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Phenotype characterization and genetic analysis of brittle culm mutants of IR64 and TNG67 varieties
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水稻（Oryza sativa L.）為世界上重要的糧食作物，亦是功能性基因體學研究的單子葉模式作物。作物的機械強度（mechanical strength）為一重要的農藝性狀，機械強度不足的作物容易造成倒伏或折斷，嚴重影響作物生長、發育與產量。本研究依手折植株葉片及稻稈的斷裂程度建立水稻脆度判定系統，將脆性程度區分為不脆（0）、微脆（1）、中脆（3）與極脆（5）四個等級。在M1到M4世代由疊氮化鈉（NaN3）誘變處理秈稻IR64品種的突變體庫，篩選出13株具脆性的突變體。在M9世代脆性突變庫中，共有9個品系具有等級5的脆度，2個品系脆度3，2個品系具有輕微脆度。以M5世代純化的兩個脆性品系IRB3（株型正常、脆度3）與IRB12（株型矮小、脆度5）之葉片及稻稈為材料，以石蠟切片進行分析，脆性突變體IRB3、IRB12之葉脈維管束間距離較IR64短，bulliform細胞寬度亦較IR64短；稻稈橫切結果顯示，IR64稻稈維管束間距離及稻稈厚度皆比脆性品系大。以非脆性品種TNG67為母本，與具脆性之父本IRB3雜交，進行遺傳分析，以SSR多型性分子標誌確認F1真偽。F1皆不具明顯脆性，在最高分蘗期進行F2族群之稻稈及葉片的脆度判定，將脆度觀測值進行卡方檢定，χ2值分別為1.673及0.956，不脆:脆之分離比均符合3：1，說明突變品系IRB3之脆性性狀由單一隱性基因控制。
The mechanical strength of crop is an important agronomic trait, less mechanical strength makes lodging or breaking of plants and results in tremendous yield loss. Four brittle levels of leaves or culms were defined as 0 (non-brittleness), 1 (slight), 3 (moderate) and 5 (extreme) according to the cracking point by hand-bending in this study. Accordingly, 13 brittle mutants were screened during M1 to M4 generations from the indica type mutant pool of IR64 variety developed by sodium azide mutagenesis. Finally, 9 mutants with level 5, 2 mutant with level 3, and 2 mutants with level 1 brittleness respectively, were obtained at the M9 generation. Two pure lines, IRB3 (normal plant type, level 3 brittleness) and IRB12 (dwarf, level 5 brittleness), were selected in M5 for anatomical analysis. The results showed that the distance between vascular bundles of mutants were shorter than of IR64 ones and the brittle mutants also had less width in bulliform cell in leaf. Similarly, in the culm of brittle mutants, shorter distance between vascular bundles and thickness of cell wall were also observed than of IR64 ones. The IRB3 mutant was crossed with TNG67 to investigate the inheritance of brittleness. Their hybrids were screened by polymorphic SSR markers and no brittleness was found in F1. The brittleness of leaf and culm in the F2 population were determined and Chi-squared analysis analyzed, the results displayed that the brittleness of IRB3 was governed by a single recessive gene.
Twelve pure brittle mutants were selected from a japonica mutant pool of TNG67 variety. Four lines, SA1067, SA1551, SA1566 and SA1610, were crossed with TNG67 and processed to F2. The genetic analysis suggested that the brittleness of these mutants were controlled by a single recessive gene. Allelic test was conducted by crossing between mutants and the brittleness of plants was investigated. All the F2 of SA1551/SA1610 showed brittle culms indicated that they were mutated at the same locus. However, the non-brittle vs. brittle ratio of F2 populations in the crosses of SA1067/SA1551, SA1067/SA1610, SA1551/SA1566 and SA1566/SA1610 fitted 9:7 suggested that the brittleness trait of these mutants were mutated or controlled by different genes. Molecular investigation of these mutants will provide valuable information for sodium azide mutagenesis as well as the applications.
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