摘""要:本研究旨在探究不同施肥水平下第2年宿根蔗根圍土壤叢枝菌根(arbuscular"mycorrhizal,"AM)真菌的多樣性,為AM真菌在甘蔗上的應(yīng)用提供理論依據(jù)。本研究以甘蔗品種桂糖29號(hào)(GT29)和桂糖58號(hào)(GT58)為試驗(yàn)材料,設(shè)4個(gè)施肥水平:0.0"kg/hm2(A)、562.5"kg/hm2(B)、1125.0"kg/hm2(C)、2250.0"kg/hm2(D)。研究結(jié)果表明:不同施肥水平對(duì)GT29的根系菌根侵染率有顯著影響,而對(duì)GT58影響不顯著。隨著施肥水平的提高,AM真菌多樣性和豐富度均顯著下降。不同施肥水平的GT29根圍土壤中共獲得185個(gè)OTUs,GT58根圍土壤中共獲得175個(gè)OTUs。不同施肥水平下2個(gè)甘蔗品種根圍土壤的優(yōu)勢(shì)AM真菌均為球囊霉屬(Glomus)。在OTU水平對(duì)不同施肥水平甘蔗根圍土壤AM真菌群落進(jìn)行差異物種分析,發(fā)現(xiàn)GT29的2個(gè)差異物種均為球囊霉屬,而GT58中的3個(gè)差異物種均為近明球囊霉屬(Claroideoglomus)。經(jīng)Mantel檢驗(yàn),發(fā)現(xiàn)土壤中的Ca、Mg含量與4個(gè)AM真菌屬之間均呈極顯著正相關(guān),土壤pH與3個(gè)AM真菌屬間也呈顯著正相關(guān)。因此,過(guò)量施肥會(huì)使增產(chǎn)效果大幅降低,且造成土壤酸化及磷元素過(guò)剩,同時(shí)使甘蔗根圍土壤AM真菌群落組成改變,多樣性和豐富度降低,但品種間有差異;適量施肥有助于豐富土壤AM真菌的多樣性,促進(jìn)甘蔗產(chǎn)業(yè)的可持續(xù)發(fā)展。
關(guān)鍵詞:甘蔗;宿根蔗;根圍土壤;叢枝菌根真菌;多樣性中圖分類號(hào):S566.1;S154.3""""""文獻(xiàn)標(biāo)志碼:A
Study"on"the"Diversity"of"Arbuscular"Mycorrhizal"Fungi"in"the"Rhizosphere"Soil"of"the"Second"Ratoon"Sugarcane
WANG"Sheng1,2,"XU"Xinting2,3,"YANG"Chunhua2,3,"QIN"Xiaojuan2,"KANG"Yihao2,"ZHANG"Jinlian2*,"ZHAO"Lijun1*,"CHEN"Tingsu2
1."College"of"Forestry,"Guangxi"University,"Nanning,"Guangxi"530004,"China;"2."Microbiology"Research"Institute,"Guangxi"Academy"of"Agricultural"Sciences,"Nanning,"Guangxi"530007,"China;"3."College"of"Agriculture,"Guangxi"University,"Nanning,"Guangxi"530004,"China
Abstract:"This"paper"aimed"to"explore"the"arbuscular"mycorrhizal"(AM)"fungal"community"structure"in"the"rhizosphere"soil"of"ratoon"sugarcane"in"the"second"year"under"different"fertilization"levels,"and"provide"a"theoretical"basis"for"the"application"of"AM"fungi"in"sugarcane."In"this"study,"sugarcane"varieties"GT29"and"GT58"were"used"as"the"materials."Four"fertilization"levels"were"set:"0.0"kg/hm2"(A),"562.5"kg/hm2"(B),"1125.0"kg/hm2"(C),"2250.0"kg/hm2(D)."The"results"showed"that"different"fertilization"levels"had"significant"effects"on"the"root"mycorrhizal"infection"rate"of"GT29,"but"had"no"significant"effect"on"GT58."With"the"increase"of"fertilization"level,"the"diversity"and"richness"of"AM"fungi"decreased"significantly."With"the"increase"of"fertilization"level,"the"diversity"and"richness"of"AM"fungi"decreased"significantly."A"total"of"185"AM"fungi"OTUs"were"detected"in"GT29"and"175"in"GT58."The"dominant"AM"fungi"in"the"rhizosphere"soil"of"the"two"varieties"under"different"fertilization"levels"were"Glomus.nbsp;The"species"diversity"of"AM"fungal"community"in"rhizosphere"soil"under"different"fertilization"levels"was"analyzed"at"the"OTU"level."It"was"found"that"the"two"species"of"GT29"belonged"to"the"genus"Glomus,"while"the"three"species"of"GT58"belonged"to"the"genus"Claroidoglomus."Mantel"test"showed"that"the"contents"of"Ca"and"Mg"in"soil"were"significantly"positively"correlated"with"the"four"genera,"and"the"soil"pH"was"also"significantly"positively"correlated"with"the"three"genera."Therefore,"excessive"fertilization"will"significantly"reduce"the"yield"increase"effect,"cause"soil"acidification"and"excess"phosphorus,"and"change"the"composition,"diversity"and"richness"of"AM"fungi"community"in"sugarcane"rhizosphere"soil,"but"the"varieties"are"different;"Proper"fertilization"was"helpful"to"enrich"the"diversity"of"soil"AM"fungi"and"promote"the"sustainable"development"of"sugarcane"yield.
Keywords:"sugarcane;"ratoon"sugarcane;"rhizosphere"soil;"arbuscular"mycorrhizae"fungi;"diversity
DOI:"10.3969/j.issn.1000-2561.2025.02.020
甘蔗是最主要的糖料和經(jīng)濟(jì)作物,作為C4植物,因其高效的光合作用能力而被廣泛栽培于熱帶和亞熱帶地區(qū)[1]。目前,全球約有110個(gè)國(guó)家和地區(qū)種植甘蔗,全球約80%的糖和35%的乙醇以其為原料,是發(fā)展中國(guó)家重要的支柱產(chǎn)業(yè)之一。我國(guó)是世界第三大甘蔗生產(chǎn)國(guó),甘蔗主要種植區(qū)分布在廣西、云南、廣東等地[2],其中廣西的甘蔗種植面積多達(dá)84.8萬(wàn)hm2,占全國(guó)甘蔗總種植面積的65.8%(《中國(guó)統(tǒng)計(jì)年鑒2023》)。目前,廣西甘蔗生產(chǎn)中存在的主要問(wèn)題有機(jī)械化程度低、健康種苗應(yīng)用率低、過(guò)量施肥、宿根年限短等[3]。在這些問(wèn)題中,不合理施肥導(dǎo)致的生產(chǎn)問(wèn)題越來(lái)越突出,不僅造成土壤營(yíng)養(yǎng)元素失衡[4],還破壞了甘蔗根圍微環(huán)境,嚴(yán)重影響甘蔗產(chǎn)量[5]。
叢枝菌根(arbuscular"mycorrhizal,"AM)真菌是一類普遍存在于土壤環(huán)境中的微生物,能夠與超過(guò)80%的高等植物的根系形成共生關(guān)系[6]。AM真菌可以提高宿主植物的抗逆性[7],改善土壤團(tuán)粒結(jié)構(gòu),修復(fù)土壤退化[8]。廣西甘蔗根圍土壤有豐富的AM真菌資源,目前已經(jīng)在甘蔗根圍土壤中鑒定出7科12屬24種AM真菌[9]。土壤中AM真菌的孢子密度受土壤環(huán)境因子的影響[10]。研究表明,常規(guī)施肥使土壤AM真菌的侵染率降低[11],而合理施肥可以提高土壤微生物群落的穩(wěn)定性,并使深層土壤的無(wú)機(jī)氮積累有效降低[12];接種AM真菌可以顯著促進(jìn)甘蔗的生長(zhǎng),接種AM真菌的甘蔗植株在株高、綠葉數(shù)、莖徑和根系發(fā)達(dá)度等方面均表現(xiàn)出明顯的優(yōu)勢(shì)[13];大田甘蔗接種AM真菌可以提高土壤pH,促進(jìn)甘蔗根系對(duì)土壤養(yǎng)分的吸收,使甘蔗增產(chǎn)10%~24%,同時(shí)減少30%以上化肥施用量[14-16]。這為可持續(xù)農(nóng)業(yè)生產(chǎn)提供了一種有效的生物技術(shù)手段,有助于提高甘蔗產(chǎn)量和質(zhì)量,減少對(duì)化肥的依賴。
在農(nóng)業(yè)生產(chǎn)過(guò)程中,AM真菌易受耕作方式、施肥等因素的影響[17-18],但有關(guān)宿根蔗根圍土壤AM真菌與施肥水平關(guān)系的研究較少,缺乏長(zhǎng)期合理的施肥指導(dǎo)。因此,本研究旨在探究不同施肥水平對(duì)甘蔗根圍土壤AM真菌多樣性的影響,為解決過(guò)量施肥造成的養(yǎng)分資源浪費(fèi)和防治面源污染問(wèn)題,推動(dòng)蔗糖產(chǎn)業(yè)的可持續(xù)發(fā)展,利用菌根技術(shù)在甘蔗生產(chǎn)方面的應(yīng)用奠定理論基礎(chǔ)。
1.1""材料
供試甘蔗品種為甘蔗優(yōu)良品種桂糖58號(hào)(GT58)和桂糖29號(hào)(GT29)。試驗(yàn)在廣西農(nóng)業(yè)科學(xué)院科學(xué)研究基地試驗(yàn)地(23°14¢N,"108°33¢E)進(jìn)行。供試土壤基本理化性質(zhì):pH為5.1,有機(jī)質(zhì)為32.3"g/kg,全氮為2.3"g/kg,全磷為1.8"g/kg,全鉀為10.9"g/kg,水解氮為52.0"mg/kg,有效磷為73.2"mg/kg,速效鉀為110"mg/kg。
1.2""方法
1.2.1""試驗(yàn)設(shè)計(jì)""田間試驗(yàn)設(shè)計(jì)采用隨機(jī)區(qū)組設(shè)計(jì),設(shè)置6次重復(fù);5行小區(qū),行長(zhǎng)為7"m,行距為1.2"m;不同小區(qū)間種植同一品種3行甘蔗作為隔離區(qū)。由于不同甘蔗品種的分蘗有差異,特別是GT29的分蘗非常強(qiáng),按品種特性,GT58的下種量為120"000芽/hm2,GT29的下種量為75"000芽/hm2。
按廣西種植甘蔗普遍采用的施肥量為1500"kg/hm2,參考近年來(lái)甘蔗菌根的大田研究結(jié)果[14-16,"19-21],設(shè)置4個(gè)不同的施肥水平(表1):0.0"kg/hm2,"0%(A);562.5"kg/hm2,"25%(B);1125.0"kg/hm2,"50%(C);2250.0"kg/hm2,"100%(D)。緩釋復(fù)合肥N∶P∶K=17∶7∶17(河北天人化工股份有限公司)。
于2020年1月8日種植新植蔗,研究數(shù)據(jù)來(lái)源于2022年甘蔗種植第3年,即第2年宿根蔗田間試驗(yàn)。宿根蔗管理措施與常規(guī)種植相同,2022年3月15日苗期復(fù)合肥用量40%,2022年5月18日培土追肥用量60%。
1.2.2""甘蔗農(nóng)藝性狀測(cè)定""于2022年4月10日統(tǒng)計(jì)甘蔗出芽數(shù),計(jì)算出芽率;5月26日調(diào)查甘蔗分蘗數(shù),11月26日調(diào)查甘蔗有效莖數(shù),12月15日采收甘蔗稱重,并從中間3行選取20株長(zhǎng)勢(shì)相近的甘蔗測(cè)量株高、莖徑和錘度。
1.2.3""土壤和根系樣品采集""參考KANG等[21]的甘蔗土壤和根系采樣方法,于2022年12月27日以五點(diǎn)取樣法采集0~30"cm土層的甘蔗根系及根圍土壤。
1.2.4""土壤理化性質(zhì)測(cè)定""參照《土壤農(nóng)化分析》[22]測(cè)定土壤pH、有機(jī)質(zhì)(SOM)、堿解氮(AN)、有效磷(AP)、鈣(Ca)、速效鉀(AK)、鎂(Mg)、銅(Cu)、鐵(Fe)、錳(Mn)、鋅(Zn)含量等甘蔗根圍土壤的理化性質(zhì)。
1.2.5""甘蔗根系菌根侵染率測(cè)定""對(duì)甘蔗根系染色,觀察AM真菌在侵染甘蔗根系后形成的定殖結(jié)構(gòu)。參考廖楠等[23]的甘蔗根系菌根檢測(cè)方法對(duì)根系菌根進(jìn)行染色。在顯微鏡下觀察染色的根系,并記錄根系A(chǔ)M真菌菌絲、叢枝、泡囊等定殖結(jié)構(gòu),對(duì)照菌根侵染與泡囊豐度分級(jí)標(biāo)準(zhǔn),參考TROVELOT等[24]的方法,利用Mycocale軟件計(jì)算菌根侵染率、侵染強(qiáng)度、相對(duì)侵染強(qiáng)度、相對(duì)叢枝豐度和叢枝豐度。
1.2.6""土壤AM真菌高通量測(cè)序""參考KANG等[21]的方法對(duì)土壤AM真菌進(jìn)行高通量測(cè)序。
1.3""數(shù)據(jù)處理
使用Trimmomatic"0.38軟件對(duì)原始測(cè)序序列進(jìn)行質(zhì)控,使用Flash"1.2.11序列進(jìn)行雙端拼接,采用UPARSE"11工具根據(jù)97%相似度聚類成不同的操作分類單元(operational"taxonomic"unit,"OTU),獲得的OTU代表序列在MaarjAM數(shù)據(jù)庫(kù)中進(jìn)行比對(duì),以RDP"Classifier法對(duì)OTUs進(jìn)行注釋,分類置信度閾值設(shè)為0.7。剔除在門(mén)水平上未分類的序列,按最小樣本序列數(shù)抽平,使用抽平后的OTU表進(jìn)行后續(xù)數(shù)據(jù)分析。使用Mothur軟件進(jìn)行α多樣性分析,以Shannon指數(shù)和Simpson指數(shù)表征物種多樣性,以Ace指數(shù)和Chao1指數(shù)表征物種豐富度;使用Bray-Curtis距離計(jì)算β多樣性距離矩陣,以偏最小二乘法判別分析(partial"least"squares"discriminant"analysis,"PLS-DA)反映不同施肥處理對(duì)甘蔗根圍AM真菌群落的影響,以Adonis檢驗(yàn)不同施肥處理之間AM真菌群落的差異顯著性;采用單因素方差分析與Duncan’s法比較各處理間農(nóng)藝性狀的差異顯著性;采用Mantel方法分析土壤理化性質(zhì)與不同AM真菌屬之間的相關(guān)性,采用Pearson相關(guān)性分析土壤因子間的關(guān)系。
2.1""不同施肥水平對(duì)宿根蔗根圍土壤理化性質(zhì)的影響
不同施肥水平顯著影響GT29和GT58的根圍土壤pH,各處理間均存在顯著差異(Plt;0.05),隨著施肥量的增多,GT29根圍土壤pH由7.57(A29)逐漸降至4.47(D58),GT58根圍土壤pH由6.97(A58)逐漸降至4.18(D58)。不同施肥水平對(duì)GT29和GT58根圍土壤的Ca含量均有顯著影響,在GT29中,D29與B29間無(wú)顯著差異,而D29分別比A29和C29顯著降低62.10%和44.49%;在GT58中,B58、C58、D58分別比A58顯著降低36.74%、39.79%和50.68%。不同施肥水平顯著影響GT29和GT58根圍土壤的Mg含量,在GT29中,A29與C29間無(wú)顯著差異,B29和D29分別比A29顯著降低29.70%和51.75%;在GT58中,B58、C58、D58分別比A58顯著降低31.54%、32.06%和43.42%。不同施肥水平顯著影響GT29根圍土壤的堿解氮、速效鉀和Mn含量,D29的堿解氮含量比A29顯著提高97.29%,C29的速效鉀含量比B29顯著提高56.50%,C29、D29的速效鉀含量分別比B29顯著降低52.99%和60.76%;而不同施肥水平對(duì)GT58堿解氮、速效鉀含量無(wú)顯著影響。不同施肥水平對(duì)GT58根圍土壤的有效磷、Cu和Fe的影響顯著,D58的有效磷含量分別比A58、B58、C58顯著提高387.39%、318.64%和151.82%,C58的Cu含量比A58顯著降低4.32%,D58的Fe含量分別比A58、B58、C58顯著提高56.11%、36.78%和50.50%;而不同施肥水平對(duì)GT29有效磷、Cu、Fe含量無(wú)顯著影響。不同施肥水平對(duì)GT29和GT58根圍土壤的有機(jī)質(zhì)、Zn含量無(wú)顯著影響(表2)??傮w而言,過(guò)量施肥使甘蔗土壤pH降低,Ca和Mg含量顯著降低,有效磷含量嚴(yán)重過(guò)剩。
2.2""不同施肥水平對(duì)宿根蔗農(nóng)藝性狀和產(chǎn)量的影響
施肥水平的提高能夠在一定程度上增加甘蔗產(chǎn)量和有效莖數(shù),但當(dāng)過(guò)量施肥(D處理)時(shí),與C處理相比甘蔗產(chǎn)量增加不顯著,即D29(99.86"t/hm2)與C29(94.93"t/hm2)、D58(120.12"t/hm2)與C58(110.44"t/hm2)間的甘蔗產(chǎn)量均無(wú)顯著差異;不同施肥水平顯著影響GT58
的莖徑,C58的莖徑比B58降低6.01%,而C58與A58、D58間無(wú)顯著差異;施肥處理(B、C、D)的甘蔗分蘗數(shù)顯著高于未施肥(A)甘蔗的分蘗數(shù);不同施肥水平對(duì)GT29和GT58的出芽率、株高、錘度無(wú)顯著影響(表3)。因此,適當(dāng)施肥可以提高甘蔗產(chǎn)量,但是過(guò)量施肥(D)使生產(chǎn)的投入產(chǎn)出比大幅降低,不適宜甘蔗的可持續(xù)生產(chǎn)。
2.3""不同施肥水平對(duì)宿根蔗根系A(chǔ)M真菌侵染情況的影響
不同施肥水平顯著影響GT29的根系菌根侵染率,B29、C29比A29和D29顯著提高5.00%;不同施肥水平對(duì)GT29和GT58的根系菌根侵染強(qiáng)度、相對(duì)侵染強(qiáng)度、相對(duì)叢枝豐度和叢枝豐度均無(wú)顯著影響(表4)。說(shuō)明施肥量對(duì)不同甘蔗品種根系A(chǔ)M真菌的影響有一定差異。
2.4""不同施肥水平對(duì)宿根蔗根際土壤AM真菌群落結(jié)構(gòu)的影響
對(duì)第2年宿根蔗土壤根圍樣品進(jìn)行測(cè)序并初步篩選,共獲得1"512"798條序列,各樣本序列數(shù)在36"391~74"687之間,平均長(zhǎng)度為217"bp。以序列數(shù)34"561進(jìn)行抽平,各樣本稀釋曲線均達(dá)到平臺(tái)期,表明各處理的土壤樣本測(cè)序數(shù)據(jù)合理。以抽平后的序列劃分OTUs,第2年宿根蔗土壤根圍AM真菌隸屬11科13屬34種。其中,在屬水平主要為球囊霉屬(Glomus)、近明球囊霉屬(Claroideoglomus)、無(wú)梗囊霉屬(Acaulospora)、盾巨孢囊霉屬(Scutellospora)(圖1)。在2個(gè)甘蔗品種的不同施肥水平中,優(yōu)勢(shì)屬均為球囊霉屬,并且隨著施肥水平的提高,球囊霉屬的豐度不斷增加,在D29和D58水平下的豐度最高,分別達(dá)95.11%和97.39%。
2.5""不同施肥水平對(duì)宿根蔗根圍土壤OTUs分布的影響
不同施肥水平的GT29根圍土壤中,共獲得185個(gè)OTUs,所有處理共有OTUs數(shù)量為49個(gè),占總數(shù)的26.49%,A29、B29、C29和D29各處理獨(dú)有的OTU數(shù)量分別60、6、5、4個(gè)(圖2A)。不同施肥水平的GT58根圍土壤中,共獲得175個(gè)OTUs,所有處理共有的OTUs數(shù)量為51個(gè),占總數(shù)的29.14%,A58、B58、C58和D58各處理獨(dú)有的OTUs數(shù)量分別32、8、12、12個(gè)(圖2B)。
2.6""不同施肥水平對(duì)宿根蔗根圍土壤AM真菌多樣性的影響
不同施肥水平顯著影響甘蔗根圍土壤AM真菌群落的α多樣性。在GT29中,C29、D29水平下Ace指數(shù)、Chao1指數(shù)均極顯著低于A29,C29水平下Shannon指數(shù)顯著低于A29,D29水平下Shannon指數(shù)極顯著低于A29,而不同施肥水平下Simpson指數(shù)差異不顯著(圖3);在GT58中,D58處理的Ace指數(shù)、Chao1指數(shù)均顯著低于A58,C58處理的Ace指數(shù)也顯著低于A58,而不同施肥水平的Shannon指數(shù)和Simpson指數(shù)均差異不顯著(圖4)??傊?,甘蔗未施肥處理的AM真菌多樣性和豐富度最高,且隨著施肥水平的提高,多樣性與豐富度呈逐漸下降的趨勢(shì),過(guò)量施肥使2個(gè)品種甘蔗根圍土壤的AM真菌多樣性和豐富度均顯著降低。
2.7""不同施肥水平對(duì)宿根蔗根圍土壤AM真菌群落組成的影響
對(duì)GT29根圍土壤AM真菌群落進(jìn)行PLS-DA分析表明,第1主成分(COMP1)解釋了變量方差的23.92%,第2主成分(COMP2)解釋了變量方差的11.36%;A29組內(nèi)AM真菌物種組成差異較大,B29、C29與D29組內(nèi)AM真菌物種組成較相似;A29的4個(gè)重復(fù)在X軸與Y軸上均有
較大偏移,表明該組內(nèi)各樣本間物種組成存在差異,而C29與D29組間差異較?。▓D5A)。
對(duì)GT58根圍土壤AM真菌群落進(jìn)行PLS-DA分析表明,第1主成分(COMP1)解釋了變量方差的16.65%,第2主成分(COMP2)解釋了變量方差的11.61%;A58與B58組內(nèi)AM真菌物種組成差異較大,C58與D58組內(nèi)AM真菌物種組成相似;A58有1個(gè)重復(fù)在X軸和Y軸上與其余3個(gè)重復(fù)出現(xiàn)偏移,表明出現(xiàn)偏移的重復(fù)與組內(nèi)的其他3個(gè)重復(fù)之間的物種組成存在差異;B58的4個(gè)重復(fù)均出現(xiàn)偏移,表明該組內(nèi)物種組成存在差異;C58與D58組間差異也較?。▓D5B)。
2.8""不同施肥水平宿根蔗根圍土壤差異物種分析
對(duì)不同施肥水平下GT29與GT58第2年宿根蔗的根圍AM真菌群落從門(mén)水平到OTU水平上進(jìn)行線性判別分析,尋找不同處理中與造成AM真菌群落差異顯著相關(guān)的物種。結(jié)果表明,不同施肥水平下GT29的根圍AM真菌群落在OTU水平上的2個(gè)差異物種為OTU139、OTU177(圖6),均屬于球囊霉屬。在OTU139中,A29(0.20%)與B29(1.76%)間呈顯著差異(Plt;0.05);在OTU177中,A29(5.71%)與C29(49.14%)、D29(49.14%)間呈極顯著差異(Plt;0.01)。不同施肥水平下GT58的根圍AM真菌群落在OTU水平上的3個(gè)差異物種為OTU94、OTU175和OTU193,均屬于近明球囊霉屬。OTU94僅存在于A58(2.08%)中;在OTU175中,A58(25.97%)與D58(1.19%)間呈顯著差異(Plt;0.05);在OTU193中,A58(6.13%)與D58(0.51%)間呈顯著差異(圖7)。
2.9""土壤環(huán)境因子與"AM真菌群落的相關(guān)性分析
將根圍土壤AM真菌群落中的4個(gè)屬(球囊霉屬、近明球囊霉屬、盾巨孢囊霉屬、梗囊霉屬)與所有土壤環(huán)境因子進(jìn)行Mantel檢驗(yàn)(圖8),結(jié)果表明,土壤中的Ca、Mg含量與4個(gè)AM真菌屬之間均呈極顯著正相關(guān),土壤pH與3個(gè)AM真菌屬間呈顯著正相關(guān)。其中,球囊霉屬與土壤pH、Ca、Mg、Fe含量呈極顯著正相關(guān)(Plt;0.01);近明球囊霉屬與土壤有機(jī)質(zhì)和Fe含量呈顯著正相關(guān)(Plt;0.0.5),與土壤Ca、Mg、Mn含量呈極顯著正相關(guān)(Plt;0.01);盾巨孢囊霉屬和無(wú)梗囊霉屬與土壤pH、AP、Fe含量呈顯著正相關(guān)(Plt;0.0.5),與土壤Ca、Mg、Mn含量呈極顯著正相關(guān)(Plt;0.01)。
由于甘蔗具有生長(zhǎng)周期長(zhǎng)、營(yíng)養(yǎng)需求量大的特點(diǎn),合理施肥對(duì)甘蔗的生長(zhǎng)至關(guān)重要。肥料中適當(dāng)?shù)牡?、磷、鉀比例可以促進(jìn)甘蔗的光合作用,從而促進(jìn)糖分的積累[25]。在前期的新植甘蔗研究中,GT29在不同施肥水平下的產(chǎn)量無(wú)明顯差異,而GT58在C58和D58處理的產(chǎn)量顯著高于A58,C58與D58處理間的產(chǎn)量無(wú)顯著差異[20]。已有研究表明,肥料達(dá)到最佳配比前,每增加1"kg肥料,甘蔗產(chǎn)量可增加62.7"kg,但過(guò)量施肥后增產(chǎn)效果明顯降低[26],與本研究結(jié)果一致。本研究發(fā)現(xiàn)隨著施肥水平的提高,GT29和GT58的產(chǎn)量均表現(xiàn)出不同程度的增長(zhǎng),B29和C29處理的產(chǎn)量分別為84.18"t/hm2和94.93"t/hm2,C58和D58處理的產(chǎn)量分別為110.44"t/hm2和120.12"t/hm2,但當(dāng)施肥量超過(guò)一定限度后,即使顯著增加施肥量,甘蔗產(chǎn)量并無(wú)顯著性差異,這說(shuō)明過(guò)量施肥后甘蔗對(duì)肥料的利用率大幅下降,同時(shí)過(guò)量的營(yíng)養(yǎng)元素?zé)o法被充分利用,不僅浪費(fèi)了資源,還造成了土壤酸化、環(huán)境污染。孔垂豹等[27]研究表明,當(dāng)施肥量減半時(shí),AM真菌對(duì)甘蔗的促生作用顯著提高。綜上,由于甘蔗對(duì)營(yíng)養(yǎng)元素的利用率不同,長(zhǎng)期過(guò)量施用復(fù)合肥,造成田間土壤營(yíng)養(yǎng)元素比例失衡[28],從而使甘蔗產(chǎn)量不及預(yù)期。
不同施肥水平顯著影響土壤理化性質(zhì),從而間接影響土壤AM真菌群落[29]。本研究發(fā)現(xiàn),不同施肥水平影響GT29的根系菌根侵染率,B29和C29處理的菌根侵染率顯著高于A29和D29處理,而對(duì)GT58的菌根侵染率卻無(wú)顯著影響,說(shuō)明不同施肥水平對(duì)甘蔗根系菌根侵染率的影響程度因品種而異。本研究中隨著施肥量的增加,2個(gè)甘蔗品種根圍土壤AM真菌多樣性和豐富度均呈逐漸下降的趨勢(shì),最高施肥水平(D29,"D58)的AM真菌多樣性和豐富度顯著低于未施肥水平(A29,"A58),表明不同施肥量可以改變土壤AM真菌的多樣性,長(zhǎng)期施用復(fù)合肥能夠顯著降低AM真菌的多樣性[30-31]。此外,與GT29相比,GT58在B58和A58處理的AM真菌多樣性和豐富度并無(wú)顯著性差異,說(shuō)明這種變化趨勢(shì)也會(huì)因甘蔗品種而不同。這種差異可能是由于宿主植物不同,引起AM真菌對(duì)施肥水平表現(xiàn)出不同的敏感程度[32]。
土壤AM真菌群落組成會(huì)因施肥水平的不同而發(fā)生改變。本研究通過(guò)物種組成分析發(fā)現(xiàn),不同施肥水平下2個(gè)甘蔗品種根圍土壤AM真菌群落的優(yōu)勢(shì)菌屬均為球囊霉屬。試驗(yàn)前期研究的新植甘蔗根圍土壤AM真菌群落的優(yōu)勢(shì)屬也為球囊霉屬,相對(duì)豐度均達(dá)93%以上[21]。李錦婷[33]的研究也同樣發(fā)現(xiàn),在喀斯特53個(gè)土壤樣品中,AM真菌的優(yōu)勢(shì)屬均為球囊霉屬。球囊霉屬能夠在農(nóng)田土壤的AM真菌群落中占據(jù)優(yōu)勢(shì),是因?yàn)槠淇梢詮木z體碎片或菌根中定殖,能夠適應(yīng)多變的外界環(huán)境,且產(chǎn)孢率高于其他屬,在競(jìng)爭(zhēng)中具有優(yōu)勢(shì)[34-35]。本研究還發(fā)現(xiàn),隨著施肥水平的提高,球囊霉屬在AM真菌群落中的占比也逐漸增加,在最高施肥水平D29和D58中分別達(dá)到95.11%和97.39%。球囊霉屬占比的這種變化趨勢(shì)與馮重陽(yáng)[11]的研究結(jié)果相同,是因?yàn)榕c其他AM真菌相比,球囊霉屬對(duì)土壤養(yǎng)分的提高更加敏感。土壤中的球囊霉屬真菌能夠促進(jìn)植物對(duì)氮、磷的吸收,其分泌的球囊霉素還可以增加土壤有機(jī)質(zhì)含量,并改善土壤孔隙水氣情況[36],這可能受球囊霉屬真菌占比提高的影響,增強(qiáng)了甘蔗對(duì)土壤中氮、磷養(yǎng)分的吸收,促進(jìn)了甘蔗的增產(chǎn)。本研究對(duì)不同施肥水平下甘蔗根圍土壤AM真菌的差異物種進(jìn)行分析,發(fā)現(xiàn)GT29中的差異物種均為球囊霉屬,而GT58中的差異物種均為近明球囊霉屬,說(shuō)明不同品種甘蔗也會(huì)對(duì)AM真菌群落組成產(chǎn)生不同的影響。
通過(guò)對(duì)甘蔗根圍土壤和AM真菌群落的相關(guān)性分析發(fā)現(xiàn),土壤中的Ca、Mg含量與AM真菌的4個(gè)優(yōu)勢(shì)屬之間均呈極顯著正相關(guān),說(shuō)明土壤中的Ca、Mg含量很大程度上決定著優(yōu)勢(shì)屬的占比,從而對(duì)AM真菌群落組成產(chǎn)生影響。劉憶等[37]也研究發(fā)現(xiàn),接種AM真菌能夠顯著增加植物對(duì)Ca、Mg的吸收。此外,本研究中的土壤pH與AM真菌的優(yōu)勢(shì)屬間也呈顯著正相關(guān),與李佳雨等[38]的研究結(jié)果一致。廣西大部分蔗區(qū)經(jīng)過(guò)30多年的連作,由于南方紅壤普遍為酸性,加之甘蔗種植過(guò)程中存在過(guò)量施用化肥,大部分蔗區(qū)土壤pH在4.0~5.5之間[39]。本研究施肥處理前的土壤pH為7.0~7.5(預(yù)試驗(yàn)測(cè)定),而過(guò)量施化肥處理(D處理),在經(jīng)過(guò)3"a的種植后,D29和D58的pH分別降至4.47和4.18,說(shuō)明連續(xù)過(guò)量施化肥使土壤酸化。相關(guān)研究還發(fā)現(xiàn),AM真菌可以通過(guò)多種途徑調(diào)節(jié)土壤pH,使酸性土壤pH提高[40]。說(shuō)明土壤pH的降低,是由增加化肥施用量和減少土壤AM真菌多樣性與豐富度共同作用引起的。
試驗(yàn)前期對(duì)新植甘蔗的研究結(jié)果表明,無(wú)梗囊霉屬和盾巨孢囊霉屬能夠?qū)Ω收岬霓r(nóng)藝性狀產(chǎn)生積極影響,GT29在不同施肥水平下的產(chǎn)量和根圍土壤AM真菌多樣性與豐富度并未表現(xiàn)出顯著差異[21],而GT29宿根二年蔗的上述指標(biāo)均表現(xiàn)出顯著差異。說(shuō)明GT29的根圍土壤AM真菌在試驗(yàn)初期還能夠在一定水平下保持穩(wěn)定,但隨著施肥年限的增加,土壤環(huán)境發(fā)生較大變化后,這種維持穩(wěn)定的能力受到破壞,從而影響群落結(jié)構(gòu),進(jìn)而對(duì)甘蔗產(chǎn)量產(chǎn)生影響。具體的作用機(jī)制還需要進(jìn)一步研究。
綜上所述,不同施肥量通過(guò)影響土壤的理化性質(zhì),進(jìn)而對(duì)土壤微生物群落產(chǎn)生影響。過(guò)量施用化肥使土壤酸化,pH大幅降低,土壤磷元素累積過(guò)剩,影響根圍土壤AM真菌豐度占比,降低AM真菌多樣性和豐富度,不利于甘蔗生長(zhǎng)。因此,甘蔗生產(chǎn)上適當(dāng)施用化肥不僅可以提高產(chǎn)量,還可以在保證甘蔗產(chǎn)量的前提下,維持根圍土壤環(huán)境和AM真菌群落的相對(duì)穩(wěn)定,有利于甘蔗產(chǎn)業(yè)的可持續(xù)發(fā)展。
致謝""感謝廣西農(nóng)業(yè)科學(xué)院甘蔗研究所楊榮仲研究員和周會(huì)研究員對(duì)文稿的修改。
參考文獻(xiàn)
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