• 作者： 成游貴; 黃真生; 賴光隆
• 作者服務機構： 台灣省畜產試驗所飼料作物系; 台灣省農業試驗所農藝系; 國立台灣大學農藝學系
• 中文摘要： 本實驗之目的在於研究不同細胞質對於花莉誘導癒合組織及分化成植株能力之影響，並進而探討細胞質雄不稔性花粉異常之原因，所用之材料為Reimei-A-line〔（O. rufipogon×Reimei)×Reimei/10〕及臺中65號-A-line〔（O. rufipogon x TC65）x TC65/10〕以及其維持親 B-line，培養基為改良MS培養基。 由花葯誘導形成癒合組織之頻度，因品種而異，Reimei A與B-line之頻度很低，而臺中65OAR. B-line各為10．8%R12.9,%'，因此，對於培養基之反應，因品種而異，而細胞質並無明顯之影響。 由培養後不同時間的花葯組織切片及外部形態觀察結果得知，位於營養層細胞上之球狀體為一明顯之特徵，可用來確認這些癒合組織均來自於小抱子而非葯壁之組織細胞。癒合組織外部細胞分裂旺盛，細胞內充滿細胞質及胞器，愈趨於癒合組織中央的細胞，則液泡化明顯，細胞基質少。A-line與B-line癒合組織之微細構造，無明顯差異。 由臺中65號A-line誘導之209個癒合組織，其分化率為76.6%，其中71.8%為綠苗，28.2%為白苗，臺中65號 B-line之分化率為83.1%，其中33.8%為綠苗，66.2%為白苗，因而A與B-line分化成綠苗比率有明顯之差異，可能與細胞質有關。分化之成株以二元體最多，單元體次之，多元體則比較少。成熟二元體植株，由A-line而來者，亦為細胞質雄不稔。 由以上結果獲知，雄不稔花葯亦如正常花葯，可經誘導而長出癒合組織並能分化出植株。由花葯培養方法雖不能證明雄不稔性花粉異常之原因，但綜合本研究之結果與前報 組織解剖研究之結果，可推論細胞質雄不稔花粉異常之原因，可能是由於營養層細胞未能充分供給小抱子發育所需之養分，而導致花粉不稔。
• 英文摘要： This experiment was conducted to inves-tigate the effect of cytoplasm on callusformation and plant regeneration, and also toclarify the cause of pollen abortion in cyto-plasmic male sterile (CMS) rice. Materials used were Reimei A-linederived from O. rufipogon/Reimei , Taichung65 A-line from O. rufipogon/Taichung 65 ,and their B lines. The medium was amodified MS medium (Murashige and Skoog, 1962) in which the major salts (except Fe-EDTA) were reduced to half of their originalstrengths and 2 mg/liter kinetin, 4 mg/literα-naphthaleneacetic acid (NAA), and 2 mg/liter 2,4-D were added. Cultures wereincubated at 25°C 1°C under 16-hour dailyillumination with cool white fluorescent light. For plant generation, 4-week-old callus weretransferred to full-strength MS medium with3% sucrose, 4 mg/liter kinetin and 1 mg/liter NAA and maintained in the samecondition as for callus induction. Frequency of callus formation was low inReimei A-line and B-line，but was 10.8% for Taichung 65 A-line and 12.9% for theB-line. Response of anthers on the mediumfor callus formation did not differ significantlybetween cytoplasms but differed significantlybetween varieties. Histological and externally morphologicalobservations of cultured anther at differentstages with light and electron microscopiesshowed that the initial callus in anthers wasfound disconnected with the inner wall ofanther. Especially, the ubisch body on thetapetum was still existed which helped toidentify the callus being originated frommicrospores，not from anther walls. Veryfast cell division was found at the peripheralregion of callus. These cells were dense incytoplasm and with numerous organelles. The closer to the interior part of callus, themore vacuolate and lesser cytoplasm the cellbecame. There were no significant differencesin callus fine structures between A-line andB-line. Among 209 callus cultures from Taichung65 A-line, 76.6% produced plants, amongthem, 71.8% were green and 28.2% werealbino. The frequency of plant regenerationin Taichung 65 B-line was 83.1%, amongthem,33.8% were green and 66.2% werealbino. The difference in albino productionfrequencies between A and B line might becaused by different cytoplasmic effects of O.sativa and O. rufipogbn. Plants obtained fromanther culture were mostly diploids, followedby haploids and polyploids in the order. Themature diploid plants derived from A-linewere also cytoplasmic male sterile. The foregoing results showed that themale sterile anthers, just like the normalones，could be induced to form callus whicheventually differentiated into plantlets．Though anther culturing method did notexplain how the abnormal pollen of cytopla-smic male sterile rice is formed. The resultsfrom this study and the previous histological/morphological investigations might suggestthat the tapetal cell's failing to supply thenutrients needed by microspores for develo-ping was the cause which rendered the pollenabnormal and sterile.
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