Effect of head removal on leaf senescence of sunflower

Greenhouse and field studies examined the effect of flower or seedhead removal on leaf senescence and associated changes in sunflower (Helianthus annuus L.) plants. At intervals during seed development, selected leaves (leaves 6 through 8 from the top in the greenhouse and leaf 7 from the top in the field) were harvested and analyzed for chlorophyll, specific leaf weight, N, P, soluble protein, and electrophoretic gel profiles of soluble polypeptides. In both the greenhouse and the field, the leaves of headless plants retained or accumulated more N, P, soluble protein, and dry weight than leaves of plants with heads. Obviously, head removal affected the partitioning of these metabolites during seed development. None of the treatments resulted in the formation of new polypeptides (electrophoretic gel profiles). Comparisons of the rates and extent of loss of chlorophyll, soluble protein, and polypeptide bands (especially ribulose 1,5-bisphosphate carboxylase) from the leaves of headed and deheaded plants showed that head removal delayed the rate of development of leaf senescence for the greenhouse-grown but had much less effect on field-grown plants. These findings illustrate the variability in different parameters commonly associated with the leaf senescence processes of headed and deheaded sunflower plants grown under different environments.     

Pathways of soil organic matter formation from above and belowground inputs in a Sorghum bicolor bioenergy crop

When aboveground materials are harvested for fuel production, such as with Sorghum bicolor, the sustainability of annual bioenergy feedstocks is influenced by the ability of root inputs to contribute to the formation and persistence of soil organic matter (SOM), and to soil fertility through nutrient recycling. Using ¹³C and ¹⁵N labeling, we traced sorghum root and leaf litter‐derived C and N for 19 months in the field as they were mineralized or formed SOM. Our in situ litter incubation experiment confirms that sorghum roots and leaves significantly differ in their inherent chemical recalcitrance. This resulted in different contributions to C and N storage and recycling. Overall root residues had higher biochemical recalcitrance which led to more C retention in soil (27%) than leaf residues (19%). However, sorghum root residues resulted in higher particulate organic matter (POM) and lower mineral associated organic matter (MAOM), deemed to be the most persistent fraction in soil, than leaf residues. Additionally, the overall higher root‐derived C retention in soil led to higher N retention, reducing the immediate recycling of fertility from root as compared to leaf decomposition. Our study, conducted in a highly aggregated clay‐loam soil, emphasized the important role of aggregates in new SOM formation, particularly the efficient formation of MAOM in microaggregate structures occluded within macroaggregates. Given the known role of roots in promoting aggregation, efficient formation of MAOM within aggregates can be a major mechanism to increase persistent SOM storage belowground when aboveground residues are removed. We conclude that promoting root inputs in S. bicolor bioenergy production systems through plant breeding efforts may be an effective means to counterbalance the aboveground residue removal. However, management strategies need to consider the quantity of inputs involved and may need to support SOM storage and fertility with additional organic matter additions.     

Characterization of bovine placental lactogen as a glycoprotein with N-linked and O-linked carbohydrate side chains

In a previous report we showed that purified bovine placental lactogen (bPL) exists in two isoforms in the 31,000-33,000 Mr range, each with at least five isoelectric variants differing in approximately 2 orders of magnitude in isoelectric points (pI) 4-6. The multiple isoelectric variants are unique to the bovine hormone. In an effort to determine the nature of these variants endo- and exoglycohydrolase digestions were conducted to determine if this hormone was glycosylated. Analysis of peptide/N-glycosidase F and endoglycosidase F digests of radioiodinated bPL on one-dimensional gel electrophoresis showed a Mr decrease from 31,000 to 24,000 and 33,000 to 26,000 for the two isoforms. Digestion with a mixture of neuraminidase plus mixed exoglycosidases resulted in a Mr decrease of 4,000. Digestion with neuraminidase resulted in a Mr decrease of 2,000. Further analysis of peptide/N-glycosidase F- and neuraminidase-treated bPL by two-dimensional gel electrophoresis showed the isoelectric variants shifted from pI 4.4-6.3 to 4.9-8.0. The sialic acid residues on the N-linkage are responsible for the pronounced acidic character of bPL, but do not account for the residual charge heterogeneity as the different isoelectric variants persist after sialic acid removal. The apparent Mr of the protein after removal of N-linked carbohydrate residues is similar to that of PRL and GH. These enzymatic digestion results demonstrate the presence of N-linked complex oligosaccharide residues attached to the beta-amide group of an asparagine residue. Analyses of the sugar content of the molecule were consistent with the presence of one biantennary N-linked and two O-linked carbohydrate chains.     

Positive interactions between leafrollers and other arthropods enhance biodiversity on hybrid cottonwoods

We examined the potential of a common herbivore to indirectly influence other diverse community members by providing habitat. Larvae of the leafroller Anacampsis niveopulvella commonly construct shelters by rolling leaves of cottonwood trees. These leaf rolls are later colonized by other arthropods. We first documented 4 times greater species richness and 7 times greater abundance on cottonwood shoots that contained a rolled leaf compared to adjacent shoots without leaf rolls. Second, with both removal and addition experiments, we showed that leaf rolls are responsible for these differences in arthropod assemblages. Leaf roll removal caused a 5-fold decline in richness and a 7-fold decline in abundance; leaf roll addition resulted in a 2.5-fold increase in richness and a 6-fold increase in abundance. Third, to determine whether rolled leaves are colonized for food or for shelter, we compared colonization of natural and artificial leaf rolls. Both richness and abundance were approximately 2-fold greater in artificial leaf rolls, indicating that leaf rolls are colonized primarily for shelter. Fourth, in a natural hybrid zone we found that leafroller densities were 2-fold greater on backcross hybrids than on F₁ hybrids. These differences are likely associated with genetically-based differences in leaf morphology and/or leaf chemistry. Ultimately, plant genotype affects positive indirect interactions that have the potential to affect community structure. This study and others demonstrate that shelter builders (i.e., leafrollers and gall formers) enhance biodiversity, while free-feeders are more likely to negatively affect biodiversity. Received: 19 April 1999 / Accepted: 8 November 1999     

The aspartic proteinase of barley is a vacuolar enzyme that processes probarley lectin in vitro.

Previous work suggested that the aspartic proteinase from Hordeum vulgare (HvAP) would be a vacuolar protein in plant cells. Based on N-terminal sequencing we show that the in vitro-translated protein was translocated into the lumen of microsomal membranes, causing a concomitant removal of 25 amino acid residues from the protein. Vacuoles were purified from barley leaf protoplasts and were shown to contain all of the aspartic proteinase activity found in the protoplasts. This vacuolar localization of HvAP was confirmed with immunocytochemical electron microscopy using antibodies to HvAP in both barley leaf and root cells. In an attempt to discern a function for this protease, we investigated the ability of HvAP to process the C-terminal proregion of barley lectin (BL) in vitro. Prolectin (proBL), expressed in bacteria, was processed rapidly when HvAP was added. Using several means, we were able to determine that 13 amino acid residues at the C terminus of proBL were cleaved off, whereas the N terminus stayed intact during this incubation. Immunohistochemical electron microscopy showed that HvAP and BL are co-localized in the root cells of developing embryos and germinating seedlings. Thus, we propose that the vacuolar HvAP participates in processing the C terminus of BL.     

Study of the role of lysine residues of the cholesterol hydroxylating cytochrome P-450 by a method of chemical modification

Chemical modification of cytochrome P-450scc by lysine-specific reagents has been performed. Modification of the hemoprotein was shown to result in the loss of its ability to interact with adrenodoxin. With a view of identifying lysine residues involved in the interaction with adrenodoxin, cytochrome P-450scc was modified by succinic anhydride in the presence of adrenodoxin. After the removal of ferredoxin, the modification was performed with the use of a radioactively labeled reagent. Subsequent hydrolysis of the succinic hemoprotein by chymotrypsin and separation of the peptides obtained by high pressure liquid chromatography resulted in the isolation of seven chymotryptic peptides containing labeled lysine residues. These amino acid sequences were identified. The role of lysine residues of cytochrome P-450scc in complex formation with adrenodoxin is discussed.     

Nitrogen Uptake and Plant Growth I. Effect of Nitrogen Removal on Growth of Polygonum cuspidatum

Polygonun cuspidatum was grown hydroponically to examine theeffect of nitrogen removal from the nutrient solution upon plantgrowth and the partitioning of dry matter and nitrogen amongorgans. Nitrogen removal reduced the growth rate mainly dueto the reduced growth of leaf area. Accelerated root growthwas observed only in plants which earlier had received highlevels of nitrogen. Nitrogen removal caused almost exclusiveallocation of available nitrogen to root growth. Nitrogen fluxfrom the shoot to the root occurred in plants which had receivedlow nitrogen. Not only was net assimilation rate (NAR) littleaffected by nitrogen removal, but it also was not correlatedwith the concentration of leaf nitrogen on an area basis. Light-saturatedCO₂ exchange rate (CER) was highly correlated with the concentrationof leaf nitrogen. Nitrogen use efficiency (NUE) in CER (CERdivided by leaf nitrogen) remained constant against leaf nitrogen,indicating efficient use of nitrogen under light saturation,while NUE in terms of NAR decreased with higher concentrationof leaf nitrogen. Polygonum cuspidatum Sieb. et Zuce., CO₂ exchange rate, growth analysis, leaf nitrogen, net assimilation rate, nitrogen use efficiency, partitioning of dry matter and nitrogen     

Staining Paraffin Embedded Sections of Scald of Barley before Paraffin Removal

Staining of paraffin embedded sections with periodic acid-Schiff reagent and fast green before paraffin removal resulted in differentiation of barley seed and leaf tissue from fungal structures of Rhynchosporium secalis. Crystal violet, toluidine blue O and aniline blue also successfully stained fungal structures of R. secalis in barley leaf tissues. Staining of embedded sections before paraffin removal allows simple processing of a series of sections, saves time and reduces solvent consumption.     

The alkaloidal responses of wild tobacco to real and simulated herbivory

Summary I compared the induced alkaloidal response in undamaged leaves of plants subjected to herbivory by the larvae of Manduca sexta and to different simulations of this herbivory; all herbivory treatments removed similar amounts of leaf mass. Although larval feeding induced a significant increase (2.2x) in alkaloid concentrations compared to undamaged plants, the alkaloid responses to larval feeding were significantly lower than the responses to an herbivory simulation (4x controls) which involved removing the same amount of leaf area from the same positions on the leaf, over a similar time period. Moreover, another herbivory simulation, identical in amount of leaf mass removed and duration of damage to the larval feeding, but without regard to spatial array of leaf damage, resulted in an alkaloidal response (5.5x controls) higher still than the previous herbivory simulation. In a second experiment the importance of leaf vein damage on the induced alkaloidal response was examined. Here, leaf removal that involved cutting leaf tissues from between secondary veins before removing the midrib, resulted in alkaloidal responses that were significantly lower (1.7x controls) than responses from leaf removal that involved cutting both veins and midribs along with the intervein tissues (2.6x controls). Vein damage alone did not produce a significant response. These results indicate that herbivory is difficult to simulate: that how a leaf is damaged can be as important as the magnitude of leaf damage in determining a plant's response to damage.     

The carboxyl-terminal region of human interferon gamma is important for biological activity: mutagenic and NMR analysis.

Deletion of nine amino acids from the carboxyl terminus of human IFN gamma (residues 138--146; LFRGRRASQ) resulted in a 7-fold increase in specific antiviral activity. Similar increases in receptor binding affinity were seen. Deletion of residues 136 and 137 (QM) had little additional effect, but removal of Ser135 resulted in a sharp drop in antiviral activity. Further removal of residues 133 and 134 (KR) lowered antiviral activity to 1% of the peak value. Comparison of the proton NMR spectra of selected deletions down to residue 132 showed that there was no significant change in the core protein structure. Deletions down to residue 125 had the same antiviral activity as those to 132, but changes could now be seen in the aromatic proton NMR spectrum of this shorter derivative. Substitution of the homologous murine sequence between residues 124 and 130 (human SPAAKTG; murine LPESSLR) resulted in only a small decrease in antiviral activity, further suggesting that the precise sequence in this region was not critical for activity. Ser135 was substituted with a number of other amino acids with little or no change in activity. The importance of the residues between 131 and 134 for biological activity was corroborated by mutagenesis, although some substitutions in this region were tolerated.     

顶空气相色谱法检测苦荞麦总黄酮提取物的有机溶剂残留量

为控制使用AB-8型大孔树脂后苦荞麦总黄酮提取物的质量,采用顶空气相色谱法,对提取物中正己烷、苯、甲苯、对二甲苯、间二甲苯、邻二甲苯、乙苯和苯乙烯等8种残留物进行检测。精密度RSD均小于6％,峰面积与浓度均有良好的线性关系,最低检测限为0．016—0．0640g／mL,回收率均符合要求。试验结果表明,方法稳定、操作简便、准确可靠,适用于使用AB-8型大孔树脂后苦荞麦总黄酮提取物中有机溶剂残留量的检测。     

Efficacy and uptake of soil-applied imidacloprid in the control of Asian citrus psyllid and a citrus leafminer, two foliar-feeding citrus pests

The systemic neonicotinoid insecticide imidacloprid, Admire Pro, was applied to 3- and 4-yr-old nonbearing 'Rio Red' grapefruit, Citrus x paradisi Macfad., trees in 2006 and 2007, respectively, to determine its effects in the control of two major citrus pests, the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), and a citrus leafminer Phyllocnistis citrella Stainton (Lepidoptera: Gracillariidae). Young flush shoots were randomly collected weekly for 13 and 11 wk in 2006 and 2007, respectively, to determine the infestation levels and densities of immature stages of both Asian citrus psyllid and P. citrella. Additional flush shoot samples were collected in 2007 and titers of imidacloprid in leaf tissue were determined using an enzyme-linked immunosorbent assay. Soil application of imidacloprid significantly reduced the infestation levels and densities of both pests on flush shoots, starting from the second week post application. The effects of the neonicotinoid insecticide were similar in both years. Analysis of imidacloprid concentration in leaf tissue showed a gradual increase during the first 3 wk, and titers remained well above 200 ppb for 11 wk postapplication. Significant positive correlations were obtained between imidacloprid titers in leaf tissue and the percentage of control levels achieved for both pests. A high level of suppression of both P. citrella and Asian citrus psyllid populations on citrus trees was associated with imidacloprid titer in leaf tissue >200 ppb, which was reached 2 wk after soil treatment. Although soil application of imidacloprid did not provide rapid knockdown of Asian citrus psyllid and P. citrella populations, it resulted in chronic residues in leaf tissue and long-term suppression of both pests.     

Combined effects of girdling and leaf removal on fluorescence characteristic of Alhagi sparsifolia leaf senescence

Plant senescence is largely influenced by carbohydrate content. In order to investigate the impact of carbohydrate content on leaf senescence and photosystem II (PSII) during the senescence process, phloem girdling (PG), leaf removal (LR) and a combination of phloem girdling and leaf removal (GR) were performed on Alhagi sparsifolia (Fabaceae) at the end of the growing season. The results showed that during senescence, leaf soluble sugar content, starch content, the energy absorbed by the unit reaction centre (ABS/RC) increased; whereas, leaf photosynthetic rate, photosynthetic pigment content, maximum photochemical efficiency (φ_Po) and energy used by the acceptor site in electron transfer (ET_o/RC) decreased. The degree of change was PG> GR> CK (control)> LR. The results of the present work implied that phloem girdling (PG) significantly accelerated leaf senescence, and that single leaf removal (LR) slightly delayed leaf senescence; although leaf removal significantly delayed the senescence process on the girdled leaf (GR). Natural or delayed senescence only slightly inhibited the acceptor site of PSII and did not damage the donor site of PSII. On the other hand, induced senescence not only damaged the donor site of PSII (e.g. oxygen‐evolving complex), but also significantly inhibited the acceptor site of PSII. In addition, leaf senescence led to an increase in the energy absorbed by the unit reaction centre (ABS/RC), which subsequently resulted in increasing excitation pressure in the reaction centre (DI_o/RC), as well as additional saved Car for absorbing residual light energy and quenching reactive oxygen species during senescence.     

Vascular development of the grapevine (<Emphasis Type="Italic">Vitis vinifera</Emphasis> L.) inflorescence rachis in response to flower number,plant growth regulators and defoliation

The grapevine inflorescence is a determinate panicle and as buds emerge, shoot, flower and rachis development occur simultaneously. The growth and architecture of the rachis is determined by genetic and environmental factors but here we examined the role of flower and leaf number as well as hormones on its elongation and vascular development. The consequences of rachis morphology and vascular area on berry size and composition were also assessed. One week prior to anthesis, Merlot and Cabernet Sauvignon field vines were exposed to manual flower removal, exogenous plant growth regulators or pre-bloom leaf removal. Manual removal of half the flowers along the vertical axis of the inflorescence resulted in a shorter rachis in both cultivars. Conversely, inflorescences treated with gibberellic acid (GA₃) and the synthetic cytokinin, 6-benzylaminopurine (BAP) resulted in a longer rachis while pre-bloom removal of all leaves on the inflorescence-bearing shoot did not alter rachis length relative to untreated inflorescences. Across the treatments, the cross-sectional areas of the conducting xylem and phloem in the rachis were positively correlated to rachis girth, flower number at anthesis, bunch berry number, bunch berry fresh mass and bunch sugar content at harvest. Conversely, average berry size and sugar content were not linked to rachis vascular area. These data indicate that the morphological and vascular development of the rachis was more responsive to flower number and plant growth regulators than to leaf removal.     

Extraction and retrieval of potassium from water hyacinth (Eichhornia crassipes)

Studies were carried out on extraction and retrieval of potassium from water hyacinth (Eichhornia crassipes). The stem and leaf were subjected to 13 treatments. The highest rate of K removal following HCl treatment was 69.7% K. Most effective removal of suspended organic substances, Ca2+ and Mg2+ were achieved at pH approximately 13, when 88.0% of K remained in filtrate. Maximum K in precipitate following this step was achieved with tartaric acid additions at n(C4H6O6)/n(K+) of 1.72 when precipitating at 4 degrees C for 3h, which resulted in 72.3% of K removal from the solution. Over the entire process, 44.3% of K in the dried stem-leaf sample of water hyacinth was retrieved in the form of KC4H5O6. This process demonstrated the potential for use of water hyacinth as a resource of potassium to produce potassium salts and provide a valuable end use for the plant, which could be highly invasive in aquatic ecosystems.     

PATTERNS OF LEAF AND FLOWER REMOVAL: THEIR EFFECT ON FRUIT GROWTH IN CHAMAENERION ANGUSTIFOLIUM (FIREWEED)

The response of Chamaenerion angustifolium (fireweed) plants to different patterns of artificial leaf and flower herbivory was examined to determine if the effects of localized herbivory were confined within vertical sectors of the plant. The effect of leaf and flower removal on fruit development was compared for removals within sectors and distributed among sectors. Fruit development did not differ in plants subjected to different patterns of leaf and flower removal. These results suggest that in disturbed plants carbohydrate movement is not confined within vertical sectors.     

Phosphorylation of vimentin head domain inhibits interaction with the carboxyl-terminal end of alpha-helical rod domain studied by surface plasmon resonance measurements

The amino-terminal head domain of vimentin is the target site for several protein kinases and phosphorylation induces disassembly of the vimentin intermediate filaments in vivo and in vitro. To better understand molecular mechanisms involved in phosphorylation-dependent disassembly, we examined domain interactions involving the head domain and the effect of phosphorylation on the interaction, using surface plasmon resonance. We observed that the head domain binds to the carboxyl-terminal helix 2B in the rod domain, under physiological ionic strength. This interaction was interfered with by A-kinase phosphorylation of the head domain. Deletion of the carboxyl-terminal 20 amino acids of helix 2B resulted in loss of the interaction. Furthermore, peptide representing the carboxyl-terminal 20 residues of helix 2B had a substantial affinity with the head domain but not with the phosphorylated one. These findings support the idea that the interaction between the head domain and the last 20 residues of helix 2B is essential for association of vimentin tetramers into the intermediate filaments and that the phosphorylation-dependent disassembly is the result of loss of the interaction.     

Yield response of pumpkin and winter squash to simulated cucumber beetle (Coleoptera: Chrysomelidae) feeding injury

Field studies were conducted to characterize the yield responses of pumpkin, Cucurbita pepo L., and wintersquash, Cucurbita maxima Duch., to simulated striped cucumber beetle, Acalymma vittatum (F.), feeding damage. Preliminary studies in pumpkin ('Spookie') were conducted in 1993, and more complete studies in 1994 and 1995 in pumpkin (Spookie), and in 1995 and 1996 in winter squash ('Waltham'). Plants were artificially injured 1 time at the cotyledon, 1st, 2nd, or 3rd true leaf stages by clipping specified percentages of leaf area from all leaves on the plant. Treatments consisted of a control treatment with no simulated injury and 4 levels of simulated injury. Pumpkin treatments were 10, 20, 40, and 80% leaf area removal in 1993 and 1994 and 20, 40, 60, and 80% in 1995. Winter squash treatments were 20, 40, 60, and 80% leaf area removal in 1995, and 10, 20, 40, and 80% in 1996. In 1993, 5 wk after planting, mean leaf, stem, and total dry weight were significantly lower on pumpkin plants subjected to > or = 20% simulated leaf injury regardless of the growth stage when the injury occurred. Injury at the 3rd leaf stage resulted in a significant reduction in the mean leaf and total dry weight of plant, across all levels of injury. At 2-3 wk before crop maturity, fruit weight and number of fruit per plant were significantly lower with 80% leaf area removal. In 1994 and 1995, and when pumpkins were grown to full maturity and apparently able to compensate for the simulated damage, neither number nor weight of marketable fruit per plant were affected significantly by simulated injury at any stage of development nor level of simulated injury. In contrast, > or = 20% and 80% leaf area removal to winter squash resulted in significant reductions in the weight of marketable fruit and/or number of marketable fruit, in 1995 and 1996, respectively. The results of this study indicate that pumpkins can tolerate relatively high levels of simulated defoliation, whereas winter squash is less tolerant. Variable results across years in winter squash may indicate an interaction between injury and plant stress caused by abiotic factors such as soil moisture availability.     

The toxicity of aphicide residues to beneficial invertebrates in cereal crops

The toxicity of dimethoate, deltamethrin and pirimicarb residues to Bembidion lampros and Coccinella septempunctata was evaluated by confining groups of insects to winter wheat foliage and soil for 24 h at different times after treatment in the field. Flag leaf residues were found to be more toxic than first leaf residues: soil residues were the least toxic with pirimicarb showing virtually no soil toxicity. In general, dimethoate and deltamethrin showed similar levels of foliar toxicity with flag leaf toxicity on the first day after treatment being in the range 60–80% for B. lampros; deltamethrin was however, less toxic than dimethoate at ground level. Both of these products were more toxic than pirimicarb. The long-term exposure of insects, surviving the 24 h bioassays, to treated soil at different times following application resulted in further mortality and provided estimates of the maximum levels of mortality that populations of predators might suffer migrating into the crop at different times following application. Dimethoate was shown to be particularly harmful at the current recommended field application rate and reduced doses were proposed to limit the severity of the initial effects.     

A biological assay technique for the assessment and comparison of systemic insecticide residues

A technique is described using mortality of aphids on excised leaf samples to assess residual systemic insecticides within the leaf or on its surface. Residues in crops or soil can also be assessed by keeping aphids on leaf samples treated with solvent extracts of contaminated material. Little or no pre-treatment for the removal of potentially interfering materials co-extracted from the crop is required. Comparison of mortalities caused by unknown residues with mortalities caused by known amounts of insecticide enables the technique to be used quantitatively. The sensitivity of the technique varies with the toxicity of the compound being assayed. With most of the insecticides tested, extract solutions down to about 10 ppm may be assessed directly; weaker solutions must first be concentrated.