research article mitigating nitrous oxide emissions from...

Research ArticleMitigating Nitrous Oxide Emissions from Tea Field Soil UsingBioaugmentation with a Trichoderma viride Biofertilizer

Shengjun Xu1 Xiaoqing Fu2 Shuanglong Ma1 Zhihui Bai1

Runlin Xiao2 Yong Li2 and Guoqiang Zhuang1

1 Research Center for Eco-Environmental Sciences Chinese Academy of Sciences Beijing 100085 China2 Institute of Subtropical Agriculture Chinese Academy of Sciences Changsha 410125 China

Correspondence should be addressed to Zhihui Bai zhbairceesaccn and Guoqiang Zhuang zhxu63sinacom

Received 25 November 2013 Accepted 5 February 2014 Published 6 April 2014

Academic Editors H M Baker C Cameselle G El-Chaghaby and C Waterlot

Copyright copy 2014 Shengjun Xu et al This is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

Land-use conversion from woodlands to tea fields in subtropical areas of central China leads to increased nitrous oxide(N2O) emissions partly due to increased nitrogen fertilizer use A field investigation of N

2O using a static closed chamber-gas

chromatography revealed that the average N2O fluxes in tea fields with 225 kgNhaminus1 yrminus1 fertilizer application were 94plusmn 62

times higher than those of woodlands Accordingly it is urgent to develop practices for mitigating N2O emissions from tea fields

By liquid-state fermentation of sweet potato starch wastewater and solid-state fermentation of paddy straw with application ofTrichoderma viride we provided the tea plantation with biofertilizer containing 24 t C haminus1 and 587 kgNhaminus1 Compared to use ofsynthetic N fertilizer use of biofertilizer at 225 kgNhaminus1 yrminus1 significantly reduced N

2O emissions by 333ndash718 and increased

the tea yield by 162ndash622Therefore the process of bioconversionbioaugmentation tested in this study was found to be a cost-effective and feasible approach to reducing N

2O emissions and can be considered the best management practice for tea fields

1 Introduction

Three typical land-uses occur in most areas of southernsubtropical China woodlands in the mountains uplandson slopes and paddy fields in the lowlands [1] In China13Mha of tea plantations has been established during thelast 50 yrs due to rapid economic development [2] Mostof these recent plantations occupy large areas that wereoriginally woodlands and are continuing to expand [3] Land-use conversion is typically characterized by frequent tillageand use of large amounts of nitrogenous fertilizer whichcontributes significantly to losses of N as nitrous oxide (N

2O)

emissions to the atmosphere [4] N2O emissions are on

average gt10 times higher from tea fields than from forestsN2O has an ozone-depleting potential similar to that of

hydrochlorofluorocarbons and a global warming potential300 times that of CO

2[3 5 6]

In China annual production of rice straw was about 174times 108 t in 2000 containing about 25 minus 40 times 104 t N [7]Due to lack of feasible conversion technologies until the

2000s on-site burning was traditionally used to dispose ofcrop residues resulting in severe environmental pollutionincluding greenhouse gases and nitrogen oxides [8] Recentlystraw return has become the preferred method of rice strawdisposal because of its positive effects on crop yield and soilproperties such as increased soil organic carbon contentHowever straw decomposition in soils takes time over theshort-term decomposition straw incorporation can hampercrop root penetration causing N deficiencies and contribut-ing to disease and weed problems [9 10] directly limitingits application in agriculture Therefore it is important todevelop a method for enhancing straw decomposition beforeor after incorporation

Many studies have attempted to hasten straw decompo-sition via microbial processes Trichoderma spp is the best-known cellulolytic fungi that can accelerate the process of ricestraw decomposition and produce more soil-available nutri-ents such as labile organicmatter and inorganicNPS [11ndash13]However a major challenge in implementing bioconversionof rice straw is identifying a suitable carrier substrate to

Hindawi Publishing Corporatione Scientific World JournalVolume 2014 Article ID 793752 9 pageshttpdxdoiorg1011552014793752

2 The Scientific World Journal

amplify the target organism promote survival ofTrichodermaspp in the rice straw and enhance colonization in the soilnear plant roots Fortunately sweet potato starch wastewater(SPSW) from local starch production plants could be used asan appropriate culture medium for microbial growth becauseof its high organic content A few studies have been publishedon production of microbial biomass through starch industrywastewater with variousmicrobes such as Trichoderma viride[14]

Compared to chemical fertilizers and protectants somestrains of Trichoderma spp provide additional benefits interms of plant growth and productivity such as systemicresistance to disease and abiotic stresses such as water deficitsand salt stress [15 16] If Trichoderma spp colonize the rootsthey strongly affect plant physiology by changing plant geneexpression providing season-long benefits to the plants [1718] Another major benefit is induction of increased N useefficiency (NUE) in plants [19] potentially reducing the Nfertilizer application rate by 30ndash50 through bioconver-sionbioaugmentation with no reduction in yield Thereforethe lower N inputs to soil and higher NUE may reduce N

2O

emissionsThe objectives of this study were to (1) develop a cost-

effective technology for using SPSW and rice straw forlarge-scale production of Trichoderma viride biofertilizer (2)evaluate the feasibility of biofortification of crops throughapplication of T virideN fertilizers to reduce N

2O emissions

and (3) minimize synthetic N fertilizer input via recycling ofN to achieve a sustainable agricultural system

2 Methods and Materials

21 Site Description The field experiment was carried outat the Changsha Research Station for Agricultural andEnvironmental Monitoring (CRSAEM) Changsha Hunanin subtropical central China (28∘3210158405010158401015840N 113∘1010158405810158401015840E) Theregion has a subtropicalmonsoon climatewith amean annualair temperature of 175∘C and a mean annual precipitation of1330mm (1979ndash2010) The red soils of this area are classifiedas ultisols (USDA soil taxonomy) [20] The selected siteis a typical hilly agricultural catchment with pine forests(woodlands) paddy fields (lowland) and tea fields (upland)as the three primary land-use types accounting for 655251 and 34 of the total catchment area (135 km2 Figure 1)respectively The other small upland areas in the catchmentare cropped sweet potato (lt05) The land-use distribu-tion in 1990 shows substantial conversion from woodlandsdominated by Masson pine and bamboo to large-scale teaplantations (Camellia sinensis L Figure 1) Each land-usetype is fertilized other than the woodlands The averageannual application rate of N fertilizers was 450 300 and250 kg Nhaminus1 yrminus1 for tea fields paddy fields and sweetpotato fields respectively

22 Preparation of Biofertilizer

221 RawMaterials and Starter Inoculum SPSW containingapproximately 25 g Lminus1 COD and 1 g Lminus1 total N was obtained

from a local sweet potato starch production company(Xiangfeng Corp Hunan China) as a by-product of sweetpotato starch processing Rice straw (total organic carbon(TOC) 565 total nitrogen (TN) 046) was collectedafter harvest from local paddy fields and ground to 5ndash8mmlengths for the solid fermentation culture of Trichodermaviride

The strain T viride EBL13 was isolated from the soiland was found to be active against phytopathogenic fungiin our laboratory [21] The starter cultures were obtained byinoculating 500 120583L of a spore suspension at 108 spores mLminus1into 20mL potato dextrose broth (PDB Sigma-Aldrich StLouis MO) and incubating at 25∘C in orbital agitation at150 rpm in the dark for 48 h

222 Mass Production of T viride Mycelium The starter cul-tures (2mL) were transferred to a 500-mL Erlenmeyer flaskcontaining 150mL sterile sweet starch industry wastewater(121 plusmn 1∘C for 15min) and were incubated in a rotary shakerat 28 plusmn 1∘C and 200 plusmn 5 rpm for 48 h The mycelium wascollected by centrifugation at 4000 g for 10min the biomassfresh weight was determined and then the fungal biomasswas prepared for solid-state fermentation (SSF) with ricestraw

223 Production of Biofertilizer by Solid-State FermentationThe homogenized mycelial mat (10 g dewatered myceliumwith 10mL fermentation supernatant) was transferred to a500-mL Erlenmeyer flask containing 10 g sterile rice straw(121 plusmn 1∘C for 15min) Afterwards the flasks were incubatedat 28plusmn1∘C for 148 h until mass production of T viride conidiaoccurred The conidia concentration was measured as cfugminus1 air-dried cotton stalk substrate using a modified conidiaassessment method [22]

23 Biofertilizer Application to the Tea Plantation Biofer-tilizer was compared with chemical fertilizer (N) using acompletely randomized design in an ongoing trial of about3-year-old tea (Camellia sinensis cv) from November 2010to May 2012 at a tea plantation in Jinjing There werefour treatments in the field experiment (1) unfertilized(CK0) (2) chemical fertilizer (CNH450 urea applied at450 kg Nhaminus1 yrminus1 CNL225 fertilizer applied at 225 kgNhaminus1 yrminus1) (3) biofertilizer (BFH225 biofertilizer appliedat 50000 kg haminus1 yrminus1 with the total N adjusted to that ofCNL225 BFL113 biofertilizer applied at 25000 kg haminus1 yrminus1with the total N adjusted to half that of CNL225) and (4)raw materials (RMH225 sweet starch industry wastewaterand rice straw at 50000 kg haminus1 yrminus1 for fermentation ofbiofertilizer RML113 sweet starch industry wastewater andrice straw at 25000 kg haminus1 yrminus1) The fertilizer was applied inthree stages each year 20 of the total N on November 1260 onMarch 1 and 20 onOctober 1 tilled 10ndash15 cm underthe soil surface The experimental plot was 60m2 (10 times 6m2)for each fertilizer treatment

The Scientific World Journal 3

24 Field Measurement of N2O Fluxes Soil N

2O fluxes were

measured using a static closed chamber and gas chromatog-raphy (GC) as described by Li et al [3] and Zheng et al[23] from March 2011 to April 2012 The closed mini-chambers were constructed of polyvinylchloride 015m indiameter and 018m in length with sharpened ends andscrew lids fitted with rubber septa for gas sampling Thechambers were gently inserted vertically into the soil to adepth of 005m using the sharpened ends Headspace gassamples were collected from 0930 to 1030 am once a weekduring tea harvest seasons (spring tea mid-March to mid-April autumn tea mid-September tomid-October) For eachtreatment 3 replicate gas samples were collected from theheadspace into preevacuated 12-mL vials (Exetainers LabcoHigh Wycombe UK) 0 and 30min after the lid was closedAfter gas sampling the air temperature in each chamber wasmeasured for subsequent correction of the flux calculationThe N

2O concentrations in the gas samples were analyzed

using a gas chromatograph (Agilent 7890A Agilent SantaClara CA) fitted with a 63Ni-electron capture detector and anautomatic sample injector system [3] N

2O fluxes (FLUX30 g

Nhaminus1 dminus1) were calculated using the following equation

FLUX30 = (11988830minus 1198880) sdot

119872N2O

1198810

sdotℎ

Δ119905

sdot1198790

1198790+ 119879air

sdot10000

1000

sdot 24 sdot2 sdot 119872N119872N2O

(1)

where 11988830minus 1198880is the difference in the N

2O concentration

in the headspace of the mini-chamber 0 and 30min afterthe lid was closed (ppmv)119872N

2O is the molecular weight of

N2O (gmolminus1) 119881

0is the molecular volume of N

2O under

standard conditions (temperature = 273K and pressure =1013 hPa) 224 times 10minus3m3119879

0= 273K119872N is the atomic weight

of nitrogen (g molminus1) ℎ is the chamber height (m) Δ119905 is theincubation period (05 h)119879air is the air temperature inside themini-chamber (∘C) and 10000 1000 and 24 are conversionfactors for m2 to ha mg to g and h to days respectively

25 Auxiliary Field Measurements In addition to measuringgas fluxes ammonium-N and nitrate-N in the soil weremeasured along with tea plant growth Four 0ndash20 cm soilsamples were collected from random locations in each plotusing a soil auger during each field measurement of N

2O

fluxes Each fresh soil sample was manually homogenizedand analyzed for soil ammonium-N (NH

4

+-N) nitrate-N(NO3

minus-N) and total N using an automated flow injectionanalyzer (FIAStar 5000 Foss Tecator Hoganas Sweden) asdescribed by Liu et al [24] Tea plant growth was monitoredby recording the weight of a fresh tea bush (bud with twoleaves) during the tea harvest seasons (spring tea mid-Marchto mid-April autumn tea mid-September to mid-October)

26 Statistical Analyses All statistical analyses were con-ducted using SPSS 120 (SPSS China Beijing China) andOri-gin 80 (Origin Lab Ltd Guangzhou China) The statisticalsignificance of the results was determined using Duncanrsquos

multiple-range test (119875 lt 005) Simple correlation coefficientsbetween soil N

2O flux and NH

4

+-N and NO3

minus-N contentswere calculated using the same statistical package

3 Results and Discussion

31 Effects of Land-Use Changes on N2O Emissions from Hilly

Areas of Subtropical Central China Figure 1 depicts changesin land-use distribution patterns using a geographic informa-tion system since 1955 In the late 1950s the land-use patternwas simple and mainly comprised of woodlands and paddyfields with 6305 and 2938 of the total land respectivelyWith rapid economic development and population increasesrapid land-use and land-cover changes have taken place inthese areas beginning in the 1990s there has been a largedecline in forest cover due to tea plantation expansionOverall 398 ha of woodland areas has been converted to teacrop cultivation about 298 of the total area of the districtWoodland cover (603) has decreased by 430 comparedto 1955 By 2012 tea plantation land had expanded to 342 ofthe total area of the district a marked increase of 152 from1990 to 2012

Under the typical fertilization model for tea plan-tations N

2O fluxes associated with high-N treatment

(450 kgNhaminus1 yrminus1) in the spring and autumn of 2011and the spring of 2012 were 248ndash490 152ndash402 and286ndash376 gNhaminus1 dminus1 respectively For low-N fertilization(225 kgNhaminus1 yrminus1) N

2O fluxes during the same periods

were 677ndash184 135ndash282 and 107ndash152 gNhaminus1 dminus1 respec-tively However for woodlands in the same district N

2O

fluxes during the same periods were minus821ndash111 085ndash212 and199ndash239 gNhaminus1 dminus1 respectivelyTherefore theN

2Ofluxes

from tea plantations with high-N (450 kgNhaminus1 yrminus1) andlow-N (225 kgNhaminus1 yrminus1) application were 177 plusmn 34 and94 plusmn 62 times higher than those from woodlands respec-tively Many studies have reported that land-use changescan impact N

2O emissions and that N

2O emissions signifi-

cantly increase when woodlands are converted into pastures[25] orchards [26] or cropland [27] Merino et al [28]measured N

2O releases from cropland and a pasture (740

and 1315 gNhaminus1 dminus1) and found that they were 3 and 6times higher than those from a forest (219 gNhaminus1 dminus1)respectively We have previously reported the high annualvariability of N

2O emissions as well as its spatial variability

and distribution from a tea field in 2010 [3 29] Many studieshave shown that agricultural land has much higher N

2O

emissions than forests because of generally higher N in soilsthat may be further enhanced through intensification of theN cycle through application of synthetic N fertilizer [30 31]

32 Bioconversion of Sweet Potato StarchWastewater and RiceStraw into Biofertilizer by Trichoderma viride In the studiedcatchment paddy fields accounting for 251 of the total landarea produced 302 times 104 t yrminus1 straw containing 907 times 103 t Cand 121 times 102 t N (Table 1) Sweet potatoes were cultivated inthe upland area making up 025 of the total land area ofthe catchmentThe sweet potatoes were processed to produce


1955 20121990

Tea field

S

N

W E

Paddy fieldResidential area

WoodlandRoadWater body

0 25(km)

5

Figure 1 Land-use distributions during 1955ndash2012

Table 1 Process parameters for bioconversion of SPSW and rice straw into biofertilizer by T viride

Parameters Raw materials T viride BiofertilizerRice straw SPSW Mycelium

Water content () 53 978 81 483Total organic carbon () 565 21 144 184Total nitrogen () 04 008 065 045Dry mycelium weight (g Lminus1) mdash mdash 896 mdashConidia concentration (cfu gminus1) mdash mdash mdash 32 times 10

10

Note ldquomdashrdquo means not test

150 t yrminus1 refined starch during which about 12 times 104 t SPSWwas produced containing 252 t C and 96 tN (Table 1) TheSPSW was found to be suitable for cultivation of T viridewith a dry mycelium weight of about 896 g Lminus1 About736 of the C and 813 of the N in the wastewater weretransferred into the mycelium To optimize conditions andenhance straw decomposition the mass T viride myceliumwas mixed with rice straw by solid-state fermentation toproduce biofertilizer with a maximum conidia concentrationof 32 times 1010 cfu gminus1 By liquid-state fermentation of the SPSWand solid-state fermentation of the paddy straw the tea plan-tation can be provided with 11 times 103 t C yrminus1 and 270 t N yrminus1or 24 t C haminus1 yrminus1and 587 kgNhaminus1 yrminus1 over the 460 hacatchment area (calculated from Table 1) Harman reportedthat Trichoderma strains can stabilize soil nutrients enhancenutrient uptake promote root development increase roothair formation and induce systemic resistance to bioticstresses (diseases) and abiotic stresses (water deficits) [1516 18 21] The biological activity of Trichoderma spp couldreplace some functions of synthetic nitrogen to minimizesynthetic inputs of fertilizer in tea cultivation

33 Mitigation Options for Reducing N2O Emissions Table 2

and Figure 2 show that application of synthetic N fertilizersignificantly increased tea yields and also N

2O emissions

compared to unfertilized fields Addition of synthetic Nfertilizer as CNL225 and CNL450 increased the average teayields by 91 and 396 (spring tea 2011) 399 and 1118(autumn tea 2011) and 642 and 1033 (spring tea 2012)respectively compared to the unfertilized treatment (CK0)At the same time fluxes of N

2O were significantly affected

by the rate of synthetic N fertilization Based on measure-ments in 2011-2012 the fertilization treatments CNL225 andCNL450 significantly stimulated N

2O emissions by 849

and 2881 (spring tea 2011) 3686 and 4731 (autumn tea2011) and 3889 and 10250 (spring tea 2012) respectivelyThese results are consistent with those of previous studiesindicating that nitrogen fertilizer application enhances emis-sions of N

2O from agricultural fields [26 32ndash34] In addition

the magnitude of the N2O emissions from our tea fields

was much higher than previously reported values for paddyfields and uplands with N fertilization Accordingly it isurgent to establish technologies and management practices


Table2Prod

uctiv

ityof

teafi

elds(fre

shteakg

haminus1)w

ithvario

usfertilizertreatments

2011-2012

Treatm

ent

2011sprin

gtea(

kghaminus1)

2011autumntea(

kghaminus1)

2012

sprin

gtea(

kghaminus1)

March

5March

12March

19Octob

er15

Octob

er22

Octob

er29

April1

April

8Ap

ril15

CK0

1254plusmn156

a3066plusmn112

a3773plusmn354

a2698plusmn159

a2554plusmn654

a2314plusmn453

a2072plusmn109

a2859plusmn499

a5082plusmn544

a

RMH2251696plusmn238

bc3473plusmn519

ab5458plusmn496

c4134plusmn146

bc3712plusmn248

bc313plusmn370

b2895plusmn163

a3017plusmn369

ab4578plusmn363

a

RML113

1379plusmn213

ab2392plusmn415

ab3947plusmn403

a3555plusmn283

b3189plusmn283

b2926plusmn196

ab2559plusmn572

a2687plusmn555

ab4687plusmn137

a

BFH225

2029plusmn407

c4263plusmn740

b7845plusmn446

d6203plusmn396

d5874plusmn433

d4635plusmn391

c4004plusmn249

c5284plusmn870

c7839plusmn1477

b

BFL113

1422plusmn125

ab2927plusmn95

ab5214plusmn312

bc4539plusmn347

c3875plusmn83

c3271plusmn347

b2823plusmn549

a3646plusmn402

b5735plusmn347

ab

CNH450

2020plusmn49

c3493plusmn459

ab5421plusmn168

c5755plusmn625

d5329plusmn305

d494plusmn460

c5303plusmn379

c6335plusmn2771198886727plusmn1213

b

CNL2

251456plusmn124

ab3093plusmn569

ab4166plusmn302

ab3888plusmn670

bc3503plusmn44

bc3203plusmn369

b448plusmn343

b4872plusmn228

b5389plusmn453

ab

Notein

each

row

means

follo

wed

bythes

amelettera

reno

tsignificantly

different

at119875lt005


0

10

20

30

40

50

60

5 M

ar20

11

12 M

ar20

11

19 M

ar20

1115

Oct

2011

22 O

ct20

1129

Oct

2011

1 Ap

r20

128

Apr

2012

15 A

pr20

12

Sampling dateCK0RMH225RML113BFH225

BFL113CNH450CNL225WDL

minus10

FLU

X30

(g N

haminus1

dminus1)

Figure 2 Variations in soil N2O flux in tea fields during 2011-2012 (CK0 unfertilized RMH225 raw materials applied at 225 kg Nhaminus1 yrminus1

RMH113 raw materials applied at 113 kg Nhaminus1 yrminus1 BFH225 biofertilizer applied at 225 kg Nhaminus1 yrminus1 BFH113 biofertilizer applied at113 kgNhaminus1 yrminus1 CNH450 urea applied at 450 kg Nhaminus1 yrminus1 and CNH225 urea applied at 225 kg Nhaminus1 yrminus1 WDL-woodland)

for mitigating N2O emissions from tea fields while sustaining

or increasing tea productionImproved management using biofertilizer instead of syn-

thetic N fertilizer significantly decreased N2O emissions

(Figure 2) The BFH225 treatment reduced N2O emissions

by 333ndash718 and increased the tea yield by 162ndash622compared to the CNL225 treatment depending on the sea-son The RMH225 treatment also reduced N

2O emissions by

436ndash800 however the yield of 2012 spring tea decreasedand some tea plants died There have been contradictoryreports on the effects of organic waste materials on N

2O

emissions Some studies have shown an inhibitory effect [35ndash37] while others have reported stimulation [38 39] Based onour results direct application of the organic waste materials(rice straw and starch industry wastewater) to the tea fieldcarried significant risks to the tea plants However transfer-ring these waste materials into biofertilizer by fermentationwas a feasible approach for decreasing N

2O fluxes from tea

fieldsIn addition compared to the high application rate of

chemical N fertilizer (CNL450) the BFH225 treatment thatreduced the annual N fertilization rate by 50 did not affectthe tea yield (Table 2) but significantly decreased averageN

2O

emissions by 716 (Figure 2)This decrease may be ascribedto three possible mechanisms First because Trichodermaviride could promote plant growth mineral nitrogen in thesoil may be more quickly taken up by the vigorously growingtea plants in the presence of Trichoderma spp [19] therebyreducing the N in the substrate available for N

2Oproduction

Second application of biofertilizer with a high CN ratio(gt30) results in temporarily improving soil N immobilizationand thus a low availability of soil N for N

2O emission [40]

Finally the buffering and loosening effect of the organicmatter in the biofertilizer resulting in higher pH and oxygenin the soilsmay have favoredN

2as an end product rather than

N2O in denitrification [41]

34 Effects of Soil 1198731198674

+ and 1198731198743

minus Concentrations on N2O

Production N fertilizer application significantly increasedthe soil (0ndash20 cm)NH

4

+ andNO3

minus concentrations over thoseof unfertilized soil (Figure 3) For CNL450 and CNL225 theNH4

+ concentrations varied between 170 and 921mg Nkgminus1soil (dw) withmean concentrations as high as 561plusmn196 and295plusmn119mgNkgminus1 respectivelyTheNO

3

minus concentrationsranged from 66 to 313mgNkgminus1 and averaged 206plusmn61 and122 plusmn 34mg Nkgminus1 for CNL450 and CNL225 respectivelyFor BFH225 and BFH113 the NH

4

+ concentration rangedfrom 36 to 172mgNkgminus1 with lower mean concentrations of114 plusmn 34 and 68 plusmn 17mg Nkgminus1 throughout the tea seasonNO3

minus concentrations varied between 09 and 80mg Nkgminus1withmean concentrations of 42plusmn25 and 21plusmn10mgNkgminus1 N concentrations in the CNL450 and CNL225 treatmentsdecreased rapidly after fertilization however in the BFH225and BFH113 treatments it remained comparatively stableIn addition the N concentrations in the BFH113 treatmentwere lower than in the BFH225 treatment with more inten-sive fertilization Organic nitrogen slowly released from thebiofertilizermay have been responsible for this phenomenon

A significant positive relationship was found when N2O

emissions were linearly regressed against soil N concen-trations across the various treatments For all N fertilizertreatments a significant positive correlation existed betweenN2O fluxes and soil NH

4

+-N contents (y = 049x + 090 1199032


0102030405060708090

100

1 D

ec20

105

Mar

2011

12 M

ar20

1119

Mar

2011

15 O

ct20

1122

Oct

2011

29 O

ct20

111

Apr

2012

8 Ap

r20

1215

Apr

2012

Sampling date

NH

4+

-N co

ncen

trat

ion

(mg k

gminus1)

CK0RMH225RML113BFH225

BFL113CNH450CNL225

(a)

0

5

10

15

20

25

30

35

40

1 D

ec20

105

Mar

2011

12 M

ar20

1119

Mar

2011

15 O

ct20

1122

Oct

2011

29 O

ct20

111

Apr

2012

8 Ap

r20

1215

Apr

2012


BFL113CNH450CNL225

NO3minus

-N co

ncen

trat

ion

(mg k

gminus1)

(b)

Figure 3 Variations in soil concentrations of (a) NH4

+-N and (b) NO3

minus-N in tea fields during 2010-2012 (CK0 unfertilizedRMH225 raw materials applied at 225 kgNhaminus1 yrminus1 RMH113 raw materials applied at 113 kgNhaminus1 yrminus1 BFH225 biofertilizer applied at225 kgNhaminus1 yrminus1 BFH113 biofertilizer applied at 113 kgNhaminus1 yrminus1 CNH450 urea applied at 450 kgNhaminus1 yrminus1 and CNH225 urea appliedat 225 kgNhaminus1 yrminus1)

0

10

20

30

40

50

0 20 40 60 80 100

FLU

X30

(g N

haminus1

dminus1)

NH4+-N (mg Lminus1)

y = 049x + 090

(r2 = 072 P lt 001)

(a)

0

10

20

30

40

50

0 10 20 30 40NO3

minus-N (mg Lminus1)

FLU

X30

(g N

haminus1

dminus1)

y = 124x + 096

(r2 = 067 P lt 001)

(b)

Figure 4 Relationships between N2O emissions from the tea field and soil concentrations of (a) NH

4

+-N and (b) NO3

minus-N

= 072 and 119875 lt 001 Figure 4(a)) and a similar effect wasobserved between N

2O fluxes and soil NO

3

minus-N contents (y =124x + 096 1199032 = 067 and 119875 lt 001 Figure 4(b))

In several previous studies soil temperature soil mois-ture and NH

4

+-N or NO3

minus-N contents were identified asthe main environmental drivers of N

2O fluxes [26 34 42]

affecting either nitrification or denitrification and thus N2O

productionThe temperature and humidity of the soil whichwere significantly correlated with N

2O fluxes are controlled

by natural factors such as seasons and precipitationHoweveranother key factor affecting N

2O flux the N content in the

soil can be controlled by changing the type of fertilizerapplied the fertilization rate and the timing of applicationproviding a feasible method of controlling the N

2O flux

4 Conclusions

Land-use conversion from woodlands to tea fields in sub-tropical areas of central China leads to increased N

2O

emissions partly due to increased N fertilizer use By liquid-state fermentation of SPSW and solid-state fermentation ofpaddy straw by T viride tea plantations can be provided


with biofertilizer that can replace some functions of syn-thetic N minimizing synthetic inputs of fertilizer in teacultivation Improved management using biofertilizer ratherthan synthetic N fertilizer can significantly decrease N

2O

emissions while sustaining or increasing tea productionThus the process described here using SPSW and ricestraw for mass-scale production of T viride biofertilizer is afeasible and cost-effective approach for minimizing syntheticinputs of fertilizer reducing cumulative N

2O emissions and

developing the best management practices for N in soils

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publishing of this paper

Acknowledgments

This work was supported by the Key Project of ChineseAcademy of Sciences (nos KZZD-EW-11-3 and KZZD-EW-09-3) and the National Key Technology RampD Program (no2013BAD11B03)

References

[1] Y Li J S Wu S L Liu et al ldquoIs the CNP stoichiometry in soiland soil microbial biomass related to the landscape and land usein southern subtropical Chinardquo Global Biogeochemical Cyclesvol 26 pp 1ndash14 2012

[2] S Li X Wu H Xue et al ldquoQuantifying carbon storage for teaplantations in Chinardquo Agriculture Ecosystems amp Environmentvol 141 no 3-4 pp 390ndash398 2011

[3] Y Li X Q Fu X L Liu et al ldquoSpatial variability anddistribution of N

2O emissions from a tea field during the dry

season in subtropical central Chinardquo Geoderma vol 193 pp 1ndash12 2013

[4] J N Galloway A R Townsend J W Erisman et al ldquoTrans-formation of the nitrogen cycle recent trends questions andpotential solutionsrdquo Science vol 320 no 5878 pp 889ndash8922008

[5] A R Ravishankara J S Daniel and R W Portmann ldquoNitrousoxide (N

2O) the dominant ozone-depleting substance emitted

in the 21st centuryrdquo Science vol 326 no 5949 pp 123ndash125 2009[6] S J Hall and P A Matson ldquoNitrogen oxide emissions after

nitrogen additions in tropical forestsrdquoNature vol 400 no 6740pp 152ndash155 1999

[7] Y Xu L Nie R J Buresh et al ldquoAgronomic performanceof late-season rice under different tillage straw and nitrogenmanagementrdquo Field Crops Research vol 115 no 1 pp 79ndash842010

[8] D C Edmeades ldquoThe long-term effects of manures and fertilis-ers on soil productivity and quality a reviewrdquo Nutrient Cyclingin Agroecosystems vol 66 no 2 pp 165ndash180 2003

[9] O C Devevre and W R Horwath ldquoDecomposition of ricestraw and microbial carbon use efficiency under different soiltemperatures andmoisturesrdquo Soil Biology and Biochemistry vol32 no 11-12 pp 1773ndash1785 2000

[10] H Shindo and T Nishio ldquoImmobilization and remineralizationof N following addition of wheat straw into soil determinationof gross N transformation rates by 15N- ammonium isotope

dilution techniquerdquo Soil Biology and Biochemistry vol 37 no3 pp 425ndash432 2005

[11] W Han and M He ldquoThe application of exogenous cellulase toimprove soil fertility and plant growth due to acceleration ofstraw decompositionrdquo Bioresource Technology vol 101 no 10pp 3724ndash3731 2010

[12] M Schmoll and A Schuster ldquoBiology and biotechnology ofTrichodermardquo Applied Microbiology and Biotechnology vol 87no 3 pp 787ndash799 2010

[13] R L Yadav A Suman S R Prasad and O Prakash ldquoEffectof Gluconacetobacter diazotrophicus and Trichoderma viride onsoil health yield andN-economyof sugarcane cultivation undersubtropical climatic conditions of Indiardquo European Journal ofAgronomy vol 30 no 4 pp 296ndash303 2009

[14] M Verma S K Brar R D Tyagi R Y Surampalli and J RValero ldquoStarch industry wastewater as a substrate for antag-onist Trichoderma viride productionrdquo Bioresource Technologyvol 98 no 11 pp 2154ndash2162 2007

[15] G E Harman C R Howell A Viterbo I Chet and MLorito ldquoTrichoderma speciesmdashopportunistic avirulent plantsymbiontsrdquo Nature Reviews Microbiology vol 2 no 1 pp 43ndash56 2004

[16] G E Harman ldquoTrichoderma-not just for biocontrol anymorerdquoPhytoparasitica vol 39 no 2 pp 103ndash108 2011

[17] G Alfano M L Lewis Ivey C Cakir et al ldquoSystemic modu-lation of gene expression in tomato by Trichoderma hamatum382rdquo Phytopathology vol 97 no 4 pp 429ndash437 2007

[18] G E Harman ldquoMultifunctional fungal plant symbionts newtools to enhance plant growth and productivityrdquo New Phytol-ogist vol 189 no 3 pp 647ndash649 2011

[19] M Shoresh G E Harman and F Mastouri ldquoInduced systemicresistance and plant responses to fungal biocontrol agentsrdquoAnnual Review of Phytopathology vol 48 pp 21ndash43 2010

[20] M Zhang andZHe ldquoLong-term changes in organic carbon andnutrients of an Ultisol under rice cropping in southeast ChinardquoGeoderma vol 118 no 3-4 pp 167ndash179 2004

[21] Z Bai B Jin Y Li J Chen and Z Li ldquoUtilization of winerywastes for Trichoderma viride biocontrol agent production bysolid state fermentationrdquo Journal of Environmental Sciences vol20 no 3 pp 353ndash358 2008

[22] M Verma S K Brar R D Tyagi R Y Surampalli and JR Valero ldquoIndustrial wastewaters and dewatered sludge richnutrient source for production and formulation of biocontrolagent Trichoderma viriderdquo World Journal of Microbiology andBiotechnology vol 23 no 12 pp 1695ndash1703 2007

[23] X Zheng B Mei Y Wang et al ldquoQuantification of N2O

fluxes from soil-plant systems may be biased by the applied gaschromatograph methodologyrdquo Plant and Soil vol 311 no 1-2pp 211ndash234 2008

[24] C Liu KWang SMeng et al ldquoEffects of irrigation fertilizationand crop straw management on nitrous oxide and nitric oxideemissions fromawheat-maize rotation field in northernChinardquoAgriculture Ecosystems amp Environment vol 140 no 1-2 pp226ndash233 2011

[25] S J Livesley R Kiese P Miehle C J Weston K Butterbach-Bahl and S K Arndt ldquoSoil-atmosphere exchange of green-house gases in a Eucalyptus marginatawoodland a clover-grasspasture and Pinus radiata and Eucalyptus globulus plantationsrdquoGlobal Change Biology vol 15 no 2 pp 425ndash440 2009

[26] S Lin J Iqbal R Hu et al ldquoDifferences in nitrous oxidefluxes from red soil under different land uses inmid-subtropical


Chinardquo Agriculture Ecosystems amp Environment vol 146 no 1pp 168ndash178 2012

[27] S S Malhi and R Lemke ldquoTillage crop residue and N fertilizereffects on crop yield nutrient uptake soil quality and nitrousoxide gas emissions in a second 4-yr rotation cyclerdquo Soil andTillage Research vol 96 no 1-2 pp 269ndash283 2007

[28] A Merino P Perez-Batallon and F Macıas ldquoResponses of soilorganic matter and greenhouse gas fluxes to soil managementand land use changes in a humid temperate region of southernEuroperdquo Soil Biology and Biochemistry vol 36 no 6 pp 917ndash925 2004

[29] X Fu Y Li W Su et al ldquoAnnual dynamics of N2O emissions

from a tea fieldin southern subtropical Chinardquo Plant Soil andEnvironment vol 58 no 8 pp 373ndash378 2012

[30] MU F Kirschbaum S Saggar K R Tate et al ldquoComprehensiveevaluation of the climate-change implications of shifting landuse between forest and grassland New Zealand as a case studyrdquoAgriculture Ecosystems amp Environment vol 150 pp 123ndash1382012

[31] M U Kirschbaum S Saggar K R Tate K P Thakur and DL Giltrap ldquoQuantifying the climate-change consequences ofshifting land use between forest and agriculturerdquo Science of theTotal Environment vol 465 pp 314ndash324 2013

[32] Z Yao X Zheng H Dong R Wang B Mei and J Zhu ldquoA3-year record of N

2O and CH

4emissions from a sandy loam

paddy during rice seasons as affected by different nitrogenapplication ratesrdquo Agriculture Ecosystems amp Environment vol152 pp 1ndash9 2012

[33] J Ma X L Li H Xu Y Han Z C Cai and K Yagi ldquoEffectsof nitrogen fertiliser and wheat straw application on CH

4and

N2O emissions from a paddy rice fieldrdquo Australian Journal of

Soil Research vol 45 no 5 pp 359ndash367 2007[34] S Lin J Iqbal R Hu et al ldquoNitrous oxide emissions from rape

field as affected by nitrogen fertilizer management a case studyin Central Chinardquo Atmospheric Environment vol 45 no 9 pp1775ndash1779 2011

[35] Z Yao X Zheng B Xie et al ldquoTillage and crop residuemanagement significantly affects N-trace gas emissions duringthe non-rice season of a subtropical rice-wheat rotationrdquo SoilBiology and Biochemistry vol 41 no 10 pp 2131ndash2140 2009

[36] J Zou YHuang J Jiang X Zheng andR L Sass ldquoA 3-year fieldmeasurement ofmethane and nitrous oxide emissions from ricepaddies in China effects of water regime crop residue andfertilizer applicationrdquo Global Biogeochemical Cycles vol 19 no2 Article ID GB2021 2005

[37] B Chaves S De Neve M D C Lillo Cabrera P Boeckx OVan Cleemput and G Hofman ldquoThe effect of mixing organicbiological waste materials and high-N crop residues on theshort-time N

2O emission from horticultural soil in model

experimentsrdquo Biology and Fertility of Soils vol 41 no 6 pp 411ndash418 2005

[38] J Shan and X Yan ldquoEffects of crop residue returning on nitrousoxide emissions in agricultural soilsrdquoAtmospheric Environmentvol 71 pp 170ndash175 2013

[39] R Garcia-Ruiz and E M Baggs ldquoN2O emission from soil

following combined application of fertiliser-N and groundweedresiduesrdquo Plant and Soil vol 299 no 1-2 pp 263ndash274 2007

[40] J Sarkodie-Addo H C Lee and E M Baggs ldquoNitrous oxideemissions after application of inorganic fertilizer and incorpo-ration of greenmanure residuesrdquo Soil Use andManagement vol19 no 4 pp 331ndash339 2003

[41] M Simek L Jısova and D W Hopkins ldquoWhat is the so-called optimum pH for denitrification in soilrdquo Soil Biology andBiochemistry vol 34 no 9 pp 1227ndash1234 2002

[42] J Zhang and X Han ldquoN2O emission from the semi-arid

ecosystem under mineral fertilizer (urea and superphosphate)and increased precipitation in northern Chinardquo AtmosphericEnvironment vol 42 no 2 pp 291ndash302 2008

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Inorganic ChemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

International Journal ofPhotoenergy


Carbohydrate Chemistry

International Journal of


Journal of

Chemistry


Advances in

Physical Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom

Analytical Methods in Chemistry

Journal of

Volume 2014

Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

SpectroscopyInternational Journal of


The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Medicinal ChemistryInternational Journal of


Chromatography Research International


Applied ChemistryJournal of



Theoretical ChemistryJournal of


Journal of

Spectroscopy

Analytical ChemistryInternational Journal of


Journal of


Quantum Chemistry


Organic Chemistry International

ElectrochemistryInternational Journal of



CatalystsJournal of


amplify the target organism promote survival ofTrichodermaspp in the rice straw and enhance colonization in the soilnear plant roots Fortunately sweet potato starch wastewater(SPSW) from local starch production plants could be used asan appropriate culture medium for microbial growth becauseof its high organic content A few studies have been publishedon production of microbial biomass through starch industrywastewater with variousmicrobes such as Trichoderma viride[14]

Compared to chemical fertilizers and protectants somestrains of Trichoderma spp provide additional benefits interms of plant growth and productivity such as systemicresistance to disease and abiotic stresses such as water deficitsand salt stress [15 16] If Trichoderma spp colonize the rootsthey strongly affect plant physiology by changing plant geneexpression providing season-long benefits to the plants [1718] Another major benefit is induction of increased N useefficiency (NUE) in plants [19] potentially reducing the Nfertilizer application rate by 30ndash50 through bioconver-sionbioaugmentation with no reduction in yield Thereforethe lower N inputs to soil and higher NUE may reduce N

2O

emissionsThe objectives of this study were to (1) develop a cost-

effective technology for using SPSW and rice straw forlarge-scale production of Trichoderma viride biofertilizer (2)evaluate the feasibility of biofortification of crops throughapplication of T virideN fertilizers to reduce N

2O emissions

and (3) minimize synthetic N fertilizer input via recycling ofN to achieve a sustainable agricultural system

2 Methods and Materials

21 Site Description The field experiment was carried outat the Changsha Research Station for Agricultural andEnvironmental Monitoring (CRSAEM) Changsha Hunanin subtropical central China (28∘3210158405010158401015840N 113∘1010158405810158401015840E) Theregion has a subtropicalmonsoon climatewith amean annualair temperature of 175∘C and a mean annual precipitation of1330mm (1979ndash2010) The red soils of this area are classifiedas ultisols (USDA soil taxonomy) [20] The selected siteis a typical hilly agricultural catchment with pine forests(woodlands) paddy fields (lowland) and tea fields (upland)as the three primary land-use types accounting for 655251 and 34 of the total catchment area (135 km2 Figure 1)respectively The other small upland areas in the catchmentare cropped sweet potato (lt05) The land-use distribu-tion in 1990 shows substantial conversion from woodlandsdominated by Masson pine and bamboo to large-scale teaplantations (Camellia sinensis L Figure 1) Each land-usetype is fertilized other than the woodlands The averageannual application rate of N fertilizers was 450 300 and250 kg Nhaminus1 yrminus1 for tea fields paddy fields and sweetpotato fields respectively

22 Preparation of Biofertilizer

221 RawMaterials and Starter Inoculum SPSW containingapproximately 25 g Lminus1 COD and 1 g Lminus1 total N was obtained

from a local sweet potato starch production company(Xiangfeng Corp Hunan China) as a by-product of sweetpotato starch processing Rice straw (total organic carbon(TOC) 565 total nitrogen (TN) 046) was collectedafter harvest from local paddy fields and ground to 5ndash8mmlengths for the solid fermentation culture of Trichodermaviride

The strain T viride EBL13 was isolated from the soiland was found to be active against phytopathogenic fungiin our laboratory [21] The starter cultures were obtained byinoculating 500 120583L of a spore suspension at 108 spores mLminus1into 20mL potato dextrose broth (PDB Sigma-Aldrich StLouis MO) and incubating at 25∘C in orbital agitation at150 rpm in the dark for 48 h

222 Mass Production of T viride Mycelium The starter cul-tures (2mL) were transferred to a 500-mL Erlenmeyer flaskcontaining 150mL sterile sweet starch industry wastewater(121 plusmn 1∘C for 15min) and were incubated in a rotary shakerat 28 plusmn 1∘C and 200 plusmn 5 rpm for 48 h The mycelium wascollected by centrifugation at 4000 g for 10min the biomassfresh weight was determined and then the fungal biomasswas prepared for solid-state fermentation (SSF) with ricestraw

223 Production of Biofertilizer by Solid-State FermentationThe homogenized mycelial mat (10 g dewatered myceliumwith 10mL fermentation supernatant) was transferred to a500-mL Erlenmeyer flask containing 10 g sterile rice straw(121 plusmn 1∘C for 15min) Afterwards the flasks were incubatedat 28plusmn1∘C for 148 h until mass production of T viride conidiaoccurred The conidia concentration was measured as cfugminus1 air-dried cotton stalk substrate using a modified conidiaassessment method [22]

23 Biofertilizer Application to the Tea Plantation Biofer-tilizer was compared with chemical fertilizer (N) using acompletely randomized design in an ongoing trial of about3-year-old tea (Camellia sinensis cv) from November 2010to May 2012 at a tea plantation in Jinjing There werefour treatments in the field experiment (1) unfertilized(CK0) (2) chemical fertilizer (CNH450 urea applied at450 kg Nhaminus1 yrminus1 CNL225 fertilizer applied at 225 kgNhaminus1 yrminus1) (3) biofertilizer (BFH225 biofertilizer appliedat 50000 kg haminus1 yrminus1 with the total N adjusted to that ofCNL225 BFL113 biofertilizer applied at 25000 kg haminus1 yrminus1with the total N adjusted to half that of CNL225) and (4)raw materials (RMH225 sweet starch industry wastewaterand rice straw at 50000 kg haminus1 yrminus1 for fermentation ofbiofertilizer RML113 sweet starch industry wastewater andrice straw at 25000 kg haminus1 yrminus1) The fertilizer was applied inthree stages each year 20 of the total N on November 1260 onMarch 1 and 20 onOctober 1 tilled 10ndash15 cm underthe soil surface The experimental plot was 60m2 (10 times 6m2)for each fertilizer treatment



2O fluxes were




2O fluxes (FLUX30 g


FLUX30 = (11988830minus 1198880) sdot

119872N2O

1198810

sdotℎ

Δ119905

sdot1198790

1198790+ 119879air

sdot10000

1000


(1)


2O concentration





2O under





2O


4

+-N) nitrate-N(NO3




2O flux and NH

4

+-N and NO3










2O


2Ofluxes









2O




1955 20121990

Tea field

S

N

W E



0 25(km)

5





10





2O emissions




2O emissions by 849






Table2Prod

uctiv

ityof

teafi

elds(fre

shteakg

haminus1)w

ithvario


2011-2012

Treatm

ent

2011sprin

gtea(

kghaminus1)

2011autumntea(

kghaminus1)

2012

sprin

gtea(

kghaminus1)

March

5March

12March

19Octob

er15

Octob

er22

Octob

er29

April1

April

8Ap

ril15

CK0

1254plusmn156

a3066plusmn112

a3773plusmn354

a2698plusmn159

a2554plusmn654

a2314plusmn453

a2072plusmn109

a2859plusmn499

a5082plusmn544

a

RMH2251696plusmn238

bc3473plusmn519

ab5458plusmn496

c4134plusmn146

bc3712plusmn248

bc313plusmn370

b2895plusmn163

a3017plusmn369

ab4578plusmn363

a

RML113

1379plusmn213

ab2392plusmn415

ab3947plusmn403

a3555plusmn283

b3189plusmn283

b2926plusmn196

ab2559plusmn572

a2687plusmn555

ab4687plusmn137

a

BFH225

2029plusmn407

c4263plusmn740

b7845plusmn446

d6203plusmn396

d5874plusmn433

d4635plusmn391

c4004plusmn249

c5284plusmn870

c7839plusmn1477

b

BFL113

1422plusmn125

ab2927plusmn95

ab5214plusmn312

bc4539plusmn347

c3875plusmn83

c3271plusmn347

b2823plusmn549

a3646plusmn402

b5735plusmn347

ab

CNH450

2020plusmn49

c3493plusmn459

ab5421plusmn168

c5755plusmn625

d5329plusmn305

d494plusmn460

c5303plusmn379

c6335plusmn2771198886727plusmn1213

b

CNL2

251456plusmn124

ab3093plusmn569

ab4166plusmn302

ab3888plusmn670

bc3503plusmn44

bc3203plusmn369

b448plusmn343

b4872plusmn228

b5389plusmn453

ab

Notein

each

row

means

follo

wed

bythes

amelettera

reno

tsignificantly

different

at119875lt005


0

10

20

30

40

50

60

5 M

ar20

11

12 M

ar20

11

19 M

ar20

1115

Oct

2011

22 O

ct20

1129

Oct

2011

1 Ap

r20

128

Apr

2012

15 A

pr20

12



minus10

FLU

X30

(g N

haminus1

dminus1)








2O emissions by


2O


2O fluxes from tea



2O


2Oproduction


2O emission [40]





+ and 1198731198743



4

+ andNO3



3


4





4

+-N contents (y = 049x + 090 1199032


0102030405060708090

100

1 D

ec20

105

Mar

2011

12 M

ar20

1119

Mar

2011

15 O

ct20

1122

Oct

2011

29 O

ct20

111

Apr

2012

8 Ap

r20

1215

Apr

2012

Sampling date

NH

4+

-N co

ncen

trat

ion

(mg k

gminus1)


BFL113CNH450CNL225

(a)

0

5

10

15

20

25

30

35

40

1 D

ec20

105

Mar

2011

12 M

ar20

1119

Mar

2011

15 O

ct20

1122

Oct

2011

29 O

ct20

111

Apr

2012

8 Ap

r20

1215

Apr

2012


BFL113CNH450CNL225

NO3minus

-N co

ncen

trat

ion

(mg k

gminus1)

(b)


+-N and (b) NO3


0

10

20

30

40

50

0 20 40 60 80 100

FLU

X30

(g N

haminus1

dminus1)

NH4+-N (mg Lminus1)

y = 049x + 090

(r2 = 072 P lt 001)

(a)

0

10

20

30

40

50

0 10 20 30 40NO3


FLU

X30

(g N

haminus1

dminus1)

y = 124x + 096

(r2 = 067 P lt 001)

(b)


4

+-N and (b) NO3

minus-N



3



4

+-N or NO3


2O fluxes [26 34 42]







2O flux

4 Conclusions


2O




2O


2O emissions and




Acknowledgments


References










































2O and CH




4and

























Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of



2O fluxes were




2O fluxes (FLUX30 g


FLUX30 = (11988830minus 1198880) sdot

119872N2O

1198810

sdotℎ

Δ119905

sdot1198790

1198790+ 119879air

sdot10000

1000


(1)


2O concentration





2O under





2O


4

+-N) nitrate-N(NO3




2O flux and NH

4

+-N and NO3










2O


2Ofluxes









2O




1955 20121990

Tea field

S

N

W E



0 25(km)

5





10





2O emissions




2O emissions by 849






Table2Prod

uctiv

ityof

teafi

elds(fre

shteakg

haminus1)w

ithvario


2011-2012

Treatm

ent

2011sprin

gtea(

kghaminus1)

2011autumntea(

kghaminus1)

2012

sprin

gtea(

kghaminus1)

March

5March

12March

19Octob

er15

Octob

er22

Octob

er29

April1

April

8Ap

ril15

CK0

1254plusmn156

a3066plusmn112

a3773plusmn354

a2698plusmn159

a2554plusmn654

a2314plusmn453

a2072plusmn109

a2859plusmn499

a5082plusmn544

a

RMH2251696plusmn238

bc3473plusmn519

ab5458plusmn496

c4134plusmn146

bc3712plusmn248

bc313plusmn370

b2895plusmn163

a3017plusmn369

ab4578plusmn363

a

RML113

1379plusmn213

ab2392plusmn415

ab3947plusmn403

a3555plusmn283

b3189plusmn283

b2926plusmn196

ab2559plusmn572

a2687plusmn555

ab4687plusmn137

a

BFH225

2029plusmn407

c4263plusmn740

b7845plusmn446

d6203plusmn396

d5874plusmn433

d4635plusmn391

c4004plusmn249

c5284plusmn870

c7839plusmn1477

b

BFL113

1422plusmn125

ab2927plusmn95

ab5214plusmn312

bc4539plusmn347

c3875plusmn83

c3271plusmn347

b2823plusmn549

a3646plusmn402

b5735plusmn347

ab

CNH450

2020plusmn49

c3493plusmn459

ab5421plusmn168

c5755plusmn625

d5329plusmn305

d494plusmn460

c5303plusmn379

c6335plusmn2771198886727plusmn1213

b

CNL2

251456plusmn124

ab3093plusmn569

ab4166plusmn302

ab3888plusmn670

bc3503plusmn44

bc3203plusmn369

b448plusmn343

b4872plusmn228

b5389plusmn453

ab

Notein

each

row

means

follo

wed

bythes

amelettera

reno

tsignificantly

different

at119875lt005


0

10

20

30

40

50

60

5 M

ar20

11

12 M

ar20

11

19 M

ar20

1115

Oct

2011

22 O

ct20

1129

Oct

2011

1 Ap

r20

128

Apr

2012

15 A

pr20

12



minus10

FLU

X30

(g N

haminus1

dminus1)








2O emissions by


2O


2O fluxes from tea



2O


2Oproduction


2O emission [40]





+ and 1198731198743



4

+ andNO3



3


4





4

+-N contents (y = 049x + 090 1199032


0102030405060708090

100

1 D

ec20

105

Mar

2011

12 M

ar20

1119

Mar

2011

15 O

ct20

1122

Oct

2011

29 O

ct20

111

Apr

2012

8 Ap

r20

1215

Apr

2012

Sampling date

NH

4+

-N co

ncen

trat

ion

(mg k

gminus1)


BFL113CNH450CNL225

(a)

0

5

10

15

20

25

30

35

40

1 D

ec20

105

Mar

2011

12 M

ar20

1119

Mar

2011

15 O

ct20

1122

Oct

2011

29 O

ct20

111

Apr

2012

8 Ap

r20

1215

Apr

2012


BFL113CNH450CNL225

NO3minus

-N co

ncen

trat

ion

(mg k

gminus1)

(b)


+-N and (b) NO3


0

10

20

30

40

50

0 20 40 60 80 100

FLU

X30

(g N

haminus1

dminus1)

NH4+-N (mg Lminus1)

y = 049x + 090

(r2 = 072 P lt 001)

(a)

0

10

20

30

40

50

0 10 20 30 40NO3


FLU

X30

(g N

haminus1

dminus1)

y = 124x + 096

(r2 = 067 P lt 001)

(b)


4

+-N and (b) NO3

minus-N



3



4

+-N or NO3


2O fluxes [26 34 42]







2O flux

4 Conclusions


2O




2O


2O emissions and




Acknowledgments


References










































2O and CH




4and

























Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


1955 20121990

Tea field

S

N

W E



0 25(km)

5





10





2O emissions




2O emissions by 849






Table2Prod

uctiv

ityof

teafi

elds(fre

shteakg

haminus1)w

ithvario


2011-2012

Treatm

ent

2011sprin

gtea(

kghaminus1)

2011autumntea(

kghaminus1)

2012

sprin

gtea(

kghaminus1)

March

5March

12March

19Octob

er15

Octob

er22

Octob

er29

April1

April

8Ap

ril15

CK0

1254plusmn156

a3066plusmn112

a3773plusmn354

a2698plusmn159

a2554plusmn654

a2314plusmn453

a2072plusmn109

a2859plusmn499

a5082plusmn544

a

RMH2251696plusmn238

bc3473plusmn519

ab5458plusmn496

c4134plusmn146

bc3712plusmn248

bc313plusmn370

b2895plusmn163

a3017plusmn369

ab4578plusmn363

a

RML113

1379plusmn213

ab2392plusmn415

ab3947plusmn403

a3555plusmn283

b3189plusmn283

b2926plusmn196

ab2559plusmn572

a2687plusmn555

ab4687plusmn137

a

BFH225

2029plusmn407

c4263plusmn740

b7845plusmn446

d6203plusmn396

d5874plusmn433

d4635plusmn391

c4004plusmn249

c5284plusmn870

c7839plusmn1477

b

BFL113

1422plusmn125

ab2927plusmn95

ab5214plusmn312

bc4539plusmn347

c3875plusmn83

c3271plusmn347

b2823plusmn549

a3646plusmn402

b5735plusmn347

ab

CNH450

2020plusmn49

c3493plusmn459

ab5421plusmn168

c5755plusmn625

d5329plusmn305

d494plusmn460

c5303plusmn379

c6335plusmn2771198886727plusmn1213

b

CNL2

251456plusmn124

ab3093plusmn569

ab4166plusmn302

ab3888plusmn670

bc3503plusmn44

bc3203plusmn369

b448plusmn343

b4872plusmn228

b5389plusmn453

ab

Notein

each

row

means

follo

wed

bythes

amelettera

reno

tsignificantly

different

at119875lt005


0

10

20

30

40

50

60

5 M

ar20

11

12 M

ar20

11

19 M

ar20

1115

Oct

2011

22 O

ct20

1129

Oct

2011

1 Ap

r20

128

Apr

2012

15 A

pr20

12



minus10

FLU

X30

(g N

haminus1

dminus1)








2O emissions by


2O


2O fluxes from tea



2O


2Oproduction


2O emission [40]





+ and 1198731198743



4

+ andNO3



3


4





4

+-N contents (y = 049x + 090 1199032


0102030405060708090

100

1 D

ec20

105

Mar

2011

12 M

ar20

1119

Mar

2011

15 O

ct20

1122

Oct

2011

29 O

ct20

111

Apr

2012

8 Ap

r20

1215

Apr

2012

Sampling date

NH

4+

-N co

ncen

trat

ion

(mg k

gminus1)


BFL113CNH450CNL225

(a)

0

5

10

15

20

25

30

35

40

1 D

ec20

105

Mar

2011

12 M

ar20

1119

Mar

2011

15 O

ct20

1122

Oct

2011

29 O

ct20

111

Apr

2012

8 Ap

r20

1215

Apr

2012


BFL113CNH450CNL225

NO3minus

-N co

ncen

trat

ion

(mg k

gminus1)

(b)


+-N and (b) NO3


0

10

20

30

40

50

0 20 40 60 80 100

FLU

X30

(g N

haminus1

dminus1)

NH4+-N (mg Lminus1)

y = 049x + 090

(r2 = 072 P lt 001)

(a)

0

10

20

30

40

50

0 10 20 30 40NO3


FLU

X30

(g N

haminus1

dminus1)

y = 124x + 096

(r2 = 067 P lt 001)

(b)


4

+-N and (b) NO3

minus-N



3



4

+-N or NO3


2O fluxes [26 34 42]







2O flux

4 Conclusions


2O




2O


2O emissions and




Acknowledgments


References










































2O and CH




4and

























Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


Table2Prod

uctiv

ityof

teafi

elds(fre

shteakg

haminus1)w

ithvario


2011-2012

Treatm

ent

2011sprin

gtea(

kghaminus1)

2011autumntea(

kghaminus1)

2012

sprin

gtea(

kghaminus1)

March

5March

12March

19Octob

er15

Octob

er22

Octob

er29

April1

April

8Ap

ril15

CK0

1254plusmn156

a3066plusmn112

a3773plusmn354

a2698plusmn159

a2554plusmn654

a2314plusmn453

a2072plusmn109

a2859plusmn499

a5082plusmn544

a

RMH2251696plusmn238

bc3473plusmn519

ab5458plusmn496

c4134plusmn146

bc3712plusmn248

bc313plusmn370

b2895plusmn163

a3017plusmn369

ab4578plusmn363

a

RML113

1379plusmn213

ab2392plusmn415

ab3947plusmn403

a3555plusmn283

b3189plusmn283

b2926plusmn196

ab2559plusmn572

a2687plusmn555

ab4687plusmn137

a

BFH225

2029plusmn407

c4263plusmn740

b7845plusmn446

d6203plusmn396

d5874plusmn433

d4635plusmn391

c4004plusmn249

c5284plusmn870

c7839plusmn1477

b

BFL113

1422plusmn125

ab2927plusmn95

ab5214plusmn312

bc4539plusmn347

c3875plusmn83

c3271plusmn347

b2823plusmn549

a3646plusmn402

b5735plusmn347

ab

CNH450

2020plusmn49

c3493plusmn459

ab5421plusmn168

c5755plusmn625

d5329plusmn305

d494plusmn460

c5303plusmn379

c6335plusmn2771198886727plusmn1213

b

CNL2

251456plusmn124

ab3093plusmn569

ab4166plusmn302

ab3888plusmn670

bc3503plusmn44

bc3203plusmn369

b448plusmn343

b4872plusmn228

b5389plusmn453

ab

Notein

each

row

means

follo

wed

bythes

amelettera

reno

tsignificantly

different

at119875lt005


0

10

20

30

40

50

60

5 M

ar20

11

12 M

ar20

11

19 M

ar20

1115

Oct

2011

22 O

ct20

1129

Oct

2011

1 Ap

r20

128

Apr

2012

15 A

pr20

12



minus10

FLU

X30

(g N

haminus1

dminus1)








2O emissions by


2O


2O fluxes from tea



2O


2Oproduction


2O emission [40]





+ and 1198731198743



4

+ andNO3



3


4





4

+-N contents (y = 049x + 090 1199032


0102030405060708090

100

1 D

ec20

105

Mar

2011

12 M

ar20

1119

Mar

2011

15 O

ct20

1122

Oct

2011

29 O

ct20

111

Apr

2012

8 Ap

r20

1215

Apr

2012

Sampling date

NH

4+

-N co

ncen

trat

ion

(mg k

gminus1)


BFL113CNH450CNL225

(a)

0

5

10

15

20

25

30

35

40

1 D

ec20

105

Mar

2011

12 M

ar20

1119

Mar

2011

15 O

ct20

1122

Oct

2011

29 O

ct20

111

Apr

2012

8 Ap

r20

1215

Apr

2012


BFL113CNH450CNL225

NO3minus

-N co

ncen

trat

ion

(mg k

gminus1)

(b)


+-N and (b) NO3


0

10

20

30

40

50

0 20 40 60 80 100

FLU

X30

(g N

haminus1

dminus1)

NH4+-N (mg Lminus1)

y = 049x + 090

(r2 = 072 P lt 001)

(a)

0

10

20

30

40

50

0 10 20 30 40NO3


FLU

X30

(g N

haminus1

dminus1)

y = 124x + 096

(r2 = 067 P lt 001)

(b)


4

+-N and (b) NO3

minus-N



3



4

+-N or NO3


2O fluxes [26 34 42]







2O flux

4 Conclusions


2O




2O


2O emissions and




Acknowledgments


References










































2O and CH




4and

























Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


0

10

20

30

40

50

60

5 M

ar20

11

12 M

ar20

11

19 M

ar20

1115

Oct

2011

22 O

ct20

1129

Oct

2011

1 Ap

r20

128

Apr

2012

15 A

pr20

12



minus10

FLU

X30

(g N

haminus1

dminus1)








2O emissions by


2O


2O fluxes from tea



2O


2Oproduction


2O emission [40]





+ and 1198731198743



4

+ andNO3



3


4





4

+-N contents (y = 049x + 090 1199032


0102030405060708090

100

1 D

ec20

105

Mar

2011

12 M

ar20

1119

Mar

2011

15 O

ct20

1122

Oct

2011

29 O

ct20

111

Apr

2012

8 Ap

r20

1215

Apr

2012

Sampling date

NH

4+

-N co

ncen

trat

ion

(mg k

gminus1)


BFL113CNH450CNL225

(a)

0

5

10

15

20

25

30

35

40

1 D

ec20

105

Mar

2011

12 M

ar20

1119

Mar

2011

15 O

ct20

1122

Oct

2011

29 O

ct20

111

Apr

2012

8 Ap

r20

1215

Apr

2012


BFL113CNH450CNL225

NO3minus

-N co

ncen

trat

ion

(mg k

gminus1)

(b)


+-N and (b) NO3


0

10

20

30

40

50

0 20 40 60 80 100

FLU

X30

(g N

haminus1

dminus1)

NH4+-N (mg Lminus1)

y = 049x + 090

(r2 = 072 P lt 001)

(a)

0

10

20

30

40

50

0 10 20 30 40NO3


FLU

X30

(g N

haminus1

dminus1)

y = 124x + 096

(r2 = 067 P lt 001)

(b)


4

+-N and (b) NO3

minus-N



3



4

+-N or NO3


2O fluxes [26 34 42]







2O flux

4 Conclusions


2O




2O


2O emissions and




Acknowledgments


References










































2O and CH




4and

























Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


0102030405060708090

100

1 D

ec20

105

Mar

2011

12 M

ar20

1119

Mar

2011

15 O

ct20

1122

Oct

2011

29 O

ct20

111

Apr

2012

8 Ap

r20

1215

Apr

2012

Sampling date

NH

4+

-N co

ncen

trat

ion

(mg k

gminus1)


BFL113CNH450CNL225

(a)

0

5

10

15

20

25

30

35

40

1 D

ec20

105

Mar

2011

12 M

ar20

1119

Mar

2011

15 O

ct20

1122

Oct

2011

29 O

ct20

111

Apr

2012

8 Ap

r20

1215

Apr

2012


BFL113CNH450CNL225

NO3minus

-N co

ncen

trat

ion

(mg k

gminus1)

(b)


+-N and (b) NO3


0

10

20

30

40

50

0 20 40 60 80 100

FLU

X30

(g N

haminus1

dminus1)

NH4+-N (mg Lminus1)

y = 049x + 090

(r2 = 072 P lt 001)

(a)

0

10

20

30

40

50

0 10 20 30 40NO3


FLU

X30

(g N

haminus1

dminus1)

y = 124x + 096

(r2 = 067 P lt 001)

(b)


4

+-N and (b) NO3

minus-N



3



4

+-N or NO3


2O fluxes [26 34 42]







2O flux

4 Conclusions


2O




2O


2O emissions and




Acknowledgments


References










































2O and CH




4and

























Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of



2O


2O emissions and




Acknowledgments


References










































2O and CH




4and

























Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of










2O and CH




4and

























Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of










Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of

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