Cell lines and plants obtained after protoplast fusions of Rosa+Rosa,Rosa+Prunus and Rosa+Rubus     

Squirrell  J.  Mandegaran  Z.  Yokoya  K.  Roberts  A. V.  Mottley  J. 《Euphytica》2005,146(3):223-231

Summary Protoplasts of three Rosa cultivars were fused with each other, with protoplasts of Prunus `Colt' and with protoplasts of Rubus laciniatus, using polyethylene glycol 4000 as a fusogen. Protoplasts of Prunus were incapable of cell division and those of Rosa and Rubus were disabled by treatments with metabolic inhibitors, either iodoacetate (IOA) or rhodamine 6-G (R6G). Parental protoplasts were then fused in combinations that required complementation for their survival. RAPD analysis of 41 fusion-derived cell lines showed that two lines resulting from fusions of Rosa + Rosa and one from a fusion of Rosa + Prunus, contained some DNA markers from both fusion partners. The others contained markers of only one fusion parent. This showed that after protoplast fusion, the heterokaryons did not develop into cell lines with stable hybrid nuclei. Plants regenerated from cell lines derived from Rosa + Prunus and Rosa + Rubus fusions contained DNA markers of only Rosa and their DNA amounts were no greater than that of the Rosa parent. However, they differed morphologically from the Rosa parent to a remarkable degree, possibly because they inherited undetected genes of Prunus or Rubus, or because they were somaclonal variants of the Rosa parent. Alternative strategies for the production of somatic hybrids are discussed.  相似文献   

Rapid detection of foot-and-mouth disease virus using a field-portable nucleic acid extraction and real-time PCR amplification platform     

Madi M  Hamilton A  Squirrell D  Mioulet V  Evans P  Lee M  King DP 《Veterinary journal (London, England : 1997)》2012,193(1):67-72

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Studies of embryo transfer from cattle clinically affected by bovine spongiform encephalopathy (BSE)     

Wrathall AE  Brown KF  Sayers AR  Wells GA  Simmons MM  Farrelly SS  Bellerby P  Squirrell J  Spencer YI  Wells M  Stack MJ  Bastiman B  Pullar D  Scatcherd J  Heasman L  Parker J  Hannam DA  Helliwell DW  Chree A  Fraser H 《The Veterinary record》2002,150(12):365-378

Semen from 13 bulls, eight with clinical bovine spongiform encephalopathy (BSE), was used to artificially inseminate (AI) 167 cows with clinical BSE, and their resultant embryos were collected non-surgically seven days after AI. The viable and non-viable embryos with intact zonae pellucidae were washed 10 times (as recommended by the International Embryo Transfer Society) then frozen. Later, 587 of the viable embryos were transferred singly into 347 recipient heifers imported from New Zealand, and 266 live offspring were born of which 54.1 per cent had a BSE-positive sire and a BSE-positive dam. The recipients were monitored for clinical signs of BSE for seven years after the transfer, and the offspring were monitored for seven years after birth. Twenty-seven of the recipients and 20 offspring died while being monitored but none showed signs of BSE. Their brains, and the brains of the recipients and offspring killed after seven years, were examined for BSE by histopathology, PrP immunohistochemistry, and by electron microscopy for scrapie-associated fibrils. They were all negative. In addition, 1020 non-viable embryos were sonicated and injected intracerebrally into susceptible mice (20 embryos per mouse) which were monitored for up to 700 days, after which their brains were examined for spongiform lesions. They were all negative. It is concluded that embryos are unlikely to carry BSE infectivity even if they have been collected at the end-stage of the disease, when the risk of maternal transmission is believed to be highest.  相似文献