research article effect of different light qualities on growth,...

Research ArticleEffect of Different Light Qualities on Growth Pigment ContentChlorophyll Fluorescence and Antioxidant Enzyme Activityin the Red Alga Pyropia haitanensis (Bangiales Rhodophyta)

Huanyang Wu12

1College of Environment and Energy South China University of Technology Guangzhou Guangdong 510006 China2Key Laboratory of Pollution Control and Ecosystem Restoration in Industry Clusters Ministry of EducationGuangzhou Guangdong 510006 China

Correspondence should be addressed to Huanyang Wu wuhuanyang163com

Received 22 May 2016 Revised 13 July 2016 Accepted 31 July 2016

Academic Editor Rita Casadio

Copyright copy 2016 Huanyang WuThis is an open access article distributed under theCreativeCommonsAttribution License whichpermits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

Spectral light changes evoke different morphogenetic and photosynthetic responses that can vary among different algae speciesThe aim of this study is to investigate the photosynthetic characteristics of the red macroalgae grown under different spectrumenvironments In this study Pyropia haitanensis were cultured under blue red and green LED and fluorescent tubes light Thegrowth rate photopigment composition chlorophyll fluorescence and antioxidative enzymes activities in different light spectrumswere investigatedThe results revealed that growth ratewas significantly higher in the thalli grownunder blue green and fluorescenttubes light Contents of Chl a and phycobiliprotein in red light were lower among all the growth conditions Furthermore a strikingincrease in SOD and CAT activity was observed in red light treatment along with the NPQ increase The results revealed thatthe photosynthetic efficiency and increased growth rate of P haitanensis benefitted from light spectrums such as blue green andfluorescent tubes light by pigment composition and photochemical efficiency manipulation whereas red light has disadvantageouseffects Accordingly the results for improving quality and the economic yield of algae species in some extent and the combinationof different wavelengths could allow better economic resource exploitation

1 Introduction

Light characteristics (spectral quality quantity and duration)have a profound influence on plant and seaweed metabolismand development [1 2] Usually the blue-green light wave-length penetrates deepest into the water as shorter andlonger wavelengths are more absorbed by water moleculesor scattered by particles Photosynthetic organs of algae havethe various capabilities to adapt to these different light wave-lengths

Among various light spectra red and blue wavelengthsplay an important role in the photosynthesis and photomor-phogenesis thus influencing plant development and metab-olism [3] It has been reported that the bright red and bluelight can increase the ratio of fucoxanthin to Chl a and Chlc a ratio in Laminaria hyperborea [4] Furthermore Barufiet al [5] reported that growth rates were higher in Gracilaria

birdiae exposed to white light and red light could be utilizedas a reproductive inductor to produce tetraspores Green lightcould also be beneficial for algal growth ofHalymenia floresiaChl a 120572-carotene and lutein contents were induced by whiteor green light and PC and PE synthesis was excited by bluelight and blue or red light respectively [6] Red light hasthe potential role of stimulating lipid production of Ettliaoleoabundans as well [7]

Previous results suggest that red green and blue lighthave special influence on regulating algae growth and photo-synthetic pigments synthesis Although there have also beensome investigations about influences of different light wave-lengths on photosynthetic efficiency and chlorophyll contentand growth in photosynthetic prokaryotes and eukaryotes[8ndash10] the effects of light spectrum in redmacroalga researchhave been limited especially the coupling effect betweenphotosynthesis and antioxidant system

Hindawi Publishing CorporationBioMed Research InternationalVolume 2016 Article ID 7383918 8 pageshttpdxdoiorg10115520167383918

2 BioMed Research International

This study focused on the Pyropia haitanensis (BangialesRhodophyta) one species of marine red macroalga whichis an economically important species primarily due to beingan economically important seaweed used for food and isthe principal species cultivated on a commercial scale insouthern China [11] The aim of this study is to illustrate theeffect of different light spectrum on chlorophyll fluorescencecharacteristics the physiological and growth parameters of Phaitanensis and the effect of lighting on the amount of growthrate photopigment composition chlorophyll fluorescenceand antioxidative enzymes activities in a different light spec-trum These results may help us realize the physiologicalstatus of themacroalgae providing data for application of nat-ural populations as well as for the improving culture industryof this species on the coast

2 Materials and Methods

21 Seaweed Materials Pyropia haitanensis was gathered inShantou China (23∘261015840 N 116∘641015840 E) The algae were lightlyrinsed and cleared of visible epiphytes and of any accu-mulated sediments and then placed in a cooler containingsome fresh seawater during the transportation to the labo-ratory (about 3 h 4∘C) The algae were preserved in a glassaquarium tank containing filtered natural seawater (salinity3) appended with 200120583M NaNO

3and 20120583M NaH

2PO4

(final concentration) in a controlled environmental compart-ment Light availability was provided by timer-controlledfluorescent tubes (100 120583mol photos mminus2 sminus1) operated on a12 h on-and-off photoperiod Water motion was providedby aeration and culture seawater was updated every 2 daysand temperature was kept at 15 plusmn 1∘C in the incubator Thealgae were preserved for 3 days prior to the start of theexperiments Only intact and healthy thalli were selected forthe subsequent experimental treatments

22 Experimental Design 5 g fresh weight (FW) algae weretransferred to a flask containing 5 L filtered seawater (threereplicates per light treatment) for a starting biomass densityof 1 g FWLminus1 Different light quality control units wereindependently designed in growth cabinets The cultureswere irradiated at 100120583mol photons mminus2 sminus1 for all treatmentgroups using LED arrays (Edlan Lighting Technology CoChina) red (R 640sim680 nm) blue (B 420sim460 nm) andgreen (G 490sim530 nm) light and fluorescent tubes (CK)light Light apparatus were affixed to a ceramic and steelsupport to facilitate efficient heat transfer to the mountingsubstrate Each light treatment employed 3 replicates All thecultures were preserved at 15 plusmn 1∘C and a lightdark regimeof 1212 h and the seawater nutrients and frequency of waterchanges were the same as indicated above Seaweeds wereharvested and used for experimental measurement after 7days of cultivation

23 Growth Rates Biomass (freshweight FW)wasmeasuredat the end of incubation and the mean relative growth rate(RGR) was assessed according to the formula RGR ( dayminus1)

= ln(1198821199051198820)119905 times 100 where119882

0refers to the initial FW and

119882119905to the FW after 119905 days

24 Chlorophyll Fluorescence Measurements Detection ofchlorophyll fluorescence was made using a pulse modulationfluorometer (JUNIOR-PAM Walz Germany) At least fouralgal samples were used for each measurement of chlorophyllfluorescence and the algae were acclimated to dark for10min before being detected The maximum quantum yieldof photosystem (PS) II of P haitanensis was estimated as119865V119865119898 and photochemical quenching coefficient (qP) andnonphotochemical quenching coefficient (NPQ) were alsodetermined [12] The rapid light curves (RLCs) consisted ofthe fluorescence response to eight different and increasingactinic irradiance levels over the range of 0sim820120583mol pho-tons mminus2 sminus1 The parameters of the RLCs were calculatedfollowing the formula from Jassby and Platt [13] rETR(relative electron transport rate) = rETRmax times tan ℎ(120572 times 119868rETRmax) where rETRmax is the saturated maximum rETRtan ℎ is the hyperbolic tangent function 120572 is the initial slopeof the RLC (the efficiency of the electron transport) and 119868 isthe incident irradiance

25 Pigment Estimation For chlorophyll a (Chl a) and car-otenoid (Car) about 01 g (FW) of thalli was ground in 5mL95 ethanol and extracted at 4∘C in darkness for 24 h Forphycoerythrin (PE) and phycocyanin (PC) 01 g (FW) ofsamples was ground in 10mL extraction buffers (01M phos-phate buffer pH = 68) at 4∘CThe extract was centrifuged at5000timesg for 10min and then used to determine the contents ofChl a Car PE andPCusing an ultraviolet spectrophotometer(UV-180 Shimadzu Japan) Chl a and Car contents wereanalyzed according toWellburn [14] and PE and PC contentswere analyzed according to Siegelman and Kycia [15]

26 Antioxidant Enzyme Activities Superoxide dismutase(SOD) activity was examined by measuring the ability toinhibit reduction of nitro blue tetrazolium (NBT) followingthe method of Beauchamp and Fridovich [16] One unitof enzyme activity was defined as the amount of enzymerequired to inhibit the photoreduction of NBT by 50and the unit is Umg protminus1 [16] Catalase (CAT) activity(Umg protminus1) was analyzed by using a ferrothiocyanatemethod of Cohen et al [17] with somemodifications Proteincontent was analyzed by the Bradford method [18]

27 Statistical Analyses All the data were expressed as meansplusmn SD (119899 ge 3) and one-way ANOVA and Tukey tests wereused to analyze differences among treatments by using SPSS170 The significance level was set at 119875 lt 005

3 Results

31 Growth Pyropia haitanensis maintained a positivegrowth during the period of culture for all the thalli grownunder experimental conditions (Figure 1) The growth rate ofthe thalli grown atCK control showedhigher value (29dminus1)

BioMed Research International 3

b

aa

a

B RCK GTreatments

00

05

10

15

20

25

30

35RG

R (

dminus1)

Figure 1 Mean relative growth rate (RGR) of Pyropia haitanensisas a function of the different light source (comparison via ANOVA)during laboratory culture Values are mean (plusmnSD) 119899 = 3 Differentletters indicate statistical significance (119875 lt 005) Error bars arestandard deviations

Table 1 Maximum relative electron transport rates (rETRmax) andefficiency of electron transport (120572 the initial slope of the RLCs) ofPyropia haitanensis grown at different light quality (comparison viaANOVA) Data are the means plusmn SD (119899 = 9) different letters indicatesignificant difference (119875 lt 005)

Light treatments ParametersrETRmax 120572

CK 7856 plusmn 831a 017 plusmn 001a

B 8148 plusmn 955a 018 plusmn 002a

R 5003 plusmn 487b 012 plusmn 001b

G 7679 plusmn 849a 016 plusmn 002a

among all experimental conditions No significant differencewas observed amongCK B andG grown (119875 gt 005) whereasthe RGR presented the lowest value among all experimentalgroups when the thalli were grown at R light (119875 lt 005)

32 Chlorophyll Fluorescence Figure 2 and Table 1 illustratedthe maximal photochemical yield (119865V119865119898) photochemicalquenching (qP) nonphotochemical quenching (NPQ) andrapid light curves (RLCs) as a function of measuring photo-synthetic capacity for Pyropia haitanensis grown at differentlight quality treatmentsThere were no significant differencesamong 119865V119865119898 rETRmax and qP value of P haitanensis inculturesmaintained inCK B andGgrown (119875 gt 005)119865V119865119898rETRmax and qP however were significantly suppressedwhen the thalli were grown at R light control (119875 lt 005)Conversely when the thalli were grown at R light controlthe NPQ showed the highest value which was statisticallydifferent as compared with the other treatments (119875 lt 005)No significant differences of NPQwere examined among CKB and G grown (119875 gt 005) Furthermore for the parametersof RLCs thalli of P haitanensis varied clearly with differentlight quality treatments Compared with others the thalli

grown at CK B and G group had higher rETRmax and theinitial slopes of RLCs (120572) value and these two parametershowever were significantly suppressed when the algae thalliwere grown at R light surroundings (119875 gt 005)

33 Pigment Experiments Pigment contents of Pyropia hai-tanensis for different experimental condition groups are sum-marized in Figure 3 Higher Chl a contents were observed inthalli grown at CK and B control whereas the Chl a contentsdecreased when the algae thalli were grown at R light (119875 lt005) For the G control no significant differences of Chl acontents were investigated compared with others The resultsof Car were not the same as Chl a There were no significantdifferences when the algae thalli were grown in any lightconditions (119875 gt 005)

The PE levels in G light group possessed significant aver-age values with 45 higher than R light treatments signifi-cant difference values were observed between G and R lighttreatments (119875 lt 005) Moreover slight decreases of PE levelswere discovered in B light treatments compared to G lightgroup but no significant differences were detected in thesetwo treatments (119875 gt 005) Likewise for PC contents highermean value was observed when the algae thalli were grownunder B and G light However slightly higher PC levels werediscovered in B light treatments compared to the G lightgroup which is dissimilar with the PE result

34 Antioxidant Enzyme Activity The change in antioxidantenzyme activity was different according to the different lightquality cultivations (Figure 4) The SOD and CAT experi-mental results are more similar comparatively For the thalligrown at R light the SOD and CAT activity was extremelyhigh compared with other experimental treatments (119875 lt005) Additionally the maximum values of CAT activity in Rlight grown treatmentswere beyond two times that of B andGgrown algae and the effects of light quality on SOD activity ofP haitanensis thalli grown followed a similar manner as thatof CAT whereas no significant differences of SOD and CATactivity were observed in B andG light cultivation (119875 gt 005)although these two cultivation groups presented a lower valueof SOD and CAT activity

4 Discussion

The present results suggest that thalli of Pyropia haitanen-sis are able to adjust their photosynthetic presentation toaccommodate various light spectra and the results revealedthat CK B and G lighting was more efficient in promotingthe algae growth than the thalli grown under R lightingSimilarly some studies have shown that B and G light couldplay an advantageous role on red algae growth and theirdevelopment [19] Moreover Pearson et al [20] reported thatSilvetia compressa releases gametes during B or in low RBlight ratios exposure showing significant influences of lightwavelength on algal growth G light resulted in the highestalgal growth rates of Halymenia floresia [6] It has also beenreported that monochromatic B LED-light could producethe highest biomass and RGR of Botryococcus braunii [21]


b

aa

a

B R GCKTreatments

00

01

02

03

04

05

06FvF

m

0 200 400 600 800

CKB

RG

0

20

40

60

80

100

rETR

Irradiance (120583mol photonsmminus2 sminus1)

a

b

aa

B R GCKTreatments

00

02

04

06

08

10

qP

b

aa

a

B R GCKTreatments

00

01

02

03

04

05

NPQ

Figure 2 Maximal photochemical yield (119865V119865119898) photochemical quenching (qP) nonphotochemical quenching (NPQ) and rapid lightcurves (RLCs) of Pyropia haitanensis as a function of the light source (comparison via ANOVA) during laboratory culture Values are mean(plusmnSD) 119899 = 9 Different letters indicate statistical significance (119875 lt 005) Error bars are standard deviations

Additionally the enzymes activities involved in carbohydratemetabolism of microalgae could be regulated by B light [22]thus most likely influencing the growth of seaweeds

It has been shown that most green algae grow in theepilittoral or upper zone of water however brown algae growoften in deeper water and many red algae can be describedas subtidal algae [23 24] In the subtidal zone where B andG light prevails the specific photopigment of the red algaeallows efficient absorption [25] In addition several red algalspecies growth rates and photosynthesis depend on thelight quality during the culture period and on the pigmentcomposition under these conditions [19] Although the lightrequirements were very low in G and B light for all redalgae analyzed the action spectrum of growth followed thephotosynthetic action spectrum with maximum efficienciesin G and B wavebands corresponding to the spectrum distri-bution occurring in deep coastal seawater [19] Nonethelesscontrary to our results Kim et al [26] reported thatGracilariatikvahiae presented an inferior growth rate for algae grownunder B light as compared to those grown under R G

and fluorescent light This could be due to algae absorp-tion characteristics depending also on several other factorsespecially the thallus morphology thickness structure ofphotosynthetic system and so forth as it applies more tocoastal waters than oceanic waters [27]

The higher growth rate of P haitanensis in CK B and Glight compared to others can be explained to some extent bythe higher photochemical electron transport rate and therebya higher efficiency of photochemical action and photosyn-thetic efficiency The 119865V119865119898 (photosynthetic efficiency) 120572(light use efficiency the slope of the photosynthesis versusirradiance curve) rETRmax (defined as the photosyntheticcapacity) and qP (photochemical quenching) are frequentlyused to represent the maximum photochemical efficiency ofPSII and proportion of oxidized (open) reaction centers withPSII used as an index of photosynthetic efficiency [28]

In this experiment 119865V119865119898 120572 rETRmax and qP weresignificantly decreased at R light growthwith the intense sup-pression in RGR which implies that a dwindle is indicativeof the decline in photosynthetic efficiency on the growth of


b

abaa

aa

a

a

ab

a

a

b

bab a

a

B R GCKTreatments

B R GCKTreatments

B R GCKTreatments

B R GCKTreatments

00

05

10

15

20

25

30

35

PE (m

ggminus1

FW)

00

01

02

03

04

05

06

PC (m

ggminus1

FW)

0

10

20

30

40

50

60

70

Car (120583

ggminus1

FW)

0

50

100

150

200

250

300

350

400Ch

l a (120583

ggminus1

FW)

Figure 3 Chlorophyll a (Chl a) carotenoid (Car) phycoerythrin (PE) and phycocyanin (PC) of Pyropia haitanensis as a function of the lightsource (comparison via ANOVA) during laboratory culture Values are mean (plusmnSD) 119899 = 3 Different letters indicate statistical significance(119875 lt 005) Error bars are standard deviations

c

bcb

a

cc

b

a

B R GCKTreatments

B R GCKTreatments

00

03

06

09

12

15

CAT

(Umgminus

1pr

ot)

00

05

10

15

20

25

SOD

(Umgminus

1pr

ot)

Figure 4 Superoxide dismutase (SOD) and catalase (CAT) activity of Pyropia haitanensis as a function of the light source (comparison viaANOVA) during laboratory culture Values are mean (plusmnSD) 119899 = 3 Different letters indicate statistical significance (119875 lt 005) Error bars arestandard deviations


P haitanensis Furthermore the suppressed qP value demon-strated that R light irradiance diminished the reoxidizing119876119860capacity This effect may be on account of the inhibition

of Calvin cycle action as indicated by the reduction inCO2assimilation rates which suggests R light irradiance

induction pressure on PSIIThis contributed to the closure ofPSII reaction centers [29] thereby decreasing the probabilityof electron transport from photosystem II to photosystemI and beyond [30] As researchers showed B light couldaugment the number of pigment systems per electron transferchain whereas R light blocks Chl b synthesis and causes adecrease in the function of the light-harvesting system [31]

The NPQ is an important parameter of plant stressresponse which indicates the distribution and the strength ofthe intrathylakoid pH gradient and the ability of chloroplaststo dissipate excess excitation energy as heat This is similarto leaf which is linearly associated with energy dissipationin tissues and is considered as requirement in protectingthe cell from light-induced damage [32] In our studiessignificant decreases in119865V119865119898 and increases inNPQoccurredin R light treatment proposing that R light caused severeabatement of photosynthesis to P haitanensis as well as anaugmentation in thermal dissipation in PSII Furthermorethis mainly suggests that P haitanensis thalli grown at R lightwere responsive to photoinhibition and can cause the ROSgeneration

Superoxide dismutase (SOD) can catalyze the dismuta-tion of superoxide into oxygen and hydrogen peroxide whilecatalase (CAT) is accountable for degrading hydrogen per-oxide into water and oxygen These two enzymes cooperateto convert ROS to H

2O thus enabling the algae to pro-

tect themselves against oxidative stress [33] The influenceof different spectra on the antioxidant enzymes activityin algae has also been studied with high SOD and CATactivities found in the light wavelengths range 440sim480 nmfor zooxanthellae [34] In this experiment a striking rise inSOD and CAT activity was observed in R light treatmentcompared with others These results showed that the highactivity of SOD and CAT is doubtless in response to theoxygen evolving photosynthetic activity in the R light Thisis correlated with the NPQ increase and higher Car contentwhich could help to quench ROS causing less production ofROS in the G lemaneiformis [35]

Light is important for the chlorophyll synthesis and algapigment synthesis is regulated by various photoreceptors thatabsorb lighting for different wavelengths [36 37] In thisinvestigation the effects of light on the Chl a content of Phaitanensis presented a higher effect in CK B and G lighttreatment which was as marked as those on growth rateLopez-Figueroa and Niell [37] have also shown that chloro-phyll synthesis was induced by B light in red algae Corallinaelongata Ellis et Soland and Plocamium cartilagineum (L)Similarly Kim et al [26] proved that chlorophyll concentra-tion was induced under G and R + B LED-light grown in Gtikvahiae However the Car content in R light treatment wasnot significantly decreased It is possible to partially attributethis to the augmentation of the NPQ value Car have a vitalprotective role of PSII as carotenoids can deactivate tripletchlorophyll and singlet oxygen [38]

The higher PE and PC contents were examined in B andGlight grown as expected if the red algae respond to decreasedlighting intensity in the PE absorption spectrum of Pyropiaumbilicalis [39] Moreover Tsekos et al [40] Franklin etal [41] and Barufi et al [5] also found that B light couldrespectively increase the PE and PC concentration of Pyropialeucosticta Chondrus crispus and Gracilaria birdiae B andG light help promote the nitrogen accumulation and thesewavelengths could induce soluble protein and phycobilipro-teins formation [6] The manifestation of the various typesof photopigments in the photosynthesis complexes and theirarrangement in both PSI and PSII are responsible for differ-ent photosynthetic efficiencies of various light wavelengthswhich affect photosynthetic activity in macroalgae

In conclusion R light has disadvantageous effects onthe photosynthetic efficiency and increased growth rate inthalli of P haitanensis Both antioxidant enzymes activityand the NPQ coefficient were increased P haitanensis canbenefit from blue green and fluorescent tubes light Thelight wavelength treatments inmariculture of redmacroalgaecan be used to operate pigment composition and enzymesactivity and further investigations are required to disclosehow the spectral light regulates the biosynthesis process

Competing Interests

The author declares that they have no competing interests

Acknowledgments

This research was supported by the National Natural ScienceFoundation of China (41276148)

References

[1] N Su QWu Z Shen K Xia and J Cui ldquoEffects of light qualityon the chloroplastic ultrastructure and photosynthetic charac-teristics of cucumber seedlingsrdquo Plant Growth Regulation vol73 no 3 pp 227ndash235 2014

[2] T Ouzounis X Frette C-O Ottosen and E Rosenqvist ldquoSpec-tral effects of LEDs on chlorophyll fluorescence and pigmen-tation in Phalaenopsis lsquoVivienrsquo and lsquoPurple Starrsquordquo PhysiologiaPlantarum vol 154 no 2 pp 314ndash327 2015

[3] AManivannan P Soundararajan N Halimah C H Ko and BR Jeong ldquoBlue LED light enhances growth phytochemical con-tents and antioxidant enzyme activities of Rehmannia glutinosacultured in vitrordquo Horticulture Environment and Biotechnologyvol 56 no 1 pp 105ndash113 2015

[4] M Dring ldquoPigment composition and photosynthetic actionspectra of sporophytes of Laminaria (Phaeophyta) grown in dif-ferent light qualities and irradiancesrdquo British Phycological Jour-nal vol 21 no 2 pp 199ndash207 1986

[5] J B Barufi F L Figueroa and E M Plastino ldquoEffects of lightquality on reproduction growth and pigment content of Gra-cilaria birdiae (Rhodophyta Gracilariales)rdquo Scientia Marinavol 79 no 1 pp 15ndash24 2015

[6] J L Godınez-Ortega P Snoeijs D Robledo Y Freile-PelegrınandM Pedersen ldquoGrowth and pigment composition in the redalga Halymenia floresii cultured under different light qualitiesrdquoJournal of Applied Phycology vol 20 no 3 pp 253ndash260 2008


[7] Y Yang and P Weathers ldquoRed light and carbon dioxide dif-ferentially affect growth lipid production and quality in themicroalga Ettlia oleoabundansrdquo Applied Microbiology and Bio-technology vol 99 no 1 pp 489ndash499 2015

[8] F Lopez-Figueroa R Conde-Alvarez and I Gomez ldquoRelationsbetween electron transport rates determined by pulse amplitudemodulated chlorophyll fluorescence and oxygen evolution inmacroalgae under different light conditionsrdquo PhotosynthesisResearch vol 75 no 3 pp 259ndash275 2003

[9] M Kula M Rys K Mozdzen and A Skoczowski ldquoMetabolicactivity the chemical composition of biomass and photosyn-thetic activity of Chlorella vulgaris under different light spectrain photobioreactorsrdquo Engineering in Life Sciences vol 14 no 1pp 57ndash67 2014

[10] P K Boss R M Bastow J S Mylne and C Dean ldquoMultiplepathways in the decision to flower enabling promoting andresettingrdquo Plant Cell vol 16 pp S18ndashS31 2004

[11] DH Zou andK S Gao ldquoExogenous carbon acquisition of pho-tosynthesis in Porphyra haitanensis (Bangiales Rhodophyta)under emersed staterdquo Progress in Natural Science vol 14 no 2pp 138ndash144 2004

[12] U Schreiber H Hormann C Neubauer and C KlughammerldquoAssessment of photosystem II photochemical quantum yieldby chlorophyll fluorescence quenching analysisrdquo AustralianJournal of Plant Physiology vol 22 no 2 pp 209ndash220 1995

[13] A D Jassby and T Platt ldquoMathematical formulation of the rela-tionship between photosynthesis and light for phytoplanktonrdquoLimnology and Oceanography vol 21 no 4 pp 540ndash547 1976

[14] A R Wellburn ldquoThe spectral determination of chlorophylls aand b as well as total carotenoids using various solvents withspectrophotometers of different resolutionrdquo Journal of PlantPhysiology vol 144 no 3 pp 307ndash313 1994

[15] H W Siegelman and J H Kycia ldquoAlgal biliproteinsrdquo in Hand-book of Phycological Methods J A Hellebust and J S CraigieEds Cambridge University Press Cambridge UK 1978

[16] C Beauchamp and I Fridovich ldquoSuperoxide dismutase im-proved assays and an assay applicable to acrylamide gelsrdquo Ana-lytical Biochemistry vol 44 no 1 pp 276ndash287 1971

[17] G Cohen M Kim and V Ogwu ldquoA modified catalase assaysuitable for a plate reader and for the analysis of brain cellculturesrdquo Journal of Neuroscience Methods vol 67 no 1 pp 53ndash56 1996

[18] M M Bradford ldquoA rapid and sensitive method for the quanti-tation of microgram quantities of protein utilizing the principleof protein-dye bindingrdquoAnalytical Biochemistry vol 72 no 1-2pp 248ndash254 1976

[19] P Leukart and K Luning ldquoMinimum spectral light require-ments andmaximum light levels for long-termgermling growthof several red algae from different water depths and a greenalgardquo European Journal of Phycology vol 29 no 2 pp 103ndash1121994

[20] G A Pearson E A Serrao M Dring and R Schmid ldquoBlue-and green-light signals for gamete release in the brown algaSilvetia compressardquo Oecologia vol 138 no 2 pp 193ndash201 2004

[21] C Okumura A N Saffreena M A Rahman H Hasegawa OMiki and A Takimoto ldquoEconomic efficiency of different lightwavelengths and intensities using LEDs for the cultivation ofgreen microalga Botryococcus braunii (NIES-836) for biofuelproductionrdquo Environmental Progress and Sustainable Energyvol 34 no 1 pp 269ndash275 2015

[22] H C P Matthijs H Balke U M VanHes B M A Kroon L RMur and R A Binot ldquoApplication of light-emitting diodes in

bioreactors flashing light effects and energy economy in algalculture (Chlorella pyrenoidosa)rdquo Biotechnology and Bioengineer-ing vol 50 no 1 pp 98ndash107 1996

[23] TW Engelmann ldquoFarbe und assimilationrdquo Botanic Zentro vol41 pp 1ndash29 1883

[24] TW Engelmann ldquoUntersuchungen uber die quantitativen bez-iehungen zwischen absorption des lichtes und assimilation inpflanzenzellenrdquo Botanic Zentro vol 42 pp 82ndash95 1884

[25] R Biebl ldquoSeaweedsrdquo in Physiology and Biochemistry of Algae RA Lewin Ed Academic Press New York NY USA 1962

[26] J K Kim Y X Mao G Kraemer and C Yarish ldquoGrowthand pigment content of Gracilaria tikvahiae McLachlan underfluorescent and LED lightingrdquo Aquaculture vol 436 pp 52ndash572015

[27] A W D Larkum E A Drew and R N Crossett ldquoThe verticaldistribution of attached marine algae in Maltardquo Journal ofEcology vol 55 no 2 pp 361ndash371 1967

[28] H A Baumann LMorrison andD B Stengel ldquoMetal accumu-lation and toxicity measured by PAM-Chlorophyll fluorescencein seven species of marine macroalgaerdquo Ecotoxicology andEnvironmental Safety vol 72 no 4 pp 1063ndash1075 2009

[29] A Calatayud and E Barreno ldquoChlorophyll a fluorescenceantioxidant enzymes and lipid peroxidation in tomato inresponse to ozone and benomylrdquo Environmental Pollution vol115 no 2 pp 283ndash289 2001

[30] G G R Seaton and D A Walker ldquoChlorophyll fluorescence asa measure of photosynthetic carbon assimilationrdquo Proceedingsof the Royal Society B Biological Sciences vol 242 no 1303 pp29ndash35 1990

[31] H Senger K Humbeck and H Schiller ldquoLight adaptation ofthe photosynthetic apparatus of green algaerdquo in EnvironmentalSignal Processing and Adaptation D Werner and G HeldmaierEds Springer Berlin Germany 2002

[32] I Sperdouli andMMoustakas ldquoSpatio-temporal heterogeneityin Arabidopsis thaliana leaves under drought stressrdquo PlantBiology vol 14 no 1 pp 118ndash128 2012

[33] H Qian X Xu W Chen et al ldquoAllelochemical stress causesoxidative damage and inhibition of photosynthesis in Chlorellavulgarisrdquo Chemosphere vol 75 no 3 pp 368ndash375 2009

[34] O Levy Y Achituv Y Z Yacobi N Stambler and Z DubinskyldquoThe impact of spectral composition and light periodicity onthe activity of two antioxidant enzymes (SOD and CAT) in thecoral Favia favusrdquo Journal of Experimental Marine Biology andEcology vol 328 no 1 pp 35ndash46 2006

[35] M Lindahl C Funk J Webster S Bingsmark I Adamskaand B Andersson ldquoExpression of ELIPs and PS II-S proteinin spinach during acclimative reduction of the photosystem IIantenna in response to increased light intensitiesrdquo Photosynthe-sis Research vol 54 no 3 pp 227ndash236 1997

[36] F Lopez-Figueroa R Perez and F X Niell ldquoEffects of red andfar-red light pulses on the chlorophyll and biliprotein accu-mulation in the red alga Corallina elongatardquo Journal of Photo-chemistry and Photobiology B Biology vol 4 no 2 pp 185ndash1931989

[37] F Lopez-Figueroa and F X Niell ldquoEffects of light quality onchlorophyll and biliprotein accumulation in seaweedsrdquoMarineBiology vol 104 no 2 pp 321ndash327 1990

[38] J H Kim B Y Chung J S Kim and S G Wi ldquoEffects ofinplanta gamma-irradiation on growth photosynthesis andantioxidative capacity of red pepper (Capsicum annuum L)plantsrdquo Journal of Plant Biology vol 48 no 1 pp 47ndash56 2005


[39] F L Figueroa J Aguilera and F X Niell ldquoRed and blue lightregulation of growth and photosynthetic metabolism in Por-phyra umbilicalis (Bangiales Rhodophyta)rdquo European Journalof Phycology vol 30 no 1 pp 11ndash18 1995

[40] I Tsekos F X Niell J Aguilera F Lopez-Figueroa and S GDelivopoulos ldquoUltrastructure of the vegetative gametophyticcells of Porphyra leucosticta (Rhodophyta) grown in red blueand green lightrdquo Phycological Research vol 50 no 4 pp 251ndash264 2002

[41] L A Franklin G Krabs and R Kuhlenkamp ldquoBlue light andUV-A radiation control the synthesis of mycosporine-likeamino acids in Chondrus crispus (Florideophyceae)rdquo Journal ofPhycology vol 37 no 2 pp 257ndash270 2001

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Anatomy Research International

PeptidesInternational Journal of


Hindawi Publishing Corporation httpwwwhindawicom

International Journal of

Volume 2014

Zoology


Molecular Biology International

GenomicsInternational Journal of


The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014


BioinformaticsAdvances in

Marine BiologyJournal of



Signal TransductionJournal of


BioMed Research International

Evolutionary BiologyInternational Journal of



Biochemistry Research International

ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014


Genetics Research International


Advances in

Virolog y

Hindawi Publishing Corporationhttpwwwhindawicom

Nucleic AcidsJournal of

Volume 2014

Stem CellsInternational



Enzyme Research



Microbiology


This study focused on the Pyropia haitanensis (BangialesRhodophyta) one species of marine red macroalga whichis an economically important species primarily due to beingan economically important seaweed used for food and isthe principal species cultivated on a commercial scale insouthern China [11] The aim of this study is to illustrate theeffect of different light spectrum on chlorophyll fluorescencecharacteristics the physiological and growth parameters of Phaitanensis and the effect of lighting on the amount of growthrate photopigment composition chlorophyll fluorescenceand antioxidative enzymes activities in a different light spec-trum These results may help us realize the physiologicalstatus of themacroalgae providing data for application of nat-ural populations as well as for the improving culture industryof this species on the coast

2 Materials and Methods

21 Seaweed Materials Pyropia haitanensis was gathered inShantou China (23∘261015840 N 116∘641015840 E) The algae were lightlyrinsed and cleared of visible epiphytes and of any accu-mulated sediments and then placed in a cooler containingsome fresh seawater during the transportation to the labo-ratory (about 3 h 4∘C) The algae were preserved in a glassaquarium tank containing filtered natural seawater (salinity3) appended with 200120583M NaNO

3and 20120583M NaH

2PO4

(final concentration) in a controlled environmental compart-ment Light availability was provided by timer-controlledfluorescent tubes (100 120583mol photos mminus2 sminus1) operated on a12 h on-and-off photoperiod Water motion was providedby aeration and culture seawater was updated every 2 daysand temperature was kept at 15 plusmn 1∘C in the incubator Thealgae were preserved for 3 days prior to the start of theexperiments Only intact and healthy thalli were selected forthe subsequent experimental treatments

22 Experimental Design 5 g fresh weight (FW) algae weretransferred to a flask containing 5 L filtered seawater (threereplicates per light treatment) for a starting biomass densityof 1 g FWLminus1 Different light quality control units wereindependently designed in growth cabinets The cultureswere irradiated at 100120583mol photons mminus2 sminus1 for all treatmentgroups using LED arrays (Edlan Lighting Technology CoChina) red (R 640sim680 nm) blue (B 420sim460 nm) andgreen (G 490sim530 nm) light and fluorescent tubes (CK)light Light apparatus were affixed to a ceramic and steelsupport to facilitate efficient heat transfer to the mountingsubstrate Each light treatment employed 3 replicates All thecultures were preserved at 15 plusmn 1∘C and a lightdark regimeof 1212 h and the seawater nutrients and frequency of waterchanges were the same as indicated above Seaweeds wereharvested and used for experimental measurement after 7days of cultivation

23 Growth Rates Biomass (freshweight FW)wasmeasuredat the end of incubation and the mean relative growth rate(RGR) was assessed according to the formula RGR ( dayminus1)

= ln(1198821199051198820)119905 times 100 where119882

0refers to the initial FW and

119882119905to the FW after 119905 days

24 Chlorophyll Fluorescence Measurements Detection ofchlorophyll fluorescence was made using a pulse modulationfluorometer (JUNIOR-PAM Walz Germany) At least fouralgal samples were used for each measurement of chlorophyllfluorescence and the algae were acclimated to dark for10min before being detected The maximum quantum yieldof photosystem (PS) II of P haitanensis was estimated as119865V119865119898 and photochemical quenching coefficient (qP) andnonphotochemical quenching coefficient (NPQ) were alsodetermined [12] The rapid light curves (RLCs) consisted ofthe fluorescence response to eight different and increasingactinic irradiance levels over the range of 0sim820120583mol pho-tons mminus2 sminus1 The parameters of the RLCs were calculatedfollowing the formula from Jassby and Platt [13] rETR(relative electron transport rate) = rETRmax times tan ℎ(120572 times 119868rETRmax) where rETRmax is the saturated maximum rETRtan ℎ is the hyperbolic tangent function 120572 is the initial slopeof the RLC (the efficiency of the electron transport) and 119868 isthe incident irradiance

25 Pigment Estimation For chlorophyll a (Chl a) and car-otenoid (Car) about 01 g (FW) of thalli was ground in 5mL95 ethanol and extracted at 4∘C in darkness for 24 h Forphycoerythrin (PE) and phycocyanin (PC) 01 g (FW) ofsamples was ground in 10mL extraction buffers (01M phos-phate buffer pH = 68) at 4∘CThe extract was centrifuged at5000timesg for 10min and then used to determine the contents ofChl a Car PE andPCusing an ultraviolet spectrophotometer(UV-180 Shimadzu Japan) Chl a and Car contents wereanalyzed according toWellburn [14] and PE and PC contentswere analyzed according to Siegelman and Kycia [15]

26 Antioxidant Enzyme Activities Superoxide dismutase(SOD) activity was examined by measuring the ability toinhibit reduction of nitro blue tetrazolium (NBT) followingthe method of Beauchamp and Fridovich [16] One unitof enzyme activity was defined as the amount of enzymerequired to inhibit the photoreduction of NBT by 50and the unit is Umg protminus1 [16] Catalase (CAT) activity(Umg protminus1) was analyzed by using a ferrothiocyanatemethod of Cohen et al [17] with somemodifications Proteincontent was analyzed by the Bradford method [18]

27 Statistical Analyses All the data were expressed as meansplusmn SD (119899 ge 3) and one-way ANOVA and Tukey tests wereused to analyze differences among treatments by using SPSS170 The significance level was set at 119875 lt 005

3 Results

31 Growth Pyropia haitanensis maintained a positivegrowth during the period of culture for all the thalli grownunder experimental conditions (Figure 1) The growth rate ofthe thalli grown atCK control showedhigher value (29dminus1)


b

aa

a

B RCK GTreatments

00

05

10

15

20

25

30

35RG

R (

dminus1)














4 Discussion



b

aa

a

B R GCKTreatments

00

01

02

03

04

05

06FvF

m

0 200 400 600 800

CKB

RG

0

20

40

60

80

100

rETR


a

b

aa

B R GCKTreatments

00

02

04

06

08

10

qP

b

aa

a

B R GCKTreatments

00

01

02

03

04

05

NPQ








b

abaa

aa

a

a

ab

a

a

b

bab a

a

B R GCKTreatments

B R GCKTreatments

B R GCKTreatments

B R GCKTreatments

00

05

10

15

20

25

30

35

PE (m

ggminus1

FW)

00

01

02

03

04

05

06

PC (m

ggminus1

FW)

0

10

20

30

40

50

60

70

Car (120583

ggminus1

FW)

0

50

100

150

200

250

300

350

400Ch

l a (120583

ggminus1

FW)


c

bcb

a

cc

b

a

B R GCKTreatments

B R GCKTreatments

00

03

06

09

12

15

CAT

(Umgminus

1pr

ot)

00

05

10

15

20

25

SOD

(Umgminus

1pr

ot)













Competing Interests


Acknowledgments


References




















































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


b

aa

a

B RCK GTreatments

00

05

10

15

20

25

30

35RG

R (

dminus1)














4 Discussion



b

aa

a

B R GCKTreatments

00

01

02

03

04

05

06FvF

m

0 200 400 600 800

CKB

RG

0

20

40

60

80

100

rETR


a

b

aa

B R GCKTreatments

00

02

04

06

08

10

qP

b

aa

a

B R GCKTreatments

00

01

02

03

04

05

NPQ








b

abaa

aa

a

a

ab

a

a

b

bab a

a

B R GCKTreatments

B R GCKTreatments

B R GCKTreatments

B R GCKTreatments

00

05

10

15

20

25

30

35

PE (m

ggminus1

FW)

00

01

02

03

04

05

06

PC (m

ggminus1

FW)

0

10

20

30

40

50

60

70

Car (120583

ggminus1

FW)

0

50

100

150

200

250

300

350

400Ch

l a (120583

ggminus1

FW)


c

bcb

a

cc

b

a

B R GCKTreatments

B R GCKTreatments

00

03

06

09

12

15

CAT

(Umgminus

1pr

ot)

00

05

10

15

20

25

SOD

(Umgminus

1pr

ot)













Competing Interests


Acknowledgments


References




















































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


b

aa

a

B R GCKTreatments

00

01

02

03

04

05

06FvF

m

0 200 400 600 800

CKB

RG

0

20

40

60

80

100

rETR


a

b

aa

B R GCKTreatments

00

02

04

06

08

10

qP

b

aa

a

B R GCKTreatments

00

01

02

03

04

05

NPQ








b

abaa

aa

a

a

ab

a

a

b

bab a

a

B R GCKTreatments

B R GCKTreatments

B R GCKTreatments

B R GCKTreatments

00

05

10

15

20

25

30

35

PE (m

ggminus1

FW)

00

01

02

03

04

05

06

PC (m

ggminus1

FW)

0

10

20

30

40

50

60

70

Car (120583

ggminus1

FW)

0

50

100

150

200

250

300

350

400Ch

l a (120583

ggminus1

FW)


c

bcb

a

cc

b

a

B R GCKTreatments

B R GCKTreatments

00

03

06

09

12

15

CAT

(Umgminus

1pr

ot)

00

05

10

15

20

25

SOD

(Umgminus

1pr

ot)













Competing Interests


Acknowledgments


References




















































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


b

abaa

aa

a

a

ab

a

a

b

bab a

a

B R GCKTreatments

B R GCKTreatments

B R GCKTreatments

B R GCKTreatments

00

05

10

15

20

25

30

35

PE (m

ggminus1

FW)

00

01

02

03

04

05

06

PC (m

ggminus1

FW)

0

10

20

30

40

50

60

70

Car (120583

ggminus1

FW)

0

50

100

150

200

250

300

350

400Ch

l a (120583

ggminus1

FW)


c

bcb

a

cc

b

a

B R GCKTreatments

B R GCKTreatments

00

03

06

09

12

15

CAT

(Umgminus

1pr

ot)

00

05

10

15

20

25

SOD

(Umgminus

1pr

ot)













Competing Interests


Acknowledgments


References




















































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology












Competing Interests


Acknowledgments


References




















































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology














































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

research article effect of different light qualities on growth,...

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