小麦根系生长对缺磷胁迫的反应

研究了缺磷诱导小麦（Ｔｒｉｔｉｃｕｍ ａｅｓｔｉｖｕｍＬ．）根系生长的反应,小麦根轴的生长与植株内外的磷浓度均呈显著的负线性关系。分根实验证明,随着低磷营养液中根比例的增加,在供磷水平不同的分根盒侧的根轴长度的均增加,这说明根轴生长是受体内磷浓度调控的。植株体内磷浓度的处理后１ｄ开始变化,而在不同供磷水平营养液中小麦根轴长度的差异达到显著水平的时间是处理后的第８天,说明植株体内磷浓度的变化可能是小     

荞麦根吸收铝和分泌草酸的部位

荞麦根在Al诱导下分泌草酸。从根尖开始依次划分为 0～ 0 .5、0 .5～ 1.0、1.0～ 1.5、1.5～ 2 .0cm区段 ,在 0 .5～ 1.0cm区Al的吸收最多 ,草酸的分泌部位主要在 0～ 0 .5cm区 ,显示Al诱导下 ,Al的吸收和草酸的分泌不在同一部位     

Influx and Efflux of Phosphorus in Roots of Wheat Plants in Non-Growth-Limiting Concentrations of Phosphorus

The regulation of net phosphorus uptake was studied in wheatplants at ambient non-growth-limiting P-concentrations. Wheat(Triticum aestivum cv. Klein Atalaya) seedlings were grown fromgermination in culture solutions containing 0.05, 0.5 and 5.0mol m–3 phosphate. Only small increments in plant P-concentrationand specific accumulation rate for phosphorus were found whenambient P-concentration was increased 100 times. P-influx, estimatedby ³²P-uptake, was markedly greater with increased externalP-concentration, but only small changes in V_max and no changesin K_m were found. Indirect estimation of P-efflux in a time-courseof ³²P-uptake, and direct P-efflux measurements in ‘washout’ experiments indicated that P-efflux markedly increasedin higher ambient P-concentration. The increase in P-effluxalmost completely neutralized the higher P-influx observed in5.0 mol m^–3 relative to 0.05 mol m^–3 phosphate.It is postulated that in non-limiting P-concentration net P-uptakeis mainly controlled by P-efflux. Key words: Net P uptake, ³²P, kinetic parameters     

Assimilation of Phytate-phosphorus by the Extracellular Phytase Activity of Tobacco (Nicotiana tabacum) is Affected by the Availability of Soluble Phytate

Phytate, the major organic phosphorus in soil, is not readily available to plants as a source of phosphorus (P). It is either complexed with cations or adsorbed to various soil components. The present study was carried out to investigate the extracellular phytase activities of tobacco (Nicotiana tabacum variety GeXin No.1) and its ability to assimilate external phytate-P. Whereas phytase activities in roots, shoots and growth media of P_i-fed 14-day-old seedlings were only 1.3–4.9% of total acid phosphatase (APase) activities, P starvation triggered an increase in phytase secretion up to 914.9 mU mg⁻¹ protein, equivalent to 18.2% of total APase activities. Much of the extracellular phytase activities were found to be root-associated than root-released. The plants were not able to utilize phytate adsorbed to sand, except when insoluble phytate salts were preformed with Mg²⁺ and Ca²⁺ ions for supplementation. Tobacco grew better in sand supplemented with Mg-phytate salts (31.9 mg dry weight plant⁻¹; 0.68% w/w P concentration) than that with Ca-phytate salts (9.5 mg plant⁻¹; 0.42%), presumably due to its higher solubility. We conclude that insolubility of soil phytate is the major constrain for its assimilation. Improving solubility of soil phytate, for example, by enhancement of citrate secretion, may be a feasible approach to improve soil phytate assimilation.     

黑麦基因组中不同染色体在缺磷胁迫下对普通小麦根系分泌酸性磷酸酯酶(Acph)遗传效应的研究

以一整套中国春－帝国黑麦二体附加系为材料,通过在低磷胁迫下对其根系分泌Ａｃｐｈ 能力测定及同工酶等电聚焦分析证明：缺磷胁迫是Ａｃｐｈ基因表达的诱导因子,帝国黑麦不同染色体在中国春小麦背景中对其根系在低磷胁迫下 Ａｃｐｈ的分泌具不同的正效应,其中以 １Ｒ 染色体的效应最为强烈, Ａｃｐｈ等电聚焦（ＩＥＦ）的酶谱清楚地表明黑麦的１Ｒ染色体上携有在缺磷胁迫下诱导表达的Ａｃｐｈ基因。     

Removal of Vacuolar Targeting Signal from Class I Vacuolar Chitinase leads to its Extracellular Secretion in Transgenic Tobacco

Chitinase, an antifungal pathogenesis related (PR) protein is present in different isoforms. Class I basic chitinase which is generally more antifungal in nature compared to other chitinase classes, is present in vacuoles. It is speculated that extracellular secretion of this vacuolar enzyme by removing its vacuolar targeting signal at C- terminus might further increase its effectivity. Tobacco class I chitinase cDNA was earlier modified by PCR to add two stop codons before vacuolar targeting signal, so that the protein without this signal would be secreted extracellularly.Transgenic tobacco plants were raised with modified chitinase cDNA and native unmodified cDNA, both under the control of CaMV 35 S promoter, using Agrobacterium mediated transformation. Transgenic plants with unmodified class I chitinase cDNA expressed the enzyme in vacuoles and those having modified cDNA expressed the enzyme in extracellular spaces while retaining its biological activity.     

Responses of Wheat Roots to Exogenous Selenium Supply Under Enhanced Ultraviolet-B

Effects of selenium (Se) on growth and some physiological traits of roots in wheat (Triticum aestivum L. cv Han NO.7086) seedlings exposed to enhanced ultraviolet-B (UV-B) stress are reported. Responses of roots were different depending on the Se concentration. Compared with the control, root weight of wheat seedlings treated with 1.0 and 2.0 mg Se kg⁻¹ soil increased by 39.47% and 16.28%, respectively. The lower amount Se (0.5 mg kg⁻¹) and the higher amount Se treatments (3.0 mg kg⁻¹) did not significantly affect on root weight. Se treatments significantly increased root activity, flavonoids and proline content, and activities of peroxidase and superoxide dimutase in wheat roots exposed to enhanced UV-B. In addition, the treatments with 0.5, 1.0, and 2.0 mg Se kg⁻¹ significantly reduced malondialdehyde content and the rate of superoxide radical (O₂⁻) production of roots, whereas the higher amount Se treatment only induced a decrease in the rate of O₂⁻ production. The results of this study demonstrated that optimal Se supply promoted roots growth of wheat seedlings, and that optimal Se supply could reduce oxidative stress in wheat roots under enhanced UV-B radiation.     

Effects of pH Changes on Ion and 2,4-D Uptake of Wheat Roots

The quantitative relationships between pH-dependent ion and 2,4-D uptake in winter wheat seedlings (Triticum aestivum L. cv. Yubileynaya 50) have been investigated. The movement of various ions (potassium, phosphate, nitrate and ammonium) and 2,4-D across the root membranes was monitored with radioactive and stable isotope tracer methods. It was found that the H₊ ion concentration of the absorption solution strongly influences the 2,4-D uptake of the roots. Simultaneously, the 2,4-D uptake stimulates secretion of H⁺ into the absorption solution, that is, a H⁺ efflux can accompany the uptake of 2,4-D. This finding is consistent with the acid secretion theory of auxin and fusicoccin action. At pH 4 the 2,4-D uptake was much higher than at pH 6, thereby inhibiting the ion uptake and increasing the phytotoxicity in the plant. The results indicate that 2,4-D enters the root cells rapidly at the lower pH, mostly as undissociated molecules. With reference to the 2,4-D concentration in the roots at pH 4, a possible transport mechanism of the auxin herbicide is briefly discussed.     

Engineering Signal Peptides for Enhanced Protein Secretion from Lactococcus lactis

Lactococcus lactis is an attractive vehicle for biotechnological production of proteins and clinical delivery of therapeutics. In many such applications using this host, it is desirable to maximize secretion of recombinant proteins into the extracellular space, which is typically achieved by using the native signal peptide from a major secreted lactococcal protein, Usp45. In order to further increase protein secretion from L. lactis, inherent limitations of the Usp45 signal peptide (Usp45sp) must be elucidated. Here, we performed extensive mutagenesis on Usp45sp to probe the effects of both the mRNA sequence (silent mutations) and the peptide sequence (amino acid substitutions) on secretion. We screened signal peptides based on their resulting secretion levels of Staphylococcus aureus nuclease and further evaluated them for secretion of Bacillus subtilis α-amylase. Silent mutations alone gave an increase of up to 16% in the secretion of α-amylase through a mechanism consistent with relaxed mRNA folding around the ribosome binding site and enhanced translation. Targeted amino acid mutagenesis in Usp45sp, combined with additional silent mutations from the best clone in the initial screen, yielded an increase of up to 51% in maximum secretion of α-amylase while maintaining secretion at lower induction levels. The best sequence from our screen preserves the tripartite structure of the native signal peptide but increases the positive charge of the n-region. Our study presents the first example of an engineered L. lactis signal peptide with a higher secretion yield than Usp45sp and, more generally, provides strategies for further enhancing protein secretion in bacterial hosts.     

Production,Rapid Purification and Catalytic Characterization of Extracellular Phytase from Aspergillus Ficuum

Abstract

A rapid purification scheme utilizing three chromatographic steps resulted in 6 fold purification of Aspergillus ficuum phytase (myo-inositol-hexakis-phosphate 3-phosphohydrolase, EC 3.1. 3.8). At pH 5.0 and 60°C the enzyme performed acceptably for 2.0 hr with only 30% diminished catalytic rate at the end. Substrate concentration exceeding 2nM was inhibitory. The inorganic orthophosphate, the product and a weak inhibitor, exhibited a Ki of 1.9 × 10^?3M. The extracellular phytase has the potential for industrial use since it can be over produced, easily purified, remain catalytically active for a longer period and is not subjected to severe product inhibition.     

Indirect Effect of a Transgenic Wheat on Aphids through Enhanced Powdery Mildew Resistance

In agricultural ecosystems, arthropod herbivores and fungal pathogens are likely to colonise the same plant and may therefore affect each other directly or indirectly. The fungus that causes powdery mildew (Blumeria graminis tritici) and cereal aphids are important pests of wheat but interactions between them have seldom been investigated. We studied the effects of powdery mildew of wheat on two cereal aphid species, Metopolophium dirhodum and Rhopalosiphum padi. We hypothesized that aphid number and size will be smaller on powdery mildew-infected plants than on non-infected plants. In a first experiment we used six commercially available wheat varieties whereas in the second experiment we used a genetically modified (GM) mildew-resistant wheat line and its non-transgenic sister line. Because the two lines differed only in the presence of the transgene and in powdery mildew resistance, experiment 2 avoided the confounding effect of variety. In both experiments, the number of M. dirhodum but not of R. padi was reduced by powdery mildew infection. Transgenic mildew-resistant lines therefore harboured bigger aphid populations than the non-transgenic lines. For both aphid species individual size was mostly influenced by aphid number. Our results indicate that plants that are protected from a particular pest (powdery mildew) became more favourable for another pest (aphids).     

不同耐盐性小麦根Na~+和K~+的吸收特性

以耐盐小麦品种‘德抗961’和盐敏感小麦品种‘鲁麦15’为材料,研究小麦根Na+、K+吸收特性及其与耐盐性关系。结果表明,2个小麦品种根K+吸收动力学曲线均符合Michaelis-Menten方程,即V=Vmax×[S]/([S]+Km)+k×[S]。低浓度(低于25mmol·L-1)NaCl处理对根高亲和K+吸收系统转运K+具有促进作用,对耐盐品种‘德抗961’的促进作用更大。小麦根高亲和K+吸收系统是通过K+/H+同向转运,而不是K+/Na+同向转运。NaCl处理对根低亲和K+吸收系统有抑制作用,对盐敏感品种‘鲁麦15’的抑制作用更大。NaCl处理导致2个小麦品种根和叶片中的K+含量显著下降,Na+含量显著升高,但‘德抗961’根和叶片中的K+含量均显著高于‘鲁麦15’,‘德抗961’根中Na+含量显著高于‘鲁麦15’,而其叶片中Na+含量显著低于‘鲁麦15’,从而保证NaCl胁迫下其叶片较高的K+/Na+比。非选择性阳离子通道是小麦根Na+吸收的主要途径,K+通道是Na+吸收的一条重要途径。这些结果表明小麦部分通过调节根系K+吸收系统而维持叶片较高的K+/Na+比,从而提高其耐盐性。     

Zinc Mobility in Wheat: Uptake and Distribution of Zinc Applied to Leaves or Roots

Little is known about transport of Zn from leaves to other plantorgans. The present study tested a range of Zn forms appliedfoliarly for their suitability to provide adequate Zn nutritionto wheat (Triticum aestivum L.). Transport of⁶⁵Zn applied eitherto leaves or to one side of the root system was also studied.Inorganic (ZnO, ZnSO₄) and chelated sources of Zn (ZnEDTA, glycine-chelatedBiomin Zn) applied foliarly provided sufficient Zn for vigorousgrowth. Zinc concentrations in roots and shoots were in thesufficiency range, except in the -Zn control. Foliar treatmentswith ZnSO₄and chelated Zn forms resulted in shoot Zn concentrationsin 7-week-old plants being about two-fold greater than thosein plants supplied with Zn in the root environment or via foliarspray of ZnO. Adding surfactant to foliar sprays containingchelated forms of Zn did not cause negative growth effects,but surfactant added to ZnO or ZnSO₄foliar sprays decreasedshoot growth. Adding urea to the ZnO foliar spray had no effecton shoot growth. Foliarly-applied⁶⁵Zn was translocated to leavesabove and below the treated leaf as well as to the root tips.Stem girdling confirmed that⁶⁵Zn transport toward lower leavesand roots was via the phloem. Split-root experiments showedintensive accumulation of⁶⁵Zn in the stem and transport to allleaves as well as to the root tips in the non-labelled sideof the root system. Foliar application of Zn in inorganic ororganic form is equally suitable for providing adequate Zn nutritionto wheat. Phloem transport of Zn from leaves to roots was demonstrated.Copyright 2001 Annals of Botany Company Foliar spraying, phloem, surfactant, urea, xylem, wheat, zinc     

Uptake and Utilization of Selenium from Selenoprotein P

Selenoprotein P (SELENOP) is a serum glycoprotein that is required for proper selenium distribution in mammals, particularly in supplying selenium to the brain and testes. As the sole mechanism for providing essential selenium to developing spermatozoa, SELENOP metabolism is central to male fertility in all mammals. In addition, this process is important for proper brain function, especially under conditions of limited dietary selenium. Several specific and nonspecific mechanisms for SELENOP uptake in target tissues have been described, but the utilization of SELENOP as a source of selenium for intracellular selenoprotein production has not been systematically characterized. In this report, we examine the process of SELENOP uptake using a robust selenium uptake assay that measures selenium utilization in cells fed ⁷⁵Se-SELENOP. Using a series of inhibitors and modulators we have identified specific regulators of the process and found that SELENOP must be in an oxidized state for uptake. This assay also demonstrates that SELENOP uptake is not highly sequence specific as the zebrafish protein is recognized and processed by mammalian cells.     

Isolation and Identification of Actinobacteria from Surface-Sterilized Wheat Roots

This is the first report of filamentous actinobacteria isolated from surface-sterilized root tissues of healthy wheat plants (Triticum aestivum L.). Wheat roots from a range of sites across South Australia were used as the source material for the isolation of the endophytic actinobacteria. Roots were surface-sterilized by using ethanol and sodium hypochlorite prior to the isolation of the actinobacteria. Forty-nine of these isolates were identified by using 16S ribosomal DNA (rDNA) sequencing and found to belong to a small group of actinobacterial genera including Streptomyces, Microbispora, Micromonospora, and Nocardiodes spp. Many of the Streptomyces spp. were found to be similar, on the basis of their 16S rDNA gene sequence, to Streptomyces spp. that had been isolated from potato scabs. In particular, several isolates exhibited high 16S rDNA gene sequence homology to Streptomyces caviscabies and S. setonii. None of these isolates, nor the S. caviscabies and S. setonii type strains, were found to carry the nec1 pathogenicity-associated gene or to produce the toxin thaxtomin, indicating that they were nonpathogenic. These isolates were recovered from healthy plants over a range of geographically and temporally isolated sampling events and constitute an important plant-microbe interaction.     

缺磷胁迫对小麦根细胞周期蛋白基因cyc1At表达的影响

用液培方法研究了缺磷胁迫对小麦(TriticumaestivumL.)根系生长的影响。结果表明,随着介质磷水平的提高,小麦根轴长度和植株生长素浓度均降低。在低磷条件下用生长素极性运输抑制剂三碘苯甲酸(TIBA)处理后,小麦的根轴长度明显降低,表明生长素参与了缺磷小麦根轴生长的调控。缺磷小麦根部生长素浓度的提高诱导了细胞周期蛋白基因cyclAt的表达,促进了根分生组织细胞的分裂并驱动了根的生长。     

Metabolism-linked Binding of Rubidium in the Free Space of Wheat Roots and its Relation to Active Uptake

Components of the initial uptake and active transport of rubidium were studied in potassium-starved and potassium-rich wheat plants. A metabolism-linked component of the initial uptake in the root was identified by means of DNP-treatment. It was shown that there is a direct proportionality between the magnitude of the metabolism-linked component of the initial uptake and the rate of active uptake. It is suggested that, in the free space, this metabolism-linked ion binding represents an initial step of the active ion transport preceding the subsequent carrier-mediated transport across the plasmalemma.