臺灣博碩士論文加值系統

此研究的主要目的是探討不同水稻品種及土壤環境中鐵載體合成細菌之數目/組成之相關性研究及其對水稻生長之影響。我們將水稻台稉8號及台中秈糯1號品種以無滅菌的酸土、近中性土及紅土於溫室環境中栽植30天。之後，於土樣、根圈土及水稻根莖組織中分析其總菌數及鐵載體合成細菌之數目。土樣中以近中性土所含之鐵載體合成細菌比例最高(5.47%)，其次是酸土(3.58%)及紅土(0.27%)。水稻種子內的內生細菌擁有鐵載體合成能力的比例台稉8號高(31.79%)較台中秈糯1號高出約10倍(3.00%)。二水稻品種之根圈土以栽植於酸性土的總菌數最高(2.3×104 to 3.0×104 CFU g−1)，其次是紅土(1.8×104 to 2.3×104 CFU g−1)及近中性土(1.4×104 to 1.6×104 CFU g−1)。相反的，在水稻根圈域中，無論品種，以近中性土含有最高數目的鐵載體合成細菌而最低的是紅土。綜而言之，無論在何種土壤或組織中，台稉8號具有較台中秈糯1號相對高的鐵載體合成細菌數目。我們由水稻組織中挑選了八株具最強鐵載體合成能力之細菌並以16S rDNA鑑定之，發現其中四株為Pantoea菌屬且均由種子中分離而得。水稻種子在表面殺菌後分別接種此八株菌並將5天齡小苗轉植在CAS培養基中。觀察發現經SPB處理的小苗其根部周圍透明圈顯著比未接種控制組為大，台稉8號根部周圍透明圈約較未接種控制組大14% 到28%，而台中秈糯1號為13% 到34%。以台中秈糯1號及紅土做盆栽實驗，接種LS-756鐵載體合成細菌能顯著提升水稻根長、地上部長度及乾重。除此之外，接種鐵載體合成細菌(除了LS-757)的水稻植株均有顯著提升葉綠素 a/b及總葉綠素總量的現象，分別為10% 到14%, 68% 到91% 及14% 到33%。以上結果證明了水稻的鐵載體合成細菌的數目及比例會被水稻遺傳背景及土壤環境條件所影響。此研究所累積的知識可提供一清楚的證據說明鐵載體合成細菌扮演水稻吸收鐵元素及與葉綠素含量之重要性及相關性。另外，亦可協助不同植物提升抗非生物性逆境之能力

The aim of this study is to understand the number and proportion of siderophore-producing bacteria (SPB) in relation to different rice genetic backgrounds and edaphic factors, as well as their effects on rice growth. Two rice Oryza sativa cvs., TK8 and TCN1, were cultivated in greenhouse condition using non-sterile acidic, near-neutral and red soils for 30 days. Total bacteria and SPB numbers were estimated from the bulk soils, rhizospheres, seed, root and shoot tissues of two rice cultivars. The percentage of SPB (SPB/total bacteria count x 100%) was highest in the near-neutral bulk soil at 5.47%, and relatively lower in acidic and red bulk soil at 3.58% and 0.27%, respectively. The percentage of SPB endophytes in TK8 seed (31.79%) was approximately ten-fold higher than in TCN1 seed (3.00%). The near-neutral rhizospheres showed the highest number of SPB in both rice varieties (1.4x102 CFU g−1 with TK8; 9.4x10 CFU g−1 with TCN1), and the lowest data was obtained from the red rhizospheres (1.1x10 CFU g−1 with TK8 and 1.5x10 CFU g−1 with TCN1). In all soil types and rice tissue types, the TK8 cultivar accommodated SPB endophytes higher than that of TCN1. By 16S rDNA sequence, a total of eight distinct bacterial endophytes exhibiting the strongest siderophore-producing activity were identified from various tissues of two rice varieties cultivated in three different soil-types. Bacteria belong to the genera Pantoea was predominant (n=4) and mostly isolated from seeds. Under sterile condition, rice seedlings inoculated with most SPBs formed a distinct orange halo on CAS medium significantly larger (from 14% to 28% with TK8, and from 13% to 34% with TCN1) than that in non-inoculated control. In the pot experiment performed with TCN1 and red soil, the inoculation of strain LS-756 showed significant increases in plant root length, shoot length and dry weight. In addition, inoculation of all strains (except LS-757) significantly enhanced chlorophyll a, chlorophyll b and total chlorophyll contents from 10% to 14%, 68% to 91% and 14% to 33%, respectively. The results proved that the number and proportion of SPBs associated with rice were influenced by host-genotype and edaphic factors. The knowledge gained from this study provided an important link about the role of SPB on rice iron uptake, the relationship with chlorophyll contents, and could further improve abiotic stress tolerances not only in rice, but also in other plants.

Abstract...i
中文摘要 (Chinese abstract)...iii
Acknowledgements...v
Table of contents...vi
List of Tables...viii
List of Figures ...ix
List of symbols ...x
Chapter 1 - Introduction...1
1.1 Overall View...1
1.2 Research Objectives...2
Chapter 2 - Literature Review...3
2.1 Introduction and definitions...3
2.1.1 Rhizosphere and Rhizobacteria...3
2.1.2 Endosphere and Endophytic colonization...5
2.1.3 Biodiversity of bacterial endophytes...6
2.2 The role of rhizo- and endosphere bacteria on soil nutrient availability and plant nutrient uptake...8
2.3 Rhizo- and endosphere bacteria in the rice plant...11
2.4 The iron deficiency of plant and role of siderophore-producing bacteria...13
2.4.1 The role of iron and effect of iron stress on plants...13
2.4.2 Siderophore biosynthesis and functions related to PGP...14
Chapter 3 - Materials and Methods...17
3.1 Materials...17
3.1.1 Experimental equipment...17
3.1.2 Chemical compounds...17
3.1.3 Soil samples and rice varieties...19
3.1.4 Plant nutrient...20
3.2 Methods...21
3.2.1. Medium substrates...21
3.2.2 Counting bacteria in bulk soils and seeds...24
3.2.3 Greenhouse cultivation and sample preparation to count endophytic bacteria in rhizo- and endosphere...25
3.2.4 Identification endophytic bacteria that possess the strongest siderophore-producing activity from rice tissues...26
3.2.5 Evaluate the effect of endophytic bacteria that possess the strongest siderophore-producing activity on rice growth...30
3.3 Statistical analysis...32
Chapter 4- Results...33
4.1 Characteristics of bacteria in three different bulk soils ...33
4.2 The effects of genetic background and edaphic factors on the total and siderophore-producing rhizobacteria...33
4.3 The effects of genetic background and edaphic factors on the total and siderophore-producing endophytes...34
4.4 Identification of bacteria that possess the strongest siderophore-producing activity from rice tissues...35
4.5 Effect of siderophore-producing bacterial strains on rice growth...35
Chapter 5- Discussion...38
5.1 Characteristics of bacteria in three different bulk soils ...38
5.2 The effects of genetic background and edaphic factors on the total and siderophore-producing rhizobacteria...38
5.3 The effects of genetic background and edaphic factors on the total and siderophore-producing endophytes...40
5.4 Rice bacterial endophytes that possess the strongest siderophore-producing activity...44
5.5 The effect of siderophore-producing bacteria on rice growth ...45
Chapter 6 – Conclusion ...50
Limitations &Future Researches...51
Reference...52
Appendix- Tables and Figures...68
Extended abstract...80
Curriculum Vitae (CV)...86

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