原生质体

Protoplast fusion can circumvent reproductive barriers facing citrus, such as polyembryony and female and/or male sterility, and paves way for citrus cultivar improvement.

柑橘原生质体融合主要采用"叶肉原生质体+胚性愈伤组织原生质体"和"胚性愈伤组织原生质体+胚性愈伤组织原生质体"两种模式，融合方式主要有对称和非对称融合。

The results showed that the Saccharopolyspora spinosa SP06081 protoplast yield was the highest under these conditions: the collected mycelium from SP06081 grown in Tryptic Soy Broth medium with 0.2% glycin for 48 h was treated by 0.1 mg/mL lysozyme at 28oC for 20 min, then plated on the R2YE medium with sucrose as osmotic stabilizer, the number of regeneration protoplast was up to 108/mL. The protoplast-regenerated strains exhibited changes in morphology and antibiotic production, 29.3% protoplast-regenerated strains was characterized by loose mycelium and abundant broken branches as did their parent. Among them, 58.2% strains presented the trend to positive variation in spinosad yield, with the highest spinosad yield of up to 582.0 mg/L, 85.6% higher than that of their parent.

结果表明：菌体在添加0.2%的甘氨酸的TSB培养基中培养48 h收集， 0.1 mg/mL 溶菌酶， 28oC作用20 min制备原生质体，将原生质体涂布于以蔗糖为渗透压稳定剂的R2YE培养基中，原生质体再生数目最多，达108个/mL以上；原生质体再生菌株在形态和抗生素产量上产生分化， 29.3%的再生菌株形态上保持与亲本菌株一致，具有菌丝松散，断裂分枝多的特点，其中53.2%的再生菌株多杀菌素产量变异向正方向移动，最高产量达到582.0 mg/L，比亲本菌株提高85.6%。

The circumtances of the protolasts culture of leguminosae and the tisse and protoplasts culture of sainfoin has been reviewed in this paper.

本文还综述了豆科原生质体和红豆草原生质体的培养概况以及影响豆科原生质体培养的几个重要因素。

To find out the suitable medium, the optimization groups of hormone of NAA content and ZT content and the appropriate cultural density, the protoplast of two species of Jujuba trees have been cultivated by taking as subject the protoplast of non-kernel small date and egg-like dates embryonic suspension cells obtained by zymolysis, and by employing different mediums with different combination content of NAA, ZT hormones and culture density of protoplast.

以无核小枣和鸡蛋枣两个枣树品种的胚性悬浮细胞经酶解获得的原生质体为材料，采用不同种类的培养基、不同含量的NAA和ZT激素组合及不同原生质体培养密度对两个枣树品种的原生质体进行培养，以获得适宜的培养基种类、较佳NAA和ZT的激素组合及适宜的培养密度。

The results showed that:The best zymolysis conditions for BS and TV protoplasts release and regeneration through orthogonal experiment were as follows: 1mg·mL~(-1) lysozyme treated BS protoplasts for 1.5 hours at 37℃ and 4 mg·mL~(-1) lywallzyme treated TV protoplasts for 2 hours at 30℃. In these conditions, the number of protoplasts release was respectively 8.87×10~8 units/mL and 8.62×10~7units/mL and the regeneration rate was respectively 54.1% and 29.3%.

研究结果表明：通过正交试验确定BS菌和TV菌原生质体制备过程中的最佳酶解条件分别为：BS菌在37℃酶解温度下，1mg/mL溶菌酶酶解1.5h，原生质体制备个数达到8.87×10~8个/mL，再生率为54.1%；TV菌在30℃酶解温度下，4mg/mL破壁酶酶解2h，原生质体制备个数为8.62×10~7个/mL，再生率为29.3%。

The protoplast fusion between wild plant species L.fendleri of the genus Lesquerella and B.napus was studied in order to transfer the industrial oil gene in L.fendleri into B.napus.

为了把野生优质工业用油植物基因导入到普通栽培油菜品种中，本文报道了野生优质工业用油植物L 。 fendleri的原生质体培养及其原生质体与普通栽培油菜品种原生质体融合。

The best combination of isolating protoplast of Mozzie buster was the enzyme mixture containing 2.0% Cellulase R-10, 0.4% Pectinase, 0.5M mannitol as osmoticpressure, 27℃under dark 80r/m rotating enzymolysis 8h, and the highest yieldachieved to 1.67×10~6per gram, the protoplast vitality was 87.4%; The bestcombination of isolating protoplast of Ocimum basilicum was the enzyme mixturecontaining 1.6% Cellulase R-10, 0.4.% Pectinase, 0.5M mannitol as osmotic pressure,27℃under dark 80r/m rotating enzymolysis 8h, and the highest yield achieved to1.67×10~6 per gram, the protoplast vitality was 87.4%.

本研究对酶解最佳条件进行研究，得出分离驱蚊草原生质体酶解条件以CPW-13进行质壁分离1h，2.0％纤维素酶和0.4％果胶酶，0.5M甘露醇为渗透压，27℃黑暗下80r／m酶解处理8h为好，产量可达1.67×10~6个／g，原生质体活力为87.4％；分离罗勒原生质体酶解条件以CPW-13进行质壁分离1h，1.6％纤维素酶和0.4％果胶酶，0.5M甘露醇为渗透压为好，27℃黑暗下80r／m酶解处理8h，产量可达2.14×10~6个／g，原生质体活力为67.8％。

One L-isoleucine high-producing strain SW0370 was screened through protoplast fusion, shake flask compound screening, genetic marker test and single clone isolation and continuous passage cultivation.

3确定了亲株ISW330和AS1.495原生质体形成和再生及进行原生质体融合的最佳条件，采用原生质体融合技术最终筛选出一株带有目的遗传标记的L-异亮氨酸高产菌SW0370〓。

2The protoplast formation and regeneration and fusion conditions of strain ISW330 and AS 1.495 were studied in this thesis. One L-isoleucine high-producing strain SW0370 was screened through protoplast fusion, shake flask screening, genetic marker test and single clone isolation and continuous cultivation for generations.

2确定了亲株ISW330和AS1.495原生质体形成和再生及进行原生质体融合的最佳条件，采用原生质体融合技术最终筛选出一株带有目的遗传标记的L-异亮氨酸高产菌SW0370(Leu~-+Met~-+AEC~r+á-AB~r)。

Second, procedures of effective protoplasts fusion and U.V. irradiation or heat treatment protoplasts fusion were developed. Equal protoplast suspensions (containing about 108/ml protoplasts) of each strain were mixed and washed by centrifugation, then the pellets were resuspended in PEG4000 solution (35%) at 37癈 for 2min.

其次，确定了原生质体融合的基本条件及灭活原生质体融合的条件：等量混合两亲株原生质体或用热灭活的原生质体(＞10~8个／ml)，用35％的PEG4000在37℃处理2min后，稀释，涂布再生，可获得融合率在50％左右的融合子。

This one mode pays close attention to network credence foundation of the businessman very much.

这一模式非常关注商人的网络信用基础。

Cell morphology of bacterial ghost of Pasteurella multocida was observed by scanning electron microscopy and inactivation ratio was estimated by CFU analysi.

扫描电镜观察多杀性巴氏杆菌细菌幽灵和菌落形成单位评价遗传灭活率。