Transgenic plants as tools to study the molecular organization of plant genes

Transgenic plants are generated in nature by Agrobacterium tumefaciens, a pathogen that produces disease through the transfer of some of its own DNA into susceptible plants. The genes are carried on a plasmid. Much has been learned about how the plasmid is transferred, how the plasmid-borne genes are organized, regulated, and expressed, and how the bacteria's pathogenic effects are produced. The A. tumefaciens plasmid has been manipulated for use as a general vector for the transfer of specific segments of foreign DNA of interest (from plants and other sources) into plants; the activities of various genes and their regulation by enhancer and silencer sequences have been assessed. Future uses of the vector (or others like it that have different host ranges) by the agriculture industry are expected to aid in moving into vulnerable plants specific genes that will protect them from such killers as nonselective herbicides, insects, and viruses.     

Role of Extracellular Polysaccharide and Endoglucanase in Root Invasion and Colonization of Tomato Plants by Ralstonia solanacearum

ABSTRACT Ralstonia solanacearum is a soilborne plant pathogen that normally invades hosts through their roots and then systemically colonizes aerial tissues. Previous research using wounded stem infection found that the major factor in causing wilt symptoms was the high-molecular-mass acidic extracellular polysaccharide (EPS I), but the beta-1,4-endoglucanase (EG) also contributes to virulence. We investigated the importance of EPS I and EG for invasion and colonization of tomato by infesting soil of 4-week-old potted plants with either a wild-type derivative or genetically well-defined mutants lacking EPS I, EG, or EPS I and EG. Bacteria of all strains were recovered from surface-disinfested roots and hypocotyls as soon as 4 h after inoculation; that bacteria were present internally was confirmed using immunofluorescence microscopy. However, the EPS-minus mutants did not colonize stems as rapidly as the wild type and the EG-minus mutant. Inoculations of wounded petioles also showed that, even though the mutants multiplied as well as the wild type in planta, EPS-minus strains did not spread as well throughout the plant stem. We conclude that poor colonization of stems by EPS-minus strains after petiole inoculation or soil infestation is due to reduced bacterial movement within plant stem tissues.     

Landscape analysis of plant diversity

Studies to identify gaps in the protection of habitat for speciesof concern have been inconclusive and hampered by single-scale orpoor multi-scale sampling methods, large minimum mapping units(MMU's of 2 ha to 100 ha), limited and subjectively selected fieldobservations, and poor mathematical and ecological models. Weovercome these obstacles with improved multi-scale samplingtechniques, smaller MMU's (< 0.02 ha), an unbiased sampling designbased on double sampling, improved mathematical models includingspecies-area curves corrected for habitat heterogeneity, andgeographic information system-based ecological models. We applythis landscape analysis approach to address resource issues inRocky Mountain National Park, Colorado. Specifically, we quantifythe effects of elk grazing on plant diversity, identify areas ofhigh or unique plant diversity needing increased protection, andevaluate the patterns of non-native plant species on thelandscape.Double sampling techniques use satellite imagery,aerial photography, and field data to stratify homogeneous andheterogeneous units and keystone ecosystems (ecosystems thatcontain or support a high number of species or have distinctivespecies compositions). We show how a multi-scale vegetationsampling design, species-area curves, analyses of within- andbetween-vegetation type species overlap, and geographic informationsystem (GIS) models can be used to quantify landscape-scalepatterns of vascular plant diversity in the Park.The new multi-scale vegetation plot techniques quickly differentiated plantspecies differences in paired study sites. Three plots in the OuzelBurn area (burned in 1978) contained 75 plant species, while only17 plant species were found in paired plots outside the burn.Riparian areas contained 109 plant species, compared to just 55species in paired plots in adjacent forests. However, plant speciesrichness patterns inside and outside elk exclosures were morecomplex. One elk exclosure contained more species than its adjacentopen range (52 species inside and 48 species outside). Two elkexclosures contained fewer species inside than outside (105 and 41species inside and 112 and 74 species outside, respectively).However, there was only 26% to 48% overlap (using Jaccard'sCoefficient) of plant species composition inside and outside theexclosures. One elk exclosure had 13% cover of non-indigenousspecies inside the exclosure compared to 4% outside, butnon-indigenous species cover varied by location.We compared plantdiversity patterns from vegetation maps made with 100 ha, 50 ha, 2ha, and 0.02 ha MMU's in the 754 ha Beaver Meadows study area usingfour 0.025 ha and twenty-one 0.1 ha multi-scale vegetation plots.Preliminary data suggested that the 2 ha MMU provided an accurateestimate of the number of plant species (–14%) for a study area,but the number of habitats (polygons) was reduced by 67%, andaspen, a unique and important habitat type, was missed entirely. Wedescribe a hypothesis-driven approach to the design andimplementation of geospatial databases for local resourcemonitoring and ecosystem management.     

利用转座子Tn5诱变植物青枯菌获得胞外蛋白输出功能丧失突变体

 通过Tn5诱变技术,分离了植物青枯菌生理小种1号、3号的胞外蛋白输出功能丧失突变体。由于Tn5在其基因组单一的eep位点的插入,突变体失去了向培养滤液分泌产生胞外酶和胞外蛋白质的能力。用碱性磷酸酯酶基因PhoA作为报导基因,研究这些胞外蛋白穿越突变体细胞内膜,结果表明,这些胞外蛋白质可以穿过其内膜,但失去了穿越其外膜的能力。胞外多糖在植物体内和体外的产生没有受到eep基因位点突变的影响。该突变体失去了对番茄植株的致萎能力。     

Boar Sperm Encapsulation Reduces In Vitro Polyspermy

A boar sperm encapsulation technology in barium alginate has been developed to enhance reproductive performances and spermatozoa preservation time; aim of this work was to evaluate the effect of in vitro sperm encapsulation on polyspermy as a function of storage time at 18°C. A total number of 40 in vitro fertilization (IVF) tests were performed using encapsulated or diluted spermatozoa (20 IVF each treatment). Overall, 1288 in vitro matured oocytes were fertilized with spermatozoa stored at 24, 48 or 72 h at 18°C for both treatments polyspermy and normospermy, and the non‐penetration rates were assessed by optical microscopy. Results indicate a significant reduction in risk of polyspermic oocytes when spermatozoa are preserved in barium alginate membranes (incidence risk ratio: 0.766 with respect to diluted); such enhancement could be explained by lesser damage of sperm membranes achieved by encapsulation technology.     

Pleistocene extinction of genyornis newtoni: human impact on australian megafauna

More than 85 percent of Australian terrestrial genera with a body mass exceeding 44 kilograms became extinct in the Late Pleistocene. Although most were marsupials, the list includes the large, flightless mihirung Genyornis newtoni. More than 700 dates onGenyornis eggshells from three different climate regions document the continuous presence of Genyornis from more than 100,000 years ago until their sudden disappearance 50,000 years ago, about the same time that humans arrived in Australia. Simultaneous extinction of Genyornis at all sites during an interval of modest climate change implies that human impact, not climate, was responsible.     

Endoscopy of the upper respiratory tract during treadmill exercise: a clinical study of 100 horses

SUMMARY: Endoscopy of the upper respiratory tract was performed in 100 horses during high speed treadmill exercise. Reasons for endoscopy were a history of an abnormal noise during exercise in 75 horses, poor performance in 17 horses and to evaluate the results of upper respiratory tract surgery in 8 horses. Of the 75 horses with a history of an abnormal noise during exercise the cause was determined in 67 (89%). Endoscopic abnormalities were detected at rest in 40 of these 75 horses (53%). In these 40 horses, a similar diagnosis as to the cause of the abnormal noise was made at rest and during exercise on the treadmill in 19 cases, while in the remaining 21 the endoscopic findings during exercise varied from that seen at rest. This included 3 horses in which a diagnosis was made at rest but no abnormalities were detected during exercise. Some of the findings during treadmill endoscopy included laryngeal dysfunction, grades 3, 4 and 5 (22 cases), dorsal displacement of the soft palate (20), epiglottic entrapment (8), epiglottic flutter (4), aryepiglottic fold flutter (4), pharyngeal collapse (3), arytenoiditis (3), vocal cord flutter (3), false nostril noise (2), pharyngeal lymphoid hyperplasia (2), soft palate haemorrhage (1) and positional arytenoid collapse (1). More than one abnormality was observed during exercise in 7 horses. A complete and correct diagnosis based on the resting endoscopy findings alone was made in 19 (25%) of these 75 cases. In the 17 horses examined because of poor performance, no abnormalities were detected during treadmill endoscopy that were not evident at rest. None of these 17 horses presented with a history of an abnormal respiratory noise, although one, diagnosed as having grade 4 laryngeal function at rest and exercise, did make a characteristic inspiratory noise during treadmill exercise. Eight horses were evaluated after surgery for correction of laryngeal hemiplegia, as the post-operative performance or the amount of respiratory noise present was considered unsatisfactory. Of these, 3 were found to have a satisfactory airway during exercise and other reasons for poor performance were detected; 3 had insufficient abduction; and 2 had intermittent dorsal displacement of the soft palate. Endoscopy of the upper respiratory tract was found to be a useful technique for evaluating the cause of abnormal respiratory noise in most cases. We concluded that treadmill endoscopy in horses presented for poor performance, without a history of an abnormal respiratory noise, was of little value. The technique, in conjunction with arterial blood gas measurements, was useful in determining the efficacy of surgical treatment of laryngeal hemiplegia.     

Acute and Chronic Effects of a Contraceptive Compound RTI‐4587‐073(l) on Testicular Histology and Endocrine Function in Miniature Horse Stallions

The objective of this study was to evaluate acute endocrine effects as well as histological changes in testicular parenchyma induced by the contraceptive compound RTI‐4587‐073(l). Six miniature stallions were used in this experiment. The treatment group (n = 3) received one oral dose of 12.5 mg/kg of RTI‐4587‐073(l), and the control group (n = 3) received placebo only. The stallions' baseline parameters (semen, testicular dimensions, endocrine values) were collected and recorded for 5 weeks before treatment and for 6 weeks after treatment. Multiple blood samples were collected for endocrine analysis. Testicular biopsies were obtained before treatment, 1 day after treatment and every other week after treatment. Ultrasound exams were performed to monitor the dimensions of the stallions' testes. All stallions were castrated 6 weeks after treatment. Sperm numbers, motility and percentage of morphologically normal sperm decreased (p < 0.05), while the number of immature germ cells increased in ejaculates from treated animals (p < 0.05). Serum concentrations of inhibin and follicle‐stimulating hormone did not change. Testosterone concentrations initially transiently decreased (p < 0.05) after administration of RTI‐4587‐073(l), and increased several days later (p < 0.05). Testicular content of testosterone and estradiol 17‐β was lower in treated stallions than in control stallions on Day 1 after treatment (p < 0.05). Severe disorganization of the seminiferous tubules, significant loss of immature germ cells and complete depletion of elongated spermatids were observed in testicular biopsies obtained from treated stallions 1 day, 2 and 4 weeks after treatment. These changes were still present in the testicular samples taken from treated stallions after castration. The results of this study confirmed that RTI‐4587‐073(l) has antispermatogenic effects in stallions. Furthermore, we concluded that this compound causes acute sloughing of immature germ cells from the seminiferous tubules. RTI‐4587‐073(l) has significant but transient effects on Leydig cell function in stallions.