毛蓮英 李海碧 桂意云 張榮華 楊榮仲 周會 韋金菊 劉昔輝
摘要:【目的】探究甘蔗間作貓豆對甘蔗根際土壤微生物多樣性的影響,為改善甘蔗根際土壤微生物群落結(jié)構(gòu)及促進甘蔗生長提供理論參考?!痉椒ā坎捎脝我蛩卦囼炘O(shè)計,分析單作甘蔗(CK)和甘蔗間作貓豆(T處理)2種種植方式對甘蔗根際土壤養(yǎng)分的影響,并采用Illumina HiSeq高通量測序技術(shù),在門和屬水平上分析甘蔗根際土壤微生物群落結(jié)構(gòu),在種水平進行主成分(PCA)分析。【結(jié)果】T處理的根際土壤中全氮、全鉀、水解性氮、有效磷和有機質(zhì)含量均大于CK。T處理的根際土壤中細菌的豐富度和多樣性高于真菌的豐富度和多樣性;T處理的根際土壤中細菌豐富度高于CK,但差異不顯著(P>0.05,下同),其多樣性顯著高于CK(P<0.05,下同);T處理的根際土壤中真菌的豐富度和多樣性均顯著高于CK。CK和T處理土壤樣品的主要優(yōu)勢細菌門為變形菌門(Proteobacteria)、放線菌門(Actinobacteria)、綠彎菌門(Chloroflexi)和酸桿菌門(Acidobacteria),其相對豐度均大于10.00%,優(yōu)勢細菌屬分別為水恒桿菌屬(Mizugakiibacter)、游動四孢屬(Luedemannella)、乳桿菌屬(Acidothermus)和布氏桿菌屬(Bryobacte),但T處理根際土壤樣品中水恒桿菌屬、游動四孢屬和乳桿菌屬的相對豐度分別較CK下降2.47%、1.78%和0.68%,其他細菌屬分別增加0.10%~0.86%。CK和T處理根際土壤中優(yōu)勢真菌門為子囊菌門(Ascomycota),分別為80.00%和92.00%,其次是擔(dān)子菌門(Basidiomycota),分別為8.00%和6.00%,排名前3的優(yōu)勢真菌屬均為戴氏霉屬(Taifanglania)、毛殼屬(Chaetomium)和鐮刀菌屬(Fusarium)。CK和T處理的6份根際土壤樣品在種水平上的主成分(PCA)分析結(jié)果顯示,細菌群落PC1的變異為29.37%,PC2的變異為20.26%,二者的總貢獻率為49.63%;真菌群落PC1的變異為33.44%,PC2的變異為27.73%,二者的總貢獻率為56.17%。CK與T處理的細菌群落分別屬于不同象限,CK位于第1和第3象限,而T處理位于第2和第4象限?!窘Y(jié)論】甘蔗間作貓豆可改善甘蔗根際的土壤養(yǎng)分狀況,改變甘蔗根際微生物的群落結(jié)構(gòu),提高細菌和真菌的豐富度和多樣性,但未影響優(yōu)勢菌門的排序。
關(guān)鍵詞: 甘蔗;貓豆;間作;根際;土壤微生物;高通量測序;群落結(jié)構(gòu)
中圖分類號: S556.106.1? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ?文獻標(biāo)志碼: A 文章編號:2095-1191(2021)02-0332-09
Abstract:【Objective】The effects of Mucuna pruriens var. utilis intercropping with sugarcane on the microbial diversity in the rhizosphere of sugarcane were investigated,and a theoretical foundation was laid for improving the soil microbial community structure of M. pruriens intercropping with sugarcane and promoting the growth of sugarcane. 【Method】A single-factor design experiment was conducted to analyze the microbial community structure of sugarcane rhizosphere soil at the phylum and genus levels by Illumina HiSeq high-throughput sequencing technology under the two planting methods of sugarcane monoculture(CK) and sugarcane intercropping M. pruriens(T treatment),and principal component analysis (PCA) to determine the effect of intercropping M. pruriens on sugarcane rhizosphere soil nutrients. 【Result】The contents of total nitrogen,total potassium,hydrolyzable nitrogen,available phosphorus and organic matter in rhizosphere soil of T treatment were higher than those of CK treatment. The richness and diversity of bacteria in the rhizosphere soil of T treatment was higher than that of fungi. The bacterial richness of? bacteria in the rhizosphere soil of Ttreatment was higher than that of CK,but the difference was not significant(P>0.05, the same below), its diversity was significantly higher than that of CK(P<0.05, the same below). The abundance and diversity of bacteria in the rhizosphere soil under T treatment were significantly higher than that of CK.The dominant phylum of bacteria in CK and T treatments were Proteobacteria,Actinobacteria,Chloroflexi and Acidobacteria,and their relative abundances were more than 10.00%. The dominant bacteria genera were Mizugakiibacter,Luedemannella,Actinothermus and Bruyobacte,but the relative abundances of Mizugakiibacter,Luedemannella,Actinothermus decreased by 2.47%, 1.78% and 0.68%, other bacteria increased by 0.10%-0.86% compared to CK under T treatment.? The dominant fungal phyla in the rhizosphere soil of CK and T treatments was Ascomycota,80.00% and 92.00% respectively,followed by Basidiomycota,8.00% and 6.00%,the top three dominant fungi genus all belonged to the genus Taifanglania,Chaetomium and Fusarium. Principal component analysis(PCA) of six root soil samples treated with CK and T showed that the variation of bacterial community PC1 was 29.37%,the variation of PC2 was 20.26%,the total contribution rate of both was 49.63%; the variation of fungal community PC1 was 33.44%,the variation of PC2 was 27.73%,the total contribution rate of both was 56.17%. The bacterial communities of CK and T treatment belonged to different quadrants,respectively,CK was located in the 1st and 3rd quadrants,and T treatment was located in the 2nd and 4th quadrants. 【Conclusion】The results show that intercropping sugarcane with M. pruriens improves the soil nutrient status,changes the microbial community structure,and increases the richness and diversity of bacteria and fungi in sugarcane rhizosphere, but does not affect the sequence of dominant microflora.
Key words: sugarcane; Mucuna pruriens var. utilis; intercropping; rhizosphere; soil microbe; high-throughput sequencing; community structure
Foundation item:National Natural Science Foundation of China(31860350); National Sugar Industry Technology System Project(CARS-170105); Central Guides Local Project(Guike ZY20198005); Basic Research Project of Guangxi Academy of Agricultural Sciences(Guinongke 2021YT006)
0 引言
【研究意義】甘蔗(Saccharum officinarum)為全球重要的糖類和能源作物,是種植行距較寬的作物,目前種植行距多為1.2 m,常與生長快速的作物間作或與生長期短的作物套種可有效改善土壤酶活性,提高土壤微生物數(shù)量和多樣性,充分提升養(yǎng)分利用效率及經(jīng)濟效益(張愛加等,2013;彭東海等,2014;沈雪峰等,2014)。貓豆為一年生豆科黎豆屬草質(zhì)藤本植物,多分支,根系較發(fā)達,葉蔓攀高,覆蓋面廣,有利于保持水土,其根部富含根瘤菌,可有效提高土地肥力(秦祖臻等,2012;韋道鞍,2013;李經(jīng)成等,2016),適合與其他作物間種。土壤微生物多樣性不僅代表微生物群落的穩(wěn)定性,同時還反映土壤的生態(tài)機制和土壤脅迫對微生物群落的影響,是導(dǎo)致微生物代謝方式和生理功能多樣化的直接原因(彭東海等,2014;張晟等,2019)。因此,探究甘蔗間作貓豆對甘蔗根際微生物多樣性的影響,對改善甘蔗根際土壤菌群結(jié)構(gòu)及促進甘蔗生長具有重要意義。【前人研究進展】據(jù)國內(nèi)外研究報道,甘蔗間作模式多種多樣,主要有甘蔗—蔬菜(如洋蔥、馬鈴薯等)(Bokhtiar et al.,2003)、甘蔗—豆科植物(大豆)(Kamruzzaman and Hasanuzzama,2007)和甘蔗—禾本科植物(玉米)(鄭亞強等,2018)等。研究發(fā)現(xiàn),甘蔗間作大豆后能顯著增加其根際土壤中的真菌和放線菌數(shù)量,土壤微生物群落多樣性豐富,蔗田土壤得到有效改善(洪德星,2011;Li et al.,2013;彭東海等,2014),且甘蔗間作大豆后在甘蔗苗期和分蘗期蔗根的內(nèi)生細菌多樣性均大于單作處理(楊建波,2014);甘蔗間作玉米后甘蔗根際微生物的群落多樣性和土壤酶活性均高于單作甘蔗(鄭亞強等,2016,2018);甘蔗與大豆和花生間作能提高根際土壤微生物多樣性和土壤酶活性(邢德峰和任南琪,2006;Vo?í?ková and Baldrian,2013)。上述研究主要利用傳統(tǒng)培養(yǎng)方法或變性梯度凝膠電泳(DGGE)技術(shù)探討甘蔗與不同作物間作對其根際土壤微生物的影響,然而傳統(tǒng)方法培養(yǎng)的微生物無法全面準確地反映根際土壤微生物的多樣性特征(張洪霞等,2009),尤其是DGGE技術(shù)僅能分析有限的優(yōu)勢微生物類群,存在高估物種豐富度及低估微生物群落大小和多樣性的可能,很難檢測出低豐度的土壤微生物類群(Tan et al.,2015)。近年來,高通量測序已成為定量和鑒別微生物群落演替研究的重要工具,尤其是二代高通量測序技術(shù)具有測序通量高、試驗過程簡化、速度快和準確率高等優(yōu)點,能更真實地揭示原位環(huán)境中微生物群落的復(fù)雜性和多樣性(李慶崗和陶立,2012),已廣泛用于生態(tài)系統(tǒng)土壤微生物多樣性研究(Will et al.,2010;井趙斌等,2013)。張東艷等(2017)采用Illumina HiSeq高通量測序技術(shù)研究玄參與煙草間作后根際土壤細菌群落結(jié)構(gòu),結(jié)果發(fā)現(xiàn),間作降低了煙草根際土壤細菌豐富度和多樣性,但提高了玄參根際土壤細菌多樣性?!颈狙芯壳腥朦c】目前鮮見有關(guān)高通量測序技術(shù)對甘蔗間作貓豆后甘蔗根際土壤微生物多樣性進行分析的文獻報道?!緮M解決的關(guān)鍵問題】利用Illumina HiSeq高通量測序技術(shù)分析甘蔗間作貓豆后甘蔗根際土壤微生物群落結(jié)構(gòu)及多樣性,為改善甘蔗根際土壤菌群結(jié)構(gòu)及促進甘蔗生長提供理論參考。
1 材料與方法
1. 1 試驗地概況
試驗在廣西農(nóng)業(yè)科學(xué)院甘蔗研究所隆安縣丁當(dāng)基地進行,前茬作物為單作甘蔗。廣西隆安縣位于廣西西南部,屬南亞熱帶季風(fēng)氣候。土壤基本理化性質(zhì):pH 4.9,有機質(zhì)26.1 g/kg,水解性氮108 mg/kg,有效磷5.4 mg/kg,速效鉀213.0 mg/kg,全氮11.7 g/kg,全磷4.1 g/kg,全鉀22.2 g/kg。
1. 2 試驗材料
參試甘蔗品種為桂糖46號,貓豆為從斐濟引進品種。2×Taq MasterMix、RNase-Free water、瓊脂糖、TruSeq? DNA PCR-Free Sample Preparation Kit建庫試劑盒購自北京康為世紀生物科技有限公司;引物由南寧國拓生物科技有限公司合成。主要儀器設(shè)備:普通PCR儀(Biometra,德國)、Thermo Scientific NanoDrop 2000c微量分光光度計(上海奧陸生物科技有限公司)、DYY-7C核酸電泳儀(北京六一儀器廠)、GS-800凝膠掃描儀(Bio-Rad,美國)、Illumina HiSeq2500 PE250高通量測序儀(Illumina,美國)。
1. 3 試驗方法
1. 3. 1 試驗設(shè)計及樣品采集 設(shè)甘蔗單作和甘蔗間作貓豆2個處理,每處理設(shè)3次重復(fù),共種植6個小區(qū),每小區(qū)面積為100 m2。采用五點取樣法從各試驗小區(qū)采集深度0~20 cm的根際土壤樣品,將五點的土樣混勻,記作1份根際土壤樣品。單作甘蔗為對照(CK),3個生物學(xué)重復(fù)分別標(biāo)記為CK1、CK2和CK3,甘蔗間作貓豆標(biāo)記為T處理,3個生物學(xué)重復(fù)分別標(biāo)記為T1、T2和T3。將6份土壤樣品低溫保存帶回實驗室,去除植物碎葉等雜物。每份土壤樣品再平均分成2份,分別用于土壤理化性質(zhì)和土壤微生物測定。
1. 3. 2 根際土壤養(yǎng)分含量測定 土壤養(yǎng)分含量按照《土壤分析技術(shù)規(guī)范》進行測定。
1. 3. 3 基因組DNA提取及PCR擴增 采用十六烷基三甲基溴化銨(CTAB)法提取土壤樣品的基因組DNA,并利用2%瓊脂糖凝膠電泳進行檢測。以稀釋后的基因組DNA為模板進行PCR擴增,所用的引物為細菌16S rDNA的V4區(qū)引物(515F和806R)和真菌ITS的ITS 1區(qū)引物(ITS5-1737F和ITS2-2043R)(李善家等,2020)。將PCR產(chǎn)物進行等量混樣,充分混勻后使用2%瓊脂糖凝膠電泳進行檢測,并利用膠回收試劑盒回收目的條帶。
1. 3. 4 文庫構(gòu)建及測序數(shù)據(jù)處理 用建庫試劑盒構(gòu)建文庫,基于Illumina HiSeq測序平臺進行測序。對原始數(shù)據(jù)進行拆分、截去引物序列,利用FLASH軟件對每個數(shù)據(jù)進行拼接(Mago? and Salzberg,2011)。經(jīng)過嚴格過濾處理得到高質(zhì)量的Tags數(shù)據(jù)(Clean tags)(Bokulich et al.,2012)。使用Qiime V1.7.0的Tags質(zhì)控流程得到最終的有效數(shù)據(jù)(Effe-ctive tags)(Caporaso et al.,2010)。
1. 3. 5 OTU聚類和物種注釋 利用QIIME V1.7.0對所有樣品的Effective tags進行聚類分析,同時依據(jù)其算法原則,篩選出作為OTUs的代表序列,并進行物種注釋分析(黃萍等,2020)。參照龔賽(2017)的方法進行后續(xù)Alpha多樣性和Beta多樣性分析。
1. 4 統(tǒng)計分析
采用Qiime Version 1.7.0計算Unifrac距離,并構(gòu)建非加權(quán)組平均法(UPGMA)聚類樹。使用R軟件(Version 2.15.3)繪制PCA、PCoA和NMDS圖。
2 結(jié)果與分析
2. 1 單作甘蔗與甘蔗間作貓豆的根際土壤養(yǎng)分含量比較結(jié)果
由表1可知,T處理的根際土壤中全氮、全鉀、水解性氮、有效磷和有機質(zhì)含量均高于CK的根際土壤,其中,有效磷和有機質(zhì)含量分別比CK的根際土壤顯著增加86.7%和16.0%(P<0.05,下同),表明甘蔗間作貓豆有利于提高甘蔗根際土壤的全氮、全鉀、水解性氮、有效磷和有機質(zhì)含量。
2. 2 單作甘蔗和甘蔗間作貓豆的甘蔗根際土壤微生物高通量測序結(jié)果
將在Illumina HiSeq測序平臺得到的測序數(shù)據(jù)進行拼接和過濾處理,得到Clean tags,再經(jīng)嵌合體過濾后得到可用于后續(xù)分析的Effective tags,對所有樣品的Effective tags進行聚類分析,以97%的一致性(Identity)將其聚類成為OTUs。6份根際土壤樣品的細菌16S rDNA測序結(jié)果(表2)顯示,CK的平均Effe-ctive tags為156376個,聚類成3185個OTUs,T處理的平均Effective tags為186446個,聚類成3184個OTUs。6份根際土壤樣品的真菌ITS測序結(jié)果(表2)顯示,CK的平均Effective tags為194892個,聚類成751個OTUs,T處理的平均Effective tags為213566個,聚類成889個OTUs。說明甘蔗間作貓豆后根際土壤中細菌的不相似序列數(shù)量與單作甘蔗無顯著差異(P>0.05,下同),真菌的不相似序列數(shù)量顯著高于單作甘蔗。
稀釋曲線可反映樣品的取樣深度,用以評價測序量覆蓋所有類群的程度。從圖1和圖2可看出,土壤樣品的微生物稀釋曲線基本趨于平緩,但仍未達到飽和,說明取樣基本合理,微生物群落結(jié)構(gòu)的置信度較高,能較真實地反映土壤微生物群落構(gòu)成。
2. 3 單作甘蔗和甘蔗間作貓豆的甘蔗根際土壤微生物多樣性指數(shù)分析結(jié)果
土壤細菌和真菌群落多樣性指數(shù)測定結(jié)果如表3和表4所示。6個土壤樣品的覆蓋率指數(shù)均大于0.990,表明樣品中序列未被測到的概率較低,測序結(jié)果已覆蓋測試樣品中的絕大部分物種。CK和T處理的細菌平均ACE指數(shù)和Chao1指數(shù)均較大,且無顯著差異,說明CK和T處理的甘蔗根際土壤中細菌多樣性均非常豐富。與CK甘蔗根際土壤相比,T處理的甘蔗根際土壤中細菌和真菌的豐富度均有所增加。T處理的細菌和真菌平均Shannon指數(shù)顯著高于CK,說明甘蔗間作貓豆會增加土壤微生物均勻度??傮w來看,CK和T處理的甘蔗根際土壤中,細菌的豐富度均高于真菌;T處理的細菌豐富度高于CK,但差異不顯著,T處理的細菌多樣性顯著高于CK;T處理的真菌豐富度和多樣性顯著高于CK。
2. 4 單作甘蔗和甘蔗間作貓豆的甘蔗根際土壤門水平的群落結(jié)構(gòu)分析結(jié)果
由于土壤樣品中所檢測出的微生物種類繁多,許多種類含量極少,不能與含量多的物種表示在同一個圖中,故選擇排名前10名的細菌門、相對豐度大于0.001%的真菌門,以相對豐度為縱坐標(biāo),繪制柱形圖。由圖3可知,CK和T處理根際土壤的前10個細菌門總相對豐度分別占土壤細菌門總數(shù)的95.71%和96.27%;CK和T處理根際土壤中的主要優(yōu)勢細菌門為變形菌門(Proteobacteria)、放線菌門(Actinobacteria)、綠彎菌門(Chloroflexi)和酸桿菌門(Acidobacteria),其相對豐度均大于10.00%,這4個細菌門總相對豐度分別占土壤細菌門總數(shù)的81.36%和85.61%;其次為芽單胞細菌門(Gemmatimonadetes)、厚壁菌門(Firmicutes)、硝化螺旋菌門(Nitrospirae)、擬桿菌門(Bacteroidetes)、軟壁菌門(Tenericutes)和奇古菌門(Thaumarchaeota),這6個細菌門總相對豐度分別占土壤細菌門總數(shù)的14.35%和10.66%;與CK根際土壤相比,T處理根際土壤中排名前3的變形菌門、放線菌門和綠彎菌門,分別降低1.49%、1.95%和2.21%,其他細菌門分別增加0.17%~1.58%。
由圖4可知,CK和T處理根際土壤中優(yōu)勢真菌門為子囊菌門(Ascomycota),分別占土壤真菌門總數(shù)的80.00%和92.00%,其次是擔(dān)子菌門(Basidiomycota),分別占土壤真菌門總數(shù)的8.55%和6.22%,以接合菌門(Zygomycota)、壺菌門(Chytridiomycota)和球囊菌門(Glomeromycota)相對豐度量較少;與CK土壤樣品相比,T處理根際土壤中優(yōu)勢真菌門子囊菌門增加12.00%,擔(dān)子菌門下降2.33%。
2. 5 單作甘蔗和甘蔗間作貓豆的甘蔗根際土壤屬水平的群落結(jié)構(gòu)分析結(jié)果
對根際土壤前10個優(yōu)勢菌屬相對豐度進行分析,結(jié)果如圖5所示。CK和T處理根際土壤中的優(yōu)勢細菌屬分別為水恒桿菌屬(Mizugakiibacter)、游動四孢屬(Luedemannella)、乳桿菌屬(Acidothermus)和布氏桿菌屬(Bryobacte)。但與CK根際土壤相比,T處理根際土壤樣品中水恒桿菌屬、游動四孢屬和乳桿菌屬的相對豐度分別下降2.47%、1.78%和0.68%,其他細菌屬分別增加0.10%~0.86%。
對根際土壤優(yōu)勢真菌屬相對豐度進行分析,結(jié)果如圖6所示。CK和T處理根際土壤中排名前3的優(yōu)勢真菌屬均為戴氏霉屬(Taifanglania)、毛殼屬(Chaetomium)和鐮刀菌屬(Fusarium)。且CK與T處理根際土壤中相對豐度相差較大(1.12%~2.05%)的真菌屬為小蘑菇屬(Micropsalliota)、鬼筆屬(Phallus)、黑團孢屬(Periconia)、圓孢霉屬(Staphylotrichum)和裸節(jié)菌屬(Talaromyces)。
2. 6 單作甘蔗和甘蔗間作貓豆的甘蔗根際土壤微生物群落的主成分分析結(jié)果
對CK和T處理的6份根際土壤樣品細菌群落進行種水平上的主成分(PCA)分析,結(jié)果如圖7所示。細菌群落PC1的變異為29.37%,PC2的變異為20.26%,二者的總貢獻率為49.63%。如圖8所示,真菌群落PC1的變異為33.44%,PC2的變異為22.73%,二者的總貢獻率為56.17%。CK與T處理的細菌群落分別屬于不同象限,CK位于第1和第3象限,而T處理位于第2和第4象限,表明甘蔗間作貓豆改變了根際土壤微生物群落結(jié)構(gòu)。
3 討論
甘蔗間作大豆可提高甘蔗的株高和葉長,根際土壤中碳、氮和磷含量大于單作甘蔗(卜俊瑤等,2020)。間作大豆改變土壤的pH,使其接近于中性,更適于甘蔗生長,還會增加土壤中氮的含量(農(nóng)寧娟,2015)。甘蔗間作花生后0~20 cm土壤中全氮、全鉀、有效磷、有機質(zhì)和pH均大于單作甘蔗(唐秀梅等,2020)。宿根蔗與大豆間作可增加甘蔗有效莖數(shù),提高甘蔗產(chǎn)量(覃劉東等,2019)。本研究結(jié)果也發(fā)現(xiàn),甘蔗間作貓豆可有效提高甘蔗根際土壤中的全氮、全鉀、水解性氮、有效磷和有機質(zhì)含量,與上述研究結(jié)果相似。
土壤微生物是植物—土壤生態(tài)系統(tǒng)的重要組成成分,其數(shù)量及多樣性對維持土壤生態(tài)系統(tǒng)的穩(wěn)定具有重要作用,并在一定程度上體現(xiàn)土壤的肥力和酶活性(徐麗慧等,2017;丁麗等,2020)。其中,細菌占整個土壤微生物群落的70%~90%,真菌在土壤微生物群落中所占比例僅次于細菌(任奎瑜等,2020)。與從功能入手的傳統(tǒng)微生物研究方法相比,高通量測序能更全面準確地從遺傳物質(zhì)上識別微生物的種屬。本研究對甘蔗根際土壤細菌16S rRNA基因V4區(qū)域和真菌ITS 1區(qū)域進行高通量測序,結(jié)果發(fā)現(xiàn),甘蔗間作貓豆后甘蔗根際土壤的Shannon和ACE指數(shù)均較單作甘蔗高,且細菌的豐富度遠大于真菌的豐富度;在這2種種植模式下甘蔗根際土壤中細菌主要門類為變形菌門、放線菌門、綠彎菌門和酸桿菌門,真菌主要門類為子囊菌門和擔(dān)子菌門;細菌主要優(yōu)勢菌屬為水恒桿菌屬、游動四孢屬、乳桿菌屬和布氏桿菌屬,真菌主要優(yōu)勢菌屬為戴氏霉屬、毛殼屬和鐮刀菌屬。彭東海等(2014)研究表明,甘蔗間作大豆后甘蔗根際土壤固氮細菌的Shannon指數(shù)高于單作甘蔗的根際土壤,可提高固氮細菌的多樣性和某些固氮細菌的優(yōu)勢度,但對群落物種的優(yōu)勢度影響較小。本研究也發(fā)現(xiàn),甘蔗間作貓豆后甘蔗根際土壤中細菌和真菌在門、屬水平上排名靠前的優(yōu)勢菌群的相對豐度略微降低,而其余的優(yōu)勢菌略微增加,說明甘蔗間作貓豆使根際土壤中的微生物發(fā)生趨向性改變,且使各菌群結(jié)構(gòu)變得均勻。目前關(guān)于間作對作物根際土壤微生物影響變化的觀點不盡相同。Wang等(2012)研究發(fā)現(xiàn),小麥間作豆科植物不影響小麥根際土壤微生物的群落結(jié)構(gòu);張曉崗等(2020)研究證明馬鈴薯間作玉米栽培后能有效改善真菌菌群結(jié)構(gòu),有害致病真菌菌屬消失或比例下降,隨之出現(xiàn)一些有益于作物生長的功能真菌;Wang等(2007)研究認為,在酸性土壤地上小麥間作蕓薹屬植物可改變小麥根際微生物群落結(jié)構(gòu),土壤細菌和放線菌的豐富度減少,真菌的豐富度增加。本研究中,甘蔗間作貓豆后甘蔗根際土壤中細菌和真菌的豐富度有所增加,且土壤微生物群落結(jié)構(gòu)發(fā)生改變,與甘蔗間作玉米的研究結(jié)果(鄭亞強等,2016)相似。根際土壤微生物群落結(jié)構(gòu)改變的具體原因尚待研究。
4 結(jié)論
甘蔗間作貓豆可改善甘蔗根際的土壤養(yǎng)分狀況,改變甘蔗根際微生物的群落結(jié)構(gòu),提高細菌和真菌的豐富度和多樣性,但未影響優(yōu)勢菌門的排序。
參考文獻:
卜俊瑤,Abdullah Khan,蔣雨珂,王梓軒. 2020. 甘蔗與大豆間作的根際促生菌篩選及促生效應(yīng)研究[J]. 鄉(xiāng)村科技,(1):92-95. doi:10.19345/j.cnki.1674-7909.2020.01.044. [Bu J Y,Abdullah K,Jiang Y K,Wang Z X. 2020. Scree-ning of rhizosphere growth promoting bacteria in sugarcane soybean intercropping and their growth promo-ting effects[J]. Rural Science and Technology,(1):92-95.]
丁麗,冀玉良,李懿. 2020. 不同林齡油松根際土壤微生物群落多樣性及其影響因子[J]. 水土保持研究,27(4):184-191. doi:10.13869/j.cnki.rswc.2020.04.025. [Ding L,Ji Y L,Li Y. 2020. Soil microbial diversity and its influen-cing factors in rhizosphere and non-rhizosphere in the stands of pinus tabuliformis with different ages in Minjiang river valley[J]. Research of Soil and Water Conservation,27(4):184-191.]
龔賽. 2017. 林下大球蓋菇栽培對土壤養(yǎng)分及微生物的影響[D]. 泰安:山東農(nóng)業(yè)大學(xué). [Gong S. 2017. The influnce on soil nutrients and microbe of cultivation of Stropharia rugosoannulata in the forest[D]. Taian:Shandong Agricultural University.]
洪德星. 2011. 不同套種作物對甘蔗生長和土壤理化性質(zhì)的影響[D]. 福州:福建農(nóng)林大學(xué). [Hong D X. 2011. Effect of two different interplanting crops on the sugarcane growth and soil physical and chemical properties of cane field[D]. Fuzhou:Fujian agriculture and Forestry University.]
黃萍,王楠,周紫羽,王婷,袁志良,常介田,葉永忠. 2020. 白云山落葉闊葉林土壤細菌群落結(jié)構(gòu)及環(huán)境因子的相關(guān)性分析[J]. 河南農(nóng)業(yè)大學(xué)學(xué)報,54(3):415-421. doi:10.16445/j.cnki.1000-2340.2020.03.005. [Huang P,Wang N,Zhou Z Y,Wang T,Yuan Z L,Chang J T,Ye Y Z. 2020. Soil bacterial community structure and correlation analysis of environmental factors in deciduous broad-leaved forest of Baiyun Mountain,54(3):415-421.]
井趙斌,程積民,張寶泉,李紅紅. 2013. 基于454焦磷酸測序法的典型草原土壤真核生物多樣性[J]. 草業(yè)科學(xué),30(11):1690-1697. [Jing Z B,Cheng J M,Zhang B Q,Li H H. 2013. Eukaryote diversity of typical grassland soil based on 454 pyrosequencing[J]. Pratacultural Science,30(11):1690-1697.]
李經(jīng)成,江洪平,羅高玲,陳燕華,蔡慶生. 2016. 貓豆種質(zhì)資源研究與綜合利用[J]. 中國種業(yè),(4):75-76. doi:10. 19462/j.cnki.1671-895x.2016.04.037. [Li J C,Jiang H P,Luo G L,Chen Y H,Cai Q S. 2016. Research and comprehensive utilization of catbean germplasm resources[J]. China Seed Industry,(4):75-76.]
李慶崗,陶立. 2012. 高通量測序技術(shù)及其在生命科學(xué)中的應(yīng)用[J]. 畜牧與飼料科學(xué),33(2):25-28. doi:10.16003/j.cnki.issn1672-5190.2012.02.003. [Li Q G,Tao L. 2012. High-throughput sequencing technology and its application in life science[J]. Animal Husbandry and Feed Scien-ce,33(2):25-28.]
李善家,王輝,茍偉,伍國強,蘇培璽. 2020. 混生荒漠植物葉片功能性狀與其根際微生物多樣性的關(guān)系[J]. 生態(tài)環(huán)境學(xué)報,29(9):1713-1722. doi: 10.16258/j.cnki.1674-5906. [Li S J,Wang H,Gou W,Wu G Q,Su P X. 2020. Relationship between leaf functional traits of mixed de-sert plants and microbial diversity in rhizosphere[J]. Eco-logy and Environmental Sciences,29(9):1713-1722.]
農(nóng)寧娟. 2015. 間作大豆對甘蔗根際土壤細菌及固氮菌多樣性的影響[J]. 南方農(nóng)業(yè),9(15):67. doi:10.19415/j.cnki. 1673-890x.2015.15.039. [Nong N J. 2015. Effects of intercropping with soybean on bacterial and nitrogen-fixing bacterial diversity in the rhizosphere of sugarcane[J]. South China Agriculture,9(15):67.]
彭東海,楊建波,李健,邢永秀,覃劉東,楊麗濤,李楊瑞. 2014. 間作大豆對甘蔗根際土壤細菌及固氮菌多樣性的影響[J]. 植物生態(tài)學(xué)報,38(9):959-969. [Peng D H,Yang J B,Li J,Xing Y X,Qin L D,Yang L T,Li Y R. 2014. Effects of intercropping with soybean on bacterial and nitrogen-fixing bacterial diversity in the rhizosphere of sugarcane[J]. Chinese Journal of Plant Ecology,38(9):959-969.]
覃劉東,楊建波,彭東海,楊麗濤,李楊瑞. 2019. 間種大豆對宿根甘蔗的影響[J]. 廣西糖業(yè),(3):10-12. [Qin L D,Yang J B,Peng D H,Yang L T,Li Y R. 2019. Effects of intercropping soybean on growth and yield in ratoon su-garcane[J]. Guangxi Sugar Industry,(3):10-12.]
秦祖臻,韋培炎,楊武興,陳仁欽,黃家訓(xùn). 2012. 石山地區(qū)春玉米套種貓豆模式栽培技術(shù)[J]. 廣西農(nóng)學(xué)報,(1):73-74. [Qin Z Z,Wei P Y,Yang W X,Chen R Q,Huang J X. 2012. Cultivation techniques of spring maize intercro-pping with cat bean in Shishan area[J]. Journal of Guangxi Agriculture,(1):73-74.]
任奎瑜,趙久成,郭霜紅,王帥帥,張傳進,龐師禪,楊尚東. 2020. 紅椎林中正紅菇生境的土壤肥力及真菌多樣性特征[J]. 西南農(nóng)業(yè)學(xué)報,33(1):109-116. doi:10.16213/j.cnki.scjas.2020.1.018. [Ren K Y,Zhao J C,Guo S H,Wang S S,Zhang C J,Pang S C,Yang S D. 2020. Chara-cteristics of soil biological properties and fungal diversity of russula vinosa in castanopsis hystrix forest[J]. Southwest China Journal of Agricultural Sciences,33(1):109-116.]
沈雪峰,方越,董朝霞,陳勇. 2014. 甘蔗/花生間作對土壤微生物和土壤酶活性的影響[J]. 作物雜志,(5):55-58. doi:10.16035/j.issn.1001-7283.2014.05.014. [Shen X F,F(xiàn)ang Y,Dong C X,Chen Y. 2014. Effects of sugarcane/peanut intercropping on soil microbes and soil enzyme activities[J]. Crops,(5):55-58.]
唐秀梅,蒙秀珍,蔣菁,黃志鵬,吳海寧,劉菁,賀梁瓊,熊發(fā)前,鐘瑞春,韓柱強,何龍飛,唐榮華. 2020. 甘蔗間作花生對不同耕層土壤微生態(tài)的影響[J]. 中國油料作物學(xué)報,42(5):713-722. doi:10.19802/j.issn.1007-9084.2019318. [Tang X M,Meng X Z,Jiang J,Huang Z P,Wu H N,Liu J,He L Q,Xiong F Q,Zhong R C,Han Z Q,He L F,Tang R H. 2020. Effects of sugarcane/peanut intercro-pping on soil microenvironment in different plough layer[J]. Chinese Journal of Oil Crop Sciences,42(5):713-722.]
韋道鞍. 2013. 石山地區(qū)玉米套種貓豆栽培技術(shù)[J]. 現(xiàn)代農(nóng)業(yè)科技,(14):32-33. doi:10.3969/j.issn.1007-5739. [Wei D A. 2013. Cultivation techniques of maize interplanting with Mucuna pruriens var. Utilis in Shishan area[J]. Mo-dern Agricultural Science and Technology,(14):32-33.]
邢德峰,任南琪. 2006. 應(yīng)用DGGE研究微生物群落時的常見問題分析[J]. 微生物學(xué)報,46(2):331-315. doi:10.13343/j.cnki.wsxb.2006.02.034. [Xing D F,Ren N Q. 2006. Common problems in the analyses of microbial community by denaturing gradient gel electrophoresis(DGGE)[J]. Acta Microbiologica Sinica,46(2):331-315.]
徐麗慧,曾蓉,高士剛,戴富明. 2017. 土壤真菌多樣性對土傳病害影響的研究進展[J]. 上海農(nóng)業(yè)學(xué)報,33(3):161-165. doi:10.15955/j.issn1000-3924.2017.03.30. [Xu L H,Zeng R,Gao S G,Dai F M. 2017. Review on the effect of soil fungal communities on soil-borne diseases[J]. Acta Agriculturae Shanghai,33(3):161-165.]
楊建波. 2014. 間作大豆對甘薦生長及內(nèi)生細菌、固氮細菌多樣性的效應(yīng)[D]. 南寧:廣西大學(xué). [Yang J B. 2014. Effects of intercropping with soybean on growth of sugarcane and diversities of endophytic baderia and endophytic nitrogen-fixing bacteria[D]. Nangning:Guangxi University.]
張愛加,周明明,林文雄. 2013. 不同種植模式對甘蔗根際土壤生物學(xué)特性的影響[J]. 植物營養(yǎng)與肥料學(xué)報,19(6):1525-1532. [Zhang A J,Zhou M M,Lin W X. 2013. Effects of different cultivation patterns on microorganism of soil rhizosphere in sugarcane fields[J]. Journal of Plant Nutrition and Fertilizers,19(6):1525-1532.]
張東艷,王軍,楊水平,張雪,劉京,趙建,何大敏,楊紅軍,莫靜靜,茍劍渝,趙新梅,蔣衛(wèi),丁偉,陳大霞. 2017. 玄參與煙草間作對土壤細菌群落結(jié)構(gòu)的影響[J]. 草業(yè)學(xué)報,26(6):120-130. doi:10.11686/cyxb2016507. [Zhang D Y,Wang J,Yang S P,Zhang X,Liu J,Zhao J,He D M,Yang H J,Mo J J,Gou J Y,Zhao X M,Jiang W,Ding W,Chen D X. 2017. Influence of Scrophularia ningpoensis-tobacco intercropping on bacterial community structure in soil[J]. Pratacultural Science,26(6):120-130.]
張洪霞,譚周進,張祺玲,盛榮,肖和艾. 2009. 土壤微生物多樣性研究的DGGE/TGGE技術(shù)進展[J]. 核農(nóng)學(xué)報,23(4):721-727. [Zhang H X,Tan Z J,Zhang Q L,Sheng R,Xiao H A. 2009. Advances of DGGE/TGGE technique for soil microbial diversity study[J]. Journal of Nuclear Agricultural Sciences,23(4):721-727.]
張晟,張徐潔,趙遠,張玉虎,胡茜,荊玉林,符菁. 2019. 不同溫度制備的水稻秸稈生物炭對稻田土壤固碳減排及微生物群落結(jié)構(gòu)的影響[J]. 江蘇農(nóng)業(yè)學(xué)報,35(5):1102-1111. doi:10.3969/j.issn.1000-4440.2019.05.015. [Zhang S,Zhang X J,Zhao Y,Zhang Y H,Hu Q,Jing Y L,F(xiàn)u J. 2019. Effects of rice straw biochar prepared at different pyrolysis temperatures on carbon sequestration and mitigation and microbial community structure in paddy soil[J]. Jiangsu Journal of Agricultural Sciences,35(5):1102-1111.]
張曉崗,劉萍,馬琨,王娜. 2020. 間作栽培對寧夏南部山區(qū)馬鈴薯根際土壤真菌菌群結(jié)構(gòu)的影響[J]. 西北農(nóng)業(yè)學(xué)報,(12):1-8. doi:10.7606/j.issn.1004-1389.2020.12.013. [Zhang X G,Liu P,Ma K,Wang N. 2020. Effect of potato intercropping on fungal community composition of rhizosphere soil in southern mountainous area of Ningxia[J]. Acta Agriculturae Boreali-occidentalis Sinica,(12):1-8.]
鄭亞強,杜廣祖,李亦菲,陳斌,李正躍,肖關(guān)麗. 2018. 間作甘蔗對玉米根際微生物功能多樣性的影響[J]. 生態(tài)學(xué)雜志,37(7):2013-2019. doi:10.13292/j.1000-4890.201807. 008. [Zheng Y Q,Du G Z,Li Y F,Chen B,Li Z Y,Xiao G L. 2018. Effects of intercropping sugarcane on functional diversity of maize rhizosphere microor-ganisms[J]. Chinese Journal of Ecology,37(7):2013-2019.]
鄭亞強,張立敏,楊進成,楊堅,高銳,陳亮新,董雪梅,孫繼紅,肖關(guān)麗,李正躍,陳斌. 2016. 甘蔗間作玉米對甘蔗根際微生物代謝功能多樣性的影響[J]. 中國生態(tài)農(nóng)業(yè)學(xué)報,24(5):618-627. doi:10.13930/j.cnki.cjea.151149. [Zheng Y Q,Zhang L M,Yang J C,Yang J,Gao R,Chen L X,Dong X M,Sun J H,Xiao G L,Li Z Y,Chen B. 2016. Effects of sugarcane and maize intercropping on sugarcane rhizosphere microbe metabolic function diversity[J]. Chinese Journal of Eco-Agriculture,24(5):618-627.]
Bokhtiar S M,Hossain M S,Mahmud K,Paul G C. 2003. Site specific nutrient management for sugarcane potato and sugarcane-onion intercropping systems[J]. Asian Journal of Plant Sciences,2(17):1205-1208. doi:10.3923/ajps.2003. 1205.1208.
Bokulich N A,Subramanian S,F(xiàn)aith J J,Gevers D,Gordon J I,Knight R,Mills D A,Caporaso J G. 2012. Quality-filtering vastly improves diversity estimates from Illumina amplicon sequencing[J]. Nature Methods,10(1):57-59. doi:10.1038/nmeth.2276.
Caporaso J G,Kuczynski J,Stombaugh J,Bittinger K,Bushman F D,Costello E K,F(xiàn)ierer N,Pe?a A G,Goodrich J K,Gordon J I. 2010. Qiime allows analysis of high-throughput community sequencing data[J]. Nature Methods,7(5):335-336.
Kamruzzaman M,Hasanuzzama M. 2007. Factors affecting profitability of sugarcane production as monoculture and as intercrop in selected areas of Bangladesh[J]. Agricultural Research,32(3):433-444. doi:10.3329/bjar.v32i3. 545.
Li X P,Mu Y H,Cheng Y B,Liu X,Hai N. 2013. Effects of intercropping sugarcane and soybean on growth,rhizosphere soil microbes,nitrogen and phosphorus availability[J]. Acta Physiologiae Plantarum,35(4):1113-1119. doi:10.1007/s11738-012-1148-y.
Mago? T,Salzberg S L. 2011. Flash:Fast length adjustment of short reads to improve genome assemblies[J]. Bioinforma-tics,27(21):2957-2963. doi:10.1093/bioinformatics/btr507.
Tan Y,Chen Q,Liu H J,Song S D,Yu X M,Dong Z H,Tang X,Zhong Y Z. 2015. Bacteria community in different aged Coptis chinensis planting soil revealed by PCR-DGGE analysis[J]. China Journal of Chinese Materia Medica,40(16):3147. doi:10.4268/cjcmm20151606.
Vo?í?ková J,Baldrian P. 2013. Fungal community on decomposing leaflitter undergoes rapid successional changes[J]. Isme Journal,7(3):477-486. doi:10.1038/ismej.2012.116.
Wang D W,Marschner P,Solaiman Z,Rengel Z. 2007. Belowground interactions between intercropped wheat and Brassicas in acidic and alkaline soils[J]. Soil Biology and Biochemistry,39(4):961-971. doi:10.1016/j.soilbio.2006. 11.008.
Wang Y,Marschner P,Zhang F. 2012. Phosphorus pools and other soil properties in the rhizosphere of wheat and legumes growing in three soils in monoculture or as a mixture of wheat and legume[J]. Plant and Soil,354(1-2):283-298. doi:10.1007/s11104-011-1065-7.
Will C,Thrmer A,Wollherr A,Thurmer A,Wollherr A,Nacke H,Herold N,Schrumpf M,Gutknecht J,Wobet T,Buscot F,Daniel R. 2010. Horizon-specific bacterial community composition of German grassland soils,as revealed by pyrosequencing-based analysis of 16S rRNA genes[J]. Applied and Environmental Microbiology,76(20):6751-6759. doi:10.1128/AEM.01063-10.
(責(zé)任編輯? 陳 燕)