSD 法） 、相关性分析（ Pearson 法）和差异显著性检验。 表 1 土壤养分含量分级标准 Table 1 Classification standard of soil nutrient content 丰 缺程度 Abundance 有机质 Organic matter /g·kg-1 碱解氮 Available N /mg·kg-1 速效钾 Available K /mg·kg-1 速效磷 Available P /mg·kg-1有效铁 Available Fe /mg·kg-1有效锰 Available Mn /mg·kg-1有效锌 Available Zn /mg·kg-1硼 Boron /mg·kg-1 有效钙 Available Ca /g·kg-1 有效镁 Available Mg /mg·kg-1极缺乏 Extremely lacking 1.2 180 很丰富 Very rich 40 150 200 40 20 30 3.0 2.0 2 结果与分析 2.1 连作土壤的理化性质 港上镇、石榴镇和白马镇的表层土壤（ 020 cm）有机质含量（表 2）分别为 18.40±5.42 g·kg-1（丰缺程度属缺乏） 、 18.26±1.28 g·kg-1（缺乏）和 28.63±8.04 g·kg-1（适中） 。随着土层深度的增加，石榴镇和白马镇的土壤有机质含量呈减少趋势，而港上镇的土壤有机质含量则呈先增后减趋势（图 1） ；港上镇、石榴镇和白马镇的表层土壤碱解氮含量分别为 75.37±1.24 mg·kg-1（缺乏） 、 81.25±3.09 mg·kg-1（缺乏）和 113.75±14.21 mg·kg-1（适中） ， 3 个地区的土壤碱解氮都主要集中在 020 cm 土层，深土层的碱解氮含量减少了 3345。港上镇、石榴镇和白马镇的表层土壤有效锰含量分别为 3.76±0.56 mg·kg-1（缺乏） 、 44.15±3.20 mg·kg-1（很丰富） 和 13.06±0.85 mg·kg-1（适中） ； 港上镇、 石榴镇和白马镇的表层连作土壤硼含量分别为 0.63±0.27 mg·kg-1（适中）、 0.51±0.03 mg·kg-1（适中）、 0.25±0.03 mg·kg-1（很缺乏）；港上镇、石榴镇和白马镇的表层连作土壤有效钙含量分别为 1.88±0.21 g·kg-1（丰富）、 0.97±0.03 g·kg-1（丰富）、 2.15±0.11 g·kg-1（适中）；三个地区连作土的表层土壤有效镁含量都较丰富，有效铁、速效磷、速效钾和有效锌含量都很丰富；根据所测土壤 EC 值，港上镇和白马镇的连作土壤有轻度盐渍化（ 0.550.90 mS·cm-1），石榴镇土壤有中度盐渍化（ 0.901.80 mS·cm-1）；根据所测土壤 pH 值，港上镇和石榴镇的土壤属于中性土壤（ 6.67.3），而白马镇土壤属于微碱性土壤（ 7.37.8）。 图 1 各地区不同深度草莓连作土层的养分含量变化 Fig.1 Nutrient Content changes of continuous cropping soil among various depth in different strawberry production areas 2.2 各地区连作土壤微生物群落结构 由表 3 可知，各地区连作土的微生物都以细菌为主，真菌的数量最低。随着土层深度的增加，港上镇、石榴镇和白马镇的土壤放线菌数量都呈减少趋势；港上镇和石榴镇的土壤细菌数量呈减少趋势，白马镇则呈增加趋势；港上镇的真菌数量呈先降后增趋势，石榴镇呈降低趋势，而白马镇呈增高趋势。 表 3 三个地区草莓连作土壤的微生物群落结构 Table 3 Microbial community structure of continuous cropping soil in three areas 地区 Areas 土层 Soil layer/cm 真菌 Fungi/103CFU·g-1 放线菌 Actinomycetes/105CFU·g-1 细菌 Bacteria/106CFU·g-1邳州港上镇 Gangshang, Pizhou 010 4.65±0.84 c 6.07±0.17 de 2.56±0.59 bc 1020 3.97±0.51 d 6.63±0.17 de 3.43±0.42 ab 2030 5.25±0.18 c 2.56±0.17 f 1.90±0.17 c 东海石榴镇 Shiliu, Donghai 010 10.12±0.84 ab 26.19±3.37 a 2.74±0.17 bc 1020 12.17±1.87 a 13.26±1.70 c 2.39±0.30 bc 2030 9.17±0.50 b 8.81±1.68 d 1.67±0.33 d 溧水白马镇 Baima, Lishui 010 6.75±0.68 c 22.17±0.34 b 3.25±0.51 abc 1020 6.05±1.57 c 7.90±1.40 de 2.60±0.87 bc 2030 10.38±0.53 ab 4.88±0.53 e 4.13±1.24 a 2.3 土壤微生物群落结构与土壤理化性质的相关性分析 三个地区的连作土壤中，细菌数量与土壤有效钙和有效镁含量呈显著正相关（表 4） ；真菌数量与土壤有效铁、有效锰含量和土壤 EC 值呈显著正相关，与土壤有效锌、速效钾和速效磷含量呈极显著正相关；放线菌数量与土壤有效钙含量呈极显著负相关，与土壤有效铁和有效锰含量呈极显著正相关，与土壤有效锌、速效磷含量和土壤 EC 值呈显著正相关，与土壤有效镁含量呈显著负相关。 表 4 土壤微生物数量与土壤理化性质的相关性分析Table 4 Correlation between soil microorganisms and soil physical and chemical properties 菌 种 Strains 有效钙 Available Ca 有效镁 Available Mg 有效铁 Available Fe 有效锰Available Mn 有效锌 Available Zn 硼 Boron 有机质Organic matter 碱解氮 Available N 速效钾 Available K 速效磷 Available P pH EC 细 菌 Bacteria 0.475*0.586*-0.043 -0.178 -0.249 -0.064 0.432 0.464 0.087 -0.224 0.467 -0.195真 菌 Fungi -0.392 -0.125 0.508* 0.488* 0.759** -0.230 0.180 0.317 0.699** 0.794** 0.444 0.545*放 线菌 Actinomycetes -0.702**-0.561*0.708** 0.713** 0.491* 0.039 -0.310 -0.231 0.340 0.568* 0.395 0.549*注表中数据为相关性系数，正负符号分别代表正相关与负相关，* 表示显著相关，** 表示极显著相关。 Note The data in the table are correlation coefficients, positive and adverse represent positive correlation and adverse correlation. respectively. * represents significant correlation, and ** represents extremely significant correlation. 表 2 三个地区 020cm 土层土壤的平均养分含量 Table 2 The average nutrient content of soil in 020cm soil layers in three regions 地 区 Areas 有效钙 Available Ca/g·kg-1 有效镁 Available Mg/mg·kg-1 有效铁 Available Fe/mg·kg-1 有效锰 Available Mn/mg·kg-1有效锌 Available Zn/mg·kg-1 硼 Boron/ mg·kg-1 有机质 Organic matter/g·kg-1碱解氮 Available N/mg·kg-1 速效钾 Available K/mg·kg-1 速效磷 Available P/mg·kg-1 pH EC /mS·cm-1 邳州港上镇 Gangshang, Pizhou 1.88±0.21 b 345.97±22.45 b 35.18±4.51 b 3.76±0.56 c 3.79±0.50 c 0.63±0.27 a 18.40±5.42 b 75.37±1.24 b 272.92±13.12 b 88.22±6.37 c 6.97±0.11 c 0.87±0.18 b 东海石榴镇 Shiliu, Donghai 0.97±0.03 c 240.61±4.06 c 51.70±3.51 a 44.15±3.20 a 12.19±0.44 a 0.51±0.03 a 18.26±1.28 b 81.25±3.09 b 404.82±42.61 a 258.34±14.21 a 7.28±0.05 b 1.24±0.11 a 溧水白马镇 Baima, Lishui 2.15±0.11 a 529.21±19.36 a 32.64±5.09 b 13.06±0.85 b 4.96±0.56 b 0.25±0.03 b 28.63±8.04 a 113.75±14.21 a 424.35±133.34 a 124.55±20.23 b 7.50±0.04 a 0.68±0.26 b 注表中数据为平均值±标准差，不同小写字母表示同列数据在 P0.05 显著差异。下同。 Note The data in the table is the mean ± standard deviation. Different lowercase letters indicate that the same column of data is significantly different at P0.05. Same as below. 表 5 三个地区草莓植株在连作土与对照中的生长状况Table 5 Growth status of strawberry plants cultivated in continuous cropping soil and control soil in three regions 地区 Areas 连作植株的 相对死亡率 Relative mortality of continuous cropping/ 茎粗 Stem diameter/mm 植株鲜重 Plant fresh weight/g 株高 Plant height/cm 根重 Root weight/g 根长 Root length/cm 连作 Continuous cropping 对照 Control 连作 Continuous cropping 对照 Control 连作 Continuous cropping 对照 Control 连作 Continuous cropping 对照 Control 连作 Continuous cropping 对照 Control 邳州港上镇 Gangshang, Pizhou 20 8.04±2.03 9.79±1.70 8.50±2.75 12.77±2.87 9.17±2.36 10.33±2.08 3.33±0.91 5.17±1.60 9.17±1.04* 12.33±0.58 东海石榴镇 Shiliu, Donghai 30 6.05±1.18* 9.30±1.44 7.77±2.05* 18.67±1.00 8.63±1.31* 11.00±1.00 3.80±1.18* 8.50±2.89 9.47±0.50* 15.83±1.26 溧水白马镇 Baima, Lishui 1 7.78±0.62 8.45±1.38 9.57±0.85 10.47±4.19 9.33±1.53 11.00±1.00 3.50±0.56 3.77±1.15 10.67±2.08 9.33±1.76 注* 表示同一地区的连作土植株与对照植株在 P0.05 水平上差异显著。为方便比较，定义连作土处理与对照处理的植株死亡率之差为 连作植株的相对死亡率。 Note * Indicates that the continuous cropping soil and control plants in the same region showed significant differences at the P0.05 level, respectively. For convenience, the difference between the mortality rates of the continuous cropping and the control treatments was defined as the relative mortality of continuous cropping.2.4 各地区连作土对植株的影响 三个地区连作土植株的死亡率均高于对照， 石榴镇的相对死亡率最高为 30， 白马镇的相对死亡率较低，仅为 1（表 5） ；三个地区的连作土对植株生长都有抑制作用，经显著性差异分析，石榴镇连作土植株的茎粗、植株鲜重、株高、根重和根长都与对照植株在 p 0.05 水平上显著减少了 21.5558.38；港上镇的连作土植株除根长与对照差异显著外，其余指标均显示差异不显著；白马镇的所有指标
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