Cytological studies on the incompatibility of protoplast fusion in crops

1.对绿豆叶肉原生质体和红豆细胞悬浮液原生质体进行了电融合，并进行了电镜观察。绿豆叶绿体和红豆原质体在融合产物中分别聚为一类，并定位于各自的原边。在部分子细胞中观察到叶绿体的消失.研究了碘乙酰胺（IOA）对红豆原生质体的影响。IOA诱导粗面内质网的过度形成，随后是细胞质的塌陷。而将IOA处理的绿豆原生质体与未处理的绿豆原生质体融合，超微结构损伤得到减轻.将烟草叶肉原生质体与大豆细胞悬浮液原生质体融合，并在不同的培养基中进行培养。在有利于大豆原生质体分裂的培养基中，子细胞中叶绿体的去除率最高，而在有利于烟草原生质体分裂的培养基中，子细胞中叶绿体的去除率不高.以鸡冠花（Celosia cristata，C_3）和苋菜（Amaranthus tricolor，C_4）为材料，通过原生质体融合获得了属间体细胞杂种愈伤组织。酸性磷酸酶同工酶分析证实了愈伤组织的杂种性，说明即使在远缘种间原生质体融合的早期过程中，也没有观察到不亲和反应。细胞器的消除似乎是逐渐发生的，由于细胞分裂过程中细胞器的不平等分离。

1. Mesophyll protoplasts of mung bean and cell suspension-derived protoplasts of azuki bean were electrofused and examined by electron microscopy. Chloroplasts from mung bean and proplastids from azuki bean were grouped respectively and located on their original sides in fusion products. Elimination of chloroplasts was observed in some daughter cells.2. Effects of iodoacetamide (IOA) on azuki bean protoplasts were examined. IOA induced an overformation of rough endoplasmic reticulum which was followed by the collapse of cytoplasm. However, ultrastructural damages were mitigated if the IOA-treated protoplasts were fused with untreated mung bean protoplasts.3. Tobacco mesophyll protoplasts and soybean cell suspension-derived protoplasts were fused and cultured in various media in which division activity of parental protoplasts was differently affected. Elimination of tobacco chloroplasts from daughter cells was the highest in the medium profitable to division of soybean protoplasts but was not observed in the medium profitable to tobacco protoplasts.4. Intergeneric somatic hybrid calli were obtained through protoplast fusion between Celosia cristata (C_3 species) and Amaranthus tricolor (C_4 species). Isozyme analysis of acid phosphatase confirmed the hybridity of calli.In conclusion, incompatibility reaction was not observed in early processes of protoplast fusion even between remote species. Elimination of organelles seems to occur gradually due to unequal segregation of organelles during cell divisions.