本文選題：淹水　切入點(diǎn)：小麥　出處：《華中農(nóng)業(yè)大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：前期研究表明,小麥胚乳發(fā)育需經(jīng)歷編程性細(xì)胞死亡(Programmed cell death,PCD)過程,淹水可加速其進(jìn)程(Fan et al 2013)。本研究以耐濕品種華麥8號(hào)和不耐濕品種華麥9號(hào)為材料,探討了淹水脅迫下小麥胚乳PCD進(jìn)程提前的生理及細(xì)胞學(xué)機(jī)制。主要結(jié)果如下:1.Evans Blue染色結(jié)果顯示,正常生長(zhǎng)的華麥8號(hào)和華麥9號(hào)胚乳均在花后15 d可檢測(cè)到PCD現(xiàn)象。淹水處理后,同期生長(zhǎng)的胚乳著色更深,PCD進(jìn)程加劇;與華麥8號(hào)相比,華麥9號(hào)胚乳PCD進(jìn)程提前。半薄切片染色結(jié)果顯示,正常生長(zhǎng)的兩品種小麥胚乳細(xì)胞中淀粉粒和蛋白體均隨發(fā)育天數(shù)增加逐漸增多,淹水可加速淀粉體和蛋白體在胚乳細(xì)胞中積累。正常生長(zhǎng)的兩品種胚乳細(xì)胞核分別在花后12 d和15 d發(fā)生降解;淹水可加劇同期胚乳核降解過程,并能使華麥9號(hào)胚乳細(xì)胞核降解提前發(fā)生。2.淹水脅迫后,華麥8號(hào)穎果含水率下降,胚乳中MDA、H2O2和O2-含量上升,早期發(fā)育的胚乳中可溶性糖、可溶性蛋白和脯氨酸含量增加,SOD和CAT活性上升,SOD同工酶譜上新增了SOD1帶,CAT同工酶譜上新增了CAT2、CAT5帶,且各種CAT同工酶活性均增強(qiáng);華麥9號(hào)胚乳中MDA、O2-、H2O2和可溶性蛋白含量顯著上升,SOD和POD活性略有升高,而CAT活性隨發(fā)育天數(shù)的增加呈“降-升-降”的趨勢(shì),SOD同工酶變化不大,僅SOD2活性有所上升,而各CAT同工酶活性均上升。3.微管蛋白免疫熒光及免疫電鏡結(jié)果顯示,隨著胚乳發(fā)育的進(jìn)行,細(xì)胞中微管蛋白熒光信號(hào)逐漸增強(qiáng)。膠體金統(tǒng)計(jì)分析結(jié)果顯示,淹水后早期發(fā)育的胚乳淀粉質(zhì)體周圍和細(xì)胞質(zhì)囊泡中微管蛋白膠體金數(shù)目顯著高于對(duì)照;花后12 d的胚乳中微管蛋白膠體金顆粒增加,主要分布在淀粉質(zhì)體周圍;花后15 d的胚乳線粒體和淀粉質(zhì)體周圍微管蛋白數(shù)量增加,細(xì)胞質(zhì)中分布較少。微絲蛋白免疫熒光及免疫電鏡結(jié)果顯示,正常發(fā)育的胚乳細(xì)胞中微絲蛋白熒光信號(hào),隨發(fā)育天數(shù)的增加逐漸增強(qiáng)。膠體金統(tǒng)計(jì)分析結(jié)果顯示,淹水處理后,早期發(fā)育的胚乳淀粉質(zhì)體周圍和細(xì)胞質(zhì)中微絲蛋白膠體金顆粒顯著增加;花后12 d的胚乳線粒體周圍微絲蛋白膠體金數(shù)目減少;花后15 d,細(xì)胞質(zhì)中仍然分布大量微絲蛋白。綜上所述,淹水處理后,華麥8號(hào)和華麥9號(hào)胚乳中MDA及活性氧含量均上升,造成胚乳PCD進(jìn)程加劇;早期發(fā)育的華麥9號(hào)胚乳中活性氧上升幅度大,而其清除能力弱,PCD進(jìn)程提前。胚乳細(xì)胞中養(yǎng)分和淀粉質(zhì)體迅速積累,細(xì)胞骨架可能作為運(yùn)輸軌道,協(xié)助胚乳細(xì)胞提前完成生命過程,以減輕淹水造成的傷害,其還可能參與淹水脅迫的應(yīng)急反應(yīng),調(diào)控胚乳細(xì)胞PCD進(jìn)程。
[Abstract]:Preliminary study shows that the wheat endosperm undergoing programmed cell death (Programmed cell, death, PCD), flooding can speed up the process (Fan et al 2013). In this study, moisture resistant varieties and Huamai 8 moisture resistant varieties Huamai No. 9 as the material, discusses the physiological and cytological mechanism of wheat in advance the PCD process of waterlogging stress. The main results are as follows: 1.Evans Blue staining showed that the normal growth of Huamai No. 8 and No. 9 in the endosperm Huamai 15 d after flowering could be detected by PCD. After flooding, the growth period of endosperm color deeper, increased compared with PCD process; Huamai No. 8, No. 9 Mr endosperm the PCD process in advance. Semi thin sections. Results showed that two varieties of wheat endosperm cell normal growth of starch grains and protein bodies were decreased with the increase in the number of days of development gradually increased, flooding can accelerate starch bodies and protein bodies accumulated in endosperm cells in normal life. Two varieties of endosperm nucleus length are 12 d after flowering and 15 d degradation; flooding may exacerbate the earlier process of endosperm degradation, and can make Huamai No. 9 endosperm nuclei occurred earlier.2. degradation after waterlogging stress, Huamai No. 8 caryopsis water content decreased, endosperm MDA, H2O2 increased and the content of O2-, soluble in early development the endosperm of sugar, soluble protein and proline content increased, the increase of SOD and the activity of CAT, SOD isozymes were added on the SOD1 band, CAT isozyme added CAT2, CAT5, and CAT isoenzyme activity were enhanced; Huamai No. 9 endosperm MDA, O2-, H2O2 increased significantly and the content of soluble protein, SOD and the POD activity increased, while CAT activity increased with the growing days of "down up down" trend, SOD isozyme changed little, only the activity of SOD2 increased, while the CAT isozyme activity increased.3. tubulin immunofluorescence and immunoelectrodes. Microscope results showed that with the development of endosperm cells, tubulin fluorescence signal gradually. Colloidal gold statistical analysis results showed that the number of gold endosperm amyloplast around and cytoplasmic vesicles in tubulin colloid early development after flooding was significantly higher than that of control after anthesis; microtubule associated protein 12 d colloidal gold particles increased in endosperm, mainly the distribution of amyloplasts in the number of flowers around; after 15 d of the mitochondrial and endosperm amyloplast around tubulin increased, less distributed in the cytoplasm. The microfilament protein by immunofluorescence and immunoelectron microscopy showed that the actin protein fluorescence signal of normal development of endosperm cells, increased gradually with the increase of development days. Colloidal gold statistical analysis showed that flooding after treatment, the early endosperm starch plastid development around the microfilaments in the cytoplasm and protein colloidal gold particles increased significantly; 12 d endosperm mitochondria weeks after flower Wai microfilament protein decreased the number of colloidal gold; 15 d after flowering, still a lot of microfilaments in the cytoplasm. The protein distribution in summary, after flooding, the contents of MDA and reactive oxygen species in Huamai No. 8 No. 9 and Mr endosperm were increased, resulting in increased endosperm PCD activity in early development process; Mr No. 9 in endosperm oxygen increase. And the scavenging ability is weak, the PCD process in advance. The rapid accumulation of nutrients and amyloplast in endosperm cells, the cytoskeleton may serve as transport track, assist in endosperm cell life early, in order to reduce the damage caused by flooding, which also may be involved in emergency response to flooding stress, regulation of endosperm cell PCD process.

【學(xué)位授予單位】：華中農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S512.1

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