氮肥和鈣肥用量對旺長期烤煙葉片抗氧化酶系統(tǒng)及活性氧的影響

鄒文桐，陳　盛，滕家財

(福建師范大學 福清分校 海洋與生化工程學院，福建 福清 350300)

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氮肥和鈣肥用量對旺長期烤煙葉片抗氧化酶系統(tǒng)及活性氧的影響

鄒文桐，陳盛，滕家財

(福建師范大學 福清分校 海洋與生化工程學院，福建 福清 350300)

氮肥;鈣肥;烤煙;抗氧化酶;活性氧

1　材料與方法

1.1材料

試驗于2012-12-2013-07在福建師范大學福清分校后山盆栽房進行，其中2012-12-2013-02是肥料固定期，2013-03-07是煙葉生長期。供試烤煙品種為G80(福建省煙草公司提供)。供試土壤來源于福建師范大學福清分校周邊水稻田，其基本理化性狀為：pH 6.750，有機質含量41.76 g/kg，堿解氮含量74.50 mg/kg，速效磷含量53.43 mg/kg，速效鉀含量123.3 mg/kg，交換性鈣含量1 553 mg/kg，交換性鎂含量63.48 mg/kg。

1.2方法

1.2.1處理方法試驗采用雙因素(3×4)隨機區(qū)組設計，因素A設N1(CK)、N2、N33個供氮水平，施氮量(以純氮計)分別為0.00，0.12和0.20 g/kg；因素B設 Ca1(CK)、Ca2、Ca3、Ca44個水平，供鈣量(以純鈣計)分別為0.00，0.40，1.00和2.00 g/kg。試驗每盆裝風干土10 kg，栽植烤煙1株，盆距為50 cm，每處理重復4盆，共48盆。氮肥由硝酸銨和硝酸鈣組成，鈣肥由硝酸鈣、氫氧化鈣和磷酸二氫鈣組成。試驗中m(N)∶m(P2O5)∶m(K2O)=1.0∶0.8∶2.5，以常規(guī)施純氮0.12 g/kg來確定各處理的純P2O5用量為0.096 g/kg，純K2O用量為0.30 g/kg，試驗同時施純鎂0.045 g/kg，硼砂6.0 mg/kg，其中磷肥由磷酸氫二銨和磷酸二氫鈣組成，鉀肥用硫酸鉀，鎂肥用硫酸鎂，硼肥用硼砂。在種植烤煙前3個月，將純氮、純磷、純鉀、純鈣等肥料轉換成每盆相應肥料的量施入土壤，保持土壤濕潤，使土壤對施用的肥料進行較充分的吸附固定。

試驗每盆栽煙1株，隨機排列，用去離子水澆灌。于烤煙旺長期(團棵到煙株現蕾這一時期，約25 d，此期是決定葉數、葉片大小、葉質量的關鍵時期，是產量、品質形成的重要階段)從每個處理的4個重復中隨機取3盆，取第17～19位葉片，備測。

1.3數據處理與分析

試驗數據采用DPS 2005軟件進行方差分析，利用Duncan’s新復極差法進行差異顯著性檢驗。

2　結果與分析

2.1氮、鈣用量對旺長期烤煙葉片POD、SOD、CAT、APX活性的影響

通過與F0.05和F0.01進行比較可知，氮肥、鈣肥以及氮肥×鈣肥互作對旺長期烤煙葉片POD、SOD、CAT和APX活性均有極顯著的影響(表1)。

表 1　氮肥、鈣肥及氮肥×鈣肥對旺長期烤煙葉片POD、SOD、CAT、APX活性影響的顯著性分析Table 1　Significant analysis on effects of nitrogen,calcium and their interaction on POD, SOD,CAT,APX activities in flue-cured tobacco leaves during fast growing period

注：表中“*”表示在P=0.05水平上有顯著差異，“**”表示在P=0.01水平上有極顯著差異。表5同。

Note:“*” and “**” mean significant difference atP=0.05 andP=0.01 level,respectively.The same table 5.

由表2可知，隨氮肥用量從N1增大到N3，旺長期烤煙葉片POD、SOD、CAT和APX活性均極顯著升高，SOD活性以N2處理的最高，較N1處理(對照)提高了11.30%，POD、CAT、APX活性均以N3處理最高，分別較N1處理(對照)提高了 33.14%，114.99%和36.91%。

表 2　氮肥用量對旺長期烤煙葉片POD、SOD、CAT和APX的影響Table 2　Effects of nitrogen on POD,SOD,CAT and APX activities in flue-cured tobacco leaves during fast growing period

注：同列數據后標不同小寫字母表示差異顯著(P<0.05)，標不同大寫字母表示差異極顯著(P<0.01)。下表同。

Note:Lowercase and uppercase letters mean significant differences atP<0.05 andP<0.01,respectively.The same below.

由表3可知，隨鈣肥施用量由0.00 g/kg(Ca1處理)增大到1.00 g/kg(Ca3處理)，旺長期烤煙葉片的POD、SOD、CAT和APX活性均極顯著提高，POD、SOD和CAT活性均以Ca3處理最高，與對照(Ca1)相比，分別提高了7.98%，19.74%和 62.48%，APX活性Ca2和Ca3處理均較高，分別較Ca1處理增加了32.92%和30.61%；隨鈣肥用量的繼續(xù)增加，4種酶活性開始呈下降的趨勢。

從表4可以看出，在N1水平下，旺長期烤煙葉片的POD、SOD和CAT活性均以Ca3處理最高，APX活性Ca2和Ca3處理均較高，表明在不施氮條件下，Ca肥用量以1.0 g/kg為宜；在N2水平下，旺長期烤煙葉片的POD、SOD、CAT和APX活性均以Ca3處理的最高，表明在氮肥用量為0.12 g/kg條件下，Ca肥用量以1.0 g/kg為宜；在N3水平下，旺長期烤煙葉片的POD和APX活性均以Ca2和Ca3處理較高，SOD和CAT活性均以Ca3處理的最高，表明在氮肥用量為0.20 g/kg條件下，Ca肥用量以0.40～1.0 g/kg為宜。

表 3　鈣肥用量對旺長期烤煙葉片POD、SOD、CAT和APX的影響Table 3　Effects of calcium on POD,SOD,CAT and APX activities in flue-cured tobacco leaves during fast growing period

表 4　不同施氮量條件下鈣對旺長期烤煙葉片POD、SOD、CAT和APX活性的影響Table 4　Effects of calcium on POD,SOD,CAT and APX activities in flue-cured tobacco leaves during fast growing period under the different nitrogen condition

通過與F0.05和F0.01進行比較可知，氮肥、鈣肥以及氮肥×鈣肥互作對旺長期烤煙葉片脯氨酸含量、MDA含量、過氧化氫含量、超氧陰離子產生速率均有極顯著的影響(表5)。

表 5　氮肥、鈣肥對旺長期烤煙葉片脯氨酸、MDA、H2O2含量產生速率影響的顯著性分析Table 5　Significant analysis on effects of nitrogen,calcium and their interaction on proline, MDA,hydrogen peroxide and producing rate in flue-cured tobacco leaves during fast growing period

表 6　氮肥用量對旺長期烤煙葉片脯氨酸、MDA、H2O2含量和產生速率的影響Table 6　Effects of nitrogen on proline,MDA,hydrogen peroxide and producing rate in flue-cured tobacco leaves during fast growing period

表 7　鈣肥用量對旺長期烤煙葉片脯氨酸、MDA、H2O2含量和產生速率的影響Table 7　Effects of calcium on proline,MDA,hydrogen peroxide and producing rate in flue-cured tobacco leaves during fast growing period

表 8　不同施氮量條件下鈣對旺長期烤煙葉片脯氨酸、MDA、 H2O2含量和產生速率的影響Table 8　Effects of calcium on proline,MDA,hydrogen peroxide and producing rate in flue-cured tobacco leaves during fast growing period under the different nitrogen condition

3　討論與結論

本試驗采用的是二因素隨機區(qū)組設計，從單因素結果來看，氮肥最佳施用量為0.20 g/kg，鈣肥最佳施用量為1.0 g/kg，但是在實際生產中必須考慮其交互作用，在0.12 g/kg施氮量條件下，配施1.0 g/kg Ca和在0.20 g/kg 氮肥用量下，配施1.0 g/kg Ca，均能夠發(fā)揮氮鈣交互作用對烤煙抗氧化酶系統(tǒng)及活性氧的影響，但綜合抗氧化酶及活性氧指標，確定氮肥和鈣肥的最佳配肥組合為N=0.20 g/kg，Ca=1.0 g/kg，此組合配肥能充分發(fā)揮烤煙生長的最大潛能和抗逆性，對提高福建煙葉的抗性和品質具有重要實踐意義。

[1]楊宇虹,馮柱安,晉艷,等.酸性土壤的煙株生長及煙葉產質量調控研究 [J].云南農業(yè)大學學報,2004,19(1):41-44.

Yang Y H,Feng Z A,Jin Y,et al.Influence of acid soil on the quality of tobacco leaf and yield and regulation to tobacco’s growth [J].Journal of Yunnan Agricultural University,2004,19(1):41-44.

[2]曹志洪.優(yōu)質烤煙的土壤與施肥 [M].南京:江蘇科技出版社,1991:101-109.

Cao Z H.Soil and fertilization of high-class flue-cured tobacco [M].Nanjing:Jiangsu Science and Technology Press,1991:101-109.

[3]張仁椒,洪曉薇,李春英,等.土壤有效氮含量對烤煙氮代謝及氮素營養(yǎng)的影響 [J].福建農林大學學報(自然科學版), 2007,36(4):342-346.

Zhang R J,Hong X W,Li C Y,et al.Effects of soil available nitrogen on metabolism and nitrogen nutrition of flue-cured tobacco [J].Journal of Fujian Agriculture and Forestry University(Natural Science Edition),2007,36(4):342-346.

[4]訾大鎮(zhèn),郭月清.煙草栽培 [M].北京:中國農業(yè)出版社,1996:113.

Zi D Z,Guo Y Q.Tobacco cultivation [M].Beijing:Chinese Agricultural Press,1996:113.

[5]Zocchi G.Calcium physiology and metabolism in fruit trees [J].Acta Horticulturae,1995,383:15-23.

[6]Ferguson I B,Brobak B K.Calcium and the regulation of plant growth and senescence [J].Hort Sci,1988,23(2):262-266.

[7]左天覺.煙草的生產、生理和生物化學 [M].朱尊權,譯.上海:上海遠東出版社,1993:367-374.

Zuo T J.Production,physiology and biochemistry of tobacco [M].Zhu Z Q,translate.Shanghai:Shanghai Far East Press,1993:367-374.

[8]唐麗娜,熊德中.酸性土壤施石灰對土壤性質與煙草品質的影響 [J].中國生態(tài)農業(yè)學報,2003,11(3):81-83.

Tang L N,Xiong D Z.Effects of applying lime on the properties of acid soil and the leaves quality in flue-cured tobacco [J].Chinese Journal of Eco-Agriculture,2003,11(3):81-83.

[9]婁春榮,韓曉日,肖千明,等.氮和鈣交互作用對番茄氮素吸收的影響 [J].應用生態(tài)學報,2004,15(4):667-672.

Lou C R,Han X R,Xiao Q M,et al.Interactive effect of N and Ca on N uptake by tomato [J].Chinese Journal of Applied Ecology,2004,15(4):667-672.

[10]李中勇,張媛,韓龍慧,等.氮鈣互作對設施栽培油桃葉片光合特性及葉綠素熒光參數的影響 [J].植物營養(yǎng)與肥料學報,2013,19(4):893-900.

Li Z Y,Zhang Y,Han L H,et al.The interactive effects of nitrogen and calcium on photosynthetic characteristics and chlorophyll fluorescence parameters of nectarine under protected culture [J].Journal of Plant Nutrition and Fertilizer,2013,19(4):893-900.

[11]趙朋,李絮花,王克安,等.氮鈣互作對日光溫室中黃瓜硝酸鹽代謝的影響 [J].山東農業(yè)科學,2008(3):68-70.

Zhao P,Li X H,Wang K A,et al.Interactive effect of N and Ca on nitrate metabolism in greenhouse cucumber [J].Shandong Agricultural Sciences,2008(3):68-70.

[12]張英鵬,李彥,張明文,等.不同氮、鈣營養(yǎng)對菠菜安全品質與抗氧化酶活性的影響 [J].植物營養(yǎng)與肥料學報,2008,14(4):754-760.

Zhang Y P,Li Y,Zhang M W,et al.Effects of nitrogen and calcium nutrition on safety quality and anti-oxidative enzyme activities of spinach [J].Plant Nutrition and Fertilizer Science,2008,14(4):754-760.

[13]Mittler R.Oxidative stress,antioxidants and stress tolerance [J].Trends in Plant Science,2002,7(9):405-410.

[14]Chen G X,Asada K.Ascorbate peroxidase in tea leaves:occurrence of two isozymes and the differences in their enzymatic and molecular properties [J].Plant Cell Physiol,1989,30(7):987.

[15]Nakano Y,Asada K.Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts [J].Plant Cell Physiol,1981,22(5):867.

[16]劉祖祺,張石城.植物抗性生理學 [M].北京:中國農業(yè)出版社,1994:369-385.

Liu Z Q,Zhang S C.Plant stress physiology [M].Beijing:Chinese Agricultural Press,1994:369-385.

[17]Giannopolitis C N,Rice S K.Superoxide dimutase purification and quantitative relationship with water-solube proteinin seedlings [J].Plant Physiol,1997,59:315-318.

[18]王晶英.植物生理生化實驗技術和原理 [M].北京:東北林業(yè)大學出版社,2003:6,83.

Wang J Y.Experimental techniques and principles for plant physiology and biochemistry [M].Beijing:Northeast Forestry University,2003:6,83.

[19]李合生.植物生理生化實驗原理和技術 [M].北京:高等教育出版社,2000:184-185.

Li H S.Experimental principles and techniques for plant physiology and biochemistry [M].Beijing:Higher EducationPress,2000:184-185.

[20]沈文飚,徐朗萊,葉茂炳,等.抗壞血酸過氧化物酶活性測定的探討 [J].植物生理學通訊,1996,32(3):203-205.

Shen W B,Xu L L,Ye M B,et al.Study on determination of ASP activity [J].Plant Physiology Communications,1996,32(3):203-205.

[21]王愛國,羅廣華.植物的超氧物自由基與羥胺反應的定量關系 [J].植物生理學通訊,1990(6):55-57.

Wang A G,Luo G H.Quantitative relation between the reaction of hydroxylamine and superoxide anion radicals in plants [J].Plant Physiology Communications,1990(6):55-57.

[22]林植芳,李雙順,林桂珠,等.衰老葉片和葉綠體中H2O2的累積與膜脂過氧化的關系 [J].植物生理學報,1988,14(1):16-22.

Li Z F,Li S S,Lin G Z,et al.The accumulation of hydrogen peroxide in senescing leaves and chloroplasts in relation to lipid peroxidation [J].Acta Phytophysiologica Sinica,1988,14(1):16-22.

[23]吳平,陳昆松.植物分子生理學進展 [M].杭州:浙江大學出版社,2000:207-215.

Wu P,Chen K S.Advances in plant molecular physiology [M].Hangzhou:Zhejiang University Press,2000:207-215.

[24]任麗花,余華,蔡南通,等.不同氮素水平對菜用甘薯葉片生理特性的影響 [J].中國園藝文摘,2011(6):3-5.

Ren L H,Yu H,Cai N T,et al.Effects of different nitrogen levels on physiological characteristics of vegetable sweet potato [J].Chinese Horticulture Abstracts,2011(6):3-5.

[25]安金花,樸世領,李云善,等.氮缺乏與過剩癥對煙草的影響 [J].延邊大學農學學報,2003,25(1):36-39.

An J H,Piao S L,Li Y S,et al.Effects of lack and surplus of nitrogen on tobacco [J].Journal of Agricultural Science Yanbian University,2003,25(1):36-39.

[26]姜照偉,林文雄,李義珍,等.不同氮肥施用量對再生稻若干生理特性的影響 [J].福建農業(yè)學報,2005,20(3):168-171.

Jiang Z W,Lin W X,Li Y Z,et al.Effects of nitrogen fertilizer rates on some physiological characteristics of ratoon rice [J].Fujian Journal of Agricultural Sciences,2005,20(3):168-171.

[27]翁伯琦,鄭向麗,趙婷,等.不同施氮量對仁稈兩用型花生生理特性及產量、品質的影響 [J].福建農業(yè)學報,2013,28(7):659-663.

Weng B Q,Zheng X L,Zhao T,et al.Effects of different nitrogen fertilizer applications on physiological characteristics,yield and quality of Dual-purpose peanut [J].Fujian Journal of Agricultural Sciences,2013,28(7):659-663.

[28]Sanders D,Brownlee C,Harper J F.Communicating with calcium [J].Plant Cell,1999,11:691-706.

[29]Bowler C,Fluhr R.The role of calcium and activated oxygens as signals for controlling cross-adaption [J].Trends P1ant Sci,2000,5:241-246.

[30]Poovaish B W.Biochemical and molecular aspects of calcium action [J].Acta Horticulturae,1993,326:139-146.

[31]Arora R,Palta J P.Invivoperturbation of membrance-associated calcium by freeze-haw stress in onion bulb cells [J].Plant Physiol,1998,87:622-628.

[32]陳立松,劉星輝.滲透脅迫下Ca2+對龍眼葉片光合作用及膜脂過氧化的影響 [J].園藝學報,1998,25(1):87-88.

Chen L S,Liu X H.Effects of Ca2+on photosynthetic pigments and lipid peroxidation of Longyan leaves under osmotic stress [J].Acta Horticulturae Sinica,1998,25(1):87-88.

[33]王敏,張連斌,張繼澍,等.采后鈣處理對冬棗生理和貯藏品質影響的研究 [J].中國食品學報,2004,4(2):78-82.

Wang M,Zhang L B,Zhang J S,et al.Study on the effect of calcium treatment to physiology and fruit quality in storage of the postharvest Dong Jujube [J].Journal of Chinese Institute of Food Science and Technology,2004,4(2):78-82.

[34]周錄英,李向東,王麗麗,等.鈣肥不同用量對花生生理特性及產量和品質的影響 [J].作物學報,2008,34(5):879-885.

Zhou L Y,Li X D,Wang L L,et al.Effects of different Ca applications on physiological characteristics,yield and quality in peanut [J].Acta Agronomica Sinica,2008,34(5):879-885.

[35]王才斌,吳正鋒,趙品績,等.調環(huán)酸鈣對花生某些生理特性和產量的影響 [J].植物營養(yǎng)與肥料學報,2008,14(6):1160-1164.

Wang C B,Wu Z F,Zhao P J,et al.Effects of prohexadione calcium on some physiological characteristics and pod yield of peanut [J].Plant Nutrition and Fertilizer Science,2008,14(6):1160-1164.

[36]李賀,劉世琦,劉中良,等.鈣對大蒜生理特性及主要礦質元素吸收的影響 [J].中國農業(yè)科學,2013,46(17):3626-3634.

Li H,Liu S Q,Liu Z L,et al.Effects of calcium on physiological characteristics and main mineral elements absorption of Garlic [J].Scientia Agricultura Sinica,2013,46(17):3626-3634.

[37]魏勝林,劉業(yè)好,屈海泳,等.氮鈣對香石竹多酚氧化酶、過氧化物酶和葉斑病的影響 [J].應用生態(tài)學報,2002,13(1):126-127.

Wei S L,Liu Y H,Qu H Y,et al.Effects of nitrogen and calcium on polyphenol oxidase,peroxidase and leaf spot in carnation [J].Chinese Journal of Applied Ecology,2002,13(1):126-127.

Effects of nitrogen and calcium on anti-oxidative enzyme system and active oxygen of flue-cured tobacco leaf during fast growing period

ZOU Wentong,CHEN Sheng,TENG Jiacai

(CollegeofOceanScienceandBiochemistryEngineering,FuqingBranchofFujianNormalUniversity,Fuqing,Fujian350300,China)

nitrogen fertilizer;calcium fertilizer;NicotianatabacumL.;anti-oxidative enzyme;active oxygen

網絡出版時間:2016-07-1208:4510.13207/j.cnki.jnwafu.2016.08.015

2015-01-06

福建省教育廳A類項目(JA12357)；福建省教育廳B類項目(JB12263)

鄒文桐 (1981-),男,福建福清人, 實驗師,碩士,主要從事植物營養(yǎng)生理生化研究。E-mail:wtz_5267370@163.com

Q945.79；S572

A

1671-9387(2016)08-0097-07

網絡出版地址:http://www.cnki.net/kcms/detail/61.1390.S.20160712.0845.030.html