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| 題 名 | 水稻原生質體之分離與融合 |
|---|---|
| 作 者 | 賴光隆; 劉麗飛; | 書刊名 | 中華農學會報 |
| 卷 期 | 93 1976.03[民65.03] |
| 頁 次 | 頁10-29 |
| 關鍵詞 | 分離; 水稻原生質體; 融合; |
| 語 文 | 中文(Chinese) |
| 中文摘要 | 本研究曾利用酵素處理之方法,自無菌培養或土耕之水稻幼苗置於各種條件,溶解組織細胞之細胞壁,使原生質體游離後,進行融合及培養之試驗。茲將試驗結果摘要分述如下: (一)水稻原生質體之分離在適當濃度的混合酵素液下確可進行,其分離之難易,主要受苗齡及材料切斷方法兩種因素之影響較鉅。於培養皿發芽10天之黃化幼苗,隨著發芽日數之增加,原生質體分離數目亦隨著增加,而本試驗範圍內以第10天之幼苗可得到最高量之原生質體之分離。供試材料中適當分化之組織最適於原生質體之分離,尚未分化之分生組織及已完全分化之葉片及葉鞘組織,均不適於用酵素方法分離原生質體。 (二)若採用橫切方式處理材料,則必須使用機械外力幫助原生質體分離,但長時間處理或較大之機械外力,均亦對原生質體造成傷害,若採用縱切方式處理材料,則不須使用機械外力,僅用靜置處理,?可顯著增加原生質體之分離。 (三)材料若在失水狀況下則不利於原生質體之分離。分離酵素液中Na-dextran sulfate 之有無原生質離劑種類,對原生質體之分離並無顯著影響。由不同組織分離之原生質體形態、大小均顯著不同,可做為融合試驗過程中,原生質體來源之識別。 (四)水稻原生質體分離過程中,部分有自然融合之發生。若以0.25M硝酸鈉溶液處理已分離之原生質體,較易誘導其融合,而硝酸鉀、硝酸鈣、白明膠、聚乙烯乙二醇等則無特殊效果。提高溫度可以促使類似融合之現象增加,但以不超過40℃為宜。pH值在4到10之範圍內對原生質體融合無特殊影響。 (五)分離之原生質體經培養後,會有類似細胞壁之構造形成,同時原生質體會有脹大之現象,細胞內容物亦會有顯著的改變。由水稻組織分離的原生質體,在培養基中至少可存活2週以上。但未觀察到明顯的細胞分裂現象。 (六)如果選擇的酵素混合液適當,水稻組織確可分離多量完整的原生質體,又初步的結果顯示,原生質體可保持一定時間之活性,如予深入探討,融合及培養均有望實現。 |
| 英文摘要 | This study was conducted to find out the effective method for enzymatic isolation of rice protoplast. The fusion and culture of the protoplast obtained were also conducted. The results obtained are summarized as follows: 1. In the process of enzymatic isolation of the protoplast, the age of the rice seedling and the isolation methods employed were the two major factors affecting on isolation of the protoplast. Within 10 days seedling, the older the seedling was, the more protoplasts were obtained. The maximum number of protoplasts can be obtained from 10-day old seedling. The tissue properly differentiated was more suitable for protoplast isolatin, but the meristematic tissue and matured leaf blade and sheath were unsuitable for the isolation of protoplast by enzymatic method. 2. The number of isolated protoplast was less if young leaf was cut in a cross way even accompanied the physical treatment of stirring or shaking during isolation. The isolation of the protoplast was enhanced if leaf was cut in a longitudinal way along the leaf axis even in stationary isolation. 3. The amount of isolated protoplast decreased when the tissue was subjected partial dehydration before the isolation treatment. It is no effect at all for the isolation of protoplast whether Na-dextran sulfate existed or not, and what kind of sugar was used as osmotic stabilizer. The morphological characteristics quite different among protoplasts originated from different tissues, may be served for descrimination the origin of protoplast during fusion. 4. The spontaneous fusion of protoplasts was observed during enzymatic isolation as well as artificially induced. The induced fusion of isolated protoplasts was achieved by 0.25M NaNO3, but not by KNO3, Ca(NO3)2, gelatin and PEG. Raising temperature to 40℃ during fusion, promoted greatly the rate of fusion. The pH value from 4 to 10 showed no difference among them. 5. The protoplasts which subjected for culture regenerated wall like structure in the spherical outer layer of the protoplast. In the cultured condition, they survived at least 2 weeks, but no apparant cell division was observed. 6. The experiments have shown that large amount of protoplasts could be isolated from rice tissue under proper enzyme solution. The isolated protoplasts could maintain their activity for a period time. Further studies for fusion and culture of isolated protoplasts, are necessary for establishment of fused cell culture. |
本系統中英文摘要資訊取自各篇刊載內容。