Phytostimulation and biofertilization in wheat by cyanobacteria

Cyanobacteria are commonly used for the phytostimulation and biofertilization of agriculture crops due to their nitrogen-fixing ability. However, the contribution by their phytohormones has been neglected. This study focuses on the screening of rhizospheric and free-living cyanobacteria for in vitro phytohormones production and growth stimulation in wheat. Selected isolates were shown to release cytokinin and indole-3-acetic acid (IAA) by using UPLC coupled with MS/MS via an electrospray interface. The maximum cytokinin and IAA concentration was 22.7 pmol mg⁻¹ ch-a and 38 pmol mg⁻¹ ch-a, respectively, in the culture medium of Chroococcidiopsis sp. Ck4 and Anabaena sp. Ck1. The growth of wheat inoculated with cyanobacterial strains was stimulated under axenic as well as field conditions. Seed germination, shoot length, tillering, number of lateral roots, spike length, and grain weight were significantly enhanced in inoculated plants. The maximum increase in grain weight (43%) was demonstrated in wheat plants inoculated with Chroococcidiopsis sp. Ck4 under natural conditions. Positive linear correlation of cyanobacterial cytokinin with shoot length (r = 0.608; P = 0.01), spike length (r = 0.682; P = 0.01), and grain weight (r = 0.0.869; P = 0.01) was recorded. Similarly, cyanobacterial IAA was correlated with the root growth parameters shoot length (r = 0.588; P = 0.01), spike length (r = 0.0.689; P = 0.01), and weight of seeds (r = 0.480; P = 0.05). The endogenous phytohormones pool of the plant was enhanced significantly as a result of the plant–cyanobacteria association in the rhizosphere. It was concluded that cyanobacterial phytohormones are a major tool for improved growth and yield in wheat.     

Rhizobacterial potential to alter auxin content and growth of <Emphasis Type="Italic">Vigna radiata</Emphasis> (L.)

Potential of non-symbiotic plant growth promoting rhizobacteria (PGPR) to influence the endogenous indole-3-acetic acid (IAA) content and growth of Vigna radiata (L.) was evaluated. The bacterial strains used belonged to Pseudomonas, Escherichia, Micrococcus and Staphylococcus genera. All strains were able to produce IAA (1.16–8.22 μg ml⁻¹) in the presence of 1,000 μg ml⁻¹ of l-tryptophan as revealed by gas chromatography and mass spectrometric (GC–MS) analysis. However, strains exhibited variable results for other growth promoting traits such as phosphate solubilization and siderophore or hydrogen cyanide production. Bacterial IAA production showed significant positive correlation with endogenous IAA content of roots (r = 0.969; P = 0.01) and leaves (r = 0.905; P = 0.01) under axenic conditions. Bacterization of V. radiata seeds significantly enhanced shoot length (up to 48.10%) and shoot fresh biomass (up to 43.80%) under fully axenic conditions. Bacterial strains applied under wire-house conditions also improved shoot length, number of pods, and grain weight up to 58, 65, and 17.15% respectively, over control. Hence, free living (non-symbiotic) PGPR have the ability to influence endogenous IAA content and growth of leguminous plants.     

Quantification of indole-3-acetic acid from plant associated <Emphasis Type="Italic">Bacillus</Emphasis> spp. and their phytostimulatory effect on <Emphasis Type="Italic">Vigna radiata</Emphasis> (L.)

Sixteen Bacillus strains isolated from rhizosphere, histoplane and phyllosphere of different plant species were identified by 16S rDNA gene sequencing and evaluated for in vitro auxin production as well as growth stimulation of Vigna radiata (L.) Wilczek. Auxin production by Bacillus spp. in L-broth medium supplemented with 1,000 μg ml⁻¹ L-tryptophan ranges from 0.60 to 3.0 μg IAA ml⁻¹ as revealed by gas chromatography and mass spectrometric (GC–MS) analysis. Rhizospheric isolates exhibit relatively more IAA synthesis than histoplane and phyllosphere isolates. Plant microbe interaction experiments conducted under gnotobiotic conditions recorded 55.55, 46.46 and 46.20% increase in shoot length with Bacillus megaterium MiR-4, B. pumilus NpR-1 and B. subtilis TpP-1, respectively, over control. Bacillus inoculations also increased shoot fresh weight with B. megaterium MiR-4 (60.94%) and B. pumilus NpR-1 (37.76%). Highly significant positive correlation between auxin production analyzed by GC–MS and shoot length (r = 0.687**, P = 0.01) and shoot fresh weight (r = 0.703**, P = 0.01) was noted under gnotobiotic conditions. Similarly, significant correlation was also found between auxin production by Bacillus spp. (GC–MS analysis) and different growth parameters such as shoot length (r = 0.495*, P = 0.05), number of pods (r = 0.498*, P = 0.05) and grain weight (r = 0.537*, P = 0.05) at full maturity under natural wire house conditions. Results showed that auxin production potential of plant associated Bacillus spp. can be effectively exploited to enhance the growth and yield of V. radiata.     

The effect of exogenous and endogenous phytohormones on the in vitro development of gametophyte and sporophyte in <Emphasis Type="Italic">Asplenium nidus</Emphasis> L.

The fern Asplenium nidus L. is in great demand as an ornamental plant. The aim of this work was to investigate the influence of phytohormones in promoting a gametophytic and sporophytic growth in homogenized sporophytes tissue. Exogenous application of 0.5 and 5 μM N ⁶-benzyladenine, 0.05 and 0.5 μM indole-3-acetic acid (IAA), and 0.3 and 3 μM gibberellic acid (GA₃) favoured sporophyte regeneration, whereas gametophyte regeneration took place when plant material was cultured in a hormone-free liquid MS medium. The endogenous contents of the auxin IAA, the cytokinins trans-zeatin, trans-zeatin riboside, dihydrozeatin, dihydrozeatin riboside, isopentenyladenine and isopentenyladenosine, and the gibberellins GA₁, GA₃, GA₄, GA₇, GA₉ and GA₂₀ in growing gametophytes and sporophytes were evaluated. Similar levels of the auxin and cytokinins and qualitative differences in the gibberellins were found between both generations.     

Rapid Determination of Cytokinins and Auxin in Cyanobacteria

Five cyanobacterial strains, Anabaena sp. Ck1, Oscillatoria sp. Ck2, Phormidium sp. Ck3, Chroococcidiopsis sp. Ck4, and Synechosystis sp. Ck5 were selected for their positive cytokinins-like activity using cucumber cotyledon bioassay and GUS assay in Arabidopsis ARR5::GUS. Classical cucumber cotyledon bioassay was modified for direct screening of cyanobacteria avoiding need for extraction and purification. Cytokinins from cyanobacteria were absorbed onto filter paper which was then assayed for cytokinins-like activity. A rapid chromatographic method was developed for the simultaneous determination of cytokinins and indole-3-acetic acid (IAA). Cyanobacterial biomass (50–100 mg) and cell-free culture filtrate were extracted in Bieleski buffer and purified by solid-phase extraction. The extract was used to determine phytohormones by ultra performance liquid chromatography and electrospray ionization-tandem mass spectrometry in positive and negative modes, respectively, with multiple reactions monitoring. Stable isotope-labeled cytokinins and IAA standards were added in the samples to follow recovery of the compounds and method validation. Five cytokinins determined in the selected strains were Zeatin (cis and trans isomers), Zeatin riboside, Dihydrozeatin riboside, and zeatin-o-glucoside. The strains were shown to accumulate as well as release the phytohormones.     

Understanding the effect of organic pollutant fluoranthene on pea in vitro using cytokinins,ethylene, ethane and carbon dioxide as indicators

Environmental contaminants like polycyclic aromatic hydrocarbons can influence many biochemical and physiological processes in plants. The effect of 0.1, 1 and 5 mg l⁻¹ of fluoranthene (FLT) in combination with indole-3-acetic acid (IAA, 0.1 mg l⁻¹) or a combination of IAA and N⁶-benzyladenine (BA, both 0.1 mg l⁻¹) on the growth and production of ethylene, ethane and CO₂ in Pisum sativum L. cultivated for 21 days in vitro was investigated. In 21 days old plants also net photosynthesis rate, content of FLT and cytokinins were evaluated. FLT 5 mg l⁻¹ significantly inhibited the growth of pea after 21 days in both IAA and IAA + BA treatments, increased production of ethylene (by 11% in IAA and 14% in IAA + BA treatments, respectively) and ethane (by 28 and 18%) and decreased production of CO₂ (by 23 and 29%). The net photosynthesis rate decreased in response to FLT concentration by up to 51% under saturating irradiation (600–1,200 μmol m⁻² s⁻¹), as found in IAA + BA + FLT 5 mg l⁻¹ treatment. The content of FLT in pea plant shoots well correlated with increasing FLT treatment in both IAA and IAA + BA medium. The content of cytokinins in pea shoots changed in response to FLT treatment. FLT 5 mg l⁻¹ caused a rise in level of trans-zeatin (by 16% in IAA and 9% in IAA + BA treatments, respectively), dihydrozeatin riboside (by 27 and 50%), benzyladenine (by 3 and 80%), benzyladenine riboside (by 44 and 17%) and meta-topolin riboside (by 139 and 214%), no change in isopentenyladenine level and a decrease in meta-topolin level (by 33% in IAA and 36% in IAA + BA treatments, respectively). Cultivation of plants in vitro allowed not only to assess their growth, photosynthetic activity and the level of cytokinins, but also to extend the knowledge about the effect of PAHs on production of gaseous stress indicators like ethylene, ethane and CO₂. Recorded changes in all studied parameters show, that persistent organic pollutants like PAHs can negatively influence plant growth and development.     

Effect of phytohormones synthesized by rhizosphere bacteria on plants

New strains of rhizosphere microorganisms Azotobacter chroococcum Az d10, Bacillus megaterium Pl-04, and Bacillus mucilaginosus B-1574 were found to be able to synthesize cytokinins (CKs) and indolylacetic acid (IAA). Three forms of CKs—dihydrozeatin riboside, isopentenyl adenosine, and trans-zeatin riboside—were identified, whose ratio was different in the three bacterial cultures. Inoculation of cucumber (Cucumis sativus L.) plants increased the content of CKs and IAA in them by 35.6 and 21.3%, respectively, and also stimulated seed germination and increased the growth rate, the biomass of shoots, the number of lateral roots, and the root hair area, which ensured better plant nutrition. The IAA/CKs ratio shifted during bacterization towards CKs due to increase in the content of riboside forms, which apparently caused growth stimulation.     

Detection of cytokinins and auxin in plant tissues using histochemistry and immunocytochemistry

We report a new method for histochemical localization of cytokinins (CKs) in plant tissues based on bromophenol blue/silver nitrate staining. The method was validated by immunohistochemistry using anti-trans-zeatin riboside antibody. Indole-3-acetic acid (auxin, IAA) was localized by anti-IAA antibody in plant tissues as a proof for IAA histolocalization. We used root sections, because they are major sites of CKs synthesis, and insect galls of Piptadenia gonoacantha that accumulate IAA. Immunostaining confirmed the presence of zeatin and sites of accumulation of IAA indicated by histochemistry. The colors developed by histochemical reactions in free-hand sections of plant tissues were similar to those obtained by thin layer chromatography (TLC), which reinforced the reactive sites of zeatin. The histochemical method for detecting CKs is useful for galls and roots, whereas IAA detection is more efficient for gall tissues. Therefore, galls constitute a useful model for validating histochemical techniques due to their rapid cell cycles and relatively high accumulation of plant hormones.     

Comparison of effects of bacterial strains differing in their ability to synthesize cytokinins on growth and cytokinin content in wheat plants

The content of cytokinins and pigments together with the morphological parameters and fresh weight were estimated in durum wheat (Triticum durum Desf.) plants 2–4 days after introduction into their rhizosphere of an aliquot of Bacillus suspension using the strains that differed in their ability of producing cytokinins. The experiments were performed under laboratory conditions at the optimum light intensity and mineral nutrition. Inoculation with microorganisms incapable to synthesize cytokinins did not affect the total cytokinin content in the wheat plants, whereas the presence of cytokinin-producing microorganisms in the rhizosphere was accompanied by a considerable increase in the total cytokinin content and the accumulation of individual hormones. On the second day after inoculation, a dramatic increase in zeatin riboside and zeatin O-glucoside contents was observed in the roots, and at the next day the accumulation of zeatin riboside and zeatin was registered in the shoots of treated plants. The increase in cytokinin content promoted plant growth (the increased leaf length and width and a faster accumulation of plant fresh and dry weight). Plant treatment with a substance obtained from microorganisms incapable to synthesize hormones resulted in the insignificant growth stimulation. Plant treatment with a substance obtained from cytokinin-producing microorganisms increased leaf chlorophyll content; in this case, the level of chlorophylls was comparable to that observed in the plants treated with a synthetic cytokinin benzyladenine. The role of cytokinins of microbial origin as a factor providing for growth-stimulating effect of bacteria on plants is discussed.     

Effects of elevated carbon dioxide and/or ozone on endogenous plant hormones in the leaves of <Emphasis Type="Italic">Ginkgo biloba</Emphasis>

Four-year-old Gingko (Ginkgo biloba L.) trees were exposed to ambient and elevated concentrations of CO₂ and O₃, and their combination for 1 year, using open-top chambers in Shenyang, China in 2006. Growth parameters and endogenous plant hormones were measured simultaneously over the experiment period. Elevated CO₂ increased leaf area and leaf dry weight but had no effect on shoot length, increased indole-3-acetic acid (IAA), gibberellins A₃ (GA₃), zeatin riboside (ZR), dihydrozeatin (DHZR) and isopentenyl-adenosine (iPA) content but decreased abscisic acid (ABA) content. Elevated O₃ significantly decreased leaf area, leaf dry weight, shoot length, and decreased IAA, GA₃, ZR content but increased ABA content and had a little effect on iPA, DHZR content. Elevated CO₂ + O₃ decreased IAA, iPA and DHZR content while increased ABA and GA₃ content in the early stage of the exposure and then decreased in the late stage. The evidence from this study indicates that elevated CO₂ ameliorated the effects of elevated ozone on tree growth, and elevated CO₂ may have a largely positive impact on forest tree growth while elevated O₃ will likely have a negative impact.     

Relationships between stream habitat and breeding territory length of the Brown Dipper (Cinclus pallasii) in Taiwan

Dippers from the genus Cinclus are highly specialised predators on aquatic invertebrates, and occupy linear territories along rivers where measurements of variations in quality are relatively straightforward. For these reasons, they are ideal model species in which to examine factors affecting territory size. In this paper, we investigated the influence of stream habitats on the territory length of the Brown Dipper (Cinclus pallasii) in Taiwan. The biomass of aquatic insects and other stream habitat variables were analyzed to determine their relationships with the territory length of Brown Dippers from November 1988 to May 1989. Compared with slow-moving waters, riffle areas contained significantly greater insect biomass (paired t test, t ₁₁ = 3.49, P < 0.01), of which trichopteran larvae contributed about 70%. Dippers spent more time foraging in riffles than in slow-moving waters. In addition, dippers preferred foraging in shallow riffles, but avoided deep, slow-moving waters (G = 62.53, df = 3, P < 0.001). Territory length (1,045 ± 165 m [SE], n = 14) was negatively related to proportion of riffles (r ² = 0.5715, P < 0.01), total aquatic insect biomass (r ² = 0.5840, P < 0.01), and altitude (r ² = 0.7176, P < 0.001). In factor analysis, four factors were extracted from the 14 stream variables. However, only factor 1 was significantly related to territory length (r ² = 0.5207, P < 0.01). Factor 1 explained 42.8% of the total variance and collectively revealed the importance of high food abundance. In other words, Brown Dipper territories were the shortest along high-altitude streams with abundant riffles, fewer pools and abundant aquatic insects. These results indicate that abundant supply of accessible invertebrate prey is the most important factor affecting the territory length of Brown Dippers. This is consistent with Cinclus species elsewhere, and reveals the importance of clean, productive river ecosystems.     

Effects of early-fruit removal on endogenous cytokinins and abscisic acid in relation to leaf senescence in cotton

Numerous studies have shown that early-fruit removal enhances vegetative growth and development of cotton (Gossypium hirsutum L.). However, few studies have examined changes in leaf senescence and endogenous hormones due to fruit removal. The objective of this study was to determine the correlation between some endogenous phytohormones, particularly the cytokinins and abscisic acid (ABA), and leaf senescence following fruit removal. Cotton was grown in pots and in the field during 2005 and 2006. Two early-fruiting branches were excised from plants at squaring to form the fruit removal treatment while the non-excised plants served as control. Plant biomass, seed cotton yield, cytokinins and ABA levels in main-stem leaves and xylem sap as well as main-stem leaf photosynthetic rate (Pn) and chlorophyll (Chl) concentration were determined after removal or at harvest. Fruit removals increased the leaf area, root and shoot dry weight and plant biomass at 35 days after removal (DAR), whether in potted or field-grown cotton; under field conditions, it also improved plant biomass and seed cotton yield at harvest. The Pn and Chl concentration in excised plants were significantly higher than in control plants from 5 to 35 DAR, suggesting that fruit removal considerably delayed leaf senescence. Fruit-excised plants contained more trans-zeatin and its riboside (t-Z + t-ZR), dihydrozeatin and its riboside (DHZ + DHZR), and isopentenyladenine and its riboside (iP + iPA) but less ABA in both main-stem leaves and xylem sap than control plants from 5 to 35 DAR. These results suggest that removal of early fruiting branches delays main-stem leaf senescence, which can be attributed to increased cytokinin and/or reduced ABA. Cytokinin and ABA are involved in leaf senescence following early fruit removal.     

Biosynthesis of phytohormones from novel rhizobacterial isolates and their in vitro plant growth-promoting efficacy

The health of the plant and soil fertility is dependent on the plant–microbes interaction in the rhizosphere. Microbial life tends to endure various rhizosphere plant–microbe interactions. Phytohormones such as auxins, cytokinins, gibberellic acid, ethylene and abscisic acid are termed as the classical group of hormones. Out of the 70 rhizobacterial strains isolated from the Coleus rhizosphere, three different rhizobacterial strains Pseudomonas stutzeri MTP40, Stenotrophomonas maltophilia MTP42 and Pseudomonas putida MTP50 having plant growth-promoting attributes were isolated and characterized for its phytohormone-producing ability. The phytohormones such as indole 3-acetic acid (IAA), gibberellic acid and cytokinin (kinetin and 6-benzyladenosine) were affirmed in culture supernatant of the above isolates. IAA was detected in all the three isolates, where in highest production was found in S. maltophilia MTP42 (240?µg/mL) followed by P. stutzeri MTP40 (250?µg/mL) and P. putida MTP50 (233?µg/mL). Gibberellic acid production was found maximum in MTP40 (34?µg/mL), followed by MTP42 (31?µg/mL) and MTP50 (27?µg/mL). The cytokinin production from the isolates, namely, MTP40, MTP42 and MTP50 were 13, 11 and 7.5?µg/mL, respectively. The isolates showing the production of plant growth enhancing phytohormones can be commercialized as potent bioformulations.     

Dynamics of growth and the content of endogenous phytohormones during kidney bean scoto-and photomorphogenesis

The dynamics of growth and the contents of free and bound endogenous IAA, gibberellins (GA), cytokinins (zeatin and its riboside), and ABA in kidney bean plants (Phaseolus vulgaris L., cv. Belozernaya) grown in darkness or in the light was studied. Phytohormones were quantified in 5–15-day-old plants by the ELISA technique. Plant growth and phytohormone content were shown to depend on plant age and the conditions of illumination. During scotomorphogenesis, changes in the biomass and hypocotyl length were highly correlated with the content of GA, whereas during photomorphogeneses, these parameters were correlated with the content of zeatin. In darkness, epicotyl growth displayed a positive correlation with the content of GA, whereas in the light, the correlation was negative. Growth characteristics of the primary leaves were shown to correlate with IAA in darkness and with GA and zeatin in the light. At a low concentration of cytokinins in illuminated leaves, cell divisions occurred, whereas, at the higher cytokinin concentrations, cell expansion occurred. The highest content of GA was characteristic of leaves in the period of growth cessation. ABA accumulated during active leaf and root elongation and biomass increment in the light and during hypocotyl growth in darkness. After plant illumination, the ratio of auxins to cytokinins increased in bean roots and decreased in their epicotyls. Thus, light changed the developmental programs of bean plants, which was manifested in the changed rate and duration of growth of various organs (root, hypocotyl, epicotyl, and leaf). Some mechanisms of light action depended on the contents of IAA, ABA, GA, and cytokinins and the ratios between these phytohormones. Differences between scotonorphogenesis of mono-and dicotyledonous plants are discussed in relation to the levels of phytohormones in them.     

Morphogenesis of potato plants in vitro. II. Endogenous levels,distribution, and metabolism of IAA and cytokinins

The levels of endogenous IAA and cytokinins (zeatin, zeatin riboside, isopentenyladenine, and isopentenyladenosine) were determined in potato plants cultured in vitro under red light (R) and blue light (B) on medium with or without hormones. On medium without hormones in B, plants contained much higher cytokinin levels, particularly in leaves and roots, and also slightly elevated IAA levels. Kinetin in the medium in B changed the distribution of cytokinins and significantly increased IAA level in roots. In R, the presence of kinetin led to an increased cytokinin level in the whole plant, while the IAA level was slightly lower. IAA in the medium in B decreased cytokinin level in all plant parts, while the IAA level did not change significantly. In R, the presence of IAA in the medium led to a moderate increase of CK level and to a significant increase in IAA level, especially in roots. Uptake of 1-¹⁴C-IAA and of ³H-zeatin was generally higher in B than in R. Higher percentage of IAA taken up in B was converted to conjugates in the roots. Metabolism of ³H-zeatin was similar in R and B with only slight differences in metabolite amounts.Thus, in all experimental situations in which tuber formation was stimulated, IAA level in roots and stolons rose significantly, stressing the importance of an IAA gradient for tuber formation.     

Stem girdling influences concentrations of endogenous cytokinins and abscisic acid in relation to leaf senescence in cotton

Many studies have shown that root–shoot imbalance influences vegetative growth and development of cotton (Gossypium hirsutum L.), but few have examined changes in leaf senescence and endogenous hormones due to stem girdling. The objective of this study was to determine the correlation between some endogenous phytohormones, particularly cytokinins and abscisic acid (ABA), and leaf senescence following stem girdling. Field-grown cotton plants were girdled on the main stem 5 days after squaring (DAS), while the non-girdled plants served as control. Plant biomass, seed cotton yield, main-stem leaf photosynthetic (Pn) rate, chlorophyll (Chl) and malondialdehyde (MDA) concentrations, as well as levels of cytokinins and ABA in main-stem leaves and xylem sap were determined after girdling or at harvest. Main-stem girdling decreased the dry root weight and root/shoot ratio from 5 to 70 days after girdling (DAG) and reduced seed cotton yield at harvest. Main-stem leaf Pn and Chl concentration in girdled plants were significantly lower than in control plants. Much higher levels of MDA were observed in main-stem leaves from 5 to 70 DAG, suggesting that stem girdling accelerated leaf senescence. Girdled plants contained less trans-zeatin and its riboside (t-Z + t-ZR), dihydrozeatin and its riboside (DHZ + DHZR), and isopentenyladenine and its riboside (iP + iPA), but more ABA than control plants in both main-stem leaves and xylem sap. These results suggested that main-stem girdling accelerated leaf senescence due to reduced levels of cytokinin and/or increased ABA. Cytokinin and ABA are involved in leaf senescence following main-stem girdling.     

Analysis of correlation between blood biochemical indicators and bone mineral density of post-menopausal women

Osteoporosis is a degenerative disease of the skeletal system, and its major complication is fracture that severely influences the living quality of the middle-aged and the aged. The purpose of this study was to investigate the significance of sex hormones and some biochemical indicators related to bone metabolism in the genesis and development of osteoporosis. The plasma samples were collected from 244 post-menopausal women of Xi’an urban area, and their plasma contents of testosterone, estradiol, calcitonin, osteocalcin and N-terminal propeptide of type I procollagen were detected by ELISA. The activity of tartrate-resistant acid phosphatase was determined by spectrophotometric method, and the content of nitric oxide was measured by Griess method. Bone mineral density (BMD) in lumbar vertebrae (L1–L4) and hips was measured by QDR-2000 dual energy X-ray absorptiometry. The concentrations of the biochemical indicators were compared among the three groups (normal bone mass group, osteopenia group and osteoporosis group), and Pearson correlation analysis was used to verify the correlations between the indicators and BMD. The comparison results of blood biochemical indicators of BMD-based groups showed that the plasma contents of estradiol (P = 0.006), testosterone (P = 0.038) and calcitonin (P = 0.042) decreased more significantly in the osteoporosis group, but the content of osteocalcin (P = 0.008) increased significantly in osteoporosis group than those in the other groups. The correlation analysis between BMD of different parts and the blood biochemical indicators showed that there was a significant positive correlation between estradiol and the BMD of lumber vertebra (r = 0.200, P = 0.002), femoral neck (r = 0.160, P = 0.013), and great trochanter (r = 0.204, P = 0.001). Significant positive correlations between calcitonin and BMD of lumber vertebra (r = 0.166, P = 0.018) and femoral great trochanter (r = 0.152, P = 0.041), and between testosterone and BMD of femoral great trochanter (r = 0.158, P = 0.014) were also observed. In addition, there existed significant negative correlations between osteocalcin and BMD of lumber vertebra (r = −0.220, P = 0.001), femoral neck (r = −0.259, P < 0.000), and great trochanter (r = −0.221, P = 0.001), and between the activity of tartrate-resistant acid phosphatase and BMD of femoral great trochanter (r = −0.135, P = 0.037). The partial correlation analysis also showed that there were significant correlations between estradiol (r = 0.160, P = 0.014), calcitonin (r = 0.240, P = 0.013), osteocalcin (r = −0.226, P = 0.023) and BMD when the influence of age was excluded. The Pearson correlation analysis of biochemical indicators showed there were positive correlations between the contents of testosterone and calcitonin, testosterone and osteocalcin, calcitonin and osteocalcin, calcitonin and PINP, calcitonin and NO, osteocalcin and NO, and PINP and NO, but negative correlations between the contents of testosterone and PINP, estradiol and calcitonin, estradiol and osteocalcin, and estradiol and NO. The blood contents of sex hormones and calcitonin significantly influence BMD and osteoporosis development, and the increase of osteocalcin contents could be used as a biomarker to indicate the degree of osteoporosis in post-menopausal women.     

Ability of bacterium Bacillus subtilis to produce cytokinins and to influence the growth and endogenous hormone content of lettuce plants

Hormone production by micro-organisms selected as antagonists of pathogenic fungi and the effect of their introduction into soil on hormone content and growth of lettuce plants were studied. Hormones in bacterial cultural media and in plant extracts were immunopurified and assayed using specific antibodies to indolyl-3-acetic acid (IAA), abscisic acid (ABA), and different cytokinins (zeatin riboside (ZR), dihydrozeatinriboside (DHZR) and isopentenyladenosine (iPA)). ZR was shown to be the main cytokinin present in bacterial cultural media as a complex with a high molecular weight component. Inoculation of lettuce plants with bacteria increased the cytokinin content of both shoots and roots. Accumulation of zeatin and its riboside was greatest in roots shortly 2days after inoculation, when their content was 10 times higher than in control. Changes in the content of other hormones (ABA and IAA) were observed at the end of experiments only. Accumulation of cytokinins in inoculated lettuce plants was associated with an increase in plant shoot and root weight of approximately 30% over 8days.     

Influence of Arbuscular Mycorrhizal (AM) Fungi and Salinity on Seedling Growth, Solute Accumulation, and Mycorrhizal Dependency of Jatropha curcas L.

Production of Jatropha curcas as a biodiesel feedstock on marginal lands is growing rapidly. Biomass production on these lands is limited. Hence, the objective of this study was to evaluate the effect of arbuscular mycorrhiza (AM) fungi and salinity (0.1, 0.2, 0.3, 0.4, and 0.5% NaCl) on (1) seedling growth, leaf relative water content (RWC), lipid peroxidation, solute accumulation (proline and sugars), and photosynthetic pigments (Chl a and b) of Jatropha; (2) mycorrhizal colonization (%) and mycorrhizal dependency (MD) of Jatropha; and (3) glomalin content (Bradford reactive soil protein) in soil. Increased soil salinity significantly (P < 0.05) decreased AM root colonization (r ² = 0.98) of AM-inoculated plants and decreased survival (r ² = 0.93) and growth (shoot length, r ² = 0.89; tap root length, r ² = 0.93; shoot diameter, r ² = 0.99; shoot dry weight, r ² = 0.92; and root dry weight, r ² = 0.92) of non-AM-inoculated Jatropha. Under salt stress, AM-inoculated Jatropha plants had greater dry weight of shoots and roots, better leaf water status, less leaf membrane damage (low lipid peroxidation activity), higher solute (proline and sugars), and higher leaf chlorophyll concentrations than non-AM-inoculated plants. The mycorrhizal dependency (MD) of Jatropha increased from 12.13 to 20.84% with salinity (0–0.4% NaCl). Root AM colonization (%) and glomalin content in soil were negatively correlated with salinity (P < 0.05, r = −0.95). We conclude that inoculation with AM fungi lessens the deleterious effect of salt stress on seedling growth parameters under salt levels up to 0.5% NaCl (electrical conductivity of 7.2 dS m⁻¹). Inoculation of Jatropha seedlings with AM fungi can promote the establishment of Jatropha under NaCl-induced stress.     

Endogenous hormonal content and somatic embryogenic capacity of Corylus avellana L. cotyledons

Endogenous indole-3-acetic acid (IAA), abscisic acid (ABA) and cytokinins [zeatin (Z) zeatin riboside, dihydrozeatin, dihydrozeatin riboside, N⁶-isopentenyl adenine (iP) and N⁶-isopentenyladenine riboside] were evaluated in hazelnut (Corylus avellana L.) cotyledons of different developmental stage and genetic source for their somatic embryogenic capacity. There was an inverse correlation between the embryogenic potential of cotyledons and the degree of maturity of zygotic embryos, the first characteristic being associated with iP-type cytokinins and the second with Z-type cytokinins. Although the differences in total cytokinin, ABA and IAA contents between the cotyledons were small, the IAA/ABA and, mainly, the iP-type/Z-type cytokinin ratios were found to be two good indexes of the embryogenic competence of explants, suggesting that the endogenous hormonal balance is a very important factor defining the in vitro potential of hazelnut cotyledons. Received: 6 January 1997 / Revision received: 3 March 1997 / Accepted 1 April 1997