禽腺联病毒全基因组的克隆及感染性病毒的拯救

为了克隆禽腺联病毒(Avian adeno-associated virus,AAAV)全基因组用于构建基因转移载体研究,以鸡胚致死孤儿病毒(CELO)作为辅助病毒与AAAV共接种SPF鸡胚进行AAAV的增殖,将AAAV约4.7kb双链基因组DNA与pCR2.1载体连接,构建了含AAAV全基因组的重组质粒pAAAV并进行了测序。序列分析表明,AAAV YZ-1株的基因组为4684bp,两端具有141bp的末端倒置重复序列和Rep蛋白结合位点特征序列,与GenBank中收录的AAAV DA-1株和VR-865株的核苷酸序列同源性分别为95.0%和92.2%。将pAAAV质粒转染CELO病毒感染的鸡胚肝细胞系,获得了感染性AAAV病毒粒子,结果证明克隆的AAAV基因组中存在与病毒复制和包装相关的正确关键序列,可用于重组AAAV载体的构建。     

无核葡萄与中国野生葡萄杂种的胚挽救技术研究

 通过无核葡萄与中国野生葡萄杂交, 获得了4个杂交组合的后代植株, 确定了各杂交组合取胚珠培养的最佳时期。1Flame ×山葡萄; 2红宝石×爱莫无核; 3红宝石×北醇; 4爱莫无核×山葡萄在授粉后45 d取样培养效果较佳; 5长穗无核白×山葡萄授粉后30 d; 6无核白自交在授粉后35 d培养效果较佳。以NitSchs为基本培养基, 附加0.5 mg·L ^{- 1} 6-BA + 0.5 mg·L^-1 GA₃ + 2.5 mg·L ^{- 1} IBA + 0.1 mg·L^-1ZT, 适合于1、3、5号杂交组合; 而0.5 mg·L^-1 6-BA + 0.5 mg·L^-1 GA₃ + 2.0 mg·L^-1 IBA + 0.1 mg·L^-1ZT适合2、4、6号杂交组合; 胚萌发培养基为2.0 mg·L^-1 IBA + 1.0 mg·L^-1 6-BA +0.2 mg·L^-1 GA₃ , 适合1、2、3、5、6号杂交组合, 而爱莫×山葡萄在1.5 mg·L ^{- 1} IBA + 1.0 mg·L^-1 6-BA + 0.2 mg·L^-1 GA₃培养基上表现较佳, 生根培养基为1 /2MS基本培养基添加0.15 mg·L^-1 IBA + 0.02 mg·L^-1 6-BA, 它适合1号与5号组合, 0.10 mg·L^-1 IBA + 0.02 mg·L^-1 6-BA适合4号与6号组合。     

Production of inter-section hybrids between Primula filchnerae and P. sinensis through ovule culture

Inter-section hybrids were obtained in the reciprocal crosses between Primula filchnerae (2n = 2x = 24) of Sect. Pinnatae and P. sinensis ‘Fanfare’ (2n = 2x = 24) of Sect. Auganthus by rescuing ovules on half-strength (1/2) Murashige and Skoog's (MS) medium supplemented with 50 g l⁻¹ sucrose, 2.5 g l⁻¹ gellan gum, 0.1 mg l⁻¹ α-naphthaleneacetic acid (NAA), 0.1 mg l⁻¹ 6-benzyladenine (BA) and 50 mg l⁻¹ gibberellic acid (GA₃). In ovule culture, germination occurred with radicle elongation but no plumule was observed. The radicle kept on the initial medium showed root proliferation with callus formation. When the calluses were transferred to (1/2)MS media containing 30 g l⁻¹ sucrose and 3 g l⁻¹ gellan gum, without plant growth regulators (PGRs) or with 1 mg l⁻¹ zeatin and 0.1 mg l⁻¹ NAA, plantlets were regenerated. The plants thus obtained were confirmed to be hybrids through flow cytometry (FCM) and random amplified polymorphic DNA (RAPD) analyses. The hybrid obtained when P. filchnerae was used as the maternal parent was diploid, whereas hexaploid hybrid was obtained when using P. sinensis as the maternal parent. The hexaploid hybrid might be produced through chromosome doubling of a triploid originated from the fertilization of P. sinensis with unreduced pollen of P. filchnerae.     

冰糖橙×枸橼杂交后代的胚抢救及杂种鉴定

为获得冰糖橙×枸橼的合子苗,以抗柑橘溃疡病枸橼为父本,以高感病品种冰糖橙为母本进行杂交,利用胚抢救技术获得杂交后代,然后利用筛选出的SSR引物对亲本和子代的基因组DNA进行PCR扩增,分析并鉴定杂交后代中的合子苗。结果表明:在最终获得的359株杂交后代中,利用SSR标记能鉴定出合子苗12株。     

蟠桃新品种‘中蟠桃11号’

‘中蟠桃11号’是以‘红珊瑚’ב91-4-18’(NJN78×奉化蟠桃)杂交,结合胚挽救技术获得的蟠桃新品种。花铃形。单果质量180~200 g,果皮60%以上着鲜红色,果肉橙黄色,肉质为硬溶质。果实风味浓甜,可溶性固形物含量14%,粘核。耐运输。7月中下旬成熟,果实发育期120 d。极丰产,平均产量为38 888 kg·hm-2,     

Risk assessment of outcrossing of transgenic rapessed to related species:

Summary The risk for a gene dispersal is reported for reciprocal crosses between a transgenic rapeseed variety resistant to the herbicide phosphinotricin and five related species. The first stages after pollination were cytologically observed and fertilized ovaries were established in in vitro culture for the production of interspecific hybrids. A similar classification was observed for the index of pollination compatibility and embryo yield. From the 243 embryos produced, 109 plantlets were obtained in a greenhouse. All the interspecific combinations tested were able to produce hybrid plants. A higher number of hybrids was obtained when rapeseed was used as the female parent. The hybrids had the expected triploid structure except for two amphidiploid, B. napus × B. oleracea, and one amphidiploid, B. napus × S. arvensis, plants with 56 chromosomes. The triploid hybrids were sterile or partially fertile but two of the amphidiploid plants, B. napus × B. oleracea, were fully fertile. The cytoplasm source did not seem to affect the fertility of the hybrids.