柑桔葉片同功異構酶之研究　Ⅰ.本省重要柑桔葉片之電泳分析與葉齡對分析結果之影響

Studies on the Isoenyzmes of Citrus Leaves Ⅰ. The Gel Electrophoretic Analysis of Six Important Citrus Species in Taiwan and the Effect of Leaf Age

吳明哲(Min-Tze Wu) ； 卓俊銘(Jung-Ming Jwo)

《中華農業研究》 35卷4期 (1986/12) Pp. 484-494

https://doi.org/10.29951/JARC.198612.0008

摘要

本研究以目前本省最重要的六個柑桔栽培種－極柑、桶柑、柳橙、檸檬、葡萄柚和柚子等二年生植株之葉片爲材料，藉電泳分離技術，探討Peroxidase, Esterase, Superoxide dismutase與Glutamate oxaloacetate transaminase等四種酵素之同功異構酶基因型之差異，做爲將來育種上珠心胚苗與有性胚苗鑑定之依據，以提高育種效率。此外，由於不同成熟度葉片之生理狀況可能不同，本研究亦同時探討葉齡對於電泳分離結果之可能影響。
試驗結果顯示，Peroxidase在Rf 0.35, 0.37之條帶，Esterase在Rf 0.3之條帶Superoxide dismutase在Rf 0.29, 0.32, 0.36, 0.39之條帶均有明顯之種间遺傳差異存在，可做爲鑑別之依據。Glutamate oxaloacetate transaminase電泳分離結果，可明顯分出兩羣獨立基因座（GOT-1和GOT-2）所控制之同功異構酶，除椪柑與桶柑之類型相同外，其餘受測柑桔種間均存有不同之類型。由於受測柑桔種間有此種明顯之差異存在，且由於電泳方法的建立，具有操作簡單，快速的優點，此等技術在未來本省柑桔雜交育種上，具有實際應用於珠心胚苗與有性胚苗早期鑑定之價值。
不同葉齡的電泳分離結果顯示，Peroxidase與Superoxide dismutase以完全成熟葉，Esterase以嫩葉，Glutamate oxaloacetate transaminase以不完全成熟葉爲最清晰完整。此外，本研究結果顯示桶柑之Esterase同功異構酶基因型介於椪柑與柳橙之間，葡萄柚之Glutamate oxaloacetate transaminase同功異構酶基因型介於柚子與柳橙之間。彼等種間之血緣關係，有待進一步之探討。

並列摘要

Leaves from 2-year-old plants of six important citrus species in Taiwan…Ponkan (Citrus reticulata), Tankan (a tangor?), Liuchen (C. sinensis), Lemmon (C. Limon. var. Eureka), grapefruit (C. paradisi var. Ruby) and Wentan (C. grandis), were used as the plant materials for identification of the isoenzyme genotypes of peroxidase, esterase, superoxide dismutase and glutamate oxaloacetate transaminase. The objective of this study was to look for the genetic differences among certain citrus species. The effect of leaf age on isoenzyme banding patterns was also tested.
The results indicated that bands of Rf 0.35, 0.37 on peroxidase, band of Rf 0.3 on esterase, and bands of Rf 0.29, 0.32, 0.36, 0.39 on superoxide dismutase could be the genetic markers for distinguishing differences among certain citrus species. From the isoenzyme banding patterns of glutamate oxaloacetate transaminase, two groups of iso enzymes controlled by two genetic Loci (GOT-1 and GOT-2) were clearly separated. The GOT banding patterns of Ponkan and Tankan were the same, but those of the other 4 were different. Referring to the evidence of differences among certain citrus species in isoenzyme genotypes identified by means of the above simple and rapid electrophoretic method, it is believed that this technology can be applied to differentiate the zygotic from nucellar seedlings in the future citrus breeding work in Taiwan.
The clearness of isoenzyme banding was influenced by leaf age. Zymograms from completely mature leaves were the clearest for peroxidase and supeoxide dismutase, while those from young leaves was for esterse and those from incompletely mature leaves was for glutamate oxaloacetate transaminase. The choice of different ages of leaf samples should be emphasized when such an electrophoretic experiment was conducted. This study also showed that esterase isoenzyme banding pattern of Tankan is in between those of Ponkan and Liuchen, the glutamate oxaloacetate transaminase isoenzyme banding pattern of grapefruit in between those of Wentan and Liuchen. However, it needs more study to evidence the taxonomy of Tankan and grapefruit.