摘要
为研究3种常用除草剂(草甘膦异丙胺盐、草铵膦、精喹禾灵)对橘园土壤理化性质、土壤根际微生物及果实品质的影响,以20年生温州蜜柑结果树为对象,检测喷施除草剂前后土壤理化性质、土壤根际微生物及果实品质等指标的变化。结果显示:喷施草甘膦异丙胺盐导致橘园土壤酸化,而喷施草铵膦或精喹禾灵对土壤pH无显著影响;喷施3种除草剂均导致土壤容重增加,且对土壤酸性磷酸酶、碱性磷酸酶、脲酶及蔗糖酶活性有不同程度的激活作用;喷施3种除草剂对土壤有机质、土壤碱解氮和速效钾、速效磷等养分含量无显著影响;喷施3种除草剂导致土壤微生物群落组成显著变化和根际微生物多样性下降,导致土壤放线菌门、酸杆菌门等微生物相对丰度下降以及蓝细菌门等微生物相对丰度上升;喷施3种除草剂均导致果实单果质量、可溶性固形物、维生素C含量下降、可滴定酸含量上升并推迟了果皮转色。研究结果表明,施用除草剂对橘园土壤生态及果实品质有显著不利影响,应客观认识除草剂的作用并合理施用除草剂。
杂草防治是橘园土壤管理的重要环节。尽管人工除草是最环保的举措,但由于农村劳动力短缺、人工成本高、除草效率低等,生产上普遍通过喷施除草剂来除
施用除草剂会影响柑橘等作物的正常生长和抗性。张
土壤微生物对土壤腐殖质形成、有机质分解、土壤肥力保持以及作物生长发育具有重要作
综上,施用除草剂会对土壤理化性质和作物生长发育带来较大负面影响,但在柑橘等果树作物上尚缺乏系统研究,尤其是关于除草剂对土壤微生物的影响还不清晰甚至有一定争议。因此,本研究以柑橘主栽品种温州蜜柑为材料,调查了田间条件下草甘膦异丙胺盐、草铵膦及精喹禾灵等3种常用除草剂的施用对土壤理化性质、土壤微生物及果实品质的影响,旨在为客观认识除草剂的作用、科学施用除草剂提供依据。
选择华中农业大学柑橘试验基地温州蜜柑(国庆1号Citrus unshiu Marc.cv.Guoqing No.1)结果树16株,树龄20 a,树体长势一致。该地块5 a内未施用供试除草剂。将此16株供试植株随机分为对照组及T1、T2、T3共4组,每组4棵树。对照组采用人工刈割除草,处理组分别喷施3种不同的除草剂。T1为喷施 0.5%草甘膦异丙胺盐(商品名:农达;农药登记证号:PD73-88)、T2为喷施0.5%草铵膦(商品名:焚草;农药登记证号:PD20171993)、T3为喷施0.2%精喹禾灵(商品名:精禾草克;农药登记证号:PD20095860)。遵循组间统一施药原则,自2021年4月1日首次施药后,每当杂草生长至出现第3片真叶或生长长度近3 cm时喷施除草
于喷施除草剂后0 d、盛花期、坐果期、转色期4个时期随机选择每棵植株的4个不同方位,取表层土(0~10 cm)土样并将来自同一植株的土样混合作为1个样
果实成熟后沿树体滴水线取表层土壤(0~10 cm)。利用苯酚钠法测土壤脲酶活性,3,5-二硝基水杨酸法测土壤蔗糖酶活性,硝基酚比色法测土壤酸性和碱性磷酸酶活
分别取每棵柑橘树滴水线附近0~10 cm表层土壤或根际土壤,将同一棵树4个不同方位的采样点的土样均匀混合为1个样品。使用OMEGA土壤DNA试剂盒(M5635-02)提取土壤总基因组DNA(Omega Bio-Tek,Norcross,GA,USA),使用NanoDrop NC2000分光光度计测量提取的土壤DNA的质量和浓度(Thermo Fisher Scientific,Waltham,MA,USA)。利用正向引物338F(5′-ACTCCTACGGGAGCAGCA-3′)和反向引物806R(5′-GGACTACHVGGGTWTCTAAT-3′)PCR扩增细菌16S rRNA基因的V3-V4区域,送上海派森诺公司使用Illlumina NovaSeq测序。使用QIIME2和R(3.2.0)进行序列数据分析。随后利用QIIME2中的ASV表计算ASV水平的α多样性指数(Shannon多样性指数)以调查群落物种丰富度和多样性等,并可视化为箱线图;基于Bray-Curtis度量和UniFrac距离度量进行β多样性分析,以调查样本中微生物群落的结构变化,并通过主坐标分析(PCoA)进行可视化。
果实达到可食用品质后(11月25日)采样,每棵树在树冠外围随机采集 5个果实,施药和对照每个处理均为4棵树,每个处理各20个果实。利用电子天平称取单果质量;利用*cm-5色彩色差仪(美能达,日本)测定果皮的亮度值(
分别检测喷施除草剂0 d、盛花期、坐果期、转色期的橘园的土壤pH及容重(

图1 施用不同除草剂对橘园土壤pH(A)和容重(B)的影响
Fig.1 The effets on pH(A) and bulk density(B) in citrus orchards after spraying different herbicides
CK:对照,人工除草 Blank control,manual weeding;T1:喷施0.5%草甘膦异丙胺盐 Spray 0.5% N-(phosphonomethyl) glycine;T2:喷施0.5%草铵膦 Spray 0.5% glufosinate-ammonium;T3:喷施0.2%精喹禾灵 Spray 0.2% quizalofop;1.喷施除草剂0 d; 2.盛花期 Flowering period;3.坐果期 Fruiting period; 4.转色期 Color changing period。下同。The same as follows.
喷施除草剂后柑橘果实成熟期的土壤酶活性检测结果(

图2 施用不同除草剂对橘园土壤酶活的影响
Fig.2 The effects on soil enzyme activity of 0-10 cm soil layer after spraying different herbicides
检测结果(

图3 施用不同除草剂对橘园土壤养分的影响
Fig.3 The effects on soil nutrient elements content of 0-10 cm soil layer after spraying different herbicides
用Shannon指数表征土壤微生物的α多样性(

图4 喷施除草剂对橘园土壤微生物的影响
Fig.4 The effects on soil microbial diversity in citrus orchard after spraying different herbicides
Shannon(A)指数表征微生物α多样性;主坐标分析PCoA(B)表征土壤微生物β多样性,其中不同分组的样本用不同形状的点代表,各样本点越接近,说明各样本物种组成越相似。下同。Shannon(Fig.A) indices characterize microorganisms α diversity; Principal coordinate analysis PCoA(Fig.B) characterizes soil microorganisms β diversity,in which the samples in different groups are represented by points of different shapes.The closer the sample points are,the more similar the species composition of each sample is. The abscissa and ordinate represent the relative distance,which has no practical significance.The same as follows.
进一步分析各样品组的微生物优势物种在门水平和属水平上的相对丰度差异,结果显示,除草剂处理组与对照组在门水平上相对丰度差异显著的微生物分别是:酸杆菌门(Acidobacteria)、放线菌门(Actinobacteria)、蓝细菌门(Cyanobacteria)、厚壁菌门(Firmicutes)和匿杆菌门(Latescibacteria)(
注: 同行不同小写字母表示差异显著(P<0.05)。下同。Note:Different lowercase letters in the same line indicate significant differences(P<0.05).The same as follows.
本研究同时检测了喷施除草剂对橘园根际微生物多样性的影响(

图5 喷施除草剂对橘园根际微生物多样性的影响
Fig.5 The effects on rhizosphere soil microorganisms diversity in citrus orchard after spraying different herbicides
如

图6 喷施除草剂对柑橘产量及品质的影响
Fig.6 The effects on fruit productionand quality after spraying different herbicides
A:单果质量; B:可食率; C:亮度值(
本研究发现喷施精喹禾灵和草铵膦导致土壤容重显著增加,而喷施草甘膦异丙胺盐对土壤容重的影响较小(
前人研究表明,施用除草剂会导致土壤微生物丰度及多样性普遍降
本研究中喷施除草剂显著降低了柑橘单果质量,导致果实转色延迟、酸度增加和维生素C含量降低(
综上,在土壤肥力适中的温州蜜柑园中多次喷施草甘膦异丙胺盐等除草剂影响了橘园土壤容重、pH值及酶活等土壤理化性质,导致土壤有益微生物的多样性与丰度下降,最终导致果实产量和品质的下降。本研究主要调查了温州蜜柑在华中地区黄棕壤背景下除草剂对土壤理化性质和果实品质的影响,对橘园除草剂的科学施用具有一定的指导意义,但除草剂在不同柑橘品种和不同土壤背景下对橘园土壤和果实品质的影响仍需进一步探讨。
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