Mechanism of phosphorus-induced zinc deficiency in cotton. II. Evidence for impaired shoot control of phosphorus uptake and translocation under zinc deficiency

Cotton (Gossypium hirsutum L. cv. Deltapine 15/21) plants were precultured for 19 to 25 days under controlled climatic conditions in nutrient solutions with different levels of Zn. With the onset of visual Zn-deficiency symptoms the pH of the nutrient solution decreased from 6.0 to about 5.0. In contrast, Zn-sufficient plants raised the pH of the nutrient solution to about 7.0. In short-term studies it could be demonstrated that the Zn nutritional status of the plants remarkably influenced the uptake and translocation rates of mineral nutrients. Compared to Zn-sufficient plants, P uptake rate in severely Zn-deficient plants was increased by a factor of 2 to 3, whereas the uptake rates of K, Ca and particularly NO₃ decreased. The accumulation of P in the roots of Zn-deficient plants was either not affected or even lower than in Zn-sufficient plants. Thus, Zn deficiency had a specific enhancement effect on root to shoot transport of P. This enhancement effect of Zn deficiency on uptake and transport of P was similar at nutrient solution pH values of 7.0 and 5.8; i.e. it was not the result of acidification of the nutrient solution. After application of ³⁶CI, ⁸⁶Rb and ³²P to plant stems, basipetal transport of ³⁶CI and ⁸⁶Rb was not affected by the Zn nutritional status of the plants. However, in Zn-deficient plants, only 7.8% of the ³²P was translocated basipetally compared to 34% in the Zn-sufficient plants. A resupply of Zn for 19 h to Zn-deficient plants enhanced basipetal ³²P transport. The results indicate that a feedback mechanism in the shoots is impaired in Zn-deficient plants which controls the P uptake by roots and especially the P transport from roots to shoots. As a result of this impairment toxic concentrations of P accumulate in the leaves. The mechanism responsible is likely the retranslocation of P in the phloem from shoots to roots.     

Expression and regulation of c-Jun N-terminal kinase (JNK) in endometrial cells in vivo and in vitro

JNK(c-Jun N-terminal kinase) is one of the main types of mitogen-activated protein kinases. JNK modulates inflammation and apoptosis in response to stress. Our hypothesis is that temporal and spatial changes in JNK activity regulate inflammation in human endometrium and that fluctuation in estrogen and progesterone levels may play a role in JNK activation. Therefore, we aimed to determine total-(t-) and active-(phosphorylated, p-) JNK expression in endometrial tissues in vivo by immunohistochemistry, and in vitro by immunocytochemistry and Western blot analysis. Immunohistochemistry revealed moderate cytoplasmic and nuclear t-JNK immunoreactivity, and mostly nuclear p-JNK immunoreactivity throughout the menstrual cycle and early pregnancy. The highest p- and t-JNK immunoreactivity was detected in late secretory phase (P < 0.05). We observed that endometrial stromal cell (ESC)s showed a significant increase in p-JNK expression following 48 h of estrogen combined with progesterone (E(2) + P(4)) withdrawal from the culture conditions, compared to control and non-withdrawal groups (P < 0.05). Upon treatment with JNK inhibitor SP600125, we observed a significantly decreased interleukin (IL)-8 level (P < 0.05) in the presence and absence of E(2). These results demonstrate that JNK expression increases during the late secretory phase when the inflammatory response is highest. Inhibition of IL-8 expression by SP600125 suggests that JNK is involved in regulation of proinflammatory mediators of endometrium.     

Expression of human cytochrome p450 3A4 gene in Schizosaccharomyces pombe

Heterologous expression systems can be utilized to great advantage in the study of cytochrome P450 enzymes. P450 3A4 is one of the major forms of cytochrome P450 found in liver. It is also involved in the metabolism of numerous widely used drugs and xenobiotics. In the present study human liver cytochrome P450 3A4 gene was transferred into the fission yeast Schizosaccharomyces pombe via two different S. pombe expression vectors carrying thiamine repressible promoter — nmt1 (pREP42) and constitutive promoter — adh1 (pART1). Heterologously expressed cytochrome P450 3A4 was detected in the cells grown in minimal (EMM) or rich medium (YEL) containing 0.5% (w/v) glucose. A typical cytochrome P450 peak for 3A4 was observed at 448 nm in microsomal fraction. The presence of heterologous expression of 3A4 form was also determined by SDS-PAGE and it molecular mass was identified as 52 kDa. The enzyme activity was confirmed by HPLC analysis, using testosterone as substrate.     

Physiological root responses of iron deficiency susceptible and tolerant tomato genotypes and their reciprocal F1 hybrids

By using two tomato genotypes line 227/1 (Fe chlorosis susceptible) and Roza (Fe chlorosis tolerant) and their reciprocal F₁hybrid, some root morphological changes, pH changes of nutrient solution, reduction capacity of Fe^III and uptake and root-to-shoot translocation of ⁵⁹Fe were studied under controlled environmental conditions in nutrient solution with 3 different Fe supplies as Fe EDDHA (i.e., 10^–7 M, severe Fe deficiency; 10^–6 M, intermediate Fe deficiency; 10^–4 M, adequate Fe supply). Tolerant parent `Roza' was less affected by low Fe supply than susceptible parent `line 227/1' as judged from the severity of leaf chlorosis. Under both Fe deficient conditions there were no differences between the reciprocal hybrids concerning the appearance of chlorosis. Under intermediate Fe deficiency, reciprocal F₁ hybrids (`line 227/1 × Roza' and `Roza × line' 227/1) showed an intermediate chlorosis between tolerant and susceptible parents. However, under severe Fe deficiency the reciprocal hybrids were more chlorotic than the tolerant parent irrespective of which parent was the cytoplasm contributor. A decreased Fe supply during preculture enhanced Fe^III reduction capacities of the parents and reciprocal hybrids. Differences in the tolerance to Fe deficiency always were better correlated with Fe^III reduction capacity of the genotypes than the Fe deficiency-induced release of H⁺ ions. Under both Fe deficient conditions the tolerant parent Roza had a much higher Fe^III reduction capacity than the susceptible parent line 227/1. The reduction capacity of the hybrids `Roza × line 227/1' was very similar to the capacity of the parent Roza, but higher than the capacity of the hybrids `line 227/1×Roza' at both Fe-deficient conditions. Under both Fe deficient conditions tolerant parent had higher number of lateral roots than the susceptible parent. Among the reciprocal hybrids `Roza × line 227/1' possessed more lateral roots than the `line 227/1 × Roza' under both Fe deficient conditions. Low Fe nutritional status resulted in marked increase in root uptake of ⁵⁹Fe. At adequate Fe supply, reciprocal hybrids and their parents did not differ in uptake and root-to-shoot translocation of Fe. However, under Fe-deficient conditions uptake and root-to-shoot translocation of ⁵⁹Fe were significantly higher in the Fe chlorosis tolerant than the susceptible parent. Based on the reduction capacity of Fe^III and uptake and root-to-shoot translocation of Fe, the F₁ hybrids obtained from the cross in which the maternal genotype was Roza appeared to be more tolerant than when the maternal genotype was the susceptible line 227/1. Uptake and translocation ratio of the F₁ hybrids obtained from `Roza × line 227/1' were similar to those of the parent Roza, but higher than the F<sub>1</sub> hybrids obtained from `line 227/1 × Roza', particularly under intermediate Fe deficiency. The results indicate that Fe^III reduction show a better relationship to Fe efficiency than Fe deficiency induced release of H⁺ ions. The inheritance of Fe deficiency tolerance of Roza seems not to be simple monogenic. It might be characterised by both, nuclear and extranuclear heredity. The intermediate responses of the reciprocal hybrids of the `line 227/1 × Roza' indicates that the Fe deficiency tolerance character of Roza is transferable by nuclear heredity. The better responses of the hybrids of `Roza × line 227/1' than the hybrids of `line 227/1 × Roza' may be due to maternal transmission from the parent Roza besides the nuclear transmission.     

Formalin fixing and paraffin embedding may lead to extra band development in PCR

The molecular biological analysis of infectious agents requires the availability of a reliable source of microorganisms to be used to recover DNA. Clinical samples can be obtained directly from infected patients or can be propagated using in vitro or in vivo systems. However, repeated sampling from patients is not always possible as the procedure may be invasive or unpleasant, or it is not possible to catch the same agent at the time of second sampling. Moreover, the techniques used may also produce false-positive and false-negative results. We therefore studied the impact of formalin-fixing and paraffin embedding on tissue sampling, and the methodologies such as DNA isolation and PCR amplification of DNAs from archival materials in the diagnosis of Mycobacterium tuberculosis. PCR analyses were done according to standard methods with some modifications. Demonstration of mycobacteria was successful both in tissue sections of the formalin-fixed lymph nodes and in stained fresh materials from patients. However, the results showed the presence of two extra bands in the gel. We accounted for extra band development due to the harshness of the methodology used to isolate nucleic acids from formalin-fixed and paraffin embedded tissue samples or the nature of the fixation procedure, or because of the time passed during storage in which alteration in the chromosomal DNA would take place. Thus, if disease- and tissue specific morphological features, such as sample size, type of fixation, and intralesional heterogeneity are ignored, errors because of sampling and methodologies used may lead to false-positive and false-negative results.     

Effect of aluminium on lipid peroxidation, superoxide dismutase, catalase, and peroxidase activities in root tips of soybean (Glycine max)

Inhibition of root elongation and modification of membrane properties are sensitive responses of plants to aluminium. The present paper reports on the effect of AI on lipid peroxidation and activities of enzymes related to production of activated oxygen species. Soybean seedlings (Glycine max L. cv. Sito) were precultured in solution culture for 3–5 days and then treated for 1–72 h with Al (AICI₃) concentrations ranging from 10 to 75 μM at a constant pH of 4.1. In response to Al supply, lipid peroxidation in the root tips (< 2 cm) was enhanced only after longer durations of treatment. Aluminium-dependent increase in lipid peroxidation was intensified by Fe²⁺ (FeSO₄). A close relationship existed between lipid peroxidation and inhibition of root-elongation rate induced by Al and/or Fe toxicity and/or Ca deficiency. Besides enhancement of lipid peroxidation in the crude extracts of root tips due to Al, the activities of superoxide dismutase (EC 1.15.1.1) and peroxidase (EC 1.11.1.7) increased, whereas catalase (EC 1.11.1.6) activity decreased. This indicates a greater generation of oxygen free radicals and related tissue damage. The results suggest that lipid peroxidation is part of the overall expression of Al toxicity in roots and that enhanced lipid peroxidation by oxygen free radicals is a consequence of primary effects of Al on membrane structure.     

Differential effects of nitrogen and sulfur deprivation on growth and biodiesel feedstock production of Chlamydomonas reinhardtii

Biodiesel production from microalgae is a promising approach for energy production; however, high cost of its process limits the use of microalgal biodiesel. Increasing the levels of triacylglycerol (TAG) levels, which is used as a biodiesel feedstock, in microalgae has been achieved mainly by nitrogen starvation. In this study, we compared effects of sulfur (S) and nitrogen (N) starvation on TAG accumulation and related parameters in wild-type Chlamydomonas reinhardtii CC-124 mt(-) and CC-125 mt(+) strains. Cell division was interrupted, protein and chlorophyll levels rapidly declined while cell volume, total neutral lipid, carotenoid, and carbohydrate content increased in response to nutrient starvation. Cytosolic lipid droplets in microalgae under nutrient starvation were monitored by three-dimensional confocal laser imaging of live cells. Infrared spectroscopy results showed that relative TAG, oligosaccharide and polysaccharide levels increased rapidly in response to nutrient starvation, especially S starvation. Both strains exhibited similar levels of regulation responses under mineral deficiency, however, the degree of their responses were significantly different, which emphasizes the importance of mating type on the physiological response of algae. Neutral lipid, TAG, and carbohydrate levels reached their peak values following 4 days of N or S starvation. Therefore, 4 days of N or S starvation provides an excellent way of increasing TAG content. Although increase in these parameters was followed by a subsequent decline in N-starved strains after 4 days, this decline was not observed in S-starved ones, which shows that S starvation is a better way of increasing TAG production of C. reinhardtii than N starvation.     

Soil fertility management effects on maize productivity and grain zinc content in smallholder farming systems of Zimbabwe

Biochar is produced as a by-product of the low temperature pyrolysis of biomass during bioenergy extraction and its incorporation into soil is of global interest as a potential carbon sequestration tool. Biochar influences soil nitrogen transformations and its capacity to take up ammonia is well recognized. Anthropogenic emissions of ammonia need to be mitigated due to negative environmental impacts and economic losses. Here we use an isotope of nitrogen to show that ammonia-N adsorbed by biochar is stable in ambient air, but readily bioavailable when placed in the soil. When biochars, containing adsorbed ¹⁵N labelled ammonia, were incorporated into soil the ¹⁵N recovery by roots averaged 6.8% but ranged from 26.1% to 10.9% in leaf tissue due to differing biochar properties with plant ¹⁵N recovery greater when acidic biochars were used to capture ammonia. Recovery of ¹⁵N as total soil nitrogen (organic+inorganic) ranged from 45% to 29% of ¹⁵N applied. We provide a proof of concept for a synergistic mitigation option where anthropogenic ammonia emissions could be captured using biochar, and made bioavailable in soils, thus leading to nitrogen capture by crops, while simultaneously sequestering carbon in soils.