Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/23065
標題: 耐鹽水稻品系的誘變育種與標幟連鎖分析
Mutagenetic Breeding and Linkage-Marker Analyses of Salt-Tolerant Rice
作者: 閻菁珠
Yen, Ching-Chu
關鍵字: 水稻
Oryza sativa
誘變
香米
耐鹽
幼苗期
遺傳
數量性狀
mutated
scented
salt tolerance
seedling stage
inheritance
quantitative trait loci (QTLs)
出版社: 生命科學系所
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摘要: 中文摘要 由於耐鹽育種材料之缺乏,以及大部分的稉稻品系對鹽分敏感,只有一些傳統的秈稻Pokkali、Nona Bokra、SR26B和Kalarata等具耐鹽性,這些耐鹽品系之耐鹽性遺傳是由微效基因(polygenes)控制,而且具有植株高、感光性、米質差及低產量等不佳的農藝性狀。欲從秈稻轉移耐鹽特徵到稉稻是困難的,因為耐鹽性和不理想的農藝性狀是連鎖的,所以具實用農藝性狀的耐鹽稉稻品系之育成十分不易。故本試驗採用稉稻之突變系為材料,挑選出耐鹽品系,分析生理特性推測其耐鹽機制,並了解其遺傳控制情形及標的基因之遺傳連鎖分析,作為培育非轉殖之耐鹽、高產品系,並進一步利用耐鹽突變體之重組純系作為分子標幟輔助育種選拔、耐鹽機制及基因調控之研究。 本試驗採用台農67 號(稉稻)經疊氮化鈉誘變之香米突變品系台農嘉農育911303號再度經EMS誘變之突變系為材料,2006年一期採收1005個M6誘變品系為基礎材料進行耐鹽檢定。在3葉齡時期,利用150 mM氯化鈉溶液處理,初步挑選出11個品系,其中SM75耐鹽性表現極佳,生長分析結果亦顯示SM75較親本台農嘉農育911303號的生長狀況及復原狀況表現佳,確定SM75為一耐鹽品系。將這11個品系在2007年一期繁殖為M7誘變品系,並與台農嘉農育911303號及台農67號,在6葉齡時期,以300 mM NaCl 進行鹽分處理3天,SM61是唯一存活品系,存活率高達83.33%。以200及250 mM NaCl處理3天,SM61較Pokkali和Nona Bokra明顯耐鹽,確認SM61為一耐鹽品系。進一步將SM61和親本嘉農育911303號,以150 mM NaCl處理一週,無論是地上部和根部,SM61之氯離子、鈉離子濃度及鈉鉀比皆較台農嘉農育911303號低;以25% PEG進行處理,SM61之葉片捲曲比例(捲曲葉片數/總葉片數)明顯較TNG67小,推測SM61之耐鹽性是因累積較少的氯離子、鈉離子以及較佳的滲透潛勢調節能力所造成。 再以200、250及300 mM NaCl處理,確定SM61是一耐鹽品系,而台中秈17號為非耐鹽品系,200 mM NaCl處理可以明顯區分SM61和台中秈17號之耐鹽性差異,因此以此作為篩選這二個品系雜交後F2耐鹽植株之最適當鹽分濃度。SM61和台中秈17號雜交後,於2007年二期採收F1、2008年一期採收F2及2008年二期採收F3。首先將SM61、F1和台中秈17號進行200 mM NaCl處理五天,結果顯示F1和SM61皆是耐鹽存活植株,然而台中秈17號為不耐鹽死亡植株,而且在F2代存活植株及死亡植株經卡方分析符合3:1之分離比,故SM61之耐鹽性是由一顯性單對基因所控制。利用所挑選的F2及F3耐鹽植株進行耐鹽標的基因之連鎖標幟分析,發現SM61耐鹽誘變基因之緊密連鎖標幟RM223位於第八對染色體上,對耐鹽之相關程度接近100%,此結果與以往所發現和提升耐鹽能力相關之QTLs位置皆不同,因此本研究是首先發現顯性單對耐鹽基因和標幟緊密連鎖之報告,對未來耐鹽水稻品系之育成將帶來重大的影響。
ABSTRACT Breeding materials for the development of salt-tolerant varieties were rarely identified and most japonica rice varieties were salt-sensitive, except for some traditional indica varieties, such as Pokkali, Nona Bokra, SR26B and Kalarata. Most previous studies suggested that the inheritance of the salt-tolerance is governed by polygenes. However, the traditional indica varieties possess undesirable agronomic characters of tall stature, photosensitive trait, poor grain quality, and low yield. Salt-tolerance genes are thus linked to undesirable agronomic characters, so it is difficult to transfer salt-tolerant traits of indica varieties into japonica varieties. For this reason, advanced salt-tolerant varieties with useful agronomic characteristics have not been successfully developed in japonica rice. The purposes of the study were to identify salt-tolerance lines, to analyze their physiological traits and salt-tolerance inheritance control, and to map linkage markers of salt-tolerance gene by using the steady genetic mutated lines originating from japonica rice. The salt-tolerance lines are not fransgenic and will be the best sources for breeding lines with salt tolerance and high yielding traits. The salt-tolerant recombinant inbreed lines provided for experiment materials of breeding programs in rice by marker-assisted selection and understanding the salt-tolerance mechanisms and the gene regulators. In our study, the wild-type cultivar, TNG67, was treated with a chemical mutagen, NaN3, to produce the steady genetic line of mutated scented rice, CNY911303. EMS was subsequently applied to CNY911303 to produce the second generation of 1005 mutated lines that were developed to the sixth generation with superfine agronomic traits in the first season of 2006. These 1005 lines at the three true leaf growth stage were screened with 150 mM NaCl for three weeks and 11 lines were selected and developed to the seventh generation in the first season of 2007. SM75 performed best among the selected 11 lines. After 150 mM NaCl treatment for one week, the fresh weight, dry weight and height of SM75 were higher than those of CNY911303. These results indicated that the ability of SM75 to grow and recover was superior than that of CNY911303. Thus, SM75 is a salt-tolerant line. Next, seedlings of the 11 selected lines, CNY911303, and TNG67 at six true leaf stage, were treated with 300 mM NaCl for three days. The screen revealed that the salt-tolerant line SM61 was the only surviving line, with a surviving percentage of 83.33%. The comparison between SM61 and the traditional indica varieties, Pokkali and Nona Bokra, was subsequently made. SM61, Pokkali and Nona Bokra seedlings, were applied with 200 or 250 mM NaCl for three days, and were then transferred to the culture media without NaCl. After the treatment of 200 or 250 mM NaCl, survival and growth of SM61 were superior than those of Pokkali and Nona Bokra. Based on the results, SM61 is considered a salt-tolerant line. At the stage of six true leaves, SM61 and CNY911303 were treated with 150 mM NaCl for one week, and the Na+ content, Cl- content, and Na+/K+ ratio of SM61 in shoots and roots were lower than those of CNY911303. When treated with 25% PEG at 6 leaf stage for five days, the leaf rolling ratio (rolling leaves /total leaves) of SM61 was much lower than that of TNG67. Based on these results, it was suggested that SM61 could tolerate salinity by accumulating lower amounts of the two ions, Na+ and Cl-, with a higher osmotic regulating ability. For instance study, SM61, the highly salt-tolerant parent line, and TCS17, a test cross parent variety, were treated with 200, 250 or 300 mM NaCl for five days. The results showed that SM61 is confirmed a salt-tolerant japonica line, while TCS17 is a salt-sensitive indica variety. Among the three different concentrations of NaCl used, 200 mM NaCl was found to be the best concentration for comparing performance and appearance differences between SM61 and TCS17. SM61was hybridized with TCS17 and produced F1 seeds in the second season of 2007. The F1 plants were self-fertilized to develop F2 seeds in the first season of 2008, and F2 plants produced F3 seeds in the second season of 2008. At six true leaf stage, under the treatment of 200 mM NaCl for five days, SM61 and F1 (SM61TCS17; TCS17SM61) rice plants were salt-tolerant and surviving (R) while TCS17 rice plants were salt-sensitive and unsurviving (S). The R and S ratio of 513 F2 rice plants produced a good fit to the Mendelian 3 : 1 segregation ratio by a Chi-square test, indicating that the salt-tolerant trait of SM61 was governed by a single dominant gene. Next, 290 F2 rice plants at a six true leaf growth stage in the culture media were supplemented with 200 mM NaCl for five days, and 46 F2 salt-tolerant rice plants were selected for the identification of the linkage markers that link to the mutated tolerant gene in SM61. F3 lines, derived from the F2 salt-tolerant surviving rice plants, were used to evaluate the genotypes of the F2 rice plants. The results showd that the mutated salt-tolerant gene could explain close to 100% of the total phenotypic variation, and it is tightly linked to the RM223 marker located on chromosome 8. The results clearly differ from those of previous studies investigating the relationship of QTLs to salt tolerance in rice. Our study was the first report to map a tightly linked marker to a single dominant mutated salt-tolerant gene that is not QTLs. Results will contribute to breeding programs of salt tolerance in rice.
URI: http://hdl.handle.net/11455/23065
其他識別: U0005-1601201114294400
文章連結: http://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-1601201114294400
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