No description available.

Field performance of micropropagated (MP) and conventionally propagated (CP) red raspberry (Rubus idaeus L. cv. Comet and Festival) was examined under hedgerow and stool cane management systems for 3 seasons (1989 to 1991). All MP plants established well compared to 58% survival rates 45 days after planting and 92% survival rates after replanting for CP plants. The MP plants were more vigorous compared with the CP plants for the duration of this study as indicated by more and taller canes. MP 'Festival' in 1990 yielded 2.2 MT$ cdot$ha$ sp{-1}$, almost half the yield of established commercial plantings in Quebec, while yields from CP 'Festival' and MP and CP 'Comet' were negligible. The MP 'Festival' crop (8.42 MT$ cdot$ha$ sp{-1}$) also outyielded CP 'Festival' (6.8 MT$ cdot$ha$ sp{-1}$) and both MP (5.72 MT$ cdot$ha$ sp{-1}$) and CP (4.91 MT$ cdot$ha$ sp{-1}$) 'Comet' in the second fruiting year. Propagation method had no effects on winter hardiness, photosynthetic capacity nor leaf and stem morphology of either cultivar. The results indicated that MP plants were superior to CP plants for both nursery propagation and fruit production due to their more consistent establishment and increased vigor. Red raspberry plantlets were successfully hardened in vitro on low-sucrose or sucrose-free media through CO$ sb{2}$ enrichment (1500 ppm) and relative humidity reduction (90%) using a forced ventilation system in specially constructed plexiglass chambers. Enriched CO$ sb{2}$ significantly increased general vigor, root formation, root growth, plantlet growth and plantlet photosynthetic capacities. Sucrose in the medium promoted plantlet growth but depressed photosynthesis. In vitro relative humidity at 90% decreased stomatal apertures and improved plantlet ex vitro performance but did not affect the CO$ sb{2}$ uptake rates of cultured plantlets or ex vitro transplants. The maximum CO$ sb{2}$ uptake rates of plantlet leaves were about 52-69% that of greenhouse control pla

Tissue-cultured shoots and plantlets usually have leaves with non-functional, open stomata and little epicuticular and cuticular wax, resulting in excess evapotranspiration after transplantation. Various strategies were evaluated to decrease ex vitro acclimatization difficulties for 'Silvan' blackberry, including transplanting unrooted shoots, increasing the medium agar concentration from 6 to 9 or 12 g/l and diluting the basal medium. Increased medium agar concentrations and medium dilution did not improve survival or growth. Stomatal function resumed sooner in new leaves of plantlets than shoots. High relative humidity ($>$95%) and low light intensity (90 $ mu$mol s$ sp{-1}$ m$ sp{-2}$) negatively affected stomatal closure both on acclimatizing transplants and greenhouse-grown plants. Guard cells developed on leaves in vitro were physiologically active but had apparent anatomical abnormalities that inhibited closure. A rapid clearing and staining method was developed for examination of foliar morphology using intact in vitro blackberry (Rubus sp. 'Silvan') and strawberry (Fragaria x ananassa Duch. 'Totem') plantlets and sections of greenhouse-grown 'Silvan' and 'Totem' leaves. This method involved three steps: (1) removing the chlorophyll by autoclaving in 80% ethanol; (2) dissolution of the protoplasm using 5% NaOH at 80$ sp circ$C; (3) post-alkali treatment with 75% bleach (4.5% NaClO) at room temperature for tissue-cultured plantlets and at 55$ sp circ$C for greenhouse-grown leaves. Aqueous safranin (10 mg/l) was used for staining.

No description available.

Call number: LD2668 .T4 1986 R35 / Master of Science / Horticulture, Forestry, and Recreation Resources

Bibliography: leaves 74-80. This study shows techniques for reducing hyperhydricity in the micropropagation of Sturt's desert pea. The effects of support matrix, tube closure and cytokinin regime are examined and the anatomy of hyperhydric shoots is investigated.

No description available.

A novel technique was developed to immobilize plant cells. The cells are deposited on a surface of man-made fibrous material which provides for strong binding of the plant tissue biomass growing in the submerged culture. It was shown that the plant cells need to be fully viable for the attachment process to occur. / The scale-up of this technique to laboratory size specifically designed bioreactors was performed successfully. The cell immobilizing matrix was formed into a vertical spirally wound configuration to provide for a high immobilizing area-to-volume ratio (0.8-1.2 cm$ sp{-1}$). A modified airlift (riser-to-downcomer area ratio of 0.03 and vessel height-to-diameter (H/D ratio of 3) and a low H/D ($ sim$1.5) mechanically stirred vessel delivered the optimum bioreactor performance characterized by low foaming of the broth and highly efficient plant cell attachment and retention ($ geq$96%). / The growth of Catharantus roseus plant cells was investigated in these bioreactors. This process was found not to be mass transfer limited above minimal mild mixing and aeration levels ensuring sufficient supply of nutrients, especially oxygen (k$ sb{ rm L}$a $ sim$ 10-15 h$ sp{-1}$) to the immobilized biomass. / The gentle surface immobilization technique developed in this work did not hinder the biosynthesis potential of the SIPC. In fact, it appeared to induce a partial secretion of some valuable compounds into the culture medium. The mildness, easiness, efficiency, mass transfer characteristics, scale-up potential and biomass loading capacity (11-13 g d.w./L) of the surface immobilization technique make it superior to all other immobilization techniques used to culture plant cells. In addition, its bioreactor overall biomass concentration compares favourably to suspended plant cell concentrations attainable in bioreactors (15-20 g d.w./L).

Orientadores: Clarice Weis Arns, Fernando Rosado Spilki / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-16T04:41:41Z (GMT). No. of bitstreams: 1 Santos_MarciaMercesAparecidaBianchidos_D.pdf: 3376725 bytes, checksum: 4e17791ec1d7e56265e067f2ce1f62bf (MD5) Previous issue date: 2010 / Resumo: O Metapneumovirus aviário (aMPV) pertence à família Paramyxoviridae, subfamília Pneumovirinae, gênero Metapneumovirus. O vírus, relatado pela primeira vez no Brasil em 1995, é o agente etiológico da Rinotraqueíte em perus (TRT) e está associado também à Síndrome da Cabeça Inchada (SHS) em frangos e poedeiras comerciais. O presente estudo foi dividido em três partes. Na primeira foi avaliada a suscetibilidades de oito sistemas celulares para a propagação de amostras virais do aMPV subtipos A e B. As células chicken embryo related (CER), Vero e baby hamster kidney cells (BHK-21) demonstraram ser as mais apropriadas para a multiplicação de ambos os subtipos. Além disso, cultivo de anel de traquéia (TOC) e cultivo primário de embrião de galinha (CEF) foram permissíveis à infecção por aMPV após terem sido isolados e propagados em CER. As curvas da cinética viral foram realizadas nas três linhagens celulares e ambos os subtipos tiveram títulos mais altos em CER durante as primeiras 30 horas após a infecção. Não foram observadas diferenças significativas entre os títulos obtidos em células CER e Vero, demonstrando que as células CER são tão adequadas à propagação do aMPV quanto as células Vero. A segunda parte do trabalho consistiu em analisar a virulência de uma amostra de aMPV subtipo B após sofrer passagens seriadas em células CER. Cinco variantes provenientes das passagens em CER foram inoculadas em galinhas e a excreção viral foi analisada. Os resultados obtidos com as amostras de traquéia demonstram que a virulência do aMPV diminui gradualmente enquanto o título viral aumenta com o número de passagens. Em contrapartida, nas amostras de seio nasal foi observado aumento da carga viral, demonstrando que não houve diminuição no fitness viral para este órgão. As seqüências do gene G das amostras utilizadas para desafio foram obtidas, porém este gene parece não ser afetado pela propagação em células CER. Na terceira e última parte deste estudo, foi avaliada a proteção viral conferida por uma vacina comercial contra isolados brasileiros do aMPV subtipos A e B em frangos de corte. Para tanto, uma amostra de cada subtipo foi avaliada quanto à sua virulência. O isolado do subtipo B foi detectado em um período mais longo e em maiores quantidades quando comparado ao subtipo A. Os resultados da analise da proteção demonstram que algumas aves imunizadas receberam proteção viral completa contra o vírus virulento heterólogo. Porém, a mesma vacina forneceu proteção viral parcial quando as aves foram desafiadas com o vírus virulento homologo ao vacinal / Abstract: Avian metapneumovirus (aMPV) belongs to the Paramyxoviridae family, Pneumovirinae subfamily, within the genus Metapneumovirus. The virus, first described in Brazil in 1995, is responsible for an acute rhinotracheitis in turkeys (TRT) and is associated with swollen head syndrome in broiler chickens and layer hens. The present study is divided in three parts. In the first part, eight cell culture systems were evaluate for the propagation of aMPV subtypes A and B. The chicken embryo related (CER) cells, Vero and baby hamster kidney cells (BHK-21) cells were shown to be the most appropriate for propagation of both subtypes of aMPV. In addition, propagation of aMPV in chicken embryo fibroblasts (CEF) and tracheal organ culture (TOC) remained efficient after the primary isolation and several passages of viruses in the CER cell line. The growth curves were created using CER, Vero and BHK-21 cell lines. Compared with growth, both yielded higher titres in CER cells during the first 30 hours after infection, but no significant difference was observed in the results obtained from CER and Vero cells. This data show that CER cell are adequate for aMPV propagation, giving similar results to Vero cells. The second part of this study was conducted to analyze the virulence of an aMPV subtype B strain after serial passage in CER cells. To accomplish this, chickens were infected with 5 different variants derived from serial passage and the amount of viral shedding was determined. The results of tracheal samples showed that the virulence decreases gradually with passage, while in vitro viral titre increases. However, an increase in viral shedding was observed in sinusal samples, demonstrating no decrease in fitness for this organ. The G gene sequences of challenge samples were analyzed, however this gene appears to not be affected when aMPV is propagated in CER cells. Finally, the last part of this study aimed to examine a commercially available vaccine in broiler chickens in terms of it ability to provide virological protection against aMPV subtypes A and B. To accomplish this, the virulence of each virulent strain was analyzed. The results demonstrate that the subtype B virulent strain could be observed longer and in larger quantities compared to subtype A. A complete heterologous virological protection was provided by the subtype B vaccine; however, a lack of complete homologous virological protection was observed when chickens were challenged with the homologous subtype B strain / Doutorado / Microbiologia / Doutor em Genetica e Biologia Molecular

No description available.