灌漿初期高溫影響水稻籽粒充實和劍葉理化特性-畢業(yè)論文

灌漿初期高溫影響水稻籽粒充實和劍葉理化特性摘要：為探明灌漿初期高溫熱害降低水稻籽粒充實度的可能生理原因，本文以遺傳背景相近而灌漿期耐熱性存在顯著差異的2個水稻純系XN0437T（耐熱）和XN0437S（熱敏感）為材料，于水稻抽穗后第10天進行高溫處理，調(diào)查了水稻籽粒充實度，測定了與水稻劍葉光合效率、細胞膜透性和滲透調(diào)節(jié)有關(guān)的生理指標。結(jié)果表明，高溫處理后水稻籽粒充實度、葉綠素含量、劍葉的凈光合速率和可溶性糖含量降低，熱敏感純系的下降幅度大于耐熱純系；超氧化物歧化酶（SOD）活性、細胞膜透性、丙二醛（MDA）和脯氨酸（Pro）含量增加，其中熱敏感純系的MDA含量、細胞膜透性增加幅度大于耐熱純系，而其SOD活性、脯氨酸含量的增加幅度小于耐熱純系。灌漿初期高溫抑制水稻劍葉光合效率，增加細胞膜透性和改變細胞內(nèi)環(huán)境是高溫熱害降低水稻籽粒充實度的生理原因。關(guān)鍵詞:水稻;高溫;籽粒充實度;理化特性Effectsofhightemperatureonricegrain-fillingofcaryopsisandphysiologicalbiochemicalcharacteristicofflagleaveatearlymilkystageofriceTheaimofthisexperimentwastoascertainpossiblephysiologicalcausesofhightemperaturedecreasinggrain-fillingrateatearlymilkystageofrice.Tworiceinbredlines,namedXN0437T(heat-sensitive)andXN0437S(heat-tolerant),wereselectedandtreatedbyhightemperatureatthe10thdayafterheading.Thegrainplumpnessrate,indexesrelatedtophotosyntheticefficiency,cellmembranepermeabilityandosmoticadjustmentinflagleavesweredetectedinricelinesXN0437TandXN0437S.TheresultshowedthatthegrainplumpnessratewasreducedunderhightemperatureinbothXN0437TandXN0437Sandthereducedrangeofheat-sensitivelinewasgreaterthanthatofheat-tolerantline.Thenetphotosyntheticrate,chlorophyllcontentandsolublesugarcontentweredecreasedincomparisonwithcontrolbothinheat-sensitiveandheat-tolerantricelines,andthedecreaserangeofthesephysiologicalmattercontentsweregreaterintheheat-sensitivelinethanthatoftheheat-tolerantline.Thesuperoxidedismutase(SOD)activity,malonaldehyde(MDA)content,cellmembranepermeabilityandproline(Pro)contentwereincreasedunderhightemperature,andtheincreasedrangeofMDAcontentandmembranepermeabilityinheat-sensitivelinewerehigherthanthatinheat-tolerantline,whiletheincreasedrangeofSODactivity,Procontentinheat-sensitivelinewaslessthanthatinheat-tolerantline.Thepresentstudysuggestedthatdecreasingthephotosyntheticefficiencyofflagleaves,damagingthestructureandfunctionofcellmembrane,andresultintheintracellularenvironmentalteringwerethephysiologicalcausesofhightemperatureatearlymilkystagelimitingthegrainplumpnessofrice.Keywords:rice;hightemperature;grainplumpness;physiologicalandbiochemicalcharacteristics高溫是限制作物產(chǎn)量和品質(zhì)的重要環(huán)境因子[1-3]，不幸的是，作物不得不適應這種高溫環(huán)境，因為過去溫室氣體的排放，導致今后幾十年全球氣候?qū)⒊掷m(xù)上升；而且短期的夏季高溫現(xiàn)象將較過去出現(xiàn)得更加頻繁且持續(xù)時間更長[4、5]。水稻起源于熱帶和亞熱帶地區(qū)，對高溫氣候具有一定的忍耐能力，但是溫度超過水稻最適生長的臨界值時，便會影響其正常生長[6]。高溫熱害對水稻的全生育期都可形成危害，影響水稻產(chǎn)量最嚴重的生育時期是開花期和灌漿期；開花期遇高溫熱害影響水稻花藥開裂與散粉而造成不正常受精，最終形成未受精的空殼籽粒而造成產(chǎn)量損失[3、7-9]，而灌漿期尤其是灌漿初期（花后10-15天）遇高溫熱害則形成灌漿不充分的秕粒，降低籽粒粒重而最終造成減產(chǎn)[8、10]。在我國，夏季高溫熱害主要發(fā)生在長江流域及其以南的部分地區(qū)，一般存在兩次高溫過程，其中一次出現(xiàn)在出梅后的7月上、中旬，由于西太平洋副熱帶高壓呈東西帶狀分布，脊線穩(wěn)定在北緯28-32度，控制著長江中下游地區(qū)，于是該區(qū)域在較長時間內(nèi)維持高溫天氣，經(jīng)常連續(xù)數(shù)日日平均氣溫達35℃，有時部分地區(qū)日最高氣溫在40℃以上[11]。此時正值長江流域以南地區(qū)雙季早稻的灌漿初期，經(jīng)常形成“籽粒高溫逼熟”，造成水稻產(chǎn)量損失。而雙季早稻又是我國糧食生產(chǎn)過程中不可替代的一茬糧食作物，稻谷產(chǎn)量幾次大的波動都與早稻面積和產(chǎn)量密切相關(guān)[12]關(guān)于高溫對水稻功能葉的生理影響已有諸多報道，學者們認為高溫導致功能葉的光合效率降低，膜透性增加，葉綠素、可溶性糖、可溶性蛋白質(zhì)和游離脯氨酸的含量以及SOD活性升高[13、14]。這些研究主要探討了抽穗至開花期高溫熱害對水稻的生理影響，而關(guān)于灌漿初期高溫熱害影響水稻籽粒充實度的生理原因分析鮮見報道[8]。為此，本文以本課題組選育的遺傳背景相似（SSR分子標記檢測結(jié)果）而灌漿初期對高溫熱害忍耐力（結(jié)實率調(diào)查結(jié)果）存在顯著差異的水稻純系XN0437T（耐高溫）和XN0437S（高溫敏感）為材料[16]，在灌漿初期（抽穗后第10天）進行高溫處理，測定了水稻劍葉與光合效率和理化特性有關(guān)的生理指標，水稻成熟后調(diào)查了籽粒充實度，擬探明灌漿初期高溫熱害降低水稻籽粒充實度的可能生理原因，旨在為選育灌漿初期耐熱的水稻新品種提供數(shù)據(jù)參考。2材料與方法2.1試驗材料試驗材料為耐熱水稻純系XN0437T和熱敏感純系XN0437S，它們是協(xié)青早B/N22//協(xié)青早B的回交重組自交系，且遺傳背景相近而灌漿初期耐熱性存在顯著差異[16]。2.2水稻材料的培育水稻的培育采用桶（高為50㎝，內(nèi)徑30㎝）栽的方法，稻田土壤經(jīng)曬干、粉碎、過篩和攪拌均勻后等量裝桶，每桶20㎏，離桶內(nèi)土表15㎝層面上均勻施用1g總氮含量15%、有效磷含量4%和有效鉀含量6%的水稻專用肥、作為基肥，土壤經(jīng)自來水浸泡5d后待用。稻種播種于大田，3葉1心移栽入桶，每桶環(huán)形栽植生育進程與長勢基本一致的單本秧苗10棵；在自然條件下按照常規(guī)栽培管理方法培育至抽穗期；并標記同一天抽穗的主莖，以保證所取樣品的生長發(fā)育進程基本一致。2.3高溫處理與取樣高溫處理在人工氣候室內(nèi)進行。帶標記的主莖抽穗后第10天開始進行高溫處理，高溫處理和平行對照的溫度分別為38.0±0.5℃和25.0±0.5℃[16]，處理時間設(shè)3、6、9、24、48、72、96和120小時；光照時長14小時，相對濕度80±5%；高溫處理設(shè)置3次重復。高溫處理結(jié)束后，水稻置于自然條件下恢復生長24小時，各處理取生育進程一致的水稻劍葉約2..3水稻籽粒充實度調(diào)查高溫處理結(jié)束后，水稻移至自然條件下按常規(guī)栽培管理方法培育成熟，每處理收獲帶標記的主莖6穗，調(diào)查籽粒充實度。籽粒充實度調(diào)查參照張宏玉等的方法[17]，取樣當時將脫粒后的每穗谷粒置于自來水中漂選，下沉者為飽粒，上浮者為空粒和秕粒，曬干后用自制透光裝置將空殼粒和秕粒分開（無胚者為空殼粒，有胚者為秕粒），各處理的實粒和秕粒在統(tǒng)一平衡水分含量后分別稱重。籽粒充實度（%）=100×（處理每穗的實粒重+秕粒重）/（對照每穗的實粒重+秕粒重）。2.4水稻劍葉的凈光合作用速率和葉綠素含量（SPAD值）測定凈光合作用速率采用美國Gene公司的LI-6400型便攜式自動光合測定系統(tǒng)測定；SPAD值采用日本生產(chǎn)的SPAD-502PLUS型便攜式葉綠素含量測定儀測定；各指標每樣品重復測定3次。2.5葉片電導率測定水稻在自然條件下恢復生長24小時后，測定處理與對照水稻功能葉的電解質(zhì)滲漏率。測定方法參照張憲政方法[18]。2.6與細胞理化特性有關(guān)的生理指標測定水稻移出人工氣候室,在自然條件下恢復生長24小時后，測定各生理指標，方法參照高俊鳳主編[19]的植物生理學實驗指導進行：丙二醛（MDA）采用三氯乙酸-巴比妥酸法測定，可溶性糖采用蒽酮法測定，脯氨酸（Pro）采用磺基水楊酸-茚三酮法測定，超氧化物歧化酶（SOD）采用光照還原法測定。2.7計算與統(tǒng)計分析各測定指標的相對變化表示處理與其平行對照的差異，相對變化（%）=100×處理值/對照值；數(shù)據(jù)處理和統(tǒng)計分析采用Excel軟件和DPS統(tǒng)計分析軟件進行。3結(jié)果與分析3.1高溫降低籽粒充實度籽粒充實度是衡量劍葉光合產(chǎn)物和莖鞘貯存物向籽粒運輸和積累的重要指標[20]。由圖2可知，高溫處理后，兩純系籽粒充實度都呈現(xiàn)下降的趨勢，隨著高溫處理時間的延長，籽粒充實度的下降幅度越大；熱敏感純系XN0437S的下降幅度大于耐熱高溫。當高溫處理120小時后，XN0437S和XN0437T的籽粒充實度分別只有對照的68.7%和77.4%。圖1灌漿初期高溫降低水稻籽粒充實度變化圖注:“HT”表示“高溫處理時間”；“S”表示水稻材料“XN0437S”；“T”表示水稻材料“XN0437T”。（下同）3.2高溫影響劍葉光合效率光合作用是植物最基本的生命活動，也是植物生長發(fā)育的原動力[21]。圖2顯示，高溫處理6小時后，兩水稻純系的凈光合作用速率就有約5%的下降，隨著高溫處理時間的延長，凈光合作用速率下降幅度越大；在相同高溫處理條件下，耐敏感純系的下降幅度大于耐熱純系。高溫處理120小時后，熱敏感純系和耐熱純系的凈光合作用速率別為其平行對照的59.9%和76.1%。葉綠素含量是反映光合強度的重要指標，而SPAD值又是衡量葉片中葉綠素含量高低的可靠參數(shù)[22-24]。由圖2可以看出，高溫熱害導致水稻劍葉的SPAD值下降，耐敏感純系的下降速度大于耐熱純系，且隨著高溫處理時間的延長，SPAD值的下降幅度越大。高溫處理120小時且在自然條件下恢復生長24小時后，熱敏感純系和耐熱純系的葉綠素含量分別為其平行對照的70.1%和76.6%。圖2灌漿初期高溫降低水稻劍葉的凈光合作用速率和葉綠素含量3.3高溫影響劍葉細胞膜特性和胞內(nèi)環(huán)境3.3.1高溫熱害加強細胞膜的氧化作用SOD是植物體內(nèi)清除活性氧自由基的關(guān)鍵酶，其活性的強弱與植物抗氧化能力密切相關(guān)[25]。由圖3可知，高溫處理后，兩水稻純系劍葉的SOD活性呈現(xiàn)先增加而后稍有下降的趨勢；高溫處理6小時后，耐熱純系劍葉的SOD活性始終高于耐熱純系。熱敏感純系在高溫處理120小時后，SOD活性的增加達最大值、活性增加了49.3%；而耐熱純系XN0437T在高溫處理96小時時，SOD活性的增加達最大值、活性增加了53.5%。丙二醛(MDA)是膜脂過氧化的產(chǎn)物，也是衡量細胞膜損傷程度的標志性物質(zhì)[26]。從圖3知，高溫處理后，兩純系劍葉中的MDA含量較對照增加，隨著高溫處理時間的延長，MDA的增加幅度加大；熱敏感純系的增加幅度大于耐熱純系。高溫處理120小時，熱敏感純系的MDA含量較對照增加了29.0%，耐熱純系增加了18.2%。圖3灌漿初期高溫增加水稻劍葉的SOD酶活性和丙二醛含量3.3.2高溫熱害增加細胞膜透性處理和對照的細胞膜透性差異是衡量植物遭受逆境傷害程度的重要指標，而相對電導率是評定細胞膜透性的一種有效方法[19]。圖4顯示，高溫處理后，兩水稻純系的相對電導率與其對照相比，呈現(xiàn)上升的變化趨勢，隨著高溫處理時間的延長，相對電導率增加越大；熱敏感純系的上升幅度大于耐熱純系。高溫處理120小時后，熱敏感純系的電導率較對照增加了36.6%，耐熱純系較對照增加了23.2%。圖4高溫熱害增加劍葉細胞膜的透性3.3.3高溫熱害影響細胞內(nèi)滲透調(diào)節(jié)物質(zhì)含量脯氨酸作為重要的滲透調(diào)節(jié)物質(zhì)，其作用是保持原生質(zhì)和環(huán)境滲透平衡、阻止水分喪失、保護膜結(jié)構(gòu)的完整[27]。由圖5可知，高溫處理后，兩水稻純系的脯氨酸含量呈現(xiàn)上升的變化趨勢，隨著高溫處理時間的延長，脯氨酸含量的增加幅度越大；耐熱純系的上升幅度大于熱敏感純系。高溫處理120小時后，耐熱純系的脯氨酸含量較對照增加了52.6%，熱敏感純系增加了38.9%。可溶性糖不僅是植物體的能量物質(zhì)，也是體內(nèi)有效的滲透調(diào)節(jié)物質(zhì)，可溶性糖在細胞內(nèi)積累可以降低細胞的滲透勢以維持細胞膨壓，防止細胞被動脫水[19]。從圖5可以看出，隨著高溫處理時間的延長，兩水稻純系的可溶性糖含量較其對照都呈下降的趨勢，耐熱純系的可溶性糖含量的下降幅度小于熱敏感純系。高溫處理120小時后，熱敏感純系的可溶性糖含量為其對照的77.7%，耐熱純系的可溶性糖含量為其平行對照的83.7%圖5高溫熱害增加脯氨酸、降低可溶性糖的含量4討論水稻籽粒充實是指穎花受精后，光合產(chǎn)物和莖鞘貯存物向籽粒的運輸和積累的過程[28]，此時遇高溫熱害會嚴重影響籽粒充實度而造成產(chǎn)量損失[29]。Lin等[30]于抽穗-灌漿期采用35.0/30.0℃(白天/夜間)的溫度對水稻進行不同時長的高溫處理后，發(fā)現(xiàn)兩個水稻品種的穗重和千粒重明顯下降。本文在水稻灌漿初期對兩純系進行了不同時間長的高溫處理，發(fā)現(xiàn)水稻籽粒充實度下降，并且隨著高溫處理時間的延長，充實度下降幅度越大；在相同高溫處理條件下，熱敏感純系籽粒充實度的下降幅度大于耐熱純系。前人主要研究了抽穗-灌漿期高溫對水稻籽粒充實度或粒重的影響[30、31]綠色植物的光合作用包括光反應和暗反應兩個過程，光反應通過葉綠體完成能量的吸收、傳遞和轉(zhuǎn)換，暗反應由一系列酶催化將活躍的化學能轉(zhuǎn)變成穩(wěn)定的化學能；光合作用過程對高溫非常敏感，高溫脅迫不僅破壞參與光反應的葉綠素，也會抑制參與暗反應的酶活性，從而影響植物光合作用效率[21]。張桂蓮等[14]在水稻始穗期進行連續(xù)7天的高溫（平均33.5℃）處理，發(fā)現(xiàn)水稻劍葉光合速率和葉綠素含量在高溫處理1天后就有下降的趨勢；隨著處理時間的延長，劍葉光合速率和葉綠素含量繼續(xù)下降，且品系間存在差異。本文在水稻灌漿初期對兩純系進行高溫處理，也發(fā)現(xiàn)水稻劍葉的葉綠素含量和凈光合作用速率下降，隨著高溫處理時間的延長，二者下降幅度越大，且熱敏感純系劍葉葉綠素含量和凈光合作用速率的下降幅度大于耐熱純系。作物的產(chǎn)量只有極小部分物質(zhì)來自根部吸收的營養(yǎng)物質(zhì),而大部物質(zhì)則來自作物光合作用的產(chǎn)物，因此，本文認為高溫脅迫造成植物器官衰老或在逆境下(如高溫熱害)遭受傷害，往往發(fā)生膜脂過氧化作用，產(chǎn)生超氧陰離子自由基和丙二醛(MDA)，造成細胞膜透性增加，超氧化物歧化酶具有清除超氧陰離子自由基的作用，因此，植物體內(nèi)超氧化物歧化酶(SOD)活性和丙二醛（MDA）含量以及細胞膜滲透率可以反映植物遭受逆境傷害的程度[19]。張桂蓮等[32]在分析高溫脅迫對水稻劍葉保護酶活性的影響時，認為SOD活性在高溫脅迫后都表現(xiàn)先升后降的趨勢，耐熱品系SOD活性的增幅大于熱敏感品系；同時，水稻劍葉電解質(zhì)外滲率升高，耐熱品系電解質(zhì)外滲率小于熱敏感品系。雷東陽等[33]在抽穗期對8個耐熱性不同的雜交水稻組合進行37℃的高溫脅迫7天后，測定各組合倒1葉和倒2葉混合物的MDA含量，結(jié)果顯示各雜交水稻組合的MDA含量極顯著增加，且耐熱性強的水稻品系MDA含量增幅明顯低于熱敏感水稻。本文在水稻灌漿初期對兩純系進行了不同時間長的高溫處理后，也發(fā)現(xiàn)SOD活性、MDA含量和細胞膜滲透率增加，且隨著處理時間的延長，上述三個指標的增幅加大，且耐熱純系的SOD活性大于熱敏感純系，而熱敏感純系的MDA可溶性糖不僅為植物的各種生命活動提供了所需的能量，而且是許多物質(zhì)如纖維素、核苷酸和核酸等的組成成分，也是植物體內(nèi)有效的滲透調(diào)節(jié)物質(zhì)[19]；脯氨酸普遍存在于動植物細胞中，在細胞適應脅迫過程中表現(xiàn)為細胞內(nèi)的滲透調(diào)節(jié)劑、還原劑或能量來源、氮素儲藏物質(zhì)、羥基自由基清除劑、細胞內(nèi)酶的保護劑以及降低細胞內(nèi)酸度和調(diào)節(jié)氧化還原電勢等的作用[27、34]?；谏鲜鲎饔茫扇苄蕴呛透彼岬暮孔兓恢弊鳛樯锬湍婢车闹匾u價指標。張桂蓮等[14]研究了抽穗期受高溫脅迫的兩個耐熱性不同的水稻品種的可溶性糖和脯氨酸含量變化，認為高溫脅迫下，劍葉中的可溶性糖含量降低，脯氨酸含量增加，且隨著高溫脅迫時間延長，可溶性糖含量持續(xù)下降，劍葉中脯氨酸含量則持續(xù)上升，耐熱品系可溶性糖含量的下降幅度小于熱敏感品系，而脯氨酸含量的上升幅度則大于熱敏感品系。本研究在水稻灌漿初期對兩純系進行了不同時間長的高溫處理后，測定處理和平行對照劍葉的可溶性糖和脯氨酸含量，也得出了相同的結(jié)論。綜合兩水稻純系劍葉各生理指標在高溫與常溫下的變化情況，本文認為高溫破壞了細胞膜的結(jié)構(gòu)和功能，使細胞膜透性增加、細胞質(zhì)外滲，從而改變了細胞內(nèi)環(huán)境、影響了細胞內(nèi)物質(zhì)的合成、轉(zhuǎn)換和運輸?shù)日Ｉ顒?，導致向籽粒運輸和貯藏的物質(zhì)減少是造成水稻籽粒充實度降低，是最終造成水稻減產(chǎn)的生理原因之二。參考文獻[1] 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附件原始數(shù)據(jù)與統(tǒng)計分析附表1籽粒充實度調(diào)查原始數(shù)據(jù)與分析附表1-A秕粒重和實粒重水稻成熟后，取帶標記的水稻主穗6穗分別脫粒后，采用自來水漂選法分出秕粒和實粒，并去除空殼粒，曬干并統(tǒng)一平衡水份后按穗分別稱重水稻材料處理時間秕粒重實粒重穗1穗2穗3穗4穗5穗6穗1穗2穗3穗4穗5穗6XN0437S對照30.6250.6210.6290.6470.6180.5913.3753.3713.3793.1973.3683.34160.6270.6120.650.6270.6190.6153.4373.3623.443.3173.3693.13590.6180.6270.6840.6350.6090.6653.4683.3773.4343.3853.4593.415240.6210.6160.6740.6810.6390.6193.4713.4663.4243.4313.443.369480.6340.6930.7020.6640.6010.6383.4843.3433.3523.4143.3513.388720.6620.6310.6110.6570.6170.6053.4123.3313.3663.4073.423.355960.6460.6740.6230.6190.6430.6593.3963.3243.4133.3643.3933.4091200.5970.6340.6320.6790.6720.6273.4473.3843.3823.4293.4223.377處理30.6120.6410.6140.6340.6110.5973.3543.2973.2963.3383.223.29160.6220.6180.6250.6420.6150.593.3083.2443.2793.3573.3453.35290.7040.6910.7250.7040.6970.6933.3493.3173.3043.3183.2913.272240.710.7180.7710.7260.7010.7533.1193.0273.1693.2353.093.123480.7380.7330.7880.7940.7550.7362.8183.0142.9772.9012.9913.017720.7710.8260.8340.7990.7410.7752.6382.7562.6962.7642.832.79960.8130.7820.7620.8080.7680.7562.5712.3542.4572.2782.3292.4191200.8370.8630.8160.8120.8340.8491.8731.982.0321.9461.9571.89XN0437T對照30.6470.6560.6820.6270.6270.613.3183.3233.3563.1573.2923.25460.6230.6770.6490.6210.6110.6243.3433.3373.3713.2463.3573.25590.6410.6260.6760.6640.6220.6513.3853.2983.3933.3423.3593.353240.6570.6340.6280.6110.6770.6343.4033.3773.3373.3263.3993.3480.6530.6370.6960.6060.6110.6943.413.2823.3483.3073.2633.375720.6720.6920.6290.6110.6390.6613.3723.3283.2923.3073.3453.316960.6750.6210.6230.6610.6270.6423.3623.2513.3243.3233.3123.3391200.6840.6470.6340.6750.6420.6383.4123.3253.313.3883.353.31處理30.6820.630.6180.6620.6150.6443.2363.2573.3423.3253.2723.22960.6420.6260.640.6550.6090.6583.1983.3293.3133.2213.3443.22790.6760.660.6120.6830.6560.6863.3193.3113.1863.2493.2163.276240.7150.6690.6630.6450.7110.6693.1013.1023.2073.0783.0743.208480.6880.6720.7380.740.7160.732.9952.8862.9833.1652.8983.03720.7750.7280.7640.7450.6740.7962.8172.752.7882.8242.8292.874960.7110.7380.7420.7960.8150.7682.6862.5682.7092.6642.6932.821200.720.8820.8690.7110.8770.8732.3732.2122.262.362.2432.208

附表1-B籽粒充實度計算公式(%)=100×（對照的秕粒重+實粒重）/（對照的秕粒重+實粒重）水稻材料粒重36924487296120XN0437S對照（秕粒重+實粒重）3.9603.9684.0634.0754.0444.0124.0274.047處理（秕粒重+實粒重）3.9183.9334.0113.8573.7103.5373.1832.782籽粒充實度（%）98.999.198.794.691.788.179.068.7XN0437T對照（秕粒重+實粒重）3.9253.9524.0023.9973.9803.9773.9604.003處理（秕粒重+實粒重）3.9193.9103.9223.8073.7073.5613.4523.098籽粒充實度（%）99.898.998.095.293.189.587.277.4附表2光合作用速率附表2-A兩水稻材料的處理與對照的光合作用速率測定值水稻材料重復36924487296120XN0437S對照118.6117.6622.2722.5217.3522.5318.2122.61220.4222.2617.0316.9421.4617.8921.9919.37318.1718.118.9417.5619.919.2719.7117.96處理118.9118.2816.1815.8714.6114.6112.8311.78219.2519.0317.3415.6915.314.6513.312.13318.3117.6716.5216.3115.1113.613.812.01XN0437T對照121.3418.7622.5219.0722.5320.8218.2523.37218.3721.5819.6722.4318.4923.1322.5921.16319.4520.8618.4821.1822.1319.8723.6521.05處理119.8318.7818.4317.8117.6817.6416.4416.8218.6819.7117.8718.116.9317.7516.5916.44319.4219.6116.917.7317.7516.6317.5416.65附表2-B光合作用速率的相對變化相對變化率%=100*處理值/對照平均值水稻材料重復36924487296120XN0437S199.294.583.383.574.773.464.259.02101.098.489.382.578.273.666.660.7396.091.485.185.877.268.469.160.1平均98.794.885.984.076.771.866.659.9標準誤2.4963.5223.0721.6781.8212.9902.4290.890XN0437T1100.684.082.976.576.9294.796.688.486.680.483.477.275.2398.596.183.684.984.378.281.676.2平均97.994.987.785.682.981.578.476.1標準誤2.95562.50263.82780.93182.15952.90192.77490.8272附表3葉綠素含量（SPAD值）附表3-A兩水稻材料的處理與對照的葉綠素含量測定值水稻材料重復36924487296120XN0437S對照141.8742.3342.6542.0741.1744.6743.7145.88242.5641.1742.5343.1443.1642.8946.144.17342.6542.0143.8744.2543.2445.5245.3646.19處理141.6441.1541.1639.8836.8436.233.6732.2242.3941.2740.7840.5336.7735.9535.1331.08342.6742.0541.3839.937.2536.534.0532.17XN0437T對照142.7846.2443.9146.1444.545.1744.2444.85245.1343.1245.7844.2845.3143.6645.3346.65341.8541.4541.6743.543.8946.2847.1747.19處理142.8543.9743.2143.1740.7539.1437.5734.72243.6842.6443.2742.2841.2538.6337.236.02342.7642.5542.3542.5640.9639.8538.0135.55附表3-B葉綠素含量的相對變化相對變化率%=100*處理值/對照平均值水稻材料重復36924487296120XN0437S198.398.495.792.486.681.674.770.92100.198.694.893.986.581.078.068.43100.7100.596.292.587.682.375.670.8平均99.799.295.692.986.981.676.170.1標準誤1.2571.1680.7060.8570.6100.6211.6811.405XN0437T199.1100.898.796.791.486.982.475.12101.097.898.894.792.685.881.677.9398.997.696.795.391.988.583.476.9平均99.698.798.195.692.087.182.576.6標準誤1.1731.8241.1761.0190.5631.3610.8901.424

附表4SOD活性測定的原始數(shù)據(jù)附表4-A樣品提取液總體積V（ml）、鮮樣重量W（克）和根據(jù)標準曲線獲得的SOD活性測定值水稻材料處理提取液總體積（ml）=V樣品質(zhì)量（g）=WSOD活性測定值=C重復1重復2重復3重復1重復2重復3重復1重復2重復3XN0437S對照0.6830.610.6293.0262.7192.409870.6170.7332.9562.4332.2520.5210.5790.5942.8372.2762.792244.354.24.350.5530.5880.5322.1822.7532.70240.6550.5350.7413.0592.6933.465750.5660.4770.4652.7512.4942.3169890.730.7213.5763.5063.86410.5340.6450.5143.1653.0842.796處理0.6370.5490.5362.7792.4582.4280.410.4780.5541.8562.0862.46550.4770.560.6332.2432.8643.1150.560.5190.52.6982.7392.43640.6110.4130.773.3632.3914.4570.5250.4950.5953.6423.2713.70696444.10.7070.6930.6415.3295.3844.79712020.6410.6117.8015.5935.041XN0437T對照34.34.354.250.7080.7420.82.9693.123.25864440.7350.6910.6953.3023.0143.2290.5150.5180.5072.2212.3752.330.4390.4860.492.1622.1772.23948444.10.6260.6130.5223.042.8462.57670.6130.6150.5323.0123.0872.85990.5440.5110.5132.9862.7332.57310.7080.4380.4883.7572.4983.142處理50.6190.8050.6062.6673.5262.77550.5140.5980.5332.2892.6492.3530.5760.6090.5152.892.8072.54324520.720.7493.5543.9643.724484.3930.5030.6422.9692.8593.27370.4130.5120.5653.0783.9024.374950.4440.5010.3953.6134.2933.369150.580.4940.5845.0254.1385.073

附表4-B單位重量鮮樣SOD活力計算公式=C×(V/2)/WC為SOD活性測定值，V為提取液總體積（ml），W為鮮樣重量，2為測定體積2ml水稻材料重復36924487296120XN0437S對照19.52513.0511.4358.5829.80710.20710.89912.44729.8068.2818.8459.83210.57110.989.84610.51938.2346.1459.87111.0479.58610.83310.71811.967處理18.94310.18510.34510.35811.55914.22115.07516.95929.4029.60110.86811.34712.15813.87715.53817.88739.7399.34410.21110.71812.13613.39115.34117.326XN0437T對照19.0168.9859.05710.3429.71210.31812.07610.34829.1468.7249.3999.2959.28510.79211.76612.26238.6549.2929.8819.59610.11611.55410.78413.843處理19.0489.57510.28610.90213.09916.39618.30919.0629.0899.3039.67911.28612.50516.00417.99518.42839.2739.60210.12310.44112.4916.25718.55118.459附表4-CSOD活力的相對變化相對變化率%=100*處理值/對照平均值水稻材料36924487296120XN0437S197.3111.2102.9105.5115.7133.2143.7145.62102.3104.8108.1115.5121.7130.0148.2153.63106.0102.0101.6109.1121.5125.5146.3148.8平均101.9106.0104.2110.1119.7129.6146.1149.3標準誤4.3494.7063.4545.0973.4013.9072.2164.014XN0437T1101.2106.4108.9111.9135.0150.6158.6156.92101.7103.4102.5115.8128.9147.0155.9151.73103.7106.7107.2107.1128.7149.3160.7151.9平均102.2105.5106.2111.6130.8149.0158.4153.5標準誤1.3411.8383.3264.3423.5791.8252.4152.932

附表5丙二醛（MDA）含量測定附表5-A三種波長下的測定值水稻材料處理600nm測定值（V600）450nm測定值（V450）532nm測定值（V532）重復1重復2重復3重復1重復2重復3重復1重復2重復3XN0437S對照30.06910.07780.08340.52410.5730.54210.38380.40140.393860.07310.08160.08030.53220.53250.5640.39720.41360.378790.06810.06730.08260.51440.53170.57540.39220.38110.407240.07550.068070.08110.51860.54320.57450.39870.39320.4218480.07380.07390.08210.53850.54040.54760.38140.41380.4103720.07470.08110.06870.52080.54380.58070.3910.39710.4164960.07970.08370.07510.51680.53910.57020.42010.39810.39331200.08190.07550.08230.52440.54410.55390.43240.39790.3896處理30.07260.07250.07170.5170.4610.50080.38930.38510.386460.07510.08090.07960.48840.47760.51640.40060.41740.406190.0810.07580.07430.55460.51870.48240.42230.43050.4146240.08390.07830.07460.54620.4970.52710.44610.44090.4231480.07540.07310.08680.4940.55040.53210.44130.43640.4518720.07260.07920.08230.5160.53110.53620.44530.45270.4612960.07820.08220.08120.51450.51640.55270.470.46630.46571200.07250.08230.07740.54020.53010.51740.48390.4910.4778XN0437T對照30.05770.06040.06010.57930.53720.54180.38820.35910.386260.06190.05580.05350.56660.54110.53490.37910.39490.395790.06470.06210.06640.57660.6020.57930.41870.38960.3601240.06570.06840.06880.5230.51680.53130.39810.37320.4081480.07180.06950.06490.58250.57750.52310.40410.39240.4125720.07470.07060.0680.57030.55860.59060.42230.39950.4168960.07590.07460.07140.55460.56120.5780.39550.37950.40371200.08020.07350.07510.53530.56980.5470.39190.41820.3785處理30.07320.06790.06910.59380.64360.61280.39020.40370.39760.08720.07910.08620.59410.63220.61770.42710.4380.436690.07910.08260.07740.58040.630.61730.42250.43940.4201240.08270.08660.08480.61970.61190.58950.43950.44820.4343480.07080.08170.07630.65850.58210.61910.44530.45340.4506720.08010.07880.08070.59710.62170.65170.4640.47650.4618960.07730.08490.07580.63540.66280.61690.44760.4610.441200.08130.07460.07750.65210.61470.67730.46540.44250.4591

附表5-B丙二醛（MDA）含量計算丙二醛含量計算公式=6.45×(V532-V600)-0.56×V450水稻材料重復36924487296120XN0437S對照11.7361.7921.8021.7941.6821.7481.9061.96721.7661.8431.7261.7931.891.7341.7261.77531.6991.6091.771.8761.811.9171.7331.672處理11.7531.8261.8912.032.0832.1152.2392.35121.7581.7031.7972.062.0352.1122.1882.33931.7491.8171.7251.9532.0562.1442.1712.293XN0437T對照11.8071.7291.961.8511.8171.9231.7511.71121.6261.8841.7751.6771.7591.8091.6521.90431.81.9081.571.8911.9491.9191.821.651處理11.7121.861.8021.9542.0472.1422.0332.11221.8051.8711.8291.992.0712.2172.0552.02931.7351.9041.8651.9242.0682.0932.0042.082附表5-C丙二醛含量的相對變化相對變化率%=100*處理值/對照平均值水稻材料重復36924487296120XN0437S1101.1104.5107.1111.5116.1117.5125.2130.32101.497.4101.8113.1113.4117.4122.3129.63100.9103.997.7107.2114.6119.1121.4127.1平均101.1101.9102.2110.6114.7118.0123.0129.0標準誤0.2603.9224.7143.0311.3410.9821.9791.696XN0437T198.1101.1101.9108.2111.1113.7116.8120.32103.5101.7103.4110.2112.5117.7118.0115.6399.5103.5105.5106.5112.3111.1115.1118.6平均100.4102.1103.6108.3112.0114.2116.6118.2標準誤2.7771.2441.7871.8290.7103.3151.4692.394

附表6相對電導率測定與計算附表6-A相對電導率測定值水稻材料重復36924487296120XN0437S對照141.4440.5640.2139.7538.1339.5539.1338.23239.8539.3641.3538.7339.8337.6740.5440.12339.2942.7842.5338.9139.7538.5237.6737.88處理142.2542.5943.9541.9642.8344.1545.8246.85239.5441.1343.8740.7143.6543.2147.9749.12341.841.7543.6541.9443.8145.0845.9347.21XN0437T對照142.3543.2542.1939.7743.2839.5243.9439.78239.7441.643.941.2139.543.6644.6344.35341.839.5739.6842.7942.6944.7339.2543.69處理141.8743.1545.9846.149.7352.1455.2157.87241.143.6748.345.2547.1254.5556.1559.78341.8344.0147.3346.5450.1153.3855.7358.22附表6-B相對電導率測定值變化相對變化率%=100*處理值/對照平均值水稻材料重復36924487296120XN0437S1102.4104.1106.3107.2109.2114.4117.1120.9298.5100.6106.1104.0111.2112.0122.6126.83102.8102.1105.5107.2111.7116.8117.4121.9平均101.3102.3105.9106.2110.7114.4119.1123.2標準誤2.3781.7920.3761.8301.3402.4243.0963.149XN0437T1101.6104.0109.7111.7118.9122.3129.6135.8299.5105.3115.2109.7112.7127.9131.8140.33101.8106.1112.9112.8119.8125.2130.8136.6平均101.0105.2112.6111.4117.1125.1130.7137.6標準誤1.2631.0442.7791.5893.8922.8271.1052.387

附表7脯氨酸含量測定附表7-A提取液總體積、樣品鮮重和樣品液在520nm波長下的測定值水稻材料處理提取液總體積（ml）=V樣品質(zhì)量（g）=W520nm波長測定值重復1重復2重復3重復1重復2重復3重復1重復2重復3XN0437S對照0.7130.70.7190.13250.1260.13070.7170.7270.7230.12950.12840.1330.7170.7190.7240.13360.1330.124850.7330.7280.7220.12840.1330.130740.7150.7050.7310.12780.13540.133750.7260.7170.7050.13660.1330.129590.7190.6820.6910.13770.13540.128410.7240.7350.7040.13950.12950.1354處理0.6470.6890.6960.12760.13040.131550.6520.6880.6640.13130.13690.131992.62.452.650.6870.670.7430.13360.13660.1330.670.6290.610.14480.12590.1225482.552.52.450.6410.6330.650.15770.160.1559722.52.452.50.6350.6050.6050.17230.17760.174190.6170.6030.6510.20280.19990.19751202.452.52.550.6030.6510.6210.21390.21650.2149XN0437T對照32.52.552.650.6080.6420.680.12950.12660.131962.42.452.40.6350.6910.6950.12780.1460.14840.6150.6180.6470.12130.11780.1219242.62.552.60.6390.6860.690.12840.13130.132540.6260.6130.6220.12950.13540.1301722.62.452.50.6130.6150.6320.12950.1360.1313962.4440.6110.6130.1330.1360.13071202.6080.6380.6880.1360.13190.1354處理0.6190.6050.6060.12780.12480.122650.6140.5980.6330.12010.12040.12130.6260.6590.6150.12840.13010.127850.6520.620.6490.13780.14250.1413482.652.452.60.5930.6030.6420.13830.14180.153722.52.652.650.6130.6120.6650.17180.15360.1641950.6040.6010.6350.1840.17110.18871202.652.42.650.6180.5940.6040.18110.19630.1875

附表7-B根據(jù)標準曲線計算和520nm波長測定值計算脯氨酸測定值A(chǔ)=23.8293511*A520+0.631605919水稻材料重復36924487296120XN0437S對照13.7893.7183.8153.6913.6773.8873.9133.95623.6343.6913.8013.8013.8583.8013.8583.71833.7463.8013.6063.7463.8013.7183.6913.858處理13.6723.7603.8154.0824.3894.7375.4645.72923.7393.8943.8873.6324.4444.8645.3955.79133.7653.7753.8013.5514.3474.7805.3385.753XN0437T對照13.7183.6773.5223.6913.7183.7183.8013.87223.6484.1113.4393.7603.8583.8723.8723.77533.7754.1683.5363.7893.7323.7603.7463.858處理13.6773.4943.6913.9153.9274.7255.0164.94723.6063.5013.7324.0274.0114.2924.7095.30933.5533.5223.6773.9994.2774.5425.1285.100附表7-C單位重量鮮樣中脯氨酸含量（ug/g）脯氨酸含量=A×V/（2×m）;其中，A為根據(jù)標準曲線和520nm波長測定值計算獲得的脯氨酸測定值，V為提取液總體積ml數(shù)，2表示測定體積為2ml，m為鮮樣重量。水稻材料重復36924487296120XN0437S對照16.9087.2597.1836.7986.9437.2277.6197.64927.0096.8556.8727.0487.3887.1577.3547.33437.0346.5716.7237.3947.2797.5147.2127.672處理17.0957.2097.2197.928.7319.32610.62711.63827.0556.7927.1067.5068.7769.84810.73711.11936.7626.9646.7787.2768.1929.87710.24911.811XN0437T對照17.6436.9497.4457.517.1267.8847.238.43927.2467.2876.9556.9897.