Translocation of leaf-fed 32P in pea plants as influenced by foliar and root applications of CCC and Phosfon

In controlled environment growth chambers, the effects of foliar and root applications of 2-chloroethyltrimethylammonium chloride (CCC) and 2,4-dichlorobenzyltributylphosphonium chloride (Phosfon) on the translocation of³²P fed to leaves, were investigated. When applied to leaves or to root, CCC had no effect on the relative amounts of³²P radioactivity retained by the fed leaf 5, 20 and 80 h after feeding. At 20 and 80 h after feeding, Phosfon concentrations of 0.01 and 0.1mg l^?1 reduced retention of the applied³²P. 80 h after³²P feeding, CCC concentration of 1 mg l^?1 applied as a foliar spray or to the root enhanced the downward movement of³²P. Phosfon at low concentrations, particularly at 0.1 mg l^?1, on the other hand, favoured an upward transport of the applied³²P. Foliar applications of CCC and Phosfon at high concentrations had no significant effect on³²P transport to the root and the shoot below the fed leaf, while root applications of CCC and of Phosfon inhibited downward transport. Root applications generally caused greater alterations in³²P distribution patterns than did foliar applications. On the basis of total active ingredient uptake, Phosfon was more effective than CCC in altering translocation patterns.     

Studies on the Mode of Action of Polyoxin D

In the previous papers we reported that the antibiotic Polyoxin D induced the characteristic swelling of the mycelia of fungi,^1,2) and strongly inhibited the incorporation of ¹⁴C-glucosamine into the fungal cell wall chitin.³⁾ The present work has been conducted to further investigate the influence of this antibiotic on the fungal cell wall biosynthesis.

Polyoxin D did not inhibit the incorporation of ¹⁴C-glucose, ¹⁴C-amino acids and ¹⁴C-sodium acetate into the cell wall. In addition, UDP-N-acetylglucosamine, a precurcor of chitin biosynthesis of cell wall, was accumulated in the Polyoxin D-treated mycelia of Cochliobolus miyabeanus more than 150 to 160% of that accumulated in the untreated one.

Chitin synthetase prepared from Piricularia oryzae which is not treated with Polyoxin D was completely inhibited by the addition of 0.1 ppm of Polyoxin D. The fungitoxicity of Polyoxins A to G was positively related to their inhibition of ¹⁴C-glucosamine incorporation into the cell wall chitin of C. miyabeanus. From above results, it became evident that the antibiotic Polyoxin complex inhibited the biosynthesis of fungal cell wall chitin.     

Morphogenetic responses of tomato plants to combined and individual applications of gibberellic acid,Phosfon and B-nine

Summary Two growth retardants, B-nine (N-dimethylamino succinamic acid) and Phosfon (2,4-dichlorobenzyl-tributyl phosphonium chloride) were applied to tomato plants, either singly or in combination with gibberellic acid (GA), in order to determine various morphogenetic responses. GA (5 ml of 10^– M ⁴ per plant) and B-nine (5 ml of 1.56×10^–2 M per plant) were applied as foliar spray whereas Phosfon (1.5×10^–3 M in 10 ml of water per plant) was applied as soil amendment. Growth retardation by Phosfon persisted through the time of harvest and was somewhat neutralized by GA. Fruit set and extent of seediness of fruits were the maximum in Phosfon-treated plants compared to others. Plants receiving B-nine, however, recovered from the initial growth retardation and indicated no residual action at harvest. GA in combination with B-nine produced significantly greater vegetative growth and dry weight accumulation than did GA alone. This indicated that applied and endogenous GA responded differently to different growth retardants. None of the treatments had any noticeable effect on the time of flowering.     

Production of gibberellin-like substance in the embryo of barley during germination

Summary Embryos were excised from barley seeds, their homogenates were incubated with ficin, and their content in gibberellin-like substance was assayed by means of -amylase-producing activity, but no gibberellin-like substance could be detected.Embryos free from endosperm which were cultured for five days produced measurable amounts of gibberellin-like substance. This substance was not produced when a carbon source was absent from the medium, or when air was removed after 2 days of aerobic culture. CCC and Phosfon D (at the concentration of 2×10^-4 M and 0.8×10^-4 M, respectively) inhibited the formation of the gibberellin-like substance in cultured embryos without affecting their growth.Mevalonic acid could be used as a carbon source in the culture of the embryos. The formation of the gibberellin-like substance was in this case inhibited by 0.8×10^-4 M Phosfon D, but was not inhibited by 2×10^-3 M CCC.     

Effect of Gibberellic Acid and Phosfon on the Critical Dark Period Reqnirement for Flowering of Impatiens balsamina cv. Rose

The critical dark period requirement for flowering of Impatiens balsamina L. cv. Rose, an obligate short day plant, is about 8.5 hours. While GA₃ completely substituted for the dark period requirement, Phosfon prolonged it to 9.5 hours. GA₃ hastened and Phosfon delayed the initiation of floral buds under all photoperiods. Floral buds opened into flowers only during 8 and 14 hour photoperiods in control and Phosfon-treated plants but during all photoperiods in GA₃-treated ones. The delay in floral bud initiation and flowering was correlated with shifting up of the node bearing the first floral bud and flower respectively. While GA₃ increased the numher of floral buds and flowers in all photoperiods except 8-hour, Phosfon increased their number in the 14-hour photoperiod only. The number of flowering plants decreased with increasing photoperiod regardless of GA₃ and Phosfon application. The effect of Phosfon was completely or partially overcome, depending upon the photoperiod, by simultaneous application of GA₃.     

Uptake and translocation of root-fed32P in response to foliar and root applications of CCC and phosfon inPisum sativum

The uptake and translocation of³²P applied to nutrient solutions, as influenced by (2-chloroethyl) trimethylammonium chloride (CCC) and 2, 4-dichlorobenzyltributylphosphonium chloride (Phosfon), were investigated in growth chambers. Specific effects depended on the “Retardin”, the method of application, and the concentration. Foliar applications of CCC had no significant effect on³²P uptake 5 h after feeding. At 20 and 80h after feeding, CCC at 100 and at 1 000 mg l^?1 decreased P uptake. With root applications of CCC the same concentrations decreased P uptake as early as 5h after feeding, while the 1 mg l^?1 concentration enhanced P uptake by 40% after 20h. Both the foliar and the root applications of Phosfon influenced P uptake 5h after feeding. With foliar applications the low Phosfon concentration of 0.01 mg l^?1 increased P uptake throughout the experimental period of 80h, but the magnitudes of enhancement declined from 109% at 5h to 63% at 80h. Similarly, the enhancement of P uptake due to the 0.1 mg l^?1 foliar treatment declined from 132% at 5h to 80% at 80h. 100 mg l^?1 CCC enhanced P translocation to the shoot in 5h, with magnitudes of 29% (foliar application) and 64% (root application). At 20 and 80h the stimulations were nullified. The highest concentrations (100 mg l^?1 CCC; 10 mg l^?1 Phosfon) enhanced P translocation 80h after feeding. There were thus general enhancements of P uptake at low concentrations and suppressions at high concentrations, but these effects diminished with time, indicating transitory influences.     

Effect of Growth Retardants on α-Amylase Production in Germinating Barley Seed

The effect of growth retarding compounds, (2-chloroethyl)trimethylammonium chloride (CCC), 2-isopropyl-4-dimethylamino-5-methylphenyl-1-piperidinecarboxylate methyl chloride (AMU-1618), tributyl-2,4-dichlorobenzylphosphonium chloride (Phosfon D) and N-dimethylamino succinamic acid (B-995) on α-amylase production in germinating barley seed was studied. Seeds were germinated in growth retardants in presence and absence of gibberellic acid (GA₃). CCC, AMO-1618 and Phosfon D inhibitedα-amylase production in germinating seed and the effect was reversed by GA₃ Phosfon D and AMO-1618 were stronger inhibitors of α-amylase production than CCC. CCC was by far the strongest inhibitor of all the other analogs tested. B-995 was comparatively only slightly inhibitory. The results reported here, when viewed in light of the results of other workers, provide good evidence that CCC, AMO-1618 and Phosfon D inhibit α-amylase production by inhibiting the synthesis of gibberellin or gibberellin-like hormone(s) during germination of barley seed. Consistent with other reports, B-995 possibly acts by other mechanism (s).     

EVIDENCE ON THE SITE OF ACTION OF GROWTH RETARDANTS

1. The ability of five growth retardants to inhibit the GA-inducedand endogenous growth of Avena leaf sections has been investigated.The retardants vary in effectiveness. The order, from most effectiveto least, is Phosfon D, Amo-1618, C011, CCC and B995. 2. The inhibition of growth caused by Phosfon D and Amo-1618is not reversed by GA. It is apparent that the retardants donot compete with GA at the site of GA-action. 3. Addition of IAA will partially reverse the inhibition inducedby Phosfon D or Amo-1618. It is concluded that the retardantsact in part in Avena leaf sections by interfering with the auxinmetabolism of the tisssue. ¹ Supported in part by grants G-14578 and GB-1950 from the NationalScience Foundation ² Present address: Department of Botany, University of Washington,Seattle, Washington     

Toward Developing a Universal Treatment for Fungal Disease Using Radioimmunotherapy Targeting Common Fungal Antigens

Background

Previously, we demonstrated the ability of radiolabeled antibodies recognizing the cryptococcal polysaccharide capsule to kill Cryptococcus neoformans both in vitro and in infected mice. This approach, known as radioimmunotherapy (RIT), uses the exquisite ability of antibodies to bind antigens to deliver microbicidal radiation. To create RIT reagents which would be efficacious against all major medically important fungi, we have selected monoclonal antibodies (mAbs) to common surface fungal antigens such as heat shock protein 60 (HSP60), which is found on the surface of diverse fungi; beta (1,3)-glucan, which is a major constituent of fungal cell walls; ceramide which is found at the cell surface, and melanin, a polymer present in the fungal cell wall.

Methods

MAbs 4E12, an IgG2a to fungal HSP60; 2G8, an IgG2b to beta-(1,3)-glucan; and 6D2, an IgM to melanin, were labeled with the alpha particle emitting radionuclide 213-Bismuth (²¹³Bi) using the chelator CHXA”. B11, an IgM antibody to glucosylceramide, was labeled with the beta emitter 188-Rhenium (¹⁸⁸Re). Model organisms Cryptococcus neoformans and Candida albicans were used to assess the cytotoxicity of these compounds after exposure to either radiolabeled mAbs or controls.

Results

²¹³Bi-mAbs to HSP60 and to the beta-(1,3)-glucan each reduced the viability of both fungi by 80–100%. The ²¹³Bi-6D2 mAb to melanin killed 22% of C. neoformans, but did not kill C. albicans. B11 mAb against fungal ceramide was effective against wild-type C. neoformans, but was unable to kill a mutant lacking the ceramide target. Unlabeled mAbs and radiolabeled irrelevant control mAbs caused no killing.

Conclusion

Our results suggest that it is feasible to develop RIT against fungal pathogens by targeting common antigens and such an approach could be developed against fungal diseases for which existing therapy is unsatisfactory.

     

Li+ effect on the cell wall of the yeast Saccharomyces cerevisiae as probed by FT-IR spectroscopy

The effect of Li⁺ ions as a transformation inducing agent on the yeast cell wall has been studied. Two Saccharomyces cerevisiae strains, p63-DC5 with a native cell wall, and strain XCY42-30D(mnn1) which contains structural changes in the mannan-protein complex, were used. Fourier transform infrared (FT-IR) spectroscopy has been used for the characterization of the yeast strains and for determination of the effect of lithium cations on the cell wall. A comparison of the carbohydrate absorption band positions in the 970–1185 cm^?1 range, of Na⁺ and Li⁺ treated yeast cells has been estimated. Absorption band positions of the cell wall carbohydrates of p63-DC5 were not influenced by the studied ions. On the contrary, the treatment of XCY42-30D(mnn1) cells with Li⁺ ions shifted glucan band positions, implying that the cell wall structure of strain XCY42-30D(mnn1) is more sensitive to Li⁺ ion treatment.     

Leaf growth in the fast-growing Holcus lanatus and the slow-growing Deschampsia flexuosa: tissue maturation

Inherent variation in the relative growth rate of grasses is negatively correlated with that in leaf mass per unit leaf area (LMA). To scrutinize this correlation, the LMA of two grass species was analysed. Changes in LMA and cell wall synthesis in leaf blades of the fast-growing grass Holcus lanatus and the slow-growing grass Deschampsia flexuosa were investigated above the elongation zone of the leaf blade. After the leaf had obtained its final length, in H. lantus final LMA values of 40-44 gm^-2 were obtained at full leaf length, whereas in D. flexuosa LMA values continued to rise to 110 gm^-2. During this period of tissue maturation the LMA value doubled in H. lanatus, whereas in D. flexuosa an increase of 30% was measured. Most of the cell walls could be hydrolysed with driselase, the residue was hydrolysed with sulphuric acid. Driselase hydrolysates were identical in sugar composition, whereas the sugars released by sulphuric acid treatment changed gradually in composition as the tissue matured. The major sites of cell wall deposition during cell maturation were the outer walls of epidermal cells, fibres adjacent to the epidermis and the mestome ring around the vascular bundles. Lignin deposition was restricted to the vascular bundles and lignin levels of the leaf blade did not exceed 0.9% of the total amount of cell wall polysaccharides. Lignin accumulation occurred mainly after the increase in LMA and is unlikely to affect measurably the growth of these leaves.     

AsSlt2基因对茄链格孢细胞壁完整性的调控

【背景】由茄链格孢(Alternaria solani)引起的马铃薯早疫病被普遍认为是马铃薯生产上的第二大叶部病害,在马铃薯各产区普遍发生,给马铃薯生产造成了巨大的经济损失。【目的】明确AsSlt2基因对茄链格孢细胞壁完整性的影响。【方法】在含有刚果红、细胞壁降解酶和十二烷基硫酸钠(sodiumdodecylsulfate,SDS)等细胞壁胁迫的培养基上观察ΔAsSlt2缺失突变株的生长情况,计算相对生长抑制率;通过实时荧光定量PCR (RT-qPCR)方法检测ΔAsSlt2菌株中细胞壁合成相关基因的表达情况;进一步检测ΔAsSlt2细胞壁中几丁质的含量及胞外酶活性。【结果】ΔAsSlt2缺失突变株对SDS、刚果红、细胞壁降解酶等细胞壁胁迫的敏感性增强,在加入细胞壁降解酶后突变株原生质体释放量显著增多;ΔAsSlt2对外源氧胁迫更敏感,突变株胞外过氧化物酶和漆酶活性均显著降低;进一步研究发现,ΔAsSlt2细胞壁中几丁质含量减少,几丁质合成相关基因与漆酶合成相关基因的表达量均明显降低。【结论】AsSlt2基因在茄链格孢细胞壁的完整性及抵御外界胁迫方面发挥重要作用。     

COMPARATIVE CYTOHISTOLOGICAL STUDIES ON INHIBITION AND PROMOTION OF STEM GROWTH IN CHRYSANTHEMUM MORIFOLIUM

Sachs , R. M. (U. California, Davis), and A. M. Kopranek . Comparative cytohistological studies on inhibition and promotion of stem growth in Chrysanthemum morifolium. Amer. Jour. Bot. 50(8): 772-779. Illus. 1963.—The present study with Amo, CCC, and Phosfon,³ 3 substances which inhibit stem elongation, shows that all inhibit subapical cell expansion and division in Chrysanthemum morifolium var. ‘Indianapolis Yellow.‘ Furthermore, GA,³ in preventing the inhibition of stem elongation, maintains subapical activity at normal or greater than normal levels. For comparative purposes concentrations of the retardants and GA have been selected which completely prevent or promote the maximum rate of stem elongation. Phosfon causes complete inhibition of root growth and almost completely prevents dry matter accumulation in the tops. However, GA does not prevent such deleterious effects. Thus, GA and the growth retardants are mutually antagonistic only with respect to stem elongation and not to other aspects of growth. Furthermore, none of the retardants inhibits transverse stem growth; on the contrary transverse cell expansion and division in the subapical tissues are stimulated by the retardants, and as a result the stems of such plants are thicker than normal. GA not only prevents the thickening effect of the retardants, but, at the doses applied, GA-treated stems are considerably thinner than those of the controls, having fewer and smaller cells across the pith, cortical, and vascular tissues. Apparently, then, there is a relationship between longitudinal and transverse growth in the subapical tissues such that if one is promoted, the other is inhibited.     

Effect of Treatments with Growth Retardant Chemicals on Petiole Abscission in Coleus

A bscission of debladed petioles of Coleus was observed following spray applications of growth retardant chemicals and particularly of Phosfon D to the foliage. Sprays were applied to some branches, which were left intact (inducing branches), or to adjacent branches the leaves of which were later debladed (induced branches). In all experiments two applications of growth retardant chemicals were made, after which the induced branches were debladed. Treatments on induced branches accelerated the petiole abscission relative to the controls. Treatments on inducing branches, instead, decreased abscission speed of debladed petioles. The evidence suggests that phosfon D affects abscission by interfering with the indoleacetic acid mechanism.     

Calcium ionophore A 23187 affects localized wall secretion in the tip region of pollen tubes of Lilium longiflorum

The effects of the calcium inonophore A 23187 on growing pollen tubes of Lilium longiflorum Thunb. cv. Ace were investigated with the light and electron microscope. Tip growth is slowed down and stopped within 20 min after application of 5x10^-5 M ionophore A 23187. The main effects are the disappearance of the clear zone at the pollen tube tip and a thickening of the cell wall at the tip and at the pollen tube flanks. This effect on cell wall formation is confirmed under the electron microscope: The vesicular zone in treated pollen tubes is reduced, numerous vesicular contents are irregularly integrated in the pollen tube wall not only in the tip, but over a long distance of the pollen tube wall. In addition, effects on mitochondria and dictyosomes are observed. These results are interpreted as a disorientation of the Ca²⁺-based orientation mechanism of exocytosis after equilibration of the Ca²⁺-gradient     

Bioactive metabolites from <Emphasis Type="Italic">Penicillium</Emphasis> sp., an endophytic fungus residing in <Emphasis Type="Italic">Hopea hainanensis</Emphasis>

The metabolites of endophytic fungus Penicillium sp. from the leaf of Hopea hainanensis were reported for the first time. By bioassay-guided fractionation, the EtOAc extract of a solid-matrix steady culture of this fungus afforded six compounds, which were identified through a combination of spectral and chemical methods (IR, MS, ¹H- and ¹³C-NMR) to be monomethylsulochrin (1), rhizoctonic acid (2), asperfumoid (3), physcion (4), 7,8-dimethyl-iso-alloxazine (5) and 3,5-dichloro-p-anisic acid (6). Compounds 2, 3 and 6 were obtained from Penicillium sp. for the first time. All of the six isolates were subjected to in vitro bioactive assays including antifungal action against three human pathogenic fungi Candida albicans, Trichophyton rubrum and Aspergillus niger and cytotoxic activity against the human nasopharyngeal epidermoid tumor KB cell line and human liver cancer HepG2 cell line. As a result, compounds 2–4 and 6 inhibited the growth of C. albicans with MICs of 40.0, 20.0, 50.0 and 15.0 μg/ml, respectively and the compound 6 showed growth inhibition against A. niger with MICs of 40.0 μg/ml. In addition, compounds 1–3 and 6 exhibited cytotoxic activity against KB cell line with IC₅₀ value of 30.0, 20.0, 20.0, 5.0 μg/ml, respectively and against HepG2 cell line with IC₅₀ value of 30.0, 25.0, 15.0, 10.0 μg/ml, respectively.     

Cotton CSLD3 restores cell elongation and cell wall integrity mainly by enhancing primary cellulose production in the Arabidopsis cesa6 mutant

Key message

Overexpression of cotton cellulose synthase like D3 (GhCSLD3) gene partially rescued growth defect of atcesa6 mutant with restored cell elongation and cell wall integrity mainly by enhancing primary cellulose production.

Abstract

Among cellulose synthase like (CSL) family proteins, CSLDs share the highest sequence similarity to cellulose synthase (CESA) proteins. Although CSLD proteins have been implicated to participate in the synthesis of carbohydrate-based polymers (cellulose, pectins and hemicelluloses), and therefore plant cell wall formation, the exact biochemical function of CSLD proteins remains controversial and the function of the remaining CSLD genes in other species have not been determined. In this study, we attempted to illustrate the function of CSLD proteins by overexpressing Arabidopsis AtCSLD2, -3, -5 and cotton GhCSLD3 genes in the atcesa6 mutant, which has a background that is defective for primary cell wall cellulose synthesis in Arabidopsis. We found that GhCSLD3 overexpression partially rescued the growth defect of the atcesa6 mutant during early vegetative growth. Despite the atceas6 mutant having significantly reduced cellulose contents, the defected cell walls and lower dry mass, GhCSLD3 overexpression largely restored cell wall integrity (CWI) and improved the biomass yield. Our result suggests that overexpression of the GhCSLD protein enhances primary cell wall synthesis and compensates for the loss of CESAs, which is required for cellulose production, therefore rescuing defects in cell elongation and CWI.

     

V型羊毛硫肽雷可肽的抑菌机制

【背景】雷可肽(Lexapeptide)为首例V型羊毛硫肽家族化合物,具有较好的抗革兰氏阳性菌活性,对耐甲氧西林金黄色葡萄球菌(Methicillin-Resistant Staphylococcus aureus,MRSA)和表皮葡萄球菌(Methicillin-Resistant Staphylococcus epidermidis,MRSE)的抑制作用强于广泛应用的食品防腐剂乳酸链球菌素,其对pH和高温的稳定性也优于乳酸链球菌素,具有较好的应用前景。由于抑菌机制不明确,限制了雷可肽的开发应用。【目的】探究雷可肽抑菌作用特征以及作用机制,为雷可肽开发应用奠定基础。【方法】通过菌落计数法与Mg2+试验表征雷可肽抑菌动力学曲线;采用流式细胞仪和透射电子显微镜研究雷可肽在靶细胞表面的成孔性;利用高效液相色谱与基质辅助激光解吸电离的时间飞行质谱分析雷可肽处理对革兰氏阳性菌肽聚糖前体积累的影响。【结果】雷可肽在抑菌动力学上与乳酸链球菌素没有显著差别,但在更宽的Mg2+浓度范围内仍可保持抑菌活性。雷可肽处理后的细胞具有透过荧光染料的能力,生物型透射电镜观察到细胞发生破损。此外,在雷可肽作用后的细胞中检测到肽聚糖合成的前体尿嘧啶核苷二磷酸-N-乙酰胞壁酸五肽。【结论】雷可肽能够通过抑制细胞壁肽聚糖生物合成并造成细胞损伤进而获得通透性,以此来抑制革兰氏阳性菌生长。     

Non-Reversibility of Gibberellin-Induced Inhibition of Regeneration in Begonia Leaves

With applied to the petioles of detached Begonia x cheimantha leaves before planting, Gibberellic acid (GA₃) inhibited the formation of adventitious buds and roots ill an apparently irreversible manner. Bud formation was entirely suppressed by 10^?6M and higher concentrations and a significant inhibition was still present at 10^?9M the lowest concentration tested. Root formation was not affected by GA₃ below 10^?7M and was possible even at 10^?4 M GA₃. Petiole elongation was stimulated by GA₃ with an optimum at 10^?5M. GA₃ also blocked the action of 6-benzyiamino-purine (BAP) and 1-naphthaleneacetic acid (NAA), compounds which are potent stimulators of bud and root formation, respectively. When applied simultaneously with GA₃ they were, at their optimal concentrations, devoid of any effect in counteracting or reversing the gibberellin-induced inhibitions. Abscisic acid and the growth retardants CCC and Phosfon also were unable to restore bud and root formation. In leaves initially treated with water or 10^?5M BAP, endogenous bud and root formation as well as BAP-induced bud formation were entirety suppressed when 10^?5M GA₃ was applied 8 days after the initial treatments. Even when delayed for 14 days GA₃ treatment inhibited BAP-induced bud formation, while treatment after 21 days bad little effect on bud and root formation. Development of pre-existing, visible bud primordia was not inhibited by GA₃. BAP and NAA competitively inhibited the action of GA₃ in petiole extension growth. The results are discussed in relation to results obtained in other plant systems. It is suggested that GA₃ acts by blocking of the organized cell divisions initiating the formation of bud and root primordia.