本文選題：中國石竹 + 鹽脅迫�。� 參考：《內(nèi)蒙古農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：鹽害是植物生長面臨的重要逆境危害之一,研究耐鹽植物的耐鹽機(jī)制可為植物改良和鹽地種植技術(shù)提供理論依據(jù)。本試驗以耐鹽性觀賞植物中國石竹的幼苗為材料,通過在100mMNaCl脅迫的Oh、1h、3h、6h及去除鹽脅迫的1h、3h分別收集幼苗葉片和根,提取總RNA,然后進(jìn)行轉(zhuǎn)錄組測序、組裝、分析,從基因表達(dá)水平深入了解中國石竹的耐鹽機(jī)理,獲得耐鹽基因,進(jìn)而達(dá)到在石竹屬植物中進(jìn)行抗鹽新品種培育的目的。取得的研究成果有:1.測序、組裝后獲得 62,371 unigenes,其中 31863 unigenes(51.08%)成功注釋。其中22662個unigenes可富集到46個GO terms,1 11810個unigenes可分配到KOG的26個groups上,經(jīng)KEGG注釋,10265個unigenes共預(yù)測了 272個途徑;2.鹽脅迫下葉中共獲得9139個鹽誘導(dǎo)基因,根中共獲得19280個鹽誘導(dǎo)基因;在鹽脅迫的1h、3h和6h,根中的DEGs分別是葉中的19.90、1.25和2.6倍;分別對葉和根中的鹽誘導(dǎo)基因進(jìn)行GO富集分析,發(fā)現(xiàn)鹽脅迫影響根部的纖維素代謝,葉中比根部延后,主要影響到轉(zhuǎn)錄、翻譯過程;KEGG分析表明,鹽脅迫下根部通過應(yīng)激反應(yīng),產(chǎn)生信號物質(zhì),影響糖代謝、苯丙素代謝、脂肪代謝,葉中比根中延后,在鹽脅迫6h開始產(chǎn)生信號物質(zhì)。3.根據(jù)GO富集中的BP途徑,篩選出與氧化還原過程或細(xì)胞氧化還原平衡有關(guān)的鹽誘導(dǎo)基因,經(jīng)RT-PCR驗證,發(fā)現(xiàn)在短期鹽脅迫下葉中的DchAAO1、DchPOD1基因和根中的DchAAO2、DchPOD2基因表達(dá)量下調(diào),葉中的DchTrx基因和根中的DchGrx1、DchGrx2、DchTrx基因表達(dá)量上調(diào),去除鹽脅迫后則相反,所有的被驗證基因的表達(dá)模式與RNA-Seq的數(shù)據(jù)吻合,表明試驗結(jié)果可靠。4.延長鹽脅迫時間后,對篩選出的差異基因進(jìn)行RT-PCR驗證,發(fā)現(xiàn)中國石竹葉片中基因DchAAO1、DchPOD1、DchTrx的表達(dá)量從9h到12h下降,12h后下降平緩,且在鹽脅迫同一時間點(diǎn)下均低于對照;根中基因DchPOD2的表達(dá)量隨脅迫時間的延長變化不大,在脅迫12、24h后其表達(dá)量低于對照,基因DchAAO2的表達(dá)量從9h到12h上升,12h后下降,鹽脅迫12、24h后基因表達(dá)量高于對照,基因DchGrx1、DchGrx2的表達(dá)量從9h到12h上調(diào)且脅迫下的高于對照,12h后下調(diào)且脅迫下的低于對照,基因DchTrx的表達(dá)量呈先升高后降低趨勢,在鹽脅迫下其基因表達(dá)量均低于對照。
[Abstract]:Salt damage is one of the important stresses on plant growth. The study of salt tolerance mechanism of salt-tolerant plants can provide theoretical basis for plant improvement and salt planting techniques.In this experiment, the seedlings of Chinese carnation, a salt-tolerant ornamental plant, were collected from the leaves and roots of the plants under 100mMNaCl stress for 3 h and 1 h for 3 h, respectively, and the total RNAs were extracted, then sequenced, assembled and analyzed.The salt-tolerant genes were obtained from the level of gene expression in Chinese carnation, and the purpose of breeding new salt-tolerant varieties in Carnation was achieved.The results of the research are as follows: 1: 1.Sequencing, assembled 62371 unigenes, of which 31863 unigenes 51.08) successfully annotated.Among them, 22662 unigenes can be enriched to 46 go termsl 11810 unigenes can be distributed to 26 groups of KOG. According to KEGG annotation, a total of 272 pathways are predicted by 10265 unigenes.A total of 9139 salt-inducible genes were obtained in leaves and 19280 in roots under salt stress, and the DEGs in roots was 19.90,1.25 and 2.6-fold higher than that in leaves at 1h and 6h, respectively. Go enrichment analysis was carried out in leaves and roots, respectively.It was found that salt stress affected cellulose metabolism in roots, and that in leaves was later than that in roots, which mainly affected transcription. KEGG analysis showed that under salt stress, the roots produced signal substances, sugar metabolism and phenylpropanin metabolism through stress response.Fat metabolism, delayed in leaves than in roots, began to produce signal substance. 3 at 6 h of salt stress.According to BP pathway in go enrichment, salt-induced genes related to redox process or redox balance of cells were screened. After RT-PCR verification, it was found that DchAAO1DchPOD1 gene in leaves and DchAAO2mDchPOD2 gene in roots were down-regulated under short-term salt stress.The expression of DchTrx gene in leaves and DchGrx1 + DchGrx2DchTrx gene in root was up-regulated, but the expression pattern of all the verified genes was consistent with that of RNA-Seq after salt stress was removed, indicating that the results of the experiment were reliable. 4.After prolonging the time of salt stress, the differentially screened genes were verified by RT-PCR. It was found that the expression of DchAAO1, DchPOD1, DchTrx in Chinese carnation leaves decreased slowly from 9h to 12h, and the expression of DchPOD1DchTrx was lower than that of control at the same time point of salt stress.The expression of gene DchPOD2 in roots did not change with the prolongation of stress time. After 1224 hours of stress, the expression of gene DchPOD2 was lower than that of control. The expression of gene DchAAO2 increased from 9 h to 12 h and decreased after 12 h of salt stress, and the amount of gene expression was higher after 1224 h of salt stress than that of control.The expression of DchGrx1 and DchGrx2 was up-regulated from 9 h to 12 h, and decreased after 12 h of stress, and the expression of DchTrx increased first and then decreased. The expression of DchGrx2 was lower than that of control under salt stress.
【學(xué)位授予單位】：內(nèi)蒙古農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S681.5

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