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標題: | 外施Sodium Nitroprusside調節甘藍淹水逆境之反應 Exogenous Application of Sodium Nitroprusside Regulates Submergence Responses in Cabbage |
作者: | 嵇威壬 Wei-Jen Gi |
指導教授: | 許富鈞 Fu-Chiun Hsu |
關鍵字: | 甘藍,淹水逆境,亞硝基鐵氰化鈉,抗氧化酵素,無氧呼吸, cabbage,submergence stress,sodium nitroprusside,antioxidant enzyme,anaerobic respiration, |
出版年 : | 2022 |
學位: | 碩士 |
摘要: | 颱風及豪雨是臺灣最頻繁發生的極端氣候現象,豪雨造成之淹水為夏季甘藍(Brassica oleracea var. capitata)生產之隱患。過去許多研究致力於建立甘藍耐淹水標誌應用於品種改良。然而傳統育種曠日廢時,預措植物可作為新品種育成前過渡期之替代方案。一氧化氮(nitric oxide , NO)為植物體內調節植物生長發育的訊號之一,外施NO可提高植物對非生物性逆境的耐受性。然而預處理NO對植株全株淹沒於水層之逆境仍缺乏研究。故本研究以NO供應劑亞硝基鐵氰化鈉(sodium nitroprusside,SNP)預處理受完全淹沒逆境之甘藍品種‘初秋’探討其對淹水耐受性之影響。
本研究發現施用100 µM SNP之甘藍於淹水後具有較少之葉片損傷,並保有較高之鮮重及相對含水量,因此推測甘藍於完全淹沒前處理SNP可減緩植株所受傷害及脫水現象。為了評估淹水後氧化逆境之情形,利用組織化學染色檢測超氧陰離子及過氧化氫並分析反應細胞膜過氧化程度的指標丙二醛,發現施用SNP之甘藍累積較少超氧陰離子及過氧化氫及少丙二醛,可推測預處理SNP可減緩甘藍於完全淹沒下所受之氧化逆境。為了解NO調節活性氧(reactive oxygen species, ROS)的機制,本研究調查了抗氧化酵素活性及可使ROS含量降低之BoAOX1a 基因表現量。發現施用100 μM SNP可提高抗氧化酵素活性並具有較高表現量之BoAOX1a。接著探討於淹水逆境下產生能量之無氧呼吸基因,發現施用100 μM SNP可於淹水前誘導BoSUS1L、BoADH1及BoPDC1表現量,而於淹水24小時無氧呼吸基因表現量較對照組低,參與非共生血氧蛋白一氧化氮循環(phytoglobin/nitric oxide cycle, Pgb/NO cycle)之BoHB1基因表現量則較對照組高,顯示施用100 μM SNP可誘導PGB/NO cycle以彌補淹水時酒精發酵回收NAD+之效率。總體而言,施用100 μM SNP可提高無氧呼吸基因、BoAOX1a之表現量與抗氧化酵素活性,最終使得受淹水逆境之甘藍氧化逆境及脫水程度減緩。 Typhoons and torrential rain are the most frequent extreme weather phenomena in Taiwan. Flooding caused by heavy rain is a hidden risk for cabbage (Brassica oleracea var. capitata) production in summer. Previous studies have been devoted to establishing cabbage flooding stress tolerance markers for cultivar improvement. However, traditional breeding is time-consuming, priming can be used as an alternative to the transition period before new varieties are developed. Nitric oxide (NO) is one of the signals that regulate plant growth and development in plants. Exogenous application of NO improves plant tolerance to abiotic stress. However, there is still a lack of research on pretreatment of NO upon the submerged in the water. Therefore, this study used the NO donor agent sodium nitroprusside (SNP) to pretreat the cabbage cultivar ‘K-Y cross’, which was subjected to complete submergence, to explore its effect on submergence tolerance. This study found that cabbage treated with 100 µM SNP had less leaf damage, higher fresh weight and relative water content after flooding, speculating that the SNP treatment of cabbage before complete submergence can reduce the damage and dehydration of the plant. To assess oxidative stress after flooding. Detection of superoxide anion and hydrogen peroxide by histochemical staining and analyze malondialdehyde, an indicator that reflects the degree of cell membrane peroxidation, found that SNP pretreated cabbage accumulated less superoxide anion, hydrogen peroxide and malondialdehyde, speculating that the pretreatment of SNP could alleviate the oxidative stress of cabbage under complete submergence. In order to understand the mechanism of NO mediating reactive oxygen species (ROS), this study investigated the activity of antioxidant enzymes and the expression of BoAOX1a gene, which can reduce the content of ROS. The cabbage treated with 100 μM SNP was found to increase the antioxidant enzyme activity with a higher expressed amount of BoAOX1a.The anaerobic respiration genes that generate energy under flooding stress were then investigated, and it was found that cabbage treated with 100 μM SNP could induce the expression levels of BoSUS1L, BoADH1 and BoPDC1 before flooding. By contrast, the expression of anaerobic respiration gene at 24 hours of flooding was lower than that of the control group, and the expression of BoHB1 involved in the PGB/NO cycle was higher than that of the control group, suggesting that applying 100 μM SNP triggers PGB/NO cycle to compensated efficiency of alcoholic fermentation for recovering NAD+. Taken together, exogenous application of 100 μM SNP increased the expression of anaerobic respiration genes, BoAOX1a, and antioxidant enzyme activity. Ultimately, the oxidative stress and dehydration of the flooded cabbage were alleviated. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/86145 |
DOI: | 10.6342/NTU202203173 |
全文授權: | 同意授權(全球公開) |
電子全文公開日期: | 2027-09-12 |
顯示於系所單位: | 園藝暨景觀學系 |
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