土壤施用硒肥對獼猴桃含硒量、鎘鉛積累及品質(zhì)的影響

龍友華，張 承，吳小毛，*，李 明，姚 雪，邵 雙（.貴州大學農(nóng)學院，貴州 貴陽 550025；2.貴州大學 作物保護研究所，貴州 貴陽 550025）

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土壤施用硒肥對獼猴桃含硒量、鎘鉛積累及品質(zhì)的影響

龍友華1，2，張 承1，吳小毛1，2，*，李 明1，2，姚 雪1，邵 雙1
（1.貴州大學農(nóng)學院，貴州 貴陽 550025；2.貴州大學 作物保護研究所，貴州 貴陽 550025）

摘 要：以“貴長”獼猴桃為實驗材料，研究外源硒對獼猴桃硒含量、鎘和鉛積累和內(nèi)在品質(zhì)的影響。將亞硒酸鈉按0.00、1.25、2.50、5.00、10.00、20.00 mg/kg的劑量分別添加于水、有機肥中進行土壤穴施。結(jié)果表明：硒肥添加于水、有機肥中均能增加獼猴桃硒含量，降低鎘和鉛的積累，改善獼猴桃果實品質(zhì)；但硒肥添加于有機肥中效果更為顯著。隨著硒肥中硒含量的增加，獼猴桃果實中硒含量逐漸增加，鎘和鉛含量急劇降低。在1.25～10.00 mg/kg的亞硒酸鈉施用含量范圍內(nèi)，獼猴桃果實VC、可溶性總糖、可溶性固形物、干物質(zhì)和可滴定酸的含量得到顯著改善。生產(chǎn)上推薦使用5.00～10.00 mg/kg的亞硒酸鈉添加于有機肥中進行土壤穴施。

關(guān)鍵詞：獼猴桃；有機肥；富硒；鎘鉛積累；品質(zhì)

引文格式：

LONG Youhua， ZHANG Cheng， WU Xiaomao， et al.Application of Se-fertilizer affects selenium content， cadmium and lead accumulation and fruit quality in kiwifruits［J］.食品科學， 2016， 37（13）: 82-88.DOI:10.7506/spkx1002-6630-201613015. http://www.spkx.net.cn

LONG Youhua， ZHANG Cheng， WU Xiaomao， et al.Application of Se-fertilizer affects selenium content， cadmium and lead accumulation and fruit quality in kiwifruits［J］.Food Science， 2016， 37（13）: 82-88.（in English with Chinese abstract）

Selenium （Se） is a trace element of fundamental importance to many organisms， including humans and other animals， and a beneficial nutrient for many plants.Se deficiency may damage the development and function of the immune system， which tends to enhance susceptibility viral infections and heart problems， such as heart disease，liver cancer， hypothyroidism［1-2］.As a crucial component of glutathione peroxidase （GSH-Px） and thioredoxin educates，Se exerts positive impact on the growth improvement of plants， increase of antioxidative capacity， reduction of reactive oxygen species （ROS） and lipid peroxidation， delay of senescence， alleviate of heavy metals toxicity and enhance of photosynthesis［3］.

Cadmium （Cd） and lead （Pb） are the most dangerous and widespread environmental contaminants［4-5］， especially in agricultural soils， have a direct risk to both human and ecological receptors due to their relatively high toxicity and plant readily uptake［6］.Numerous studies have shown that Cd and Pb can do obviously harm to nervous system，bone marrow hematopoietic function， digestive system，reproductive system and other human body function，and induce the wrong expression of gene and arouse the cancer［7-8］.In plants， Cd and Pb can cause oxidative stress，inhibit photosynthesis and decrease the chlorophyll contents，destroy the cell membrane integrity， and inhibit the uptake of certain essential elements such as Zn， Fe， Cu and Mg［9］.So it is important to develop reliable approaches to prevent Cd and Pb accumulation in agricultural products.It is interesting to note that Se is one of the potential antagonists to Cd and Pb， recent publications indicate that Se addition may also alter the total content of Cd and Pb by reducing their uptake by plants such as Pteris vittata L.， Sinapis alba L.， Brassica rapa and Lactuca sativa L.［9-11］.Similar results were observed on broccoli， strawberry， sweet persimmon， watermelon， and potato［12-14］.Why Se can cause the antagonism of Cd and Pb in plants？ There are two possible mechanisms.Firstly， it may affect by regulating the uptake and redistribution of essential elements in the antioxidative systems or in maintaining the ion balance and structural integrity of the cell.Secondly，it may interfere with electron transport by affecting the assembly of the photosynthesis complexes［15］.

Organic fertilizer， a green fertilizer， not only can provide more comprehensive nutrition for crops， but increase and update organic matter of soil， promote the breeding of microorganism， improve soil physical and chemical properties， compared with chemical fertilizer.Studies have shown that organic fertilizer can improve yield and quality of crops， and increase the stability of heavy metals in the soil and reduce the enrichment of heavy metal in agricultural products［16-18］.On one hand， organic fertilizer has abundant functional groups and large specific surface area that can increase precipitation of heavy metals in the form of sulfide.On the other hand， heavy metals can generate stable clathrate with organic ligands in organic fertilizer， so as to reduce the bioavailability of heavy metal ions［19］.

As an emerging fruit of world recognized［20］， kiwifruit has a high nutritional value that derives from the fruit containing a variety of amino acids and vitamins， and is rich in minerals， which plays a key role in maintaining human health.The harvested area and yield of kiwifruit in the world continuously increased in the 21stCentury.In China， kiwifruit industry had rapid development， and cultivation area and yield were rank first in the world.However， the cultivation technique deficiency and agricultural contamination may cause the kiwifruit quality and safety problem.In view of the importance of kiwifruit and Se for human nutrition and health， the aim of this study was to investigate the potential role of exogenous Se by different application forms on the selenium-enriched，accumulation of Cd and Pb， and fruit internal quality in kiwifruit.The present study can provide a basis for developing strategies to lower risks associated with Cd and Pb toxicity and cultivating selenium-enriched plant products.

1 Materials and Methods

1.1 Study site and plant material

Experiments were carried out on kiwifruit cultivar ‘Guichang'， planted in 2003， in the kiwifruit garden in 2013 at Xiuwen county， Guiyang， China （26?48′36.1″N，106?28′25.3″E； 1 354 m）.The kiwifruit cultivar ‘Guichang' has long storage period， good quality and high yield performance， and planting area in Southwest China reached 20 000 hm2.The “T” type frame of kiwifruit， with spacing of 3.0 m × 3.0 m， the proportion of male: female vines was 1:8.Mean temperature in the kiwifruit garden was about 15－16 ℃， rainfall was 1 293 mm， and soil was loam.The soil （0－60 cm in deep） had total organic matter content of 29.54 g/kg， total nitrogen of 1.42 g/kg， alkali-hydrolyzable nitrogen of 98.45 mg/kg， available phosphorus of 4.40 mg/kg，available potassium of 0.61 mg/kg， available iron of 49.31 mg/kg， available zinc of 1.89 mg/kg， available manganese of 19.17 mg/kg ， total zinc of 50.66 mg/kg， totalchromium of 0.56 mg/kg， total Pb of 29.30 mg/kg， total Se of 77.69 μg/kg.The pH value of the planting soil was 5.82.

1.2 Chemicals and Instruments

Sodium selenite was purchased from Xiya Reagent （AR，Chengdu， China）； Standard solutions of Se， Cd and Pb were purchased from SIMT （GBW（E）， Shanghai， China）； Organic fertilizer was purchased from Dibao Biological Technology （Guiyang， China）， and its organic matter content is more than 45%， （N+P2O5+K2O） content is more than 5%； Microwave dissolver was produced by SINEO （Shanghai， China）；AF-7500 double-channel hydride atomic fluorescence photometer was obtained from East & West Analytical Instruments； TAS-986 atomic absorption spectrophotometer was obtained from Purkinje General Instrument （Beijing， China）.

1.3 Experimental design

Sodium selenite was spiked with the irrigation water and organic fertilizer at a concentration of 0.00， 1.25， 2.50，5.00， 10.00， 20.00 mg/kg.And a total of twelve treatments，forty-eight plots were arranged in a randomized block design with four replicates.Each plot consisted of six trees， only the interior four trees were used for measurements.On 29thApril and 2edAugust， six sodium selenite treatments （0.00， 1.25，2.50， 5.00， 10.00， 20.00 mg/kg as sodium selenite in water solution and organic fertilizer） were applied two times by soil deep-ditch fertilization.Soil deep-ditch was excavated by digging around and close to the plant roots which the deep and wide of circular ditch （30－50 cm）.

1.4 Plant sampling and analysis

Kiwifruit samples of 100 fruits were collected randomly from each plot on October 1st， 2013 and stored at －20 ℃until analysis.VC， total soluble solids， total soluble sugar，dry matter percentage， titratable acidity， as well as Se， Cd and Pb contents were evaluated.The VC content was analyzed by 2，6-dichlorophenolindophenol titration methods， the total soluble solid was measured with a digital refractometer.Total soluble sugar was analyzed by anthrone colorimetric method， and dry matter percentage （%） was evaluated by the difference between weighting the samples before and after 48 h in a stove at 100 ℃， while titratable acidity was analyzed by acid-base titration.

Total Se content in fruit samples was determined using atomic fluorescence spectrometry （AFS）.Before the measurement， samples of 1.000 g were digested with 6 mL of concentrated HNO3and 2 mL of 30% H2O2in a microwave digestion system.Cd and Pb contents were analyzed by the atomic absorption spectrometry （AAS） method with digestion of 1.000 g fruit samples with a mixture of 6 mL HNO3and 4 mL HClO4.After digestion， the residues were diluted to 25 mL with ultrapure water.

1.5 Statistical analyses

2 Results and Analysis

2.1 Effects of Se-fertilizer on Se content in kiwifruit

The soil deep-ditch fertilization of sodium selenite in the irrigation water and organic fertilizer were highly effective for increasing total Se content in kiwifruit， which increased gradually with the increasing input of sodium selenite （Fig.1）.While sodium selenite was added in the irrigation water， Se content in kiwifruit for treatments of 1.25， 2.50 mg/kg sodium selenite was not significantly different， compared to 0.00 mg/kg sodium selenite （Fig.1）.The total Se content in samples from sodium selenite in the irrigation water and organic fertilizer was 0.017 6－0.021 6 mg/kg and 0.017 9－0.067 4 mg/kg FW （in fresh weight）， respectively.The application of 2.50，5.00， 10.00 and 20.00 mg/kg levels of sodium selenite in organic fertilizer led to the greatest increase of Se content in kiwifruit with content values of 0.026 7， 0.037 9， 0.045 8 and 0.067 4 mg/kg FW， respectively.It was 1.46， 2.02，2.26 and 3.11 times higher than the corresponding levels of sodium selenite in irrigation water， respectively （Fig.1）.This indicates the application of sodium selenite combined with organic fertilizer can better promote the intake of selenium by kiwifruit.The possible reasons that application of organic fertilizer could quickly increase soil organic matter content and microbial quantity， enhance the activity of soil enzyme and root system［21-22］， and then promote plant intake mineral elements from soil.

Human Se intake is determined largely by food Se composition.Many areas in the world contain low Se levels，such as in Europe， Australia， New Zealand， India， Thailand and China［23］.It is estimated that approximately more than 1 billion people worldwide are Se deficient， considered as the fourth most serious deficiency of minerals in human body［24］.According to the Food and Agriculture Organization of the United Nations （FAO）， the recommended minimum daily Se intake for adults is 70 μg/d， with a maximum tolerable level of 400 μg/d.While the World Health Organization recommends 40 μg/d，supplementation at 50－200 μg/d is nontoxic and improves immune responses， reduces bacterial and viral infections，and the onset of many heart diseases［23］.Based on the present results， the Se-enriched kiwifruit could contribute to the dietary Se intake and kiwifruit commodity value.

2.2 Effects of Se-fertilizer on Cd content in kiwifruit

Cd content in kiwifruit was markedly affected by the different sodium selenite regimes （Fig.2）.Sodium selenite at all applied levels induced a significant decrease of internal Cd concentrations in kiwifruit， especially the treatments that sodium selenite was added in the organic fertilizer.The reduction in Cd content in kiwifruit at 1.25－20.00 mg/kg levels of sodium selenite in both irrigation water and organic fertilizer with values of 4.59%－11.46% and 9.72%－29.33%， respectively， as compared to 0.00 mg/kg sodium selenite of irrigation water （Fig.2）.Whereas 0.00 mg/kg and 10.00 mg/kg sodium selenite in organic fertilizer also reduced Cd content by 5.58% and 25.81%， respectively （Fig.2）.

Strategies for reducing Cd contamination and related health risks are urgently desired in recent years.The alleviated Cd toxicity by Se application related to reduce Cd intake has been reported［25-26］.Furthermore， organic matter also has become a growing concern in decrease the mobility of heavy metals by sorption and precipitation［27-29］， hindering heavy metals into biological cycles.The results by Karlsson［30］indicate that Cd could form stable clathrate with the carboxyl （RCOOH） and sulfhydryl （RSH） groups in organic matter.In this study， sodium selenite applied with combination of organic fertilizer had better effect on significant reducing Cd content in kiwifruit than single application of sodium selenite or organic fertilizer （Fig.2）.The results presented here show that the mixed fertilization of sodium selenite and organic fertilizer should have obviously synergistic effects to relieve Cd contamination in kiwifruit.

2.3 Effects of Se-fertilizer on Pb content in kiwifruit

The data presented in Fig.3 shows that 0.00 mg/kg sodium selenite in irrigation water resulted in a high Pb accumulation within fruits of kiwifruit， reaching 2.76 μg/kg FW.Pb content in 0.00 mg/kg sodium selenite treatments was 1.07－1.28 and 1.11－1.69 times higher than that of 1.25－20.00 mg/kg levels of sodium selenite in irrigation water and organic fertilizer， respectively.As indicated， the different sodium selenite regimes significantly decreased the kiwifruit plant intake of Pb from soil， which decreased with the increase of application doses， and a similar phenomenon was observed for Cd accumulation.The effect of the sodium selenite added in organic fertilizer was greater than in irrigation water.As compared with 0.00 mg/kg sodium selenite in irrigation water， 10.00 mg/kg and 20.00 mg/kg sodium selenite in organic fertilizer could reduce 33.01% and 40.66% Pb content in kiwifruit， respectively.The application of 20.00 mg/kg sodium selenite in irrigation water also could reduce 21.71% Pb content （Fig.3）.

Se can reduce Pb content in plants and antagonize Pb toxicity by preventing damage from oxygen free radicals or by forming Se-Pb protein complexes， such as reducing superoxide anion （O·） and increasing the activity and content of GSH-Px， superoxide dismutase （SOD） or thyroid stimulating hormone （TSH） etc.［31-33］.Additionally， related studies indicate that the increases of soil pH value and organic matter caused the reduction of Pb in plants［27-28，34］，while organic fertilizer could precisely increase soil organic matter content and improve soil acidity.In the present study，single fertilization of sodium selenite or organic fertilizer，and their mixed fertilization were able to reduce the content of Pb in kiwifruit （Fig.3）.Compared to alleviating Cd contamination， the synergistic effects for Pb contamination was more noticeably of sodium selenite and organic fertilizer.

2.4 Effects of Se-fertilizer on fruit quality in kiwifruit

The nutritional parameters in fruits of kiwifruit are shown in Table 1.The appropriate doses of sodium selenite treatments influenced VC， soluble total sugar， soluble solid，titratable acid， dry matter content and sugar-acid ratio of kiwifruit， and improved fruit internal quality of kiwifruit.With the increasing levels of sodium selenite to 10.00 mg/kg，all quality parameters in fruits gradually increased （Table 1）.However， these quality parameter values did not increase when the dose of sodium selenite was more than 10.00 mg/kg.The results show that after soil application with sodium selenite （from 1.25 to 10.00 mg/kg ） in irrigation water and organic fertilizer， VC， soluble total sugar， soluble solid， and dry matter content of kiwifruit was increased by 1.51%－19.93% and 9.36%－32.53%， 0.83%－15.84% and 4.82%－26.35%， 0.82%－8.05% and 2.53%－11.67%， and 0.83%－6.38% and 2.78%－13.92%， respectively， as compared to 0.00 mg/kg sodium selenite in irrigation water.While titratable acidity of kiwifruit was reduced by 1.27%－9.43% and 5.03%－12.58% （Table 1）.It indicates that soil application with sodium selenite in organic fertilizer had more effective improvement to fruit internal quality of kiwifruit.And the mixed fertilization of sodium selenite and organic fertilizer should have obviously synergistic effects.

Table 1 VC， soluble total sugar， soluble solid， titratableacid， anddrymatter contents and sugar-acid ratio in kiwifruits Treatments Addition/ （mg/kg）VC comtent/ （mg/kg FW）Total soluble sugar content/% Total soluble solid content/% Titratable acidity content/% Dry matter content/% Sugar-acidity ratio Added in the irrigation water 0.00 1 093.12±0.70fJ10.78±0.21dI14.65±0.02eI1.59±0.04aA20.54±0.91aE6.78±0.07dH1.25 1 109.59±0.11eI10.87±0.03dI14.77±0.10dH1.57±0.04abAB20.71±1.08aDE6.92±0.20dH2.50 1 157.43±1.06dH11.25±0.06cG15.04±0.06cF1.54±0.01bBC21.14±0.67aCDE7.31±0.06cFG5.00 1 245.78±0.68cF12.15±0.14bD15.31±0.02bDE1.48±0.02cDE21.33±0.77aCDE8.21±0.05bD10.00 1 311.00±0.92aD12.46±0.11aC15.83±0.05aC1.44±0.02cEFG21.85±0.43aBCD8.65±0.07aC20.00 1 289.22±0.05bE11.97±0.04bE15.35±0.03bD1.47±0.02cDEF21.35±0.33aCDE8.14±0.09bDEAdded in the organic fertilizer 0.00 1 161.86±0.13fH11.08±0.07fH14.93±0.06dG1.54±0.03aBC20.92±0.32cDE7.19±0.09eG1.25 1 195.43±0.29eG11.30±0.05eG15.02±0.09dFG1.51±0.02aCD21.11±0.28cCDE7.48±0.07dF2.50 1 246.21±0.40dF11.61±0.05dF15.24±0.06cE1.46±0.03bEF21.53±0.30cCDE7.95±0.22cE5.00 1 320.57±0.35cC12.88±0.05bB15.79±0.06bC1.43±0.03bcFG22.80±0.28abAB9.01±0.13bB10.00 1 448.70±0.32aA13.62±0.00aA16.36±0.04aA1.39±0.02cH23.40±0.44aA9.80±0.12aA20.00 1 398.61±0.16bB12.76±0.02cB16.25±0.04aB1.41±0.02cGH22.27±0.41bBC9.05±0.12bB

Se is primarily assimilated from the soil by plants as selenate （SeO） or selenite （SeO）.After intake， it has been proposed to be transported into the chloroplasts， inadvertently via sulfate transporters， metabolized it through the sulfate assimilation pathway， incorporated into multiple organic selenium compounds and dedicated vital potential role to growth of plants［3，35］.Se has two distinct effects on plants: low concentration of selenium can enhance photosynthesis and promote plant growth， lower lipid peroxidation and alleviate influences of adversity［36-37］.And other a high dose of Se may lead to plants poisoning， decrease quality and yield［3］.Studies have shown that organic fertilizer has a good role in promoting on yield formation and quality improvement of kiwifruit， also can extend kiwifruit storage period［38］.The results from this study show that organic fertilizer and appropriate doses of sodium selenite could enhance kiwifruit quality， and the effect of their mixed fertilization was better （Table 1）.However， more than 10.00 mg/kg sodium selenite may cause kiwifruit poisoning due to a high concentration of Se in kiwifruit and the nutritional parameters did not continue to increase.Therefore， 5.00－10.00 mg/kg sodium selenite in organic fertilizer was recommended for cultivation of kiwifruit.Sodium selenite was applied with combinationof organic fertilizer could be applicable to the development of excellent Se-enriched kiwifruit and remediation of heavy metals （such as Cd and Pb） contamination.

3 Conclusions

The results from a field experiment show soil fertilization of appropriate dosage of Se in both irrigation water and organic fertilizer could reliably increase Se content， notably decrease the accumulation of Cd and Pb， and effectively improve quality of kiwifruit cultivar ‘Guichang'.Furthermore， application of sodium selenite combined with organic fertilizer was more effective than with irrigation water solution.Based on results of the current study， soil application of Se with combination of organic fertilizer might provide a promising strategy for cultivating selenium-enriched kiwifruit and further reducing kiwifruit heavy metals content to guarantee quality and safety of agriculture products.

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DOI:10.7506/spkx1002-6630-201613015 10.7506/spkx1002-6630-201613015. http://www.spkx.net.cn

中圖分類號：S663.4

文獻標志碼：A

文章編號：1002-6630（2016）13-0082-07

收稿日期：2015-10-26

基金項目：貴州省科技廳農(nóng)業(yè)攻關(guān)項目（（2009）3022；（2011）3024；（2012）3010）；國家自然科學基金地區(qū)科學基金項目（21267007）；貴陽市科技技術(shù)計劃項目（（2009）2-007）；三穗縣科技成果應用及產(chǎn)業(yè)化項目（201602）

作者簡介：龍友華（1970—），男，副教授，博士，研究方向為獼猴桃栽培育種與農(nóng)產(chǎn)品安全評價。E-mail：gzlyh126@126.com

*通信作者：吳小毛（1978—），男，副教授，博士，研究方向為農(nóng)產(chǎn)品安全評價。E-mail：wuxm827@126.com

Application of Se-Fertilizer Affects Selenium Content， Cadmium and Lead Accumulation and Fruit Quality in Kiwifruits

LONG Youhua1，2， ZHANG Cheng1， WU Xiaomao1，2，*， LI Ming1，2， YAO Xue1， SHAO Shuang1
（1.College of Agriculture， Guizhou University， Guiyang 550025， China；2.Institute of Crop Protection， Guizhou University， Guiyang 550025， China）

Abstract:The aim of the present study was to evaluate the effect of exogenous selenium （Se） on Se content， cadmium （Cd）and lead （Pb） accumulation， and internal quality of the kiwifruit cultivar ‘Guichang'.The kiwifruit plants growing under conventional conditions were supplied with sodium selenite at 0.00， 1.25， 2.50， 5.00， 10.00 or 20.00 mg/kg in irrigation water and organic fertilizer， respectively， via soil deep-ditch fertilization.The results showed that the application of exogenous Se in both irrigation water and organic fertilizer could increase Se content， decrease Cd and Pb accumulation， and improve fruit quality in kiwifruits.Moreover， the supplement of sodium selenite was more effective in organic fertilizer than in irrigation water.With increasing amount of Se supplement， Se content in kiwifruits gradually increased and Cd and Pb contents in kiwifruits sharply decreased.At sodium selenite dose levels of 1.25-10.00 mg/kg， the contents of vitamin C， soluble sugar， soluble solid and dry matter were increased， and titratable acid was decreased in kiwifruit.Sodium selenite in organic fertilizer at dose levels of 5.00-10.00 mg/kg was recommended for cultivation of the kiwifruit cultivar ‘Guichang'.These findings might have practical implications for the development of selenium-enriched kiwifruits and remediation of soil heavy metal contamination.

Key words:kiwifruit； organic fertilizer； selenium-enriched； Cd and Pb accumulation； quality