Seed germination and tissue culture of Cymbidium giganteum Wall. ex Lindl.     

M.M. Hossain  Madhu Sharma  Jaime A. Teixeira da Silva  Promila Pathak 《Scientia Horticulturae》2010

Efficient protocols were established for in vitro seed germination, neo-formation of secondary (2°) protocorms from primary (1°) protocorms and multiple shoot buds and protocorm-like body (PLB) induction from pseudo-stem segments of in vitro-raised seedlings of Cymbidium giganteum. Four nutrient media, namely Murashige and Skoog (MS), Phytamax (PM), Mitra et al. (M), and Knudson ‘C’ (KC) were evaluated for seed germination and early protocorm development. In addition, the effects of peptone, activated charcoal (AC) and two plant growth regulators [6-benzylaminopurine (BAP) and 2,4-dichlorophenoxyacetic acid (2,4-D)] were also studied. Both M and PM supplemented with 2.0 g l⁻¹ peptone or 1.0 mg l⁻¹ BAP resulted in ∼100% seed germination. Media supplemented with 2.0 g l⁻¹ AC could effectively induce large protocorms (1.6 ± 0.1 mm in diameter). Neo-formation of 2° protocorms from 1° protocorms was achieved in liquid and agar-solidified PM medium fortified with different concentrations and combinations of auxins (α-naphthalene acetic acid (NAA) and 2,4-D) and cytokinins [BAP and kinetin (KN)]. The highest number of 2° protocorms was obtained in liquid medium (10.7 ± 0.9/1° protocorm) supplemented with 2.0 mg l⁻¹ BAP + 1.0 mg l⁻¹ NAA. Although protocorms proliferated profusely in liquid medium, these did not develop further unless transferred to agar-solidified medium within 6–8 weeks. Multiple shoot buds and PLBs were induced from pseudo-stem segments on agar-solidified PM medium fortified with different concentrations and combinations of BAP and NAA and the maximum number of PLBs (6.00 ± 0.20) was recorded when BAP and NAA were applied at 2.0 mg l⁻¹ each. A solid root system was induced from PLBs and shoot buds when these were transferred to half-strength PM or M media fortified with 0.5 mg l⁻¹ indole-3-acetic acid. Well-rooted plants were transferred to the greenhouse with 95% survival.  相似文献   

Water deficit and polyethylene glycol 6000 affects morphological and biochemical characters of Pelargonium odoratissimum (L.)     

Khalid A. Khalid  Jaime A. Teixeira da Silva  Weiming Cai 《Scientia Horticulturae》2010

The effects of water deficit and polyethylene glycol (PEG) 6000 on the growth and content of essential oil, photosynthetic pigments, soluble sugars, proline, macroelements (N–P–K) and microelements (Mg–Zn–Fe–Mn) of Pelargonium odoratissimum (L.) plants were investigated. Water deficits of 80% and 40% of field water capacity decreased certain growth characters (total leaf area, leaf number, and total fresh and dry mass). The photosynthetic pigments (chlorophylls a and b, total carotenoids) and mineral content (N–P–K–Mg–Zn–Fe–Mn) also decreased as water deficit level increased. Water deficit promoted the accumulation of essential oil content and its main components (methyl eugenol, isomenthone and limonene) as well as total soluble sugars and proline contents. The water deficit × PEG 6000 interaction was greater than water deficit treatments alone on all plant growth characters assessed.  相似文献   

Pathogenicity of dried-shrink disease and evaluation of resistance in a germplasm collection of sea buckthorn (Hippophae L.) from China and other countries     

Cheng-Jiang Ruan  Jaime A. Teixeira da Silva  Qun Li  He Li  Jun Zhang 《Scientia Horticulturae》2010

Sea buckthorn (Hippophae L.) is a woody, dioecious plant that can be grown in dry and poor soil conditions. It has recently received world-wide attention as a new berry crop with a very high nutritional value and unique medicinal properties, as well as a means of combating soil erosion. A major problem is, however, dried-shrink disease (DSD), which destroys sea buckthorn plants and halts commercial production. In this study, we investigated symptoms and pathogenicity of DSD, isolated and identified pathogens using a combination of morphology and ITS sequences, and evaluated the disease index (DI) of different cultivars in the field. DSD infection causes isolated yellowing of plant tissues, and as the disease develops the bark putresces, dries and shrinks, and finally the plant dies. DSD infects older plants (>3 years), often at the plant base. Death of plant parts above the infected area does not influence root vitality, and a new plant can therefore be regenerated. Combining morphological characters and ITS sequences, we isolated and identified four fungal pathogens causing DSD, including Fusarium acuminatum, F. oxysporum, Fusarium camptoceras and Phomopsis spp. DI values for 27 cultivars ranged from 0 to 10.4. About half of the cultivars were regarded as resistant (DI < 3.2) to DSD, and half as susceptible (DI > 6.7). Cultivar ‘Zhongguoshaji’ (DI = 4.2) showed an intermediate level of DSD resistance. Our data provides basic information for further studies of the mechanism of pathogenicity, and for breeding of DSD-resistant cultivars.  相似文献   

Effect of α-Tocopherol on Bovine In Vitro Fertilization     

A Marques  P Santos  G Antunes  A Chaveiro  F Moreira da Silva 《Reproduction in domestic animals》2010,45(1):81-85

The objectives of this work are to determine if exogenous supplementation with α-tocopherol increases the in vitro fertilization (IVF) rate of bovine oocytes and improves viability of selected spermatozoa after 'swim-up'. The percentage of fertilized oocytes was significantly but negatively correlated ( r  = −0.941, p < 0.01) with the concentration of α-tocopherol. The control resulted in 95% of fertilized oocytes, which decreased as follows: 25 μM α-tocopherol (α25) 86%, 50 μM α-tocopherol (α50) 74%, 100 μM α-tocopherol (α100) 66% and 200 μM α-tocopherol (α200) 56%. Relatively to sperm viability after 'swim-up' with α-tocopherol supplementation, this antioxidant proved to have a beneficial effect as its concentration increased up to α50, decreasing for the concentrations of α100 and α200. Control resulted in 83% of live cells and 16% of dead cells; α25 resulted in 88% of live cells and 12% of dead cells; α50 resulted in 91% of live cells and 9% of dead cells; α100 resulted in 67% of live cells and 33% of dead cells; and finally α200 resulted in 57% of live cells and 42% of dead cells. In summary, the present study allows to conclude that, in our conditions, supplementation with the antioxidant α-tocopherol in IVF of bovine oocytes has a detrimental effect on fertilization rates. Nevertheless, exogenous supplementation with α-tocopherol at a concentration of 50 mM in the sperm-TALP media during the 'swim-up' technique has a significant beneficial effect on the selected spermatozoa viability.  相似文献   

Generation of Buffalo (Bubalus bubalis) Transgenic Chimeric and Nuclear Transfer Embryos Using Embryonic Germ-Like Cells Expressing Enhanced Green Fluorescent Protein     

B Huang  K Cui  T Li  X Wang  F Lu  Q Liu  FM da Silva  D Shi 《Reproduction in domestic animals》2010,45(1):103-108

The possibility of producing transgenic buffalo embryos by chimera and nuclear transfer (NT) using buffalo embryonic germ (EG)‐like cells expressing enhanced green fluorescent protein (EGFP) has been explored in this study. Buffalo EG‐like cells and fibroblasts with two to eight passages were transfected with the lined plasmid (pCE‐EGFP‐IRES‐Neo‐dNdB) using Lipofectamine^TM 2000 and selected by culturing in 200 μg/ml G418 for 6–8 days. G418 resistant fibroblasts and EG‐like cells were used for embryo chimera and NT. To produce blastocysts by chimera, 8–16 cells embryos were injected with EG‐like and fibroblast cells. Then, to produce blastocysts by NT, in vitro maturated oocytes were enucleated and afterwards EG‐like/fibroblast cells transferred into the perivitelline space. No statistical differences were observed for the total blastocyst produced by the chimeric method, using EG‐like and fibroblasts as donor cells, resulting on an accomplishment of 35.6% vs 33.3%, respectively. Nevertheless, besides from the 37 blastocysts produced, 23 (62.2%) from EG‐like cells expressed EGFP, none of blastocysts from foetal fibroblasts expressed this protein. When the NT method was used, no statistical difference among different generations was observed in the percentage of oocytes fused, cleaved, and developed to blastocysts after NT for EG‐like cells. On average, the percentage of oocytes fused, cleaved, and developed to blastocysts after NT was respectively 81.8%, 67.7% and 10.7%. For the expression of EGFP, from the 12 blastocysts produced by NT, 7 of them were positive, while none of NT embryos from EGFP positive fibroblasts developed to blastocysts. Results of the present study clearly demonstrated that gene transfected buffalo EG‐like cells have the ability to form chimeric embryos after injecting into buffalo early embryos and reprogramming ability after NT, which can be employed to produce transgenic buffalos through either embryo chimera or NT.  相似文献   

Sediment,nutrient and water losses by water erosion under agroforestry systems in the semi-arid region in northeastern Brazil     

Maria Ivanilda de Aguiar  Stoécio Malta Ferreira Maia  Francisco Alisson da Silva Xavier  Eduardo de Sá Mendonça  João Ambrósio Araújo Filho  Teógenes Senna de Oliveira 《Agroforestry Systems》2010,79(3):277-289

Inadequate soil management practices adopted in the Brazilian semi-arid region contribute to erosive processes. Agroforestry systems (AFs) have been considered an alternative to reduce water erosion. This study aimed to evaluate the impact of two alternatives AFs, a traditional and an intensive cropping system on the losses of sediments, water, organic carbon and nutrients caused by water erosion in comparison to the natural vegetation (caatinga) in a semi-arid region of northeastern Brazil. The agroecosystems studied were: agrosilvopasture (AGP) which consisted of an alley cropping system, cultivated with Leucaena leucocephala and maize, within an area composed by 22% of native trees (200 native trees per hectare) which was grazed during the dry season; silvopasture (SILV) that was composed by 38% of native trees (260 trees per hectare) with a stocking rate of 20 ewes during whole year; traditional agrosilvopasture (TRAG) being managed as following: total deforestation, burning of the residues, cropped with maize for 2 years (1998 and 1999) and fallow during 8–10 years; and intensive cropping (IC) system which was deforested and burned in 1997 followed by cultivation of maize from 1998 to 2002, and thereafter by a fallow period of 8–10 years similar to TRAG. Two areas of native forest (NF1, NF2) known as ‘caatinga’, used as grassland during the dry season and as a source of wood, were selected and used as reference of steady state in the comparative study in relation to the cultivated sites. Sediment and water losses as a result of erosion were collected during two rainy seasons, i.e. 2003 and 2004, and nutrients and organic carbon contents were determined. Soil samples were collected and organic carbon, pH in water, pH in KCl, water dispersible clay (WDC) and hydraulic conductivity (K₀) were measured. In 2003, sediment and water losses did not differ significantly among all treatments. However, in 2004, TRAG (0.70 Mg ha⁻¹) and NF1 (1.37 Mg ha⁻¹) showed the highest sediment losses, whereas TRAG and IC presented the highest water losses. On average, nutrients losses in cropped areas were lower than in natural vegetation (NF1, NF2). The alternative AFs (AGP, SILV) were efficient to reduce water erosion effects when compared to the most common agricultural practices adopted in the region, being highly recommended as sustainable technical alternatives for food production in the region.  相似文献   

Factors controlling Eucalyptus productivity: How water availability and stand structure alter production and carbon allocation     

Michael G. Ryan  Jose Luiz Stape  Dan Binkley  Sebastiao Fonseca  Rodolfo A. Loos  Ernesto N. Takahashi  Claudio R. Silva  Sergio R. Silva  Rodrigo E. Hakamada  Jose Mario Ferreira  Augusto M.N. Lima  Jose Luiz Gava  Fernando P. Leite  Helder B. Andrade  Jacyr M. Alves  Gualter G.C. Silva 《Forest Ecology and Management》2010

Wood production varies substantially with resource availability, and the variation in wood production can result from several mechanisms: increased photosynthesis, and changes in partitioning of photosynthesis to wood production, belowground flux, foliage production or respiration. An understanding of the mechanistic basis for patterns in wood production within a stand and across landscapes requires a complete annual carbon budget. We measured annual carbon flows to wood production, foliage production and total belowground carbon flux (the sum of root production, root respiration, and mycorrhizal production and respiration) from ages three to five years in clonal Eucalyptus plantations at four sites in Brazil to test if fertility, water availability and stand structure changed wood production and by what mechanism. We also quantified the patterns in light interception and the efficiency of light use to provide additional mechanistic insights into growth responses and to determine if light-use efficiency was related to changes in flux and partitioning.  相似文献   

Prevalence of Leptospira interrogans antibodies in free-ranging Tayassu pecari of the Southern Pantanal, Brazil, an ecosystem where wildlife and cattle interact     

Tatiana P. Tavares de Freitas  Alexine Keuroghlian  Donald P. Eaton  Emanuel Barbosa de Freitas  Aline Figueiredo  Luciano Nakazato  Jacqueline M. de Oliveira  Flávia Miranda  Rita Cassia S. Paes  Leticia A. R. Carneiro Monteiro  José Vergílio B. Lima  Aparecida A. da C. Neto  Valéria Dutra  Julio Cesar de Freitas 《Tropical animal health and production》2010,42(8):1695-1703

We surveyed a wild population of white-lipped peccaries (Tayassu pecari) in the Brazilian Pantanal for evidence of Leptospira interrogans. Serum samples from 71 free-ranging T. pecari were obtained between 2003 and 2005 in the southern Pantanal of Mato Grosso do Sul state. We used microscopic microagglutination to test for antibodies against 14 L. interrogans serovars (antibody titers ≥1:100 were considered seropositive). Seventy percent of captured animals tested positive for leptospirosis antibodies. Antibodies against icterohaemorrhagiae and autumnalis serovars were the most prevalent. We used log-linear analyses to test for associations among seropositivity, age class, and sex of captured animals. Seropositivity was strongly associated with animal age class, but independent of sex. Forty-six percent of animals less than 2 years old, 63% of adults during peak reproductive years, and 100% of the oldest age class were seropositive. A nonparametric multivariate procedure (MRPP) showed that the composition of serovar antibody types changed with age, and ANOVA models demonstrated that antibody titers increased with age, suggesting long-term exposure to a greater number and variety (i.e., serovar types) of L. interrogans infections. This study presents the first quantitative survey of antibodies against L. interrogans serovars in a T. pecari population of the Pantanal. The high prevalence of leptospirosis antibodies in free-ranging white-lipped peccaries and the potential impacts on reproduction and population dynamics emphasize the need for further studies investigating the roles of Pantanal wildlife and livestock in the transmission and maintenance of L. interrogans in the environment.  相似文献   

Subcutaneous embryonal rhabdomyosarcoma in a dog: cytologic, immunocytochemical, histologic, and ultrastructural features     

Avallone G  Pinto da Cunha N  Palmieri C  Della Salda L  Stefanello D  Roccabianca P  Caniatti M 《Veterinary clinical pathology / American Society for Veterinary Clinical Pathology》2010,39(4):499-504

Abstract: A subcutaneous mass on the left antebrachium of an 11‐year‐old intact female English Pointer dog was evaluated presurgically by cytologic examination and immunocytochemical staining. The sample consisted of discrete, variably sized, markedly pleomorphic neoplastic cells that expressed vimentin with diffuse cytoplasmic staining, desmin with focal paranuclear staining, and myoglobin with diffuse cytoplasmic staining, consistent with a diagnosis of rhabdomyosarcoma. Lymphocytic and histiocytic markers were negative. Aspirates of the enlarged ipsilateral prescapular lymph node were positive for metastatic disease. Surgical excision of the tumor and lymph node were followed by histologic and electron microscopic examination. Histomorphologic appearance of neoplastic cells from the mass and the lymph node paralleled cytologic findings; the histologic diagnosis was round cell variant of embryonal rhabdomyosarcoma. By ultrastructural evaluation, cells contained numerous mitochondria and masses of cytoplasmic tangled myofilaments, features typical of rhabdomyoblasts. The dog received doxorubicin (30 mg/m²) every 3 weeks for 5 treatments. Local recurrence developed 6 months after resection but was not treated. Despite a guarded prognosis and untreated local recurrence, the dog was still alive 18 months after surgery. Cytologic evaluation and immunocytochemical staining were pivotal for the presurgical diagnosis of rhabdomyosarcoma.  相似文献   

Soil apparent electrical conductivity and geographically weighted regression for mapping soil     

J. M. Terrón  J. R. Marques da Silva  F. J. Moral  Alfonso García-Ferrer 《Precision Agriculture》2011,12(5):750-761

To resolve the spatial variation in soil properties intensively is expensive, but such knowledge is essential to manage the soil better and to achieve greater economic and environmental benefits. The objective of this study was to determine whether the soil apparent electrical conductivity (EC_a), alone or combined with other variables, is a useful alternative for providing detailed information on the soil in the Extremadura region of Spain. Apparent soil electrical conductivity was measured and geographically weighted regression was used to characterize the spatial variation in soil properties, which in turn can be used for soil management. This study shows that soil cation exchange capacity, calcium content, clay percentage and pH have a relatively strong spatial correlation with EC_a in the soil of the study area.  相似文献