Influence of growth rate and starvation on fluorescent in situ hybridization of Rhodopseudomonas palustris

In situ hybridization with a fluorescently labeled 16S rRNA-targeted probe was examined using Rhodopseudomonas palustris as a model organism, which had been grown at different rates and under different conditions of growth and starvation. The specific growth rate did not affect the percentage of hybridized cells in aerobically grown R. palustris cultures. However, significant changes in the percentage of hybridized cells occurred during extended periods of starvation. These changes were observed both in batch cultures grown and starved aerobically in the dark, and in cultures grown phototrophically and starved anaerobically in the dark. Aerobic growth in batch culture and subsequent starvation resulted in a complete lack of detectable hybridization after 20 days of starvation. In contrast, even after 30 days of starvation, 50% of all cells were still detectable in cultures grown aerobically at growth rates <0.06 h(-1) and then starved aerobically in the dark. The same was true for phototrophically grown cells that were starved anaerobically in the light. During starvation there was a clear, though non-linear, positive correlation between the percentage of hybridized cells and the RNA content. In contrast, no direct correlation was observed between the number of hybridized cells in a culture and the viability of this culture. Thus, in habitats with growing, non-growing, and starving bacteria, data on quantitative detection of populations based on 16S rRNA-targeted probing should be used with extreme caution as the detectability of the individual cells is strongly influenced by their physiological history and current physiological state.     

The influence of fungicides on Arthrobotrys oligospora in simulated putting green soil

Plant‐parasitic nematodes are destructive pests in bentgrass putting greens. Few chemical or nonchemical approaches for nematode management exist. Studies were conducted to determine: the in vitro tolerance of the nematophagous fungus Arthrobotrys oligospora, to the fungicides chlorothalonil and myclobutanil used to manage diseases on putting greens; the concentration of fungicides obtained from simulated putting green soil; and the ability of the fungus to reduce populations of the ring nematode, Criconemella ornata. Both fungicides reduced in vitro hyphal growth and germination of conidia above 10 mg kg^‐1. Soil concentrations of chlorothalonil were less than 5 mg kg^‐1 and concentrations of myclobutanil were below detection limits. Nematode populations were not affected by A. oligospora in simulated greens but nematode populations were lowest in pots inoculated with A. oligospora and receiving fungicide treatments. Results of these studies indicate that applications of chlorothalonil and myclobutanil used to manage fungal diseases of bentgrass may not adversely affect A. oligospora; however, the fungus may not reduce nematode populations below desired thresholds.     

Burning Influences on Wiregrass (Aristida beyrichiana) Restoration Plantings: Natural Seedling Recruitment and Survival

Regeneration and expansion of Aristida beyrichiana and Aristida stricta (wiregrass) populations in remaining fire‐maintained Pinus palustris (longleaf pine) stands of the southeastern United States has become an objective of land managers. Although growing‐season fire is required for successful wiregrass seed production, studies examining naturally occurring wiregrass seedling dynamics are few. This study investigates how seedling survivorship is affected by season of burn, seedling size, time since germination, and proximity to adult plants. Restoration at this research site was begun in 1992 with the planting of containerized longleaf pine and wiregrass seedlings. Study plots were established in November 1997 after a growing‐season prescribed fire (June 1996) that resulted in successful seed production and seedling recruitment. Burn treatment plots included (1) no burn (control), (2) fire in the dormant season of the first year after germination (March 1998), (3) fire in the growing season of the first year after germination (August 1998), and (4) fire in the growing season of the second year after germination (July 1999). Seedling mortality increased with growing season burning and close proximity to planted adults. Natural seedling recruitment continued into the second year after initial seed‐drop in all plots, which verifies that wiregrass seed banking occurs for a minimum of 2 years after seed drop. Where wiregrass management objectives include population expansion, seedling recruits should be allowed 1 to 2 years post‐germination without growing season fire for successful establishment.     

The role of oxygen in submergence-induced petiole elongation in Rumex palustris: in situ measurements of oxygen in petioles of intact plants using micro-electrodes

In a study on the mechanism of stimulated petiole elongation in submerged plants, oxygen concentrations in petioles of the flood-tolerant plant Rumex palustris were measured with micro-electrodes. Short-term submergence lowered petiole partial oxygen pressure to c . 19 kPa whereas prolonged submergence under continuous illumination depressed oxygen levels to c . 8–12 kPa after 24 h. Oxygen levels in petioles depended on the presence of the lamina, even in submerged conditions, and on available light. In darkness, petiole oxygen levels in submerged plants dropped quickly to values as low as 0.5–4 kPa. It is hypothesized that prolonged submergence in the light is accompanied by a decrease in carbon dioxide in the petiole. Submergence-enhanced petiolar elongation rate was compared with emergent plants. Peak daily elongation rates occurred at the end of the dark period in emergent plants, but in the middle of the light period in submerged plants. We suggest that this shift in daily elongation pattern is induced by dependence of growth on photosynthetically derived oxygen in submerged plants. Implications of reduced oxygen for ethylene production are raised. Levels of 1- aminocyclopropane-1-carboxylic acid synthase and 1-aminocyclopropane-1-carboxylic acid oxidase and ethylene sensitivity are cited as potential factors in hypoxia-induced ethylene release.     

Efficient selection and regeneration of creeping bentgrass transformants following particle bombardment

We have established an efficient genetic transformation system for creeping bentgrass (Agrostis palustris Huds.) using particle bombardment. The transformation was performed using the plasmid pZO1052 which contains the reporter β-glucuronidase (uidA) gene and the selectable marker hygromycin phosphotransferase (hph) gene. Transformed calli and plants were obtained via particle bombardment followed by selection of transformants on medium containing 200 mg/l of hygromycin. An average of 4.6 resistant colonies per bombardment were obtained. Southern analysis confirmed the integration of foreign genes in 19 of 21 putative transformants, indicating that selection by hygromycin was highly effective. Received: 6 February 1997 / Revision received: 16 April 1997 / Accepted: 9 May 1997     

Elongation by primary lateral roots and adventitious roots during conditions of hypoxia and high ethylene concentrations

Soil flooding results in unusually low oxygen concentrations and high ethylene concentrations in the roots of plants. This gas composition had a strongly negative effect on root elongation of two Rumex species. The effect of low oxygen concentrations was less severe when roots contained aerenchymatous tissues, such as in R. palustris Sm. R. thyrsiflorus Fingerh., which has little root porosity, was much more affected. Ethylene had an even stronger effect on root elongation than hypoxia, since very small concentrations (0.1 cm³ m^?3) reduced root extension in the two species, and higher concentrations inhibited elongation more severely than did anoxia in the culture medium. Thus, ethylene contributes strongly to the negative effects of flooding on root growth. An exception may be the highly aerenchymatous, adventitious roots of R. palustris. Aerenchyma in these roots provides a low-resistance diffusion pathway for both endogenously produced ethylene and shoot-derived oxygen. This paper shows that extension by roots of R. palustris in flooded soil depends almost completely on this shoot-derived oxygen, and that aerenchyma prevents accumulation of growth-inhibiting levels of ethylene in the root.     

Influence of atmospheric CO2 enrichment, soil N, and water stress on needle surface wax formation in Pinus palustris (Pinaceae)

Interactive effects of increasing atmospheric CO2 with resource limitations on production of surface wax in plants have not been studied. Pinus palustris seedlings were grown for 1 yr at two levels of soil N (40 or 400 kg N_ha-1_yr-1) and water stress (-0.5 or -1.5 MPa xylem pressure potential) in open-top field chambers under two levels of CO2 (365 or 720 mumol/mol). Needle surface wax content was determined at 8 mo (fall) and 12 mo (spring) and epicuticular wax morphology was examined using scanning electron microscopy (SEM) at 12 mo. Wax content expressed on both a leaf area and dry mass basis was increased due to main effects of low N and water stress. No main effects of CO2 were observed; however, a CO2 x N interaction at 12 mo indicated that under low soil N the elevated CO2 treatment had less wax (surface area or dry mass basis) compared to its ambient counterpart. Morphologically, low N needle surfaces appeared rougher compared to those of high N needles due to more extensive wax ridges. Although the main effect of water treatment on wax density was not reflected by changes in wax morphology, the CO2 x N interaction was paralleled by alterations in wax appearance. Decreases in density and less prominent epicuticular wax ridges resulting from growth under elevated CO2 and limiting N suggest that dynamics of plant/atmosphere and plant/pathogen interactions may be altered.     

长白山沼泽山雀的生态习性研究

沼泽山雀(Parus palustris brevirostris Taczanovski)是中山、低山带较常见的典型森林鸟类。关于它的生态习性,La Touche(1925—30),ⅡeMe~TKCB,r.n.等(1954),郑作新等(1958),郑作新等(1963)、范忠民等(1965)及李桂垣等(1982)均有过报道;但鉴于其生态习性的地区差异及某些环节的不足,尤其是对其生长发育的研究欠缺。为此,笔者将1963—1965,1979—1983年,在长白山北坡对该鸟生态习性的研究结果,报道如下。     

应用RT-PCR检测基因的体外转录活性

体外转录分析已广泛应用于分子生物学领域．由于体外转录所产生的ＲＮＡ的量极少,需有灵敏的方法检测生成的ＲＮＡ．本研究发展了一种基于ＲＴ－ＰＣＲ技术鉴定体外转录产物的方法．将待研究基因的启动区与任何一段已知的含转录起始位点的编码序列相连,制备成体外转录的模板．体外转录后,用ＤＮａｓｅⅠ将ＤＮＡ模板完全降解;生成的ＲＮＡ经反转录合成ｃＤＮＡ;再以ｃＤＮＡ为模板,用相应的引物进行ＰＣＲ扩增,产物用琼脂糖凝胶电泳检测．该法灵敏度高,操作简便,无需同位素,且体外转录模板制备方便．还可结合其他技术,如Ｓｏｕｔｈｅｒｎ印迹分析,能获得更高的灵敏度．