龙眼胚性悬浮细胞原生质体培养及其体胚发生再生植株     

赖钟雄  陈振光 《热带作物学报》2002,23(3):53-60

以“红核子”、“东壁”、“十二月龙眼”等龙眼品种的胚性悬浮细胞为起始材料分离原生质体,采用液体浅层培养、包埋悬浮培养等方式,进行原生质体培养与植株再生研究。研究结果表明,海藻酸钙包埋悬浮振荡培养(50r/min)是龙眼悬浮细胞原生质体培养的适宜培养方式,在含有多种生长调节剂的改良MS培养基(MP6)上,再生小克隆的形成频率可高达5%;采用龙眼胚性愈伤组织体胚诱导、成熟和萌发的优化方案,原生质体再生小克隆分化子叶形胚状体的频率达100%,萌发植株的频率一般大于45%。   相似文献   

产纤维素酶原生质体形成和再生条件选择   总被引：5，自引：0，他引：5  

尹睿  陈红漫  张成刚 《沈阳农业大学学报》2000,31(6):556-558

研究了4种营养缺陷型产纤维素酶的原生质体形成和再生条件。结果表明,它们原生质体形成条件均以0．2mol.L^-1（pH5.8)磷酸缓冲液为好。绿色木霉（Trichoderma viride)UV2-11和另一未定木霉（T．sp.)UV2-2以0．2mol.L^-1KCl的渗透稳定剂为好,菌丝菌龄分别为20,18－20h;康宁木霉（T.Toningii)UV2-15以0．6mol.L^-1NaCl的渗透稳定剂为好,菌丝菌龄均为16－20h,酶解时间均为2－3h。原生质体再生培养基以加入0．5％酵母膏和0．5％泛酸钙钙的改良Czapek培养基效果好,用0．7mol.L^-1KCl和NaCl作为渗透稳定剂。菌株原生质体的生率较高。  相似文献   

Somatic hybrids between navel orange (Citrus sinensis) and grapefruit (C. paradisi) for seedless triploid breeding     

W.W. Guo  X.X. Deng  H.L. Yi 《Euphytica》2000,116(3):281-285

Protoplasts from cell suspension cultures of ‘Bonnaza’ navel orange (Citrus sinensis (L.) Osbeck) were electrically fused with mesophyll protoplasts isolated from seedless ‘Red Blush’ grapefruit (Citrusparadisi). After 6 months of culture, a total of 20 plants were regenerated. Root tip chromosome counting revealed that 4 of them were tetraploids (2n = 4x = 36)and the rest were diploids (2n = 2x = 18) morphologically resembling the mesophyll parent. After 6 months of transplantation into the greenhouse, 4 of the diploidmesophyll regenerants unexpectedly flowered, but this phenomenon disappeared in the next year. This is the first report of precocious flowering in citrus via protoplast fusion. RAPD analysis further confirmed that the tetraploid regenerants were somatic hybrids while the diploid regenerants were mesophyll parent type. This somatic hybrid will be utilized as a possible pollen parent for improving the seedy pummelo cultivars in China by producing triploid seedless pummelo hybrid. The mechanism of early flowering was also discussed. This revised version was published online in July 2006 with corrections to the Cover Date.  相似文献   

低温锻炼期间柑桔原生质体SOD和CAT酸活性的变化   总被引：16，自引：0，他引：16  

彭昌操  孙中海 《华中农业大学学报》2000,19(4):384-387

３种抗寒性不同的柑桔胚性愈伤组织原生质体经低温锻炼后,原生质体抗寒力得到增强,并在第１２ｄ达到最大。柑桔原生质体ＳＯＤ、ＣＡＴ酶活性变化同步提高,而细胞膜脂过氧化水平随低温锻炼进程逐步下降,至第１２ｄ回升。结果表明：柑桔原生质体ＳＯＤ、ＣＡＴ酶活性与抗寒性正相关,而ＭＤＡ的含量则与抗寒性成负相关,可以利用柑桔原生质体ＳＯＤ、ＣＡＴ酸活性和ＭＤＡ的含量鉴定柑桔原生质体的抗寒性。  相似文献   

用原生质体融合进行冬菇育种的研究     

刘作喜  赵华 《中国食用菌》1999,18(5):6-7,12

用灭活的〔Flammulinasubnudus (Pr .)Sing〕双核菌株原生质体与〔Flammulinavelutipes (Fr.)Sing〕双核菌株原生质体融合 ,在 2 2℃的条件下筛选得融合子。融合频率为 0× 10 -5～ 4 1× 10 -5。融合子与双亲有明显的拮抗性。融合子的菌丝体形态 ,氨基酸含量 ,子实体的形态 ,以及酸性磷酸酶的测定都与双亲不同。  相似文献   

中国沙棘子叶叶肉细胞电流全细胞记录的研究     

朱俊英  闫慧  高荣孚  尹伟伦 《江西农业大学学报》2007,29(5):762-766

实验得出适合膜片钳实验的耐盐木本植物中国沙棘子叶原生质体的游离条件,即取当天脱壳的子叶,放入渗透浓度为0.88 mol/L,pH为5.8,酶液的组分为10 g/LCellulase R-10、0.5 g/LPectolyase Y-23、1.5 g/LMacrozymeR-10、1 g/L Hemicellulase、1 g/L BSA、1 g/L PVP-40、2 mmol/L CaCl2、10 mmol/L Mes和5 mmol/L Vc的溶液中,在26℃、黑暗条件下,静置4 h。并且利用Multiclamp 700B放大器和Clampex 9.2软件,记录到了该原生质体质膜全细胞内向K 通道电流(-459.137 pA及-290.527 pA)和非选择性阳离子通道的外向电流(8.036 pA)。这为沙棘属木本植物细胞的电生理特性及其耐盐性机制关系的研究奠定基础。  相似文献   

草地早熟禾种间原生质体电融合条件的研究     

牛奎举  张然  赵春旭  马晖玲 《草原与草坪》2017,(5):30-34,40

为改良培育草地早熟禾(Poa pratensis)新品种,拟建立草地早熟禾商用品种与具有优良性状的野生早地早熟禾种间原生质体融合体系。采用电融合法,研究了电融合仪的电场条件和原生质体密度对草地早熟禾种间原生质体融合的影响。结果表明:在交流电场强度和频率分别为20 V/cm和2000k Hz时,原生质体可在较短的时间内形成2~4个原生质体串;最适于草地早熟禾种间原生质体融合的直流脉冲强度为200 V/cm、DC幅宽为40μs、直流脉冲次数为3次,原生质体密度为3×105个/m L,在此条件下,草地早熟禾种间原生质体总融合率和一对一融合率可分别达到25.39%和12.27%。  相似文献   

鸭疫里默杆菌与大肠杆菌融合株的构建     

倪粒琼  蒋文灿  李晓艳  李木子  刘雪梅 《中国兽医学报》2011,31(12)

对鸭疫里默杆菌（YL1）和大肠杆菌（ZYD2）原生质体的制备、再生、融合条件进行优化,成功获得了64株具有双亲菌株耐药性,能够同时凝集兔抗ZYD2血清和兔抗YL1血清,且能够稳定遗传的融合菌株;通过培养特性、菌落特征、融合菌株形态染色特性、生化试验等鉴定,筛选出4株典型融合菌株DL4、DL5、DL8、DL29。攻毒试验表明,ZYD2的半数致死量为1.5×107,YL1的半数致死量为5.0×107,4株典型融合菌株的半数致死量分别为2.8×108、1.7×108、1.0×109、4.4×109,融合菌株毒性比亲本菌株毒性减弱。免疫保护试验和免疫鸭抗体水平测定结果证实,融合菌株具有良好免疫原性。  相似文献   

不同电融合参数及融合液对昆明小鼠2-细胞胚胎融合及囊胚率的影响     

周川  赵云程  黄俊成 《动物医学进展》2012,33(5):56-59

四倍体胚胎在研究哺乳动物配子发生、发育、四倍体的发生、遗传、免疫机制等方面具有重要作用,因此提高融合及发育效率至关重要。为找出适合小鼠2-细胞胚胎融合的最佳方案,试验采用两种融合液0.3mol/L甘露醇溶液和0.3mol/L蔗糖溶液,两种融合液均采用了120V/mm,20μs×2、100V/mm,40μs×2、80V/mm,80μs×2三种电融合参数。其中采用0.3mol/L甘露醇溶液作为融合液且融合参数采用80V/mm,80μs×2获得了92.5%的融合率和90.6%的囊胚率,是小鼠2-细胞胚胎融合的最佳方案。  相似文献   

Construction and Genetic Analysis of Hybrid Strains between Apple and Tomato Pathotypes of Alternaria alternata by Protoplast Fusion     

SALAMIAH  Hajime AKAMATSU  Yukitaka FUKUMASA-NAKAI  Hiroshi OTANI  Motoichiro KODAMA 《Journal of General Plant Pathology》2001,67(2):97-105

The genetic controls of host-specific toxin (HST) biosynthesis and the pathogenicity of A. alternata pathogens have been limited by the asexual nature of the life cycle of these fungi. We used a protoplast fusion system for A. alternata to analyze the genetics of HST production and its relation to the specific pathogenicity of these pathogens. Drug-resistant transformants were isolated by genetic transformation, using vectors conferring resistance to hygromycin B and geneticin, for the A. alternata apple pathotype (AM-toxin producer) and A. alternata tomato pathotype (AAL-toxin producer), respectively. Protoplasts of the respective transformants were fused by electrofusion. The majority of resultant stable fusants produced both AM- and AAL-toxins and were pathogenic to susceptible cultivars of both apple and tomato. Pulsed-field gel electrophoresis analysis demonstrated that these fusants (or hybrids) carried small 1.7-and 1.1-Mb chromosomes, characteristic of the parental strains of the apple and tomato pathotypes, respectively. Detection of the AMT gene, involved in AM-toxin biosynthesis, by polymerase chain reaction revealed that all fusants pathogenic to apple maintained this gene. Microfluorimetry analysis using propidium iodide staining suggested that the fusants might be diploid. Received 14 November 2000/ Accepted in revised form 11 December 2000  相似文献