Protoplasts were isolated from Agrobucterium rhizogenes A-transformed cell line of Medicago sativa L. The highest yield of protoplasts (4.2×106per g fresh weight) was obtained from 12-d-old calluses after being subcultured on fresh medium. The viability of protoplasts reached over 80%. Protoplasts were induced to undergo sustained divisions when cultured in Durand et al. (DPD) medium supplemented with 2mgl-1 (9.05μM)2, 4-dichlorophenoxyacetic acid. 0.2mgl-1(0.93μM)kinetin, 0.3M mannitol, 2%(w/v)sucrose, and 500mgl-1 casein hydrolyzate at a plating density of 1.0×106perml. An agarose-beads culture method was appropriate for protoplast division of transformed alfalfa. The division frequency was ablut 30%. Numerous hairy roots were induced from protocalluses on Murashige and Skoog medium without grouwth regulators. Paper electrophoresis revealed that all of the regenerated hairy roots tested synthesized the corresponding opines. This protoplast culture system would be valuable for further somatic hybridization in forage legumes.

The legume forage Alhagi pseudoalhagi was transformed by the Agrobacterium rhizogenes strain A4 using cotyledon and hypocotyl segments as infection materials. Regenerated plants were achieved from sterile calli derived from hairy roots, which occurred at or near the infection sites. The regenerated plants from hairy root were characterized by normal leaf morphology and stein growth but a shallow and more extensive root system than normal plants. Opine synthesis, PCR and Southern blot confirmed that TDNA had becn integrated into the A. pseudoalhagi genome. Acetosyringone (AS) was found to be vital for successful transformation of A. pseudoalhagi.

A reproducible release of viable protoplasts was obtained from friable calli of Astragalusudsurgens. Protoplasts underwent sustained divisions and formed cell colonies when cultured in either liquid or agarose-solidified Kao and Michayluk (1975) protoplast medium (KM8P) supplemented with 1.5 rag/1 2,4-D, 0.5 mg/I BA and 0.5 M glucose. Compared to liquid culture, agarose bead culture improved division frequency effectively, the two culture systems showing a plating efficiency of 0.8_+0.5% and 6.5_+0.7%. respectively. Upon transfer to Murashige and Skoog (1962) medium (MS) with 1-2 mg/I BA, alone or in combination with NAA or 2,4-D at 0.1 rag/l, the protoplast-derived ealli produced complete plantlets through somatic embryogenesis. The maximum percentage of calli producing somatic embryos (52.5

Protoplast fusion was induced between sainfoin and alfalfa by and improved polyethylenegylcol (PEG) method. The intergeneric somatic calluses were selected based on complementation of hydroxyproline-resistance of sainfoin and hormone autonomy growth of alfalfa transformation cell line. 17 somatic hybrid plantlets were regenerated. PEG could induce the tight agglutination of protoplasts. During diluting and washing process, cyclization of the linked membrane and formation of vesicle-like structures were observed, resulting in protoplast fusion. 5%-10% glycerol supplemented in the fusion inducing solution markedly increased the frequency of heterogeneous fusion. Better fusion results were obtained when mixed protoplast suspension was dripped in petri dishes in which PEG sclution was previously placed. Chromosome number of regenerated hybrid buds varied from 30to 60. The genome of hybrids included the small chromosome from sainfoin and two chromosomes with two clear constrictions from alfalfa. The hybridity of obtained hybrid calluses was confirmed by their isozyme banding patterns and their nopaline synthetase acivity.

Callus were induced from the segments of basal part of leaf blade of rye (Secale cereal L) on MS medium supplemented with 3 mg/L 2, 4-D. Embryogenic calli were obtained when the calli derived from leaf tissues were subcultured on N6 medium plus 500 mg/L yeast cxtract, 1-2 mg/L 2, 4-D and 1-2 mg/L kinetin. Somatic embryos frequently were produced after transfering the cultures onto MS or N6 medium with 5% sucrose without growth regulators. The embryos germinated and developed into normal plantlets on hormone-free medium. The regenerated plants were potted. Histological investigation on callus initiation from leaf tissues was carried out.

Wheat (Triticum aestivum L.) is one of the most important cereal crops in the world. Great attention has long been paid to the technique of protoplast culture of this species[1,2] Up to now, plantlet regeneration from protoplasts of anther-derived suspension cultures of wheat has been reported for the first time[3], In the present study, we have obtained numerous embryoids and plantlets from protoplasts of immature inflorescence-derived calli of spring wheat.

从继代培养欧当归（Levisticum officinale Koch）愈伤组织分离的原生质体在C81V培养基中培养，得到了较高频率的持结分裂。形成的细胞团转移到固体培养基上后，发展成了白色松软型的愈伤组织。将其转移到附加6-苄氨基嘌吟和吲丁酸的N6培养基上继续培养了2-3个月后，表面出了浅黄色、颗粒状的胚性愈伤组织。后者在附加吲丁酸和萘乙酸的MS培养上培养，可通过体细胞胚胎发生途径分化出大量小植株。对供体愈伤组织和原生质体再生植株进行了细胞学观察。

Cultures of haploid and diploid leaf protoplasts of Petunia hybrida var. Mitchell were treated with 5 virulent strains of Agrobacterium tumefaciens. Transformed clones, characterised by their hormone independent growth and nopaline production, were isolated following treatment with strain T37.

以豆科牧草沙旺为一亲本，碘乙酰胺处理的紫花苜蓿发根农植菌A4菌株转化系为另一亲本，通过PEG-高PH，高钙法诱导原生质体融合。在不加外源激素的DPD培养基上有效地筛选了杂种细胞。经培养首次得到沙打旺（+）紫花苜蓿的属间体细胞杂种。尽管双亲原生质体均已丧失分化植株的能力，但杂种细胞系R1仍得到苗的分化。杂种R1细胞的染色体数检查、冠瘿碱检测、同工酶和RAPD分析结果，都证实了其杂种特性。

Mesophyll protoplasts have been isolated from soil grown plants of Solanum melongena var depressum Bailey and protoplast-derived colonies regenerated in liquid DPD culture medium. Protoplast-derived colonies proliferated on agar-solidified MS culture medium and regenerated shoots which grew to fertile plants in potting compost.