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標題: 台灣薏苡種原農藝性狀變異及利用RAPD鑑別與親緣關係之研究
Studies on the character variation and phyletic relationship among Coix lacryma-jobi L. in Taiwan based on RAPD markers
作者: 蘇宗振
Su, Tsung-Chen
關鍵字: job''''s tear;薏苡;adlay;relationship;親緣關係
出版社: 農藝學系
一、共收集國內外66個薏苡品種(系)進行農藝性狀調查,探討薏苡種原性狀變異及品種間親緣關係。經調查14個農藝性狀結果顯示薏苡品種(系)間農藝性狀有顯著差異存在。性狀相關性分析結果顯示每株中間穗數、每株葉數、稔實粒重等三個性狀與產量呈極顯著正相關。又利用主成分分析發現抽穗日數、收穫期、株高及最低穗位高等四個性狀是種原變異的最主要來源。經歸群分析後可分為三大類群,其中以包括對照品種在內的類群最大,Indonesia sumatra san則與其他品種關係最遠。
二、本試驗調查薏仁組成分及薏仁酯含量等8個品質性狀,發現台灣現有66個薏苡品種(系),其脫殼薏仁之成分為:灰分1.7%、粗脂肪7.4%、粗蛋白13.9%、粗纖維0.16%及薏仁酯含量為27.7mg/g oil;帶殼薏仁之成分為:灰分6.7%、粗脂肪3.9%、粗蛋白8.9%、粗纖維0.29%。而脫殼薏仁之粗蛋白與帶殼薏仁粗纖維呈極顯著正相關(r=0.586);薏仁酯的含量與帶殼薏仁之粗蛋白含量呈正相關性。利用典型相關分析結果表明,影響帶殼薏仁粗纖維及脫殼薏仁粗蛋白的農藝性狀為百粒重、抽穗期、收穫期、種子寬度等,並與種皮顏色呈負相關。綜合品質以及農藝22個性狀之分析結果,將台灣薏苡種原分為三個類群,其中C類群為major變種。
三、由PCR得到41個有效引子其擴增產物表現出DNA多型性,總條帶數為258條,其中具多型性條帶78條,多型性條帶獲得比例約30%。並從41個有效引子中選擇具多型性比例高之8及10個引子來進行薏苡種原之RAPD分析。綜合遺傳距離遠近之結果,發現台灣收集之薏苡種原間尚存在著遺傳差異。且由混合植體DNA之結果綜合討論,推論常異交作物之RAPD分析中,具80%以上條帶較具代表性,其分辨major及mayuen變種之能力。比較混合植體DNA之RAPD與農藝性狀之分群結果發現大致上吻合。所以利用RAPD分析所得的結果,可以和其形態上的分群資料相佐證,但並未完全反應其在形態上的差異。採用較嚴格的譜型(III)較能有效鑑別mayuen 及major等二變種。

Coix lacryma-jobi L. is a popular crop in Taiwan. Owing to its function of curing many diseases, including cancer, allergy and other diseases, this crop is also titled as ‘the king of cereals'. Despite the identification of phenotypic characteristics through different varieties is simple and fast, little is known about the DNA markers level, such as RAPD analysis, in this crop. In this thesis, the main objectives were to study the agronomic characters, various chemical components analysis and relationships of 66 coix varieties (lines) from different countries through RAPD method.
To identify fourteen agronomic characters in 66 coix varieties (lines), we found grain yield revealed the significantly positive correlation with spikelet number, total leaf number and fertile seed weight. According to the PCA analysis, it revealed that the diversity of germplasm was caused by he traits of plant height, lowest spikelet, silking period and harvesting time. And there are three clusters had been classified using the cluster analysis. Among these three clusters, the variety of Indonesia sumatra san had the farthest distance with others.
The investigation of various chemical components from the dehulled grains of 66 varieties (lines) revealed the percentage of 1.7%, 7.4%, 13.9%, 0.16% and 27.7﹪in ash, crude fat, crude protein, crude fiber and coixenolide respectively. The same investigation from the grains with hull appeared 6.7%, 3.9%, 8.9%, 0.29% in ash, crude fat, crude protein, crude fiber each. We also discovered that the crude protein in dehulled grains showed the significantly positive correlation (r=0.586) with the crude fiber in the grains with hull. The similar result also appeared in the amount of coixenolide and crude protein in the grains with hull. Based on the typical correlation analysis, it showed that 100 grain weight, silking time, harvesting time, and grain width are the main agronomic traits to affect crude fiber in grains with hull and dehulled grains, however the traits mentioned above showed negative correlation with the color of the grains' hull. In this section, there were 3 groups which were classified through quality analysis and 22 agronomic traits analysis, and the group C was confirmed as the major sub-variety.
In the primary RAPD analysis in coix germplasm, we obtained 78 polymorphic bands from 258 bands in total with 41 valid primers (about 30% polymorphism ratio). And about 8-10 valid primers (with highly polymorphic ratio) were chosen from the former 41 ones for the subsequent RAPD analysis. Nevertheless, there are some genetic differences existed in the coix germplasm via the results of genetic distance. By using the mixed DNAs from different plants of same variety as the template, we deduced that above 80% of bands showed the representatives in often-cross-pollinated crops and the ability to distinguish between the sub-varieties of major and mayuen. And the result from the RAPD analysis is as similar as the cluster analysis of agronomic traits. Conclusion to the results showed above, the outcomes from RAPD analysis can close to the results from morphological differences, but not absolutely. The pattern III would be more efficient to identify the sub-variety of major and mayuen.
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