焦玉洁1, 杜如万2, 王剑2, 王勇2, 吴叶宽2, 刘东阳2, 袁玲1
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1.西南大学资源环境学院, 重庆 400716;
2.四川省凉山州烟草公司, 四川 西昌 615000
收稿日期:2016-05-23;修回日期:2016-09-26;网络出版日期:2016-10-09
基金项目:国家“973项目”(2013CB127405);四川省凉山州烟草公司科技项目(2014-01)
*通信作者:袁玲,Tel:+86-23-68251249;Fax:+86-23-68250444;E-mail:lingyuanh@aliyun.com
摘要: [目的]紫茎泽兰(Ageratina adenophora)内含对微生物、植物和动物有毒的化学物质,列为我国危害最严重的外侵植物,评价腐熟紫茎泽兰对土壤微生物和作物的毒性,有益于无害化处理生产有机肥。[方法]利用微生物菌剂腐熟紫茎泽兰,田间设置不施肥(CK)、单施化肥(CF)、单施紫茎泽兰有机肥(OF)、化肥配施紫茎泽兰有机肥(50%化肥+50%紫茎泽兰有机肥,CF+OF)等4种施肥处理,研究了紫茎泽兰有机肥对土壤细菌、养分和辣椒产量品质等的影响。[结果]在施用OF的土壤中,微生物碳氮高于CF、OF和CF+OF提高细菌群落的多样性指数,CF增加优势度指数。在4种施肥处理的土壤中,酸杆菌门和变形菌门均为优势门类,丰富度合计超过50%;在20种优势菌株中,有7株细菌普遍存在,6-8株细菌单独存在于不同处理土壤中。此外,在辣椒初果期,CF土壤中的有效养分含量较高;但至末果期,CF+OF处理的有效磷钾显著高于CF,碱解氮CF+OF与CF相似。施用CF+OF使辣椒吸收了较多的氮、磷、钾,辣椒产量比CF增加14.42%,并使果实游离氨基酸和维生素C含量提高,硝酸盐含量降低。[结论]紫茎泽兰有机肥兼具供肥改土作用,能提高土壤微生物生物量,丰富土壤细菌种群,增加辣椒产量,改善果实品质。
关键词: 紫茎泽兰 腐熟 辣椒 土壤 细菌
Effect of composted Ageratina adenophora on soil bacteria, nutrients, and pepper yield and quality
Jiao Yujie1, Du Ruwan2, Wang Jian2, Wang Yong2, Wu Yekuan2, Liu Dongyang2, Yuan Ling1
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1.College of Resources and Environment, Southwest University, Chongqing 400716, China;
2.Sichuan Tobacco Corporation Liangshanzhou Branch, Xichang 615000, Sichuan Province, China
Received 23 May 2016; Revised 26 September 2016; Published online 09 October 2016
*Corresponding author: Corresponding author. Tel: +86-23-68251249; Fax: +86-23-68250444;E-mail:lingyuanh@aliyun.com
Supported by the National Basic Research Program of China (2013CB127405) and by the Scientific Project of Liangshan Tobacco Company of Sichuan Province (2014-01)
Abstract: [Objective] Ageratina adenophora, a most harmful invasive plant in China, contains many toxicants to animals, plants and microbes. To understand the toxicity of its decomposed materials by microbial inoculants to soil microbes and plants is beneficial to produce organic fertilizer (OF).[Methods] A field experiment, including the control (CK), sole chemical fertilizer (CF), pure OF, and combination of CF and OF (CF+OF), was conducted to study the effect of OF on soil bacteria, and yield and quality of pepper.[Results] Providing soil with OF resulted in higher microbial biomass than CF. Both OF and CF+OF increased bacterial community indexes in contrast to CF that increased microbial dominant index in soils. Actinobacteria and Proteobacteria accounted for more than 50% of the total bacteria in each soil examined. Among top 20 predominant bacteria, 7 were ubiquitous in all studied soils and 6 to 8 were unique in each soil. Furthermore, there were higher nutrient concentrations in the soil supplied with CF in early fruit stage, whereas that with CF+OF contained higher available phosphorus and rapidly available potassium than CF, and alkaline hydrolyzed nitrogen was similar in late fruit period. Pepper plants fertilized with CF+OF absorbed more nitrogen, phosphorus, and potassium, produced 14.4% more fruits than CF and improved pepper fruit quality.[Conclusion] OF application enriched soil bacterial groups, increased soil microbial biomass, and pepper fruit yield and quality. Organic fertilizer made from A. adenophora improves soils for pepper plants.
Key words: Ageratina adenophora composted pepper soil bacteria
紫茎泽兰(Ageratina adenophora)是一种菊科多年生的恶性毒草,原产于墨西哥和南美洲,列为我国危害最严重的外侵植物。在桂、滇、黔、川、渝和西藏亚热带地区,紫茎泽兰大规模入侵农田、森林和草场。在四川省凉山州,紫茎泽兰的危害面积占幅员面积的14.1%,给生态环境,农业、林业、畜牧业造成了巨大灾难[1-2]。
紫茎泽兰通过释放化感物质排除其它植物,形成单优种群。其根、茎、叶的浸提液对植物新陈代谢、养分吸收、物质运输、光合作用、细胞结构、DNA复制与修复造成危害,抑制辣椒、番茄、茄子、黑麦草和白三叶等多种植物的种子发芽和幼苗生长,抑制作用随浓度升高而增强[3-4]。目前,人们已从紫茎泽兰体内分离出100多种化学物质,主要为单萜类、倍萜类、三萜类、苯丙素类、黄酮类及各种衍生物,大部分含有多环芳烃,具有动物、微生物和植物毒性[5-6]。但是,紫茎泽兰生长快,生物量大,养分吸收能力强,富含氮、磷、钾及微量元素[7],是一种优良的有机肥源。在农业生产中,施用有机肥提供土壤养分,提高土壤肥力和生产力,同时,秸秆还田也是增加土壤有机质的重要手段之一。研究表明,施用有机肥和秸秆还田可均衡补充氮、磷、钾及微量元素,保持或提高土壤有机质含量,改善土壤物理结构,增强微生物活性,促进土壤物质循环,提高养分生物有效性[8]。据报道,假单胞菌(Pseudomonas)、反硝化产碱菌(Alcaligenes denitrificans)、诺卡氏菌(Nocardia)、解环菌(Cycloclasticus)和多色节杆菌(Arthrobacter polychromogenes)等细菌能降解如苯、萘、菲、蒽等多环芳烃[9];产黄青霉(Penicllium chrysogenum)可降解香豆素和香草醛;黑曲霉(Aspergillus niger)和焦曲霉(Aspergilusustus)还能分解丹宁[10]。由此可知,利用微生物降解紫茎泽兰有毒物质,再经腐熟作用生产有机肥可实现无害化处理与资源化利用。但前人研究止步于描述紫茎泽兰生物学性质和对其他作物的危害[11]。另外,在紫茎泽兰的人工防除过程中,大量堆放的植物残体急需科学处理和利用。因此,很有必要选择一种生长周期完整的作物,评价腐熟紫茎泽兰的肥效,以及对土壤微生物和作物产量品质的影响。
四川省凉山州的大部分地区属于亚热带印度洋季风气候,夏季多雨,冬季温暖,既是全国重要的蔬菜生产基地,又是紫茎泽兰入侵的重灾区,紫茎泽兰侵入菜地的现象在当地十分普遍。此外,当地复种强度高,土壤深度风化,粘重板结,有机质缺乏,严重影响蔬菜生长,施用有机肥是改良土壤和蔬菜增产的重要措施。论文作者以当地广泛种植的辣椒为对象,研究了紫茎泽兰有机肥对土壤细菌群落结构、养分和辣椒产量品质的影响,旨在评估腐熟紫茎泽兰的肥效和对植物、微生物毒性,为开发有机肥源,防除紫茎泽兰,促进蔬菜生产提供有益信息。
1 材料和方法 1.1 供试材料
1.1.1 辣椒(Capsicum annuum)品种: 四川小米辣,由四川省西昌市西溪乡种子公司提供。
1.1.2 肥料: (1) 紫茎泽兰有机肥:紫茎泽兰生命力强,根、茎和种子均可再生繁殖,不能直接还田,其对部分微生物毒性抑制堆肥生化反应,自然堆腐效果差,难于有效杀灭营养繁殖器官和种子[4]。因此,于2014年7月中旬,采集试验地周围的紫茎泽兰地上部,用恶臭假单胞菌(Pseudomonas putita sp.)和热解纤维梭菌(Clostridium thermocellum sp.)组成的专用菌剂堆制腐熟(表 1)。(2) 化肥:包括尿素(N 46.4%)、过磷酸钙(P2O5 16%)和硫酸钾(K2O 50%),均购于当地农资公司。 表 1. 腐熟紫茎泽兰的化学成分(%干重) Table 1. Chemical components of decomposed A. adenophora (% dry weight)
pH | Moisture | Organic matter | Nitrogen | Phosphorus | Potassium | Humic acid |
7.63±0.21 | 30.14±0.55 | 69.47±1.55 | 2.79±0.24 | 0.80±0.05 | 2.70±0.187 | 8.47±0.23 |
Data in the table are expressed by means±SD and same below unless stated otherwise. |
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1.1.3 供试土壤: 位于四川省西昌市西溪乡(东经102.25,北纬27.72),海拔1700 m,年均气温 17.1 °C,年日照2431 h,年降水1087.5 mm,属高原亚热带印度洋季风气候。成土母质为第四纪红色坡积物,土壤类型为红壤,质地中壤,肥力中等,土壤pH 5.58、有机质22.75 g/kg、全氮 1.13 g/kg、全磷0.56 g/kg、全钾10.83 g/kg,有效氮、磷、钾依次为116.91 mg/kg、12.95 mg/kg和129.7 mg/kg。 1.2 试验设计 试验于2015年3至9月进行。设置(1) 不施肥(CK);(2) 单施化肥(CF);(3) 单施紫茎泽兰有机肥(OF);(4) 化肥配施紫茎泽兰有机肥(50%化肥+ 50%紫茎泽兰有机肥,CF+OF)等4种处理。在CF处理中,每667 m2施用尿素43.1 kg、过磷酸钙62.5 kg、硫酸钾40.0 kg,相当于20 kgN,10 kgP2O5和20 kg K2O。在CF+OF和OF处理中,有机肥用量以氮计,不足的磷钾用化肥补足,使氮、磷、钾用量与CF相等。在施肥时,有机肥和钾磷肥全部做基肥,基肥用氮量占总用量的60%,剩余的氮肥追施于初花期。每处理重复3次,小区面积30 m2,随机区组排列,常规管理,具体施肥方案见表 2。
表 2. 辣椒施肥试验方案(kg/667 m2)* Table 2. Schedules of pepper fertilization in field
Treatments | Base fertilizer | Top dressing | ||||
N fertilizer | P fertilizer | K fertilizer | Organic fertilizer | N fertilizer | ||
CK | 0 | 0 | 0 | 0 | 0 | |
CF | 19.57 | 60.00 | 30.00 | 0 | 13 | |
OF | 0 | 42.83 | 12.58 | 460 | 13 | |
CF+OF | 9.78 | 51.40 | 21.30 | 230 | 13 | |
*CK: control (without fertilizer); CF: chemical fertilizer; OF: organic fertilizer made from A. adenophora; CF+OF: chemical fertilizer plus organic fertilizer made from A. adenophora. N fertilizer: nitrogen fertilizer; P fertilizer: phosphatefertilizer; K fertilizer: potassium fertilizer. |
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1.3 测定项目与方法 在辣椒栽前、初果期、盛果期和末果期,用抖根法采集根际土壤,拣去杂物,风干,制样,常规分析碱解氮、有效磷和有效钾含量[12]。立即用液氮冷冻盛果期的另一部分土壤样品,用于测定微生物生物量(氯仿熏蒸-0.5 mol/L K2SO4提取,K2CrO7氧化法测碳和腚酚蓝比色法测氮)和细菌16S rRNA基因序列[13-14]。参照454高通量测序方法,提取、扩增、纯化、定量和均一化16S rRNA基因后,送上海美吉生物科技有限公司利用Roche Genome Sequencer FLX 平台测序进行高通量测序。然后,对有效序列进行去杂、修剪、除嵌合体等过滤处理,得到优化序列,通过聚类分析形成分类单元(Operational taxonomic units,OTUs),用BLAST程序对比GenBank (http://ncbi.nlm.nih.gov)中的已知序列,根据97%的相似度确定16S rRNA基因序列对应的细菌名称。
分批采收成熟的辣椒果实,累计产量。在辣椒盛果期,分别采取50个成熟果实,用紫外分光光度法测定硝酸盐,2,6-二氯酚靛酚滴定法测定维生素C,蒽酮比色法测定可溶性糖,考马斯亮蓝法测定蛋白质,水合茚三酮法测定游离氨基酸含量[15]。同时收获植株,105 °C杀青,(80±1) °C烘干,H2SO4-H2O2消化,蒸馏法定氮,钼锑抗比色法测磷,火焰光度法测定含钾量[12]。
1.4 数据处理 利用土壤细菌的分类单元数(OTUs)和16S rRNA序列数计算土壤细菌的种群特征值,包括多样性指数、优势度指数和丰富度等[16]。
用Excel 2003对试验数据进行基本计算,SPSS 16.0软件进行统计分析,显著水平为P<0.05。
2 结果和分析 2.1 土壤微生物生物量 微生物量碳CF+OF>OF>CF>CK,变化于50.67-217.40 mg/kg;微生物量氮OF>CF+OF>CF> CK,变化于6.49-26.42 mg/kg;微生物量碳氮比CF+OF>CF>CK>OF,变化于6.89-14.25 (表 3)。
表 3. 在不同施肥处理中,土壤微生物量的变化 Table 3. Changes of microbial biomass in soils with variable fertilizers
Treatments | MBC/(mg/kg) | MBN/(mg/kg) | MBC/MBN |
CK | 50.67±4.87 d | 6.49±1.27 c | 7.94±1.10 bc |
CF | 144.19±24.05 c | 14.27±3.07 b | 10.50±3.15 b |
OF | 180.22±17.42 b | 26.42±3.49 a | 6.89±1.04 c |
CF+OF | 217.40±19.69 a | 15.26±1.12 b | 14.25±1.09 a |
CK: control (without fertilizer); CF: chemical fertilizer; OF: organic fertilizer made from A. adenophora; CF+OF: chemical fertilizer plus organic fertilizer made from A. adenophora. MBC: microbial biomass carbon; MBN: microbial biomass nitrogen; MBC/MBN ratio: microbial biomass carbon and nitrogen ratio. In each column,means followed by different small letters are significantly different at P<0.05. |
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2.2 土壤细菌群落结构
2.2.1 细菌群落特征值: 土壤细菌的多样性指数以OF和CF+OF最高,变化于5.771-5.822,CK和CF最低,变化于5.215-5.398。优势度指数CK(0.020)> CF(0.013)>OF(0.006)和CF+OF(0.005)(表 4)。 表 4. 不同施肥处理土壤中细菌群落的特征值 Table 4. Character values of bacteria community in soils with different fertilizers
Bacterial community character values | CK | CF | OF | CF+OF |
Diversity index | 5.215±0.988 b | 5.398±1.012 b | 5.822±0.756 a | 5.771±0.687 a |
Dominant index | 0.020±0.005 a | 0.013±0.003 b | 0.006±0.002 c | 0.005±0.002 c |
CK: control (without fertilizer); CF: chemical fertilizer; OF: organic fertilizer made from A. adenophora; CF+OF: chemical fertilizer plus organic fertilizer made from A. adenophora. In each row,means followed by different small letters are significantly different at P<0.05. |
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2.2.2 稀释曲线: 随机抽取测序样品中的16S rRNA序列数(reads),以细菌分类单元数(OTUs)为纵坐标,16S rRNA读数为横坐标,获得稀释曲线(图 1)。结果表明,OUTs的峰值CF+OF>OF> CF>CK。抽样读数大约在4000以下时,OUTs随读数增加而迅速增加;读数在4000与13000之间,OUTs随读数增加而缓慢增加;读数超过13000之后,OUTs随读数增加而趋于平缓。当曲线趋向平坦时,说明增加更多的读数量不会或少量增加OTU,测序结果合理可靠。
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图 1. 不同施肥处理的土壤中细菌稀释性曲线 Figure 1. Bacteria rarefaction curves in soils with different fertilizers. |
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2.2.3 细菌门类: 在CK、CF、OF和CF+OF处理的土壤中,细菌16S rRNA序列数依次为45131,39930,51194和46557个,分别代表 889,949,986和1000 OTUs,归属于酸杆菌门(Acidobacteria)、 变形菌门(Proteobacteria)、绿弯菌门(Chloroflexi)、放线菌门(Actinobacteria)、厚壁菌门(Firmicutes)、拟杆菌门(Bacteroidetes)、浮霉菌门(Planctomycetes)、芽单胞菌门(Gemmatimonadetes)、硝化螺旋菌门(Nitrospirae)和蓝藻门(Cyanobacteria)等25个门。其中,酸杆菌门和变形菌门较多,合计超过细菌总数的50%。此外,施肥处理显著影响土壤细菌门类的丰富度。例如,CF、OF和CF+OF使酸杆菌门的丰富度增加,厚壁菌门的丰富度降低;在CF处理的土壤中,芽单胞菌门和拟杆菌门最高;但在OF处理的土壤中,浮霉菌门的丰富度最高(图 2)。
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图 2. 不同施肥处理的土壤中门类细菌的丰富度 Figure 2. The abundance of bacterial phyla in soil with different fertilizers. |
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2.2.4 优势菌属: 由表 5可见,纤线杆菌-1 (Ktedonobacteria-1)、小单孢菌-1 (Micromonosporaceae-1)、放线菌-1 (Actinobacteria-1)、鞘氨醇单胞菌-1 (Sphingomonas-1)、放线菌-2 (Actinobacteria-2)、酸杆菌-4 (Acidobacteriales-4)和慢生根瘤菌-1 (Bradyrhizobium-1)等7株细菌均存在于各处理的土壤中。在CK处理的土壤中,独有的菌株是产黄杆 菌-1 (Rhodanobacter-1)、拟无枝酸菌(Amycolatopsis)、假诺卡氏菌-1 (Pseudonocardiaceae-1)、纤线杆菌-2 (Ktedonobacteria-2)、不可培养的弗兰克氏菌(Uncultured Frankineae Bacterium)、假诺卡氏菌-2 (Pseudonocardiaceae-2)、酸杆菌-2 (Actinobacteria-2)和纤线杆菌-3 (Ktedonobacteria-3)等8种。在CF处理的土壤中,独有的菌株包括不可培养细菌-3 (Uncultured Bacterium-3)、产黄杆菌-2 (Rhodanobacter-2)、伯克氏菌(Burkholderia Bacterium)、不可培养的康奈斯氏杆菌(Uncultured Conexibacter)、绿弯菌(Chloroflexi)和不可培养的黄单胞菌(Uncultured Xanthomonas)等6种。在OF处理的土壤中,独有的菌株是假黄单胞菌(Pseudoxanthomonas)、新疆植物放线菌(Actinophytocola)、微细菌(Microbacterium)、链霉菌-2 (Streptomycetaceae-2)、交替赤杆菌(Altererythrobacter)、生丝微菌(Hyphomicrobiaceae)、鞘氨醇单胞菌-3 (Sphingomonas-3)和小单孢菌-2 (Micromonosporaceae-2)等8种。在CF+OF处理的土壤中,独有菌株包括不可培养的放线菌-1(Uncultured Actinobacterium-1)、酸热菌-2 (Acidothermaceae-2)、不可培养的放线菌-2 (Uncultured Actinobacterium-2)、酸杆菌-5 (Acidothermaceae-5)、黄色杆菌-2 (Xanthobacteraceae-2)和黄色杆菌-1 (Xanthomonadaceae-1)等6种。 表 5. 在不同施肥土壤中优势细菌属的丰富度 Table 5. The predominant bacteria in the soil with different fertilizers/%
CK | CF | OF | CF+OF | |||||||
Phylotype | Abundance | Phylotype | Abundance | Phylotype | Abundance | Phylotype | Abundance | |||
Ktedonobacteria-1 | 8.82 | Acidothermaceae-1 | 2.22 | Pseudoxanthomonas | 7.05 | Uncultured Actinobacterium-1 | 10.61 | |||
Micromonosporaceae-1 | 3.10 | Sphingomonas-1 | 1.77 | Sphingomonas-1 | 4.58 | Ktedonobacteria-1 | 2.61 | |||
Rhodanobacter-1 | 2.02 | Acidobacteriales-4 | 1.57 | Actinophytocola | 3.94 | Acidothermaceae-2 | 2.56 | |||
Amycolatopsis | 1.85 | Unknown bacteria | 1.56 | Bradyrhizobium-1 | 2.70 | Micromonosporaceae-1 | 2.36 | |||
Actinobacteria-1 | 1.81 | Uncultured bacterium-2 | 1.47 | Actinobacteria-1 | 1.92 | Uncultured bacterium-1 | 1.58 | |||
Sphingomonas-1 | 1.70 | Uncultured Burkholderiales bacterium | 1.28 | Microbacterium | 1.68 | Acidothermaceae-1 | 1.44 | |||
Actinobacteria-2 | 1.44 | Uncultured bacterium-3 | 1.27 | Streptomycetaceae-2 | 1.50 | Streptomycetaceae-1 | 1.26 | |||
Pseudonocardiaceae-1 | 1.38 | Ktedonobacteria-1 | 1.27 | Nocardioidaceae | 1.45 | Sphingomonas-1 | 1.26 | |||
Unknown bacteria | 1.36 | Sphingomonas-2 | 1.26 | Altererythrobacter | 1.35 | Acidobacteriales-4 | 1.10 | |||
Ktedonobacteria-2 | 1.34 | Actinobacteria-1 | 1.25 | Sphingomonas-2 | 1.24 | Uncultured bacterium-4 | 1.07 | |||
Acidobacteriales-2 | 1.11 | Uncultured bacterium-4 | 1.22 | Hyphomicrobiaceae | 1.18 | Bradyrhizobium-1 | 1.06 | |||
Ktedonobacteria-3 | 1.01 | Bradyrhizobium-1 | 1.20 | Uncultured bacterium-4 | 1.06 | Unknown bacteria | 1.02 | |||
Acidobacteriales-3 | 1.01 | Acidobacteriales-1 | 1.18 | Acidobacteriales-4 | 1.02 | Actinobacteria-2 | 1.00 | |||
Bradyrhizobium-1 | 0.99 | Rhodanobacter-2 | 1.17 | Actinobacteria-2 | 0.96 | Uncultured Actinobacterium-2 | 0.94 | |||
Acidobacteriales-4 | 0.98 | Micromonosporaceae-1 | 1.15 | Sphingomonas-3 | 0.89 | Acidothermaceae-5 | 0.94 | |||
Uncultured bacterium-1 | 0.94 | Burkholderia bacterium | 1.00 | Streptomycetaceae-1 | 0.88 | Xanthobacteraceae-2 | 0.93 | |||
Uncultured Frankineae bacterium | 0.93 | Uncultured Conexibacter | 0.99 | Ktedonobacteria-1 | 0.86 | Xanthomonadaceae-1 | 0.92 | |||
Uncultured bacterium-2 | 0.92 | Chloroflexi | 0.97 | Micromonosporaceae-1 | 0.82 | Acidobacteriales-3 | 0.88 | |||
Nocardioidaceae | 0.90 | Actinobacteria-2 | 0.93 | Micromonosporaceae-2 | 0.81 | Acidobacteriales-1 | 0.84 | |||
Pseudonocardiaceae-2 | 0.87 | Uncultured Xanthomonas | 0.91 | Uncultured Burkholderiales bacterium | 0.80 | Actinobacteria-1 | 0.84 | |||
Σ | 34.48 | 25.59 | 35.23 | 36.69 | ||||||
CK: control (without fertilizer); CF: chemical fertilizer; OF: organic fertilizer made from A. adenophora; CF+OF: chemical fertilizer plus organic fertilizer made from A. adenophora. |
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除独有和共有菌株之外,其余菌株共同存在于每2种或3种施肥处理的土壤中。例如,酸杆菌-3 (Acidobacteriales-3)共存于CK和CF+OF的土壤中;未知细菌-1 (Unknown Bacteria-1)则共存于CK、CF和CF+OF的土壤中。 2.3 土壤有效养分 图 3是辣椒不同生育期,土壤有效氮、磷和钾的含量情况。
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图 3. 施肥对土壤碱解氮(A)、有效磷(B)和速效钾(C)含量的影响 Figure 3. Effects of fertilization on nutriment alkali- hydrolyzable nitrogen (A),available phosphorus (B) and rapidly available potassium (C) in soil. |
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2.3.1 碱解氮: 在初果期,CF>CF+OF、OF和CK;在盛果期,CF+OF>CF>OF和CK;在末果期,CF、OF和CK之间无显著差异,但CF+OF>CK。此外,在辣椒不同生育期,CK土壤中的碱解氮无显著变化;但施肥之后,盛果期的碱解氮较高,尤以CF和CF+OF处理的土壤最为显著。
2.3.2 有效磷: 在初果期,CF高于OF和CF+OF;在盛果期,CF、OF和CF+OF之间无显著差异,但均显著高于CK;在末果期,CF+OF>OF>CF和CK。此外,在辣椒不同生育期,土壤有效磷CK无显著变化;CF和OF在盛果期上升到最高,然后降低;CF+OF在盛果期达到峰值,持续至末果期。
2.3.3 有效钾: 在初果期,CF+OF>CF>OF和CK;在盛果期,OF>CF+OF>CF>CK;在末果期,CF+OF和OF>CF和CK。在辣椒不同生育期,CK土壤在盛果期有效钾最高;CF和CF+OF呈持续降低的趋势;OF处理的土壤有效钾盛果期最高,末果期次之,初果期最低。 2.4 植株养分吸收 与CF相比,施用紫茎泽兰有机肥促进辣椒植株吸收氮、磷和钾(表 6)。吸氮量CF+OF>OF>CF>CK,高低相差约3倍;吸磷量OF>CF+OF>CF>CK,变化于0.45克/株-1.19克/株;吸钾量OF和OF+CF最高,CF次之,CK最低,变化于6.61克/株-10.07克/株。
表 6. 施肥对辣椒植株养分吸收的影响 Table 6. Effects of fertilization on nutrient uptake by pepper seedlings
Treatments | Nutrient absorption (g/plant) | ||
N | P | K | |
CK | 3.28±0.18 d | 0.45±0.09 d | 6.61±0.89 c |
CF | 5.73±0.65 c | 0.99±0.05 c | 9.13±1.45 b |
OF | 7.09±0.41 b | 1.39±0.09 a | 10.41±0.76 a |
CF+OF | 8.92±0.51 a | 1.19±0.08 b | 10.07±0.83 a |
CK: control (without fertilizer); CF: chemical fertilizer; OF: organic fertilizer made from A. adenophora; CF+OF: chemical fertilizer plus organic fertilizer made from A. adenophora. In each column,means followed by different small letters are significantly different at P<0.05. |
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2.5 辣椒产量品质 由表 7可知,辣椒产量为CF+OF>OF和CF>CK,CF+OF比CF增产14.42%。辣椒果实蛋白质含量CF+OF处理的最高,OF次之,CF和CK最低;游离氨基酸和维生素C也以CF+OF最高,但CK、CF和OF之间无显著差异;相反,硝酸盐含量CF最高,CF+OF次之,OF和CK最低;施肥对辣椒果实的可溶性糖含量无显著影响。
表 7. 施肥对辣椒产量和品质的影响 Table 7. Effects of fertilization on yield and quality of pepper fruits
Indicators | CK | CF | OF | CF+OF |
Yield/(kg/hm2) | 4327.7±254.31 c | 5355.5±332.74 b | 5511.1±269.19 b | 6127.7±127.50 a |
Protein/% | 0.42±0.09 c | 0.47±0.02 c | 0.62±0.08 b | 0.79±0.12 a |
Free amino/(mg/100g) | 82.88±12.30 b | 83.75±15.29 b | 87.93±9.01 b | 101.81±14.91 a |
Soluble sugar/% | 0.51±0.03 a | 0.51±0.06 a | 0.55±0.16 a | 0.53±0.02 a |
Vc content/(mg/100g) | 66.23±10.67 b | 71.46±8.22 b | 77.16±2.98 ab | 87.55±2.13 a |
Nitrate content/(mg/kg) | 28.82±3.43 c | 56.38±3.14 a | 28.88±4.93 c | 36.49±3.72 b |
CK: control (without fertilizer); CF: chemical fertilizer; OF: organic fertilizer made from A. adenophora; CF+OF: chemical fertilizer plus organic fertilizer made from A. adenophora. In each row,means followed by different small letters are significantly different at P<0.05. |
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3 讨论 微生物是土壤生态系统的重要组成成分,影响土壤功能、质量、肥力和生产力[17]。在OF和CF+OF处理的土壤中,微生物碳氮提高,碳氮比改变,说明微生物数量增加,组成变化。此外,OF和CF+OF处理多样性指数最大,稀释曲线的峰值和细菌OUTs 均大于CF。众所周知,多样性指数表示生物群落中的物种多寡,数值愈大物种越丰富;优势度指数愈大,生物群落中优势种群越突出。在稳定健康的生态环境中,生物物种丰富,多样性指数较高;反之则较小,优势度指数较大[18]。因此,紫茎泽兰有机肥对土壤微生物的影响类似普通有机肥,可能改善土壤生态环境,促进微生物生长繁殖,增加细菌种群多样性等作用;相反,施用化肥则降低细菌种群多样性,适合生存的细菌种群减少,优势种群突出,与许多长、短期肥料试验结果相似[19-20]。但高通量测序显示,在对照、紫茎泽兰有机肥料和化肥处理的土壤中,优势细菌组成和丰富度有所差异,加之肥料对土壤的影响是长期过程,故还需持续研究。
从土壤细菌种群结构看,酸杆菌门和变形菌门均为各处理的优势细菌,二者之和超过细菌总量的50%,说明土壤是决定微生物种群结构的主要因素之一。但在20种优势菌株中,有7株细菌普遍存在,6-8株细菌单独存在于不同处理土壤中,说明施肥不同程度地改变了细菌群落的组成,类似前人研究[21]。从土壤细菌功能方面看,酸杆菌和变形菌门的细菌种类繁多,功能多样;硝化螺旋菌参与硝化作用,将铵态氮转化为硝态氮;放线菌分解有机质,形成腐殖质,分泌抗生素和植物生长激素[22]。鞘氨醇单胞菌能降解多环芳烃等高分子有机污染物,清除农药污染[23];慢生根瘤菌和小单孢菌分别能固氮、溶磷、解钾[24-25];微杆菌能降解萘、菲、蒽、芘等有毒物质[9]。因此,土壤细菌具有多种多样的生理、生化和生态功能。
辣椒生育期长,多次结果,需要持续较长时间的养分尤其是磷钾供应[4]。在辣椒初果期,CF处理的有效养分含量较高(有效钾例外),但到末果期,CF+OF处理的有效磷钾显著高于CF,碱解氮CF+OF与CF相似,施用CF+OF的辣椒吸收了较多的氮、磷和钾。说明CF+OF的肥效速缓相济,可协调土壤养分供应,满足辣椒各生育期的养分需要,促进生长发育,提高产量品质。因此,施用CF+OF的辣椒产量显著高于CF,果实蛋白质、游离氨基酸和维生素C也以CF+OF最高,硝酸盐含量则低于CF,类似其他有机肥的增产提质作用[26]。我们曾用新鲜紫茎泽兰浸提液处理牧草和辣椒种子,导致发芽率、发芽指数和活力指数降低,种子根卷曲、发黑,甚至死亡,并抑制幼苗生长;用恶臭假单胞菌和热解纤维梭菌腐熟紫茎泽兰后,其浸提液的作用则相反[3-4]。推测在微生物腐熟紫茎泽兰的过程中,对植物有害的化感物质被降解,植物毒性消失。此外,在旱地土壤中,有机肥经微生物矿化成NH3/NH4+,再经硝化作用形成硝态氮,故有机肥释放硝酸盐平缓,很少在蔬菜体内大量积累。
4 结论 紫茎泽兰有机肥兼具供肥改土作用,能提高土壤微生物生物量,丰富土壤细菌种群,增加辣椒产量,改善果实品质。进一步充分评估微生物腐熟紫茎泽兰的肥效有益于无害化处理与资源化利用。
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