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研究生:蘇萍雅
研究生(外文):Pinyapach Sukkasem
論文名稱:1. LED 光源影響阿拉伯芥及萵苣葉組織硝酸鹽累積 2. 根際促生菌影響阿拉伯芥生長、非生物性逆境反應及硝酸鹽累積
論文名稱(外文):1.The effects of light quality on nitrate accumulation in leafy tissues of Arabidopsis and lettuce 2.The impacts of the plant growth promoting rhizobacteria on plant growth, nitrate accumulation, and abiotic stress response in Arabidopsis
指導教授:莊慧文
指導教授(外文):Chuang, Huey-wen
學位類別:博士
校院名稱:國立嘉義大學
系所名稱:農業科學博士學位學程
學門:農業科學學門
學類:一般農業學類
論文種類:學術論文
論文出版年:2019
畢業學年度:107
語文別:英文
論文頁數:106
外文關鍵詞:Nitrate contentnitrate accumulationlight qualityblue light
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Chapter 1 Abstract
Cellular processes that determine the level of nitrate in plant tissue include ion fluxes through cell membrane and enzymes activity such as nitrate reductase (NR), nitrite reductase (NIR), and glutamine synthetase (GS), are involved in nitrogen assimilation. In plants, nitrate accumulation is closely regulated with photosynthesis which provides ATP and the electrons from NAD(P)H and ferredoxin for supporting nitrogen assimilation enzymes, including NR, NIR, and GS. Light is the main source of energy which is one of the driving forces in photosynthesis. In addition, the specific wavelength of light also activates various signal transductions through photoreceptors including phytochromes, cryptochromes, and phototropins. The perception of light mediates proteins activity, including NR and GS activity. To know the effect of light spectrum on the accumulation of nitrate, this study was conducted using 3 different light sources, white light-emitting diodes (LW), red light-emitting diodes (LR), and the combination of red and blue light-emitting diodes (L5R2B). In Arabidopsis, tissues treated with LR were accumulated higher nitrate levels than that under the other treatment and the result indicated that the reduction of nitrate level might present in blue spectrum. LB treatment was conducted with LW and the result showed that LB influenced the reduction of nitrate levels. Similar LB effect on reduced nitrate accumulation was observed in lettuce. Real-time PCR transcriptionally analyzed nitrate transporters showed that LB light altered expression of nitrate transporter 1.2 (NRT1.2), chloride channel (AtCLCa), phototropin 1 (PHOT1), phosphate transporter 1 homolog 7 (PHO1-7), and sucrose-phosphate synthase (SPS). Different light sources also altered total protein accumulation including NR, GS2, ascorbate peroxidases (APX), fructose 1,6-bisphospate aldolase (ALD), cytosolic fructose 1,6 bisphosphatase (cFBPase), and phosphoenolpyruvate carboxylase (PEPC) in both Arabidopsis and lettuce tissues. In conclusion, blue light influences on decreasing of nitrate through various cellular processes associated with photosynthesis and cellular respiration.

Chapter 2 Abstract
Several evidences indicate that plant growth promoting rhizobacteria (PGPRs) enhance plant growth and development by the production of phytohormones, solubilization of nutrients and regulation of genes involved in plant metabolism. Furthermore, PGPRs support the bioactive compounds to plant for decreasing the stress from environment. Bacillus licheniformis CH102 (CH102) was identified and confirmed by using 16S rDNA fragments. The effect of CH102 on promoting plant growth, nitrate assimilation, and abiotic stress response was studied. Our results showed that CH102 promoted not only plant growth in normal condition but also under drought and high temperature. Western blot analysis showed that the isolated of bacteria altered Arabidopsis protein accumulation including enzyme involved in nitrogen assimilation and pathogenesis-related protein 1 (PR-1) protein involved in systemic resistance. Moreover, next generation sequencing (NGS) analysis showed that expression of Arabidopsis genes were up-regulated by CH102 including genes associated with signal transductions, stress responses, metabolisms, transcription factors, and phytohormones. These results indicated that Bacillus licheniformis CH102 might promote plant growth and regulate plant tolerance to both biotic and abiotic stresses.
Chapter 1 Abstract i
Chapter 2 Abstract iii
Acknowledgements iv
Table of Contents v
List of Tables ix
List of Figures x
Chapter 1 1
The effects of light quality on nitrate accumulation in leafy tissues of Arabidopsis and lettuce 1
Introduction 1
Nitrogen cycle in ecology 1
The causes of nitrate accumulation in plant tissues 2
Relationship between photosynthesis and nitrate accumulation 4
Relationship between light quality and nitrate transport / nitrate reductase activity 5
Materials and methods 8
Plant materials and growth conditions 8
Light treatments 8
Photosynthesis efficiency 8
Chlorophyll (chl) contents 9
Nitrate determination 9
DPPH radical scavenging assay 10
Protein extraction 10
Western blot analysis 11
RNA extraction 12
Real-time PCR 12
Statistical analysis 13
Results 14
Correlation between different colors light wavelengths and nitrate accumulation in Arabidopsis 14
Reduction of nitrate accumulation in Arabidopsis treated with blue light 14
Effect of blue light on expression of genes involved in nitrate transporter and photosynthesis in Arabidopsis 15
Short-term illumination effect of blue light on nitrate level and nitrate transporter gene expression in Arabidopsis 16
Effect of blue light on nitrate level in lettuce leaf 16
Regulation of blue light on proteins involved in nitrate assimilation, photosynthesis and ROS scavenging in lettuce 16
Short-term illumination effect of blue light on nitrate level, proteins involved in nitrate assimilation, photosynthesis and ROS scavenging in lettuce 17
Discussion 18
References 31
Appendix I Primer name 38
Chapter 2 39
The impacts of the plant growth promoting rhizobacteria on plant growth, nitrate accumulation, and abiotic stress response in Arabidopsis 39
Introduction 39
Direct effect of PGPRs on promoting plant growth 40
Nitrogen fixation 40
Phosphate solubilization 40
Siderophore production 41
Phytohormone production 41
Indirect effect of PGPRs on promoting plant growth 44
PGPRs as a biocontrol agent 45
Induced tolerance to abiotic stress 46
Materials and methods 51
PGPRs isolation 51
IAA quantification and phosphate solubilization analysis of bacterial isolate 51
Salt and high temperature tolerance test of bacterial isolate 52
Plant materials and growth promoting effects of CH102 analysis 53
Drought stress treatment 53
Heat stress treatment 54
Hydrogen peroxide content and antioxidant enzyme activity determination 54
Next Generation Sequencing (NGS) 55
Results 57
Identification and characterization of the isolated rhizobacteria 57
Quantitation of indole-3-acetic acid (IAA) and phosphate solubility of CH102 activity 57
The impact of CH102 on sterilized culture Arabidopsis growth and development 58
The effect of CH102 on non-sterilized culture Arabidopsis growth and development 58
CH102 up-regulated Arabidopsis genes expression involved in nutrient uptake, antioxidant activity, and defense response 59
The CH102 strain altered Arabidopsis gene expression and protein accumulation, associated with nutrient uptake, nitrate assimilation, and defense response 60
Plant inoculated with CH102 showed more tolerance after being induced to water deficit 60
Plant inoculated with CH102 showed tolerance induction to heat stress 60
The impact of CH102 strain on Arabidopsis transcriptome 61
CH102 up-regulated Arabidopsis gene expression involved in defense response and abiotic response 62
CH102 up-regulated expression of transcription factors involved in biotic abiotic stress response 62
CH102 stimulated hormone signaling pathways in Arabidopsis 63
Discussion 65
References 81
Appendix I Primer name 105
Appendix II Nucleotide sequences of Bacillus licheniformis CH102 106
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