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秸稈覆蓋還田對(duì)桑園土壤生物學(xué)性狀及其細(xì)菌群落結(jié)構(gòu)的影響

2020-01-21 14:14林剛云肖健吳銀秀黃小丹楊尚東屈達(dá)才
關(guān)鍵詞:土壤酶活性桑園

林剛云 肖健 吳銀秀 黃小丹 楊尚東 屈達(dá)才

摘要:【目的】分析秸稈覆蓋還田對(duì)桑園土壤肥力及其細(xì)菌多樣性的影響,闡明秸稈覆蓋還田技術(shù)對(duì)南方桑園土壤肥力及健康的影響機(jī)制與應(yīng)用前景,為構(gòu)建穩(wěn)定高產(chǎn)、可持續(xù)發(fā)展的桑樹(shù)栽培管理體系提供參考依據(jù)?!痉椒ā吭O(shè)秸稈覆蓋還田桑園和對(duì)照桑園(非秸稈覆蓋還田桑園)2個(gè)處理,其中,秸稈覆蓋還田處理是將水稻秸稈切碎、自然堆漚50~60 d后覆蓋于桑樹(shù)根系兩旁,覆蓋45 d后分別采集秸稈覆蓋還田桑園和對(duì)照桑園的土壤樣品,利用傳統(tǒng)的測(cè)定方法和Illumina高通量測(cè)序技術(shù)分析秸稈覆蓋還田對(duì)桑園土壤生物學(xué)性狀及土壤細(xì)菌群落結(jié)構(gòu)特征的影響?!窘Y(jié)果】秸稈覆蓋還田桑園土壤β-葡糖苷酶、氨肽酶和磷酸酶活性及土壤微生物生物量碳(MBC)、微生物生物量氮(MBN)和微生物生物量磷(MBP)含量均顯著高于對(duì)照桑園土壤(P<0.05,下同);秸稈覆蓋還田桑園土壤的菌群Chao1和Shannon指數(shù)也顯著高于對(duì)照桑園土壤。在秸稈覆蓋還田桑園和對(duì)照桑園土壤中相對(duì)豐度大于1.00%的優(yōu)勢(shì)細(xì)菌門(mén)分類數(shù)量均為11個(gè),但二者的優(yōu)勢(shì)細(xì)菌門(mén)分類組成比例存在一定差異;相對(duì)于對(duì)照桑園土壤,秸稈覆蓋還田桑園土壤中Bacteroidota的相對(duì)豐度急劇增加,而綠彎菌門(mén)(Chloroflexi)和Verrucomicrobiota的相對(duì)豐度急劇下降。在秸稈覆蓋還田桑園和對(duì)照桑園土壤中相對(duì)豐度大于1.00%的優(yōu)勢(shì)細(xì)菌屬分類數(shù)量分別為23和24個(gè);與對(duì)照桑園土壤相比,秸稈覆蓋還田桑園土壤中雖然慢生根瘤菌屬(Bradyrhizobium)、假雙頭斧形菌屬(Pseudolabrys)、Dongia、Candidatus_Udaeobacter和norank_f_JG30-KF-AS9等優(yōu)勢(shì)細(xì)菌屬部分缺失,但富集了類諾卡氏菌屬(Nocardioides)、norank_f_Methyloligellaceae、黃桿菌屬(Flavobacterium)和微枝形桿菌屬(Microvirga)等特有優(yōu)勢(shì)菌屬。秸稈覆蓋還田桑園土壤的特有細(xì)菌屬為199個(gè)、特有細(xì)菌種為390個(gè),分別是對(duì)照桑園土壤特有細(xì)菌屬和細(xì)菌種的3.75和2.52倍?!窘Y(jié)論】秸稈覆蓋還田不僅顯著提高桑園土壤肥力,還改變桑園土壤優(yōu)勢(shì)細(xì)菌不同(門(mén)、屬)分類水平的組成比例,形成更豐富多樣的土壤細(xì)菌群落結(jié)構(gòu),而有助于維護(hù)桑園土壤健康。

關(guān)鍵詞: 秸稈覆蓋還田;桑園;土壤酶活性;土壤微生物生物量;細(xì)菌群落結(jié)構(gòu)

中圖分類號(hào): S154.36? ? ? ? ? ? ? ? ? ? ? ? ? 文獻(xiàn)標(biāo)志碼: A 文章編號(hào):2095-1191(2020)10-2339-09

Effects of straw mulching on soil biological properties and bacterial community structure in mulberry plantation

LIN Gang-yun, XIAO Jian, WU Yin-xiu, HUANG Xiao-dan,

YANG Shang-dong, QU Da-cai*

(College of Agriculture, Guangxi University, Nanning? 530004, China)

Abstract:【Objective】Analyzing the effects of straw mulching on soil fertility and bacterial diversity in mulberry fields, and clarifying the mechanism and application prospects for straw mulching techniques on soil fertility and health in sou-thern mulberry fields, in order to provide a reference basis for building a stable, high-yield and sustainable mulberry cultivation system. 【Method】Two treatments were set up, including straw mulching and returning to the mulberry field and the control mulberry field(non straw mulching and returning field). The straw mulching treatment was to cut the rice straw to pieces, pile it up naturally for 50-60 d and then cover both sides with the mulberry root system, and collect the straw mulch after 45 d of covering. The soil samples of returning mulberry fields and control mulberry fields were used to analyze the effects of straw mulching on soil biological properties and soil bacterial community structure characteristics using traditional measurement methods and Illumina high-throughput sequence technology. 【Result】The activities of β-glucosidase, amino peptidase and phosphate in mulc-hing soils of mulberry plantation were all significantly higher than those of control(P<0.05, the same below), as well as the microbial biomass carbon(MBC), microbial biomass nitrogen(MBN), microbial biomass phosphorus(MBP). The Chao1 and Shannon indexes of the microbial community of the mulberry soil covered with straws were also significantly higher than those of the control mulberry soil. In both straw mulching mulberry fields and control mulberry fields, there were 11 types of dominant bacteria whose relative abundance was greater than 1.00%, but there were some differences in the composition ratio of dominant bacteria. In addition, compared to the control mulberry soil, the ratios of Bacteroidota increased sharply and Chloroflexi, Verrucomicobiota decreased sharply in mulching soils of mulberry plantation at phylum level. There were 23 and 24 dominant bacterial genera in the soil with relative abundance greater than 1.00% in straw mulching mulberry fields and control mulberry fields, respectively. Moreover, some dominant bacteria, such as Bradyrhizobium,Pseudolabrys,Dongia,Candidatus_Udaeobacter and norank_ f_JG30-KF-AS9 even though lost in mulching soil of mulberry plantation, but Nocardioides,norank_f_Methyloligellaceae,F(xiàn)lavobacterium and Microvirga accumulated in mulching soil as its unique dominant bacteria. There were 199 unique bacteria genera and 390 unique bacteria species in the soil of straw mulching mulberry field, which were 3.75 times and 2.52 times of those in the soil of control mulberry field. 【Conclusion】Straw mulching not only can significantly improve soil fertility, but also change the proportions of soil dominant bacterial community structure(phylum and genus), and form a more diverse soil bacterial community structure. It is helpful for maintaining the health of mulberry field soil.

Key words: straw mulching and returning to field; mulberry field; soil enzyme activity; soil microbial biomass; bacterial community structure

Foundation item: Special Project for Guangxi Innovation Team Construction of National Modern Agricultural Industrial Technology System(nycytxgxcxtd-02-04); University Science and Technology Innovation and Service Capacity Enhancement Project of Guangxi(Guijiaokeyan〔2020〕8)

0 引言

【研究意義】桑樹(shù)(Morus alba L.)是我國(guó)重要的多年生木本經(jīng)濟(jì)植物。據(jù)統(tǒng)計(jì),我國(guó)現(xiàn)有的桑樹(shù)種質(zhì)資源居世界第一,約3000份,分屬15個(gè)種和4個(gè)變種(蘇超和焦鋒,2011;杜偉等,2017)。廣西是我國(guó)最大的桑蠶生產(chǎn)基地,無(wú)論是桑園面積還是蠶繭產(chǎn)量均連續(xù)多年位居全國(guó)首位,2018、2019年廣西桑園面積分別為22.85萬(wàn)和19.72萬(wàn)ha,蠶繭產(chǎn)量分別達(dá)40.44萬(wàn)和37.4萬(wàn)t(林強(qiáng),2020;張志林等,2020)。但目前廣西桑園管理偏施氮肥,普遍存在氮、磷、鉀肥施用比例失調(diào)等問(wèn)題,造成土壤肥力下降進(jìn)而導(dǎo)致桑葉產(chǎn)量降低,嚴(yán)重制約了廣西桑蠶產(chǎn)業(yè)的健康發(fā)展(張志林等,2020)。因此,研究并優(yōu)化桑園施肥技術(shù)對(duì)提高桑園土壤肥力和桑葉產(chǎn)量及保護(hù)生態(tài)環(huán)境具有重要意義?!厩叭搜芯窟M(jìn)展】秸稈還田作為改善土壤環(huán)境和提高土壤肥力的一種重要方式已被廣泛認(rèn)可(王靜靜等,2019;劉明等,2020)。秸稈中的碳氮比較高,與燃燒作物秸稈相比,將秸稈還田不僅可減少CO2排放,增加土壤固碳容量(逯非等,2010;Li et al.,2018;彭廷等,2019),還能提高有機(jī)質(zhì)含量與土壤肥力,以及促進(jìn)土壤團(tuán)粒結(jié)構(gòu)形成,進(jìn)而改善土壤結(jié)構(gòu)、透氣性、持水能力和陽(yáng)離子交換量(Lenka and Lal,2013;伍佳等,2019;Su et al.,2020)。此外,在秸稈分解過(guò)程中能釋放出有利于植物吸收利用的無(wú)機(jī)磷,而提高土壤養(yǎng)分的有效性(Shan et al.,2010)。趙亞麗等(2015)研究表明,與常規(guī)耕作+無(wú)秸稈還田相比,深耕+秸稈還田、深松+秸稈還田處理的土壤有機(jī)碳含量分別提高14.8%和12.4%,土壤微生物數(shù)量分別提高45.9%和33.9%,土壤酶活性分別提高34.1%和25.2%,作物產(chǎn)量分別提高18.0%和19.3%,即土壤深松(耕)結(jié)合秸稈還田有利于作物產(chǎn)量及土壤微生物數(shù)量和酶活性的提高。王靜靜等(2019)研究證實(shí),越冬期還田水稻秸稈的腐解呈由慢到快又變慢的變化趨勢(shì),且與氣溫及降水量的變化相關(guān);秸稈還田具有培肥地力的作用,可提高土壤有機(jī)質(zhì)和鉀素含量,同時(shí)有效調(diào)節(jié)土壤pH。石含之等(2020)通過(guò)研究秸稈還田對(duì)土壤有機(jī)碳結(jié)構(gòu)的影響,發(fā)現(xiàn)秸稈還田能促使紅壤和黑土中有機(jī)碳疏水性增強(qiáng),提高土壤團(tuán)聚體穩(wěn)定性,對(duì)改善土壤結(jié)構(gòu)具有促進(jìn)作用;此外,外源秸稈的加入會(huì)引起土壤中有機(jī)碳分解,具體表現(xiàn)為:黑土發(fā)生負(fù)激發(fā)效應(yīng),其有機(jī)碳結(jié)構(gòu)趨于復(fù)雜,穩(wěn)定性增強(qiáng);而紅壤和褐土發(fā)生正激發(fā)效應(yīng),其有機(jī)碳結(jié)構(gòu)簡(jiǎn)單、穩(wěn)定性減弱。可見(jiàn),腐熟分解后的作物秸稈還田不僅能有效提高土壤速效養(yǎng)分含量及作物產(chǎn)量(Han et al.,2013),還有助于提高土壤肥力和維護(hù)土壤健康(Gaind and Nain,2007)。自20世紀(jì)90年代以來(lái),秸稈還田在我國(guó)北方小麥、玉米種植體系中得到大面積推廣應(yīng)用,其累計(jì)推廣面積已達(dá)110億ha(Qi et al.,2015)。但也有研究表明,新鮮秸稈還田會(huì)對(duì)土壤健康產(chǎn)生負(fù)面影響,可能是作物品質(zhì)和產(chǎn)量提高的限制因素(Hamada et al.,2011;Yang et al.,2011;楊麗華等,2013)。因此,科學(xué)的秸稈還田方法應(yīng)避免采用新鮮秸稈還田方式?!颈狙芯壳腥朦c(diǎn)】目前,在小麥、玉米等大宗作物生產(chǎn)中秸稈還田技術(shù)已得到廣泛應(yīng)用,但在我國(guó)南方地區(qū)鮮見(jiàn)桑園應(yīng)用秸稈還田技術(shù),尤其缺乏秸稈還田對(duì)桑園土壤肥力影響及其改良措施的相關(guān)研究?!緮M解決的關(guān)鍵問(wèn)題】基于田間定位試驗(yàn),系統(tǒng)分析秸稈覆蓋還田對(duì)桑園土壤肥力及其細(xì)菌多樣性的影響,旨在闡明秸稈覆蓋還田技術(shù)對(duì)南方桑園土壤肥力及健康的影響機(jī)制與應(yīng)用前景,為構(gòu)建穩(wěn)定高產(chǎn)、可持續(xù)發(fā)展的桑樹(shù)栽培管理體系提供參考依據(jù)。

1 材料與方法

1. 1 試驗(yàn)地概況

試驗(yàn)在廣西大學(xué)農(nóng)學(xué)院教學(xué)實(shí)驗(yàn)桑園基地(東經(jīng)108°17′14″,北緯22°51′17″)進(jìn)行,試驗(yàn)區(qū)域?qū)儆趤啛釒Ъ撅L(fēng)氣候,陽(yáng)光充足,雨量充沛;年均氣溫21.7 ℃,年均降水量1600 mm。桑園土壤類型為赤紅壤,土壤pH 5.79,有機(jī)質(zhì)含量6.75 g/kg,全氮含量0.84 g/kg,全磷含量0.53 g/kg,全鉀含量14.54 g/kg,堿解氮含量57.45 mg/kg,速效磷含量3.39 mg/kg,速效鉀含量81.77 mg/kg。

1. 2 樣品采集

試驗(yàn)設(shè)秸稈覆蓋還田桑園(SR)和對(duì)照桑園(CK,非秸稈覆蓋還田桑園)2個(gè)處理,桑樹(shù)品種均為桂桑12號(hào),樹(shù)齡6年。其中,SR處理是將水稻秸稈切碎、自然堆漚50~60 d后覆蓋于桑樹(shù)根系兩旁;CK處理除了未進(jìn)行相應(yīng)的秸稈覆蓋外,其余田間管理措施與SR處理完全相同。覆蓋秸稈45 d后(2020年6月18日)分別采集SR處理和CK處理桑園0~30 cm土層的土壤樣品。每個(gè)處理桑園隨機(jī)選擇6個(gè)土壤取樣點(diǎn),用無(wú)菌密封袋收集后裝入放有冰袋的冰盒中帶回實(shí)驗(yàn)室,將每份土壤樣品分成2份。1份在室內(nèi)風(fēng)干后過(guò)40目篩,用于土壤理化性質(zhì)測(cè)定;1份過(guò)10目篩后用于土壤生物學(xué)性狀和細(xì)菌群落結(jié)構(gòu)分析。

1. 3 土壤生物學(xué)性狀分析

土壤β-葡糖苷酶、氨肽酶和磷酸酶活性參照楊尚東等(2013)的方法進(jìn)行測(cè)定;土壤微生物生物量碳(MBC)含量采用氯仿熏蒸提取—容量分析法測(cè)定(龐師嬋等,2020),土壤微生物生物量氮(MBN)含量采用茚三酮比色法測(cè)定(Jorrgensen and Brookes,1990),土壤微生物生物量磷(MBP)含量采用磷鉬藍(lán)比色法測(cè)定(吳金水等,2003)。

1. 4 土壤細(xì)菌多樣性分析

根際土壤樣品總DNA提取、PCR擴(kuò)增及序列測(cè)定均委托上海美吉生物醫(yī)藥科技有限公司完成。具體操作:根據(jù)FastDNA? Spin Kit for Soil試劑盒(MP Biomedicals)說(shuō)明進(jìn)行總DNA提取,使用NanoDrop 2000分光光度計(jì)(Thermo Fisher Scientific)檢測(cè)其濃度和純度,同時(shí)利用1%瓊脂糖凝膠電泳檢測(cè)DNA提取質(zhì)量;然后以338F(5'-ACTCCTACGGGAGGCAG CAG-3')和806R(5'-GGACTACHVGGGTWTCTAA T-3')引物對(duì)16S rDNA序列V3~V4可變區(qū)進(jìn)行PCR擴(kuò)增,并進(jìn)行鑒定、純化及定量分析。Illumina MiSeq測(cè)序:以2%瓊脂糖凝膠電泳回收PCR擴(kuò)增產(chǎn)物,經(jīng)Extraction Kit純化、Tris-HCl洗脫及電泳檢測(cè)后,采用Quantus? Fluorometer(Promega)進(jìn)行檢測(cè)定量,以NEXTflex? Rapid DNA-Seq Kit構(gòu)建文庫(kù),然后利用Illumina測(cè)序公司的MiSeq平臺(tái)300PE完成測(cè)序。

1. 5 統(tǒng)計(jì)分析

試驗(yàn)數(shù)據(jù)采用Excel 2019和SPSS 21.0進(jìn)行整理統(tǒng)計(jì),并以上海美吉生物醫(yī)藥科技有限公司的I-Sanger云數(shù)據(jù)分析平臺(tái)進(jìn)行在線分析。

2 結(jié)果與分析

2. 1 秸稈覆蓋還田桑園土壤生物學(xué)性狀特征

由表1可知,SR處理桑園土壤β-葡糖苷酶、氨肽酶和磷酸酶活性均顯著高于CK處理(P<0.05,下同),表明秸稈覆蓋還田能顯著提高桑園土壤中涉及碳、氮、磷循環(huán)的相關(guān)酶活性。在土壤微生物生物量含量方面,SR處理桑園的土壤MBC、MBN和MBP含量也顯著高于CK處理,表明秸稈覆蓋還田能顯著提高桑園土壤中碳、氮和磷的庫(kù)容,即有助于改良桑園土壤肥力。

2. 2 秸稈覆蓋還田桑園土壤細(xì)菌OTU聚類分析結(jié)果

基于Illumina高通量測(cè)序分析,從12個(gè)桑園土壤樣品中共獲得4271個(gè)OTUs(表2),依據(jù)細(xì)菌群落組成不同分類水平可分為39門(mén)119綱283目449科820屬1620種。其中,從SR處理桑園土壤樣品中獲得3565個(gè)OTUs,其細(xì)菌群落組成依據(jù)不同分類水平可分為37門(mén)114綱271目429科767屬1465種;從CK處理桑園土壤樣品中獲得3017個(gè)OTUs,其細(xì)菌群落組成依據(jù)不同分類水平可分為34門(mén)101綱222目350科621屬1230種。說(shuō)明秸稈覆蓋還田能改變桑園土壤中不同分類水平的細(xì)菌群落組成,且與對(duì)照處理相比,秸稈覆蓋還田有效提高了桑園土壤中細(xì)菌群落不同分類水平的數(shù)量。

2. 3 秸稈覆蓋還田桑園土壤細(xì)菌Alpha多樣性分析結(jié)果

覆蓋率越高說(shuō)明樣本中目的序列被測(cè)出的概率越高,而未被測(cè)出的概率越低(孫珂岱,2015)。由表3可知,桑園土壤樣品分析覆蓋率均達(dá)98%,說(shuō)明分析數(shù)據(jù)真實(shí)可信。Alpha多樣性指數(shù)包括ACE、Chao1、Shannon和Simpson等(洪嘉煒等,2019),其中,Chao1指數(shù)表征細(xì)菌群落豐富度,Chao1指數(shù)越大表明土壤中細(xì)菌數(shù)目越多,群落豐富度越高(Chao et al.,2000);Shannon指數(shù)表征細(xì)菌群落多樣性,Shannon指數(shù)越大表明物種多樣性越豐富(張傳進(jìn)等,2020)。由表3可知,SR處理桑園土壤樣品的Chao1和Shannon指數(shù)均顯著高于CK處理,說(shuō)明秸稈覆蓋還田能顯著提高桑園土壤細(xì)菌的多樣性和豐富度。

2. 4 桑園土壤細(xì)菌群落組成分析結(jié)果

2. 4. 1 桑園土壤細(xì)菌門(mén)分類水平分析結(jié)果 在SR處理和CK處理桑園土壤中相對(duì)豐度大于1.00%的優(yōu)勢(shì)細(xì)菌門(mén)分類數(shù)量均為11個(gè)(圖1),但二者的優(yōu)勢(shì)細(xì)菌門(mén)分類組成比例存在一定差異。其中,SR處理桑園土壤優(yōu)勢(shì)細(xì)菌門(mén)的相對(duì)豐度排序?yàn)樽冃螚U菌門(mén)(Proteobacteria,26.26%)>放線菌門(mén)(Actinobacteriota,24.76%)>Acidobacteriota(14.26%)>綠彎菌門(mén)(Chloroflexi,11.54%)>Bacteroidota(5.57%)>Myxococcota(3.85%)>厚壁菌門(mén)(Firmicutes,3.55%)>Gem-matimonadota(1.82%)>Methylomirabilota(1.75%)>Planctomycetota(1.42%)>Verrucomicrobiota(1.12%),其他門(mén)類的合計(jì)相對(duì)豐度為4.10%;CK處理桑園土壤優(yōu)勢(shì)細(xì)菌門(mén)的相對(duì)豐度排序?yàn)樽冃螚U菌門(mén)(Proteobacteria,25.04%)>放線菌門(mén)(Actinobacteriota,23.71%)>綠彎菌門(mén)(Chloroflexi,15.95%)>Acidobacteriota(14.33%)>厚壁菌門(mén)(Firmicutes,3.58%)>Gemmatimonadota(3.48%)>Myxococcota(2.97%)>Bacteroidota(2.40%)>Verrucomicrobiota(1.87%)>Methylomirabilota(1.59%)>Planctomycetota(1.10%),其他門(mén)類的合計(jì)相對(duì)豐度為3.97%。由此可見(jiàn),秸稈覆蓋還田雖然未改變桑園土壤優(yōu)勢(shì)細(xì)菌門(mén)分類水平組成,但部分優(yōu)勢(shì)細(xì)菌門(mén)的相對(duì)豐度排序已發(fā)生改變。如綠彎菌門(mén)類細(xì)菌在CK處理桑園土壤中的相對(duì)豐度排在變形桿菌門(mén)和放線菌門(mén)之后,位列第三;但在SR處理桑園土壤中的相對(duì)豐度降至11.54%,相對(duì)于CK處理桑園土壤(15.95%)下降了4.44%(絕對(duì)值),排在Acidobacteriota之后,位列第四。同時(shí),Bacteroidota的相對(duì)豐度在覆蓋秸稈后急劇增加,由CK處理桑園土壤的2.40%上升至5.57%。秸稈覆蓋還田不僅改變桑園土壤中部分優(yōu)勢(shì)細(xì)菌門(mén)分類水平組成比例,還會(huì)改變桑園土壤細(xì)菌群落結(jié)構(gòu),最終改善土壤的生物學(xué)功能。

2. 4. 2 桑園土壤細(xì)菌屬分類水平分析結(jié)果 在SR處理和CK處理桑園土壤中相對(duì)豐度大于1.00%的優(yōu)勢(shì)細(xì)菌屬分類數(shù)量分別為23和24個(gè)(圖2)。其中,SR處理桑園土壤優(yōu)勢(shì)細(xì)菌屬的相對(duì)豐度排序?yàn)閚orank_f_norank_o_Vicinamibacterales(5.08%)>no-rank_ f_Vicinamibacteraceae(4.55%)>Gaiella(2.87%)> norank_f_Xanthobacteraceae(2.38%)>norank_f_no-rank_o_norank_c_KD4-96(2.37%)>norank_f_norank_ o_norank_c_MB-A2-108(1.90%)>鏈霉菌屬(Streptomyces,1.85%)>類諾卡氏菌屬(Nocardioides,1.84%)>norank_f_JG30-KF-CM45(1.79%)>norank_f_norank_ o_Gaiellales(1.66%)>芽孢桿菌屬(Bacillus,1.65%)>norank_f_67-14 (1.49%)>norank_f_SC-I-84(1.44%)>norank_f_Gemmatimonadaceae(1.43%)=norank_f_ no-rank_o_Rokubacteriales(1.43%)>norank_f_Roseiflexaceae(1.32%)>黃桿菌屬(Flavobacterium,1.31%)>norank_f_norank_o_norank_c_TK10(1.29%)>鞘脂單胞菌屬(Sphingomonas,1.25%)>norank_f_Methyloligellaceae(1.16%)>norank_f_norank_o_IMCC26256 (1.10%)>分枝桿菌屬(Mycobacterium,1.09%)>微枝形桿菌屬(Microvirga,1.02%),其他屬類的合計(jì)相對(duì)豐度為54.02%(圖2-A)。CK處理桑園土壤優(yōu)勢(shì)細(xì)菌屬的相對(duì)豐度排序?yàn)閚orank_f_Roseiflexaceae(4.74%)>norank_f_norank_o_Vicinamibacterales(4.73%)>norank_f_Xanthobacteraceae(3.82%)>no-rank_f_norank_o_Gaiellales(3.58%)>Gaiella(3.51%)>芽孢桿菌屬(Bacillus,2.97%)>norank_f_Gemmatimonadaceae(2.72%)>norank_f_Vicinamibacteraceae(2.33%)>norank_f_SC-I-84(2.30%)>norank_f_no-rank_o_norank_c_TK10(1.82%)>norank_f_67-14(1.78%)>鏈霉菌屬(Streptomyces,1.64%)>鞘脂單胞菌屬(Sphingomonas,1.60%)>norank_f_norank_o_ IMCC 26256(1.56%)>慢生根瘤菌屬(Bradyrhizobium,1.44%)>norank_f_norank_o_Rokubacteriales(1.43%)>假雙頭斧形菌屬(Pseudolabrys,1.40%)>norank_f_ norank_o_norank_c_KD4-96(1.36%)>分枝桿菌屬(Mycobacterium,1.26%)>norank_f_norank_ o_norank_c_MB-A2-108(1.18%)>norank_f_JG30-KF-AS9(1.15%)>Dongia(1.08%)>Candidatus_Udaeobacter(1.05%)>norank_f_JG30-KF-CM45(1.04%),其他屬類的合計(jì)相對(duì)豐度為46.21%(圖2-B)。

與對(duì)照相比,秸稈覆蓋還田同步改變了桑園土壤優(yōu)勢(shì)細(xì)菌屬分類水平組成及其相對(duì)豐度。在SR處理桑園土壤中,norank_f_norank_o_Vicinamibacterales的相對(duì)豐度最高(5.08%),但其在CK處理桑園土壤中的相對(duì)豐度排名第二,僅為4.73%;norank_ f_Roseiflexaceae在CK處理桑園土壤中相對(duì)豐度(4.74%)排名第一,但在SR處理桑園土壤中的相對(duì)豐度急劇下降,僅為1.32%,排在第十六位。此外,秸稈覆蓋還田還改變了桑園土壤中優(yōu)勢(shì)細(xì)菌屬的群落結(jié)構(gòu),如慢生根瘤菌屬(Bradyrhizobium)、假雙頭斧形菌屬(Pseudolabrys)、Dongia、Candidatus_Udaeobacter和norank_f_JG30-KF-AS9等均為CK處理桑園土壤中的優(yōu)勢(shì)細(xì)菌屬,但經(jīng)秸稈覆蓋還田處理后這些優(yōu)勢(shì)細(xì)菌屬缺失,同時(shí)富集了類諾卡氏菌屬(Nocardioides)、norank_f_Methyloligellaceae、黃桿菌屬(Flavobacterium )和微枝形桿菌屬(Microvirga)等細(xì)菌屬,成為SR處理桑園土壤的特有優(yōu)勢(shì)細(xì)菌屬。

2. 5 桑園土壤細(xì)菌群落結(jié)構(gòu)Venn分析結(jié)果

基于屬和種分類水平的Venn分析結(jié)果顯示,在屬分類水平(圖3-A)上,SR處理和CK處理桑園土壤的共有細(xì)菌屬為568個(gè),SR處理桑園土壤的特有細(xì)菌屬達(dá)199個(gè),CK處理桑園土壤的特有細(xì)菌屬僅為53個(gè),前者是后者的3.75倍;在種分類水平(圖3-B)上,SR處理和CK處理桑園土壤的共有細(xì)菌種為1075個(gè),SR處理桑園土壤的特有細(xì)菌種達(dá)390個(gè),CK處理桑園土壤的特有細(xì)菌種為155個(gè),前者是后者的2.52倍。表明秸稈覆蓋還田明顯改變桑園土壤中細(xì)菌屬、種分類水平上的群落結(jié)構(gòu),提高桑園土壤細(xì)菌多樣性及豐富度,而有助于提高和維護(hù)桑園土壤健康。

3 討論

桑樹(shù)隸屬于??疲∕oraceae)桑屬(Morus L.),為多年生的深根性、闊葉型、落葉性小喬木或灌木。桑樹(shù)除了是養(yǎng)蠶產(chǎn)業(yè)不可或缺的原料外,對(duì)恢復(fù)脆弱生態(tài)區(qū)植被也具有積極作用,還可作為優(yōu)質(zhì)飼料作物,具有極高的經(jīng)濟(jì)價(jià)值和應(yīng)用前景。廣西是我國(guó)桑蠶的主產(chǎn)區(qū),桑園面積約占全國(guó)桑園總面積的1/4,連續(xù)13年位居全國(guó)之首(鐘春云,2019)。近年來(lái),由于廣西大部分桑園種植年限已超過(guò)10年,部分桑農(nóng)為了獲得較高的桑葉產(chǎn)量,而無(wú)限制地加大氮肥施用量,導(dǎo)致桑園土壤有機(jī)質(zhì)含量降低、土壤板結(jié)及肥力下降,甚至發(fā)生嚴(yán)重的土傳病害等一系列問(wèn)題(祁廣軍等,2015;王帥帥等,2019),嚴(yán)重威脅著廣西桑蠶產(chǎn)業(yè)的可持續(xù)發(fā)展。秸稈覆蓋還田是將秸稈直接或堆漚后施入土壤中,具有培肥、蓄水、調(diào)溫、增產(chǎn)及減少污染的作用,能有效改善農(nóng)田生態(tài)環(huán)境、培肥地力和提高作物品質(zhì)與產(chǎn)量。因此,在我國(guó)南方桑園開(kāi)展秸稈覆蓋還田技術(shù)研究,對(duì)構(gòu)建穩(wěn)定高產(chǎn)、可持續(xù)發(fā)展的桑樹(shù)栽培管理體系具有重要意義。

土壤酶是土壤的重要組成成分,幾乎參與土壤中所有有機(jī)物和營(yíng)養(yǎng)元素的循環(huán)過(guò)程,在穩(wěn)定土壤結(jié)構(gòu)、分解有機(jī)廢棄物、有機(jī)質(zhì)形成和養(yǎng)分循環(huán)等方面發(fā)揮重要作用,其活性高低能客觀地反映土壤肥力及其健康狀況(Dick et al.,1994;林天等,2005;楊寧等,2014)。土壤微生物生物量是衡量土壤質(zhì)量、維持土壤肥力和作物生產(chǎn)力的一個(gè)重要指標(biāo),土壤微生物生物量越大,即土壤通過(guò)有機(jī)養(yǎng)分礦化提供給植物所需養(yǎng)分的能力越強(qiáng)(任奎瑜等,2020)。本研究結(jié)果表明,秸稈覆蓋還田桑園土壤中的β-葡糖苷酶、氨肽酶和磷酸酶活性均顯著高于對(duì)照桑園土壤,MBC、MBN和MBP等土壤微生物生物量也顯著高于對(duì)照桑園土壤,說(shuō)明秸稈覆蓋還田能顯著提高桑園土壤酶活性及土壤中碳、氮和磷的庫(kù)容,從而有助于提高桑園土壤肥力。

微生物是生態(tài)系統(tǒng)中功能最活躍、開(kāi)發(fā)潛力最大的生物資源庫(kù)(Vanniern et al.,2018)。土壤中豐富的微生物多樣性在陸地生態(tài)系統(tǒng)中發(fā)揮著重要的功能作用(楊尚東等,2019),其微生物群落結(jié)構(gòu)越復(fù)雜、物種多樣性越豐富時(shí),植物對(duì)抗病原菌的綜合能力越強(qiáng)(楊尚東等,2020)。本研究結(jié)果表明,在秸稈覆蓋還田桑園土壤中,指示細(xì)菌豐富度的Chao1指數(shù)和細(xì)菌多樣性的Shannon指數(shù)均顯著高于對(duì)照桑園土壤,說(shuō)明秸稈覆蓋還田有助于提高桑園土壤微生物的多樣性和豐富度。本研究還發(fā)現(xiàn),秸稈覆蓋還田改變了桑園土壤的部分優(yōu)勢(shì)細(xì)菌門(mén),如Bacteroidota的相對(duì)豐度急劇增加,而綠彎菌門(mén)(Chloroflexi)和Verrucomicrobiota的相對(duì)豐度急劇下降;此外,與對(duì)照桑園土壤相比,在秸稈覆蓋還田桑園土壤中雖然慢生根瘤菌屬(Bradyrhizobium)、假雙頭斧形菌屬(Pseudolabrys)、Dongia、Candidatus_Udaeobacter和norank_f_JG30-KF-AS9等優(yōu)勢(shì)細(xì)菌屬部分缺失,但富集了類諾卡氏菌屬(Nocardioides)、norank_f_Methyloligellaceae、黃桿菌屬(Flavobacterium)和微枝形桿菌屬(Microvirga)等特有優(yōu)勢(shì)菌屬。其中,類諾卡氏菌屬(Nocardioides)細(xì)菌不僅能利用C2~C16一系列烷烴及苯酚等作為碳源,具有降解原油、治理污水和生物防護(hù)等功能(杜慧竟等,2012),還具有溶磷、產(chǎn)鐵載體及固氮等作用(劉冰冰,2014)。黃桿菌屬(Flavobacterium)細(xì)菌是具有脫氮除磷功能的好氧反硝化微生物(楊浩等,2017)。微枝形桿菌屬(Microvirga)是根瘤菌中的一類新種屬,具有與黏細(xì)菌相似的噬大腸桿菌活性和抗馬鈴薯晚疫病菌活性,主要分布在干旱或半干旱且偏堿性的土壤中,個(gè)別類型菌株可生長(zhǎng)在50 ℃左右的地下含水層內(nèi),屬于中性嗜熱微生物,與植物共生時(shí)能產(chǎn)生明顯的結(jié)瘤,即具有顯著的固氮活性(趙璞鈺等,2017),少數(shù)微枝形桿菌屬還具有運(yùn)輸鐵離子的載體功能。

基于屬和種分類水平的Venn分析結(jié)果顯示,秸稈覆蓋還田桑園土壤的特有細(xì)菌屬為199個(gè),特有細(xì)菌種為390個(gè),分別是對(duì)照桑園土壤特有細(xì)菌屬和細(xì)菌種的3.75和2.52倍。說(shuō)明秸稈覆蓋還田不僅能改變桑園土壤細(xì)菌不同分類水平的組成比例和群落結(jié)構(gòu),還有助于富集更多的特有細(xì)菌種屬,提高桑園土壤微生物的多樣性及豐富度,而有助于維護(hù)桑園土壤健康。

4 結(jié)論

秸稈覆蓋還田不僅顯著提高桑園土壤肥力,還改變桑園土壤優(yōu)勢(shì)細(xì)菌不同(門(mén)、屬)分類水平的組成比例,形成更豐富多樣的土壤細(xì)菌群落結(jié)構(gòu),而有助于維護(hù)桑園土壤健康。

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(責(zé)任編輯 蘭宗寶)

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