Improved assessment of viability and germination of Cattleya (Orchidaceae) seeds following storage

Effective seed storage methods for orchid species provide a unique opportunity to preserve large amounts of germplasm. Early evaluation of the physiological status of seeds is useful for selecting seed lots for conservation. To score seed quality, seeds from nine Cattleya species (Cattleya granulosa, Cattleya hegeriana, Cattleya intermedia, Cattleya mossiae, Cattleya purpurata, Cattleya sanguiloba, Cattleya tenuis, Cattleya tigrina and Cattleya walkeriana) were germinated on Knudson C medium and viability was measured using a tetrazolium (TZ) test following pre-conditioning in a 10% sucrose solution, both before and after storage at −18°C for 3 mo. Tetrazolium test results were quantified using digital imaging of germinating seeds on a flatbed scanner. Seed viability (with or without pre-conditioning in sucrose), germination, Germination Velocity Index (GVI) and the correlation between viability measurements and germination were also evaluated. Both the tetrazolium test and the germination results showed that there was no loss of viability after storage, although in five seed lots, the GVI decreased. Pre-conditioning improved the correlation between viability exhibited in the tetrazolium test and seed germination. Image analysis proved to be a powerful tool for analysing these small seeds.     

EFFECTS OF ETHEPHON,ITS NONETHYLENE-GENERATING ANALOG ETHYLPHOSPHONIC ACID,AND PHOSPHOROUS ACID IN ASEPTIC CULTURE OF ORCHID SEEDLINGS

Seeds of Cattleya aurantiaca (Orchidaceae) were germinated and grown aseptically on Knudson C medium containing 2.5, 5, 10, 20, and 50 mg l^-1 of 2-chloroethylphosphonic acid (Ethephon), ethylphosphonic acid, or the inorganic acid moiety of both, phosphosphorous (phosphonic) acid. 2-Chloroethylphosphonic acid, an ethylene precursor, reduced leaf length at 5–50 mg l^-1 proportional to increasing concentrations. Seedlings produced fewer leaves within the range of 20–50 mg l^-1 Ethephon compared to lower concentrations (2.5–10 mg l^-1). Germination was not affected adversely. Cultures containing the homologous, but not ethylene-generating ethylphosphonic acid in the same concentrations brought about similar but generally less severe reductions in leaf length and number. Phosphorous acid at the same concentration levels did not reduce the number and length of leaves. Our observation suggests that the intact organic phosphonates contribute to the inhibition of leaf growth caused by ethylene, which is released slowly from Ethephon in the culture medium and/or plant tissue, following uptake.     

Effect of phosphorus and zinc on nodulation and nitrogen fixation in chickpea (Cicer arietinum L.)

A green house study was conducted on the effect of P and Zn on nodulation and N fixation in chickpea (Cicer arietinum L.) in a loamy sand (Typic Torripsamments) using treatment combinations of five levels of P (0, 25, 50, 100 and 250 ppm), and six levels of Zn (0, 5, 10, 20, 40 and 100 ppm). The number, dry matter and leghaemoglobin content of nodules, and amount of N fixed generally increased with Zn alone upto 19 ppm and P alone upto 50 ppm, and decreased with their higher levels. Application of 25 to 50 ppm P and 5 to 10 ppm Zn counteracted to a greater extent the adverse effect of 40 and 100 ppm Zn, and 250 ppm P, resp. Maximum nodulation and N fixation (91 to 145% over zero P and Zn, at maturity) was recorded with 25 to 50 ppm P applied along with 5 to 10 ppm Zn. At 64 days, depletion in soil-N was noted, particularly when P was applied, whereas at maturity there was a gain in soil-N, ranging from 10.5 to 44.5 kg/2×10⁶ kg soil depending upon P and Zn treatments. The increase in nodulation and N fixation with balanced P and Zn nutrition might be attributed to an increase in leghaemoglobin, and K and Fe concentration in nodules, and increased plant growth, resulting into enhanced activity of N fixing organisms. The results showed that balanced P and Zn nutrition is essential not only for plant growth but also for maximum activity of Rhizobium for N fixation. Work done at Harvana Agricultural University, Hissar, India.     

Effect of carbofuran on the growth and nodulation ofVigna radiata

Effect of carbofuran on the growth, nodulation, phytomass, chlorophyll and carotene ofVigna radiata Roxb., was studied. Out of different concentrations of carbofuranviz., 0, 5, 10, 25, 50 and 100 ppm, the concentration of 5 ppm carbofuran revealed no toxic effect, instead the growth, number of nodules, phytomass and chlorophyll were increased in comparison to control. However, 10, 25 and 50 ppm carbofuran proved to be toxic. The plants showed inhibition of growth. While 100 ppm of carbofuran was found to be fatal for the growth of plants.     

Effects of early plant growth regulator treatments on flavonoid levels in grapefruit

Early foliar spray treatments containing gibberellic acid(GA₃) significantly lower the concentration ofthe bitter flavonoid naringin in fruit tissues. Sprayscontaining a surfactant and different levels ofGA₃ (5, 50, 100, and 500 ppm) or abscisic acid(ABA) (5, 25, and 50 ppm) were applied to young,developing fruit on mature grapefruit (Citrusparadisi) trees during the period from April to June,beginning just after fruit set. The fruit were allowedto mature and were harvested early the following year.Harvested fruit were evaluated for weight, juicecharacteristics, and flavonoid concentrations.GA₃ application resulted in larger mature fruit,which yielded juice with the same soluble solids valueas juice from control fruit, but with slightly loweracid percentages and lower concentrations of naringin.ABA treatment had little effect on juice solublesolids, acid content and naringin content except atthe highest concentration of 50 ppm, which lowerednaringin levels slightly in juice.     

Induction of sex reversal in Oreochromis mossambicus by diethylstilbestrol

The efficacy of the synthetic estrogen, diethylstilbestrol (DES), for sex-reversal in the Java tilapia, Oreochromis mossambicus was investigated. Fry of 8–10 mm total length were fed diets containing 25, 50 and 100 ppm of DES for 30 days in plastic pools; this was followed by 45–65 days rearing in fertilized cement cisterns where hormone-free diet was given. DES at 50 and 100 ppm induced 100% sex-reversal; DES at 25 ppm resulted in only a slightly larger proportion of females to males. The untreated control group had a higher proportion of males than females. No intersex or sterile individuals were observed among the steroid-treated fish. The present investigation demonstrates that 50 ppm DES administered for 30 days is sufficient to induce a 100% sex-reversal in O. mossambicus.     

Are cadmium effects on plasma gonadotropins, prolactin, ACTH, GH and TSH levels, dose-dependent?

It is well established that cadmium affects plasma levels of the pituitary hormones studied. However, whether the effects of the metal are dose dependent needs to be clarify. This work was designed to evaluate the possible changes in plasma levels of gonadotropins, prolactin, ACTH, GH and TSH after oral cadmium exposure in adult male rats. Plasma levels of these hormones were measured in adult male rats exposed to cadmium chloride (CdCl₂) in the drinking water at the doses of 5, 10, 25, 50 or 100 ppm for one month. The lower dose of cadmium increased plasma prolactin levels and higher doses of the metal (25 or 50 ppm) decreased them. There was a continuous increase of plasma ACTH levels from the lower to 25 ppm dose of CdCl₂ and decreased them after to basal values with the highest dose. Plasma GH levels were increased with the dose of cadmium of 10 ppm, although the doses of 5, 25 and 50 ppm decreased them. Plasma LH levels were only reduced with the dose of 50 ppm of CdCl₂, whereas those of FSH increased. Plasma TSH levels were increased with the doses of 5, 25 and 100 ppm of CdCl₂. Cadmium concentration increased in pituitary with the doses of 125, 50 and 100 ppm of CdCl₂. These data suggest that cadmium differentially affects the secretory mechanisms of the pituitary hormones studied depending on the dose used. The effects of the metal on prolactin and ACTH are dose-dependent.     

Plant treatment in relation to the rhizosphere effect

Summary Unlike the usual methods of studying the rhizosphere microflora of crop plants, a new method of pretreatment of roots of one week old rice seedlings (MTU9) (grown in modified Robbins' nutrient solution) with 1 ppm and 5 ppm of patulin, griseofulvin, gibberellin and actidione as well as in 5, 10 and 50 ppm of agrimycin-100 and 0.1M urea has been tried. The effect of these substances on quality and quantity of rhizosphere microflora are discussed from the results of rhizosphere analysis obtained by routine methods.Part of Doctoral thesis, University of Madras 1960.     

Efficacy of bacterial auxin on in vitro growth of Brassica oleracea L.

Murashige and Skoog’s (MS) medium was supplemented with supernatant of Halomonas desiderata RE1 in different combinations to observe the impact of bacterial auxin on in vitro growth of Brassica oleracea L. Three groups of combinations MS + BS (Bacterial supernatant), MS + BS + 10% CW (coconut water) and MS + BS + 4 mg l⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D) were considered. Different amounts of BS used in each combination were 50, 100, 150 and 200 μl in 5 ml MS medium. Media combinations inoculated with seeds, internodal explants and callus of B. oleracea L. were incubated in a growth chamber at 25 ± 1°C and exposed to 16-h cool fluorescent light. Seeds inoculated on MS + BS and MS + BS + 10% CW, shoot elongation was observed over control whereas this response was suppressed in 2,4-D-containing media. In explants inoculated on MS + BS, MS + BS + 10% CW and MS + BS + 4 mg l⁻¹ 2,4-D different responses such as callus induction, adventitious shoot induction and hypertrophy were observed at different supernatant treatments. In callus inoculation, callus proliferation was observed in most of the treatments at different media combinations.     

Endophytic bacteria isolated from orchid and their potential to promote plant growth

Twelve endophytic bacteria were isolated from the meristem of in vitro Cymbidium eburneum orchid, and screened according to indole yield quantified by colorimetric assay, in vitro phosphate solubilization, and potential for plant growth promotion under greenhouse conditions. Eight strains with positive results were classified into the genus Paenibacillus by FAME profile, and evaluated for their ability to increase survival and promote the growth of in vitro germinated Cattleya loddigesii seedlings during the acclimatization process. The obtained results showed that all strains produced detectable indole levels and did not exhibit potential for solubilizing inorganic phosphate. Particularly, an increase of the total biomass and number of leaves was observed. Two strains of Paenibacillus macerans promoted plant growth under greenhouse conditions. None of the treatments had a deleterious effect on growth of inoculated plants. These results suggest that these bacterial effects could be potentially useful to promote plant growth during seedling acclimatization in orchid species other than the species of origin.     

Evaluating alder-endophyte (Alnus acuminata-Frankia-Mycorrhizae) interactions

The objective of this study was to compare the interaction betweenFrankia and vesicular-arbuscular mycorrhizae (VAM) on the growth and N-fixing response ofAlnus acuminata seedlings under three different phosphorus levels.A. acuminata is an actinorhizal tree, commonly associated with pastures on upland areas. Seedlings were grown in sterile vermiculite, and inoculated withFrankia strain ArI3 and/or VAM (asGlomus intra-radices) under three phosphorus levels (10, 50 and 100 ppm). After 120 days differences in growth were observed at the 50 ppm P level between nodulated and non-nodulated plants; either if inoculated withFrankia+VAM or just with VAM. Interaction betweenFrankia and VAM was positive on nodule weight at 50 ppm P level. Differences in acetylene reduction, per gram of fresh nodule, were observed between and within both groups:Frankia inoculated andFrankia+VAM inoculated seedlings. InFrankia inoculated seedlings differences were observed between seedlings receiving 50 ppm P which showed higher nitrogenase activity than seedlings treated with 100 ppm P. Plants inoculated withFrankia andGlomus intra-radices at low P level (10 ppm) showed the highest acetylene reduction. It was 150% higher than the mean of the other treatments within the group, and 87% higher than the general mean of the onlyFrankia inoculated plants.     

Interaction between Biological Activities on Rice Seedlings of Dehydroabietic Acid Derivatives and Gibberellin A3

Effects of hydrofluorene and hydrophenanthrene compounds derived from dehydroabietic acid on the second leaf sheath growth of rice seedlings were examined in the presence and absence of gibberellin A₃(GA₃). In the absence of GA₃, nineteen compounds at 100 ppm inhibited more than 20% the growth of normal rice seedlings. In the presence of GA₃ (1.5 ppm) with dwarf rice seedlings, nine compounds at 500 ppm suppressed the elongation caused by the hormone, and a compound was slightly promotive. Then, three compounds were selected and subjected to the bioassay under various conditions.     

Bioremediation of Vegetable Oil and Grease from Polluted Wastewater Using a Sand Biofilm System

Pseudomonas sp. (L1), P. diminuta(L2) were among eight bacterial strains isolated from vegetable grease and oil-contaminated industrial wastewater, four of which only were found to have the ability to degrade oil and grease. They were identified and investigated for oil and grease degradation either individually or in combinations in previous unpublished work by the authors. Since the combination M1 (Pseudomonas sp. andP. diminuta) produced the highest degradative activity, it was used in the present study in a biofilm sand filter system for vegetable oil and grease removal. This system was tested either as one unit or two units in sequence where different flow rates (30, 50, 100 ml/h) were applied compared to a control unit(s). Results showed that both biofilm systems reduced oily wastewater, even in cases of high degree of pollution (fat, oil & grease (FOG), 7535 ppm; biochemical oxygen demand (BOD₅), 525 ppm; chemical oxygen demand (COD), 1660 ppm). Results also showed a removal of FOG with efficiency at 100%; BOD₅ at 95.9% and COD at 96%, at 50 ml/h flow rate using one unit of biofilm system. On using two units in sequence, a complete removal of FOG, BOD₅ and COD with efficiency 100%, at flow rate 100 ml/h was achieved. In conclusion, the previous biofilm results indicated the efficiency of such a system in treating oily polluted wastewater (vegetable oil origin) on the basis of bacterial isolates being used, the optimum flow rate, and the number of biofilm units used in sequence to obtain the highest removal capacity of such a system. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effects of some organophosphate and carbamate insecticides on nitrification and legume growth

Summary Nine insecticides (six organophosphates and three carbamates) were tested for their effects on soil nitrification, growth of legume seedlings, and growth of four species of rhizobia bacteria. No inhibition of nitrification was found at normal field rates (5 ppm) of application. Some instances of inhibition were observed at 50 ppm and at 500 ppm. Similarly, 5 ppm applications did not inhibit growth of alfalfa or sweetclover seedlings ... with one exception. Disc inhibition tests of the rhizobia bacteria showed thatRhizobium leguminosarum andRhizobium trifolii were most sensitive to the pesticides.Rhizobium meliloti, and particularlyRhizobium japonicum, were resistant to the insecticides. No consistent correlation was observed between tests on the nodulating bacteria and the tests on legume growth.Published with the approval of the Director of the North Dakota Agricultural Experiment Station as Journal Article No.309. Portion of a thesis presented by the senior author in partial fulfillment of the requirements for the M.S. degree in bacteriology at North Dakota State University.     

Similarity of growth patterns between plantlets and seedlings of Brassica campestris L. under different in vitro environmental conditions

Explants and seeds of Brassica campestris L. were cultured on Murashige & Skoog (1962) medium with and without sucrose in a vessel with different numbers of air changes per hour under different PPF (photosynthetic photon flux) conditions. The growth and development of plantlets in the vessel were similar to those of seedlings when cultured under the same in vitro environmental conditions. The growth and development of seedlings when cultured under the same in vitro environmental conditions. The growth and development of plantlets/seedlings were greater for treatments with a higher number of air changes per hour and a higher PPF regardless of the sucrose concentration in the culture medium.The CO₂ concentration in the vessel with a lower number of air changes per hour decreased to approximately 100 ppm during the photoperiod on day 21 due to the photosynthetic activities of the plantlets/seedlings. The low CO₂ concentration, in turn, reduced the net photosynthetic rate of plantlets/seedlings in the vessel, and thus affected their growth and development.Abbreviations C_in CO₂ concentration in the culture vessel - C_out CO₂ concentration in the culture room - MS mineral composition of Murashige & Skoog (1962) medium - PPF photosynthetic photon flux     

Ultrastructural changes and diosgenin content in cell suspensions of <Emphasis Type="Italic">Trigonella foenum-graecum</Emphasis> L. by ethylene treatment

This paper studies the effect of different concentrations of ethephon (an ethylene releasing compound) on the level of the steroidal sapogenin diosgenin in cell suspensions of Trigonella foenum-graecum L. The results reveal that diosgenin synthesis and/or accumulation was stimulated by 5 ppm ethephon treatment, an increase of 126% being observed, while concentrations of 25 ppm and 50 ppm reduced the levels of this secondary compound. Changes in the isopentenyl diphosphate isomerase activity, increased cell diameter, decreased cell packing (with all the ethephon concentrations assayed), increased cytoplasmic density (5 ppm ethephon treatment) and alterations in the membrane structures (25 and 50 ppm ethephon treatments) were also observed.     

猕猴桃属种间嫁接亲和性试验研究及抗根结线虫砧木的初步筛选

以中华猕猴桃（Actinidia chinensis)武植2号（Wuzhi-2),通山5号（Tongshan-5)品种和美味猕猴桃（A.deliciosa)金魁（Jinkui)品种为接穗,采用美味猕猴桃等9种类型猕猴桃一年生实生苗为砧木进行嫁接,研究其短期嫁接亲和性,并对这9种砧木进行抗根结线虫（Meloidogyne)筛选试验。结果表明,种间的嫁接亲和性及抗根线虫性 均有显著差异：以美味猕猴桃和中华猕猴桃优良品种实生苗为砧木,与3个品种均有高的亲和性,成活率高达100％,且抗性在9种砧木中最强;其它物种作砧木均有不足,武植2号／山猴猴桃,通山5号／M16,通山5号／山梨猕猴桃3个嫁接组合的亲和性均较好,成活率在80％以上,但山梨猕猴桃较易感病且与金魁的亲和力较低,M16抗性虽较好,但与金魁,武植2号的亲和力也较低,漓江猕猴桃是抗性较强的物种,但它与3种接穗品种的亲和力都较小,其它5种砧木,对根结线虫的抗性均不强,因此结合成活率及抗根结线虫两因素,在供试材料中,美味猴猕桃和中华猕猴桃接适应性广且抗性强,是较好的砧木或选育砧木的亲本材料。     

Occurrence ofFusarium sacchari var.subglutinans and its mycotoxin production ability in broiler feed

Spores ofFusarium sacchari var.subglutinans isolated from broiler feed BR1 were obtained at an average concentration of 1.5/mg in 25% of tested samples. The spore concentration was increased from 1 to 100/mg of solid substrate (BR2; biscuit) or to 1/nL of Sabouraud broth after 3 weeks of cultivation. Mycotoxin analyses of these three substrates showed negative reactions for T-2 toxin and zearalenone but a positive reaction for deoxynivalenol (DON) which was found in concentrations of 5 ppm in Sabouraud broth, 50 ppm in BR2 and 220 ppm in biscuit. Therefore, ourF. sacchari isolate appeared to be a DON producer.     

The effect of the systemic fungicide benomyl on survival and reproduction of the bird cherry aphid (Rhopalosiphum padi)

Weight gain and intrinsic rate of population increase (r_m) of Rhopalosiphum padi were measured on wheat plants given soil drenches of benomyl at various concentrations. Reduced weight gain occurred with concentrations as low as 3.125 ppm. Mortality of aphids significantly exceeded control mortality at 25 ppm, and no aphids survived concentrations above 50 ppm. Development time was extended at 12.5 ppm and above. Reductions in fecundity and r_m became statistically significant by 25 ppm. Topical application of benomyl at 100 ppm or above increased aphid mortality, which reached 97% at 400 ppm. Weight gain of aphids was also reduced by topical application at 100 ppm. A factorial experiment showed absence of interaction between the effects on aphid weight of soil drench application to plants and topical application to aphids.     

Selenium-mediated differential response of β-glucosidase and β-galactosidase of germinatingTrigonella foenum-graecum

β-Glucosidase and β-galactosidase activity profile tested in different seeds during 24 h germination revealed reasonably high levels of activity inVigna radiata, Cicer arietinum, andTrigonella foenum-graecum. In all seeds tested, β-galactosidase activity was, in general, higher than that of β-glucosidase.T. foenum-graecum seedlings exhibited maximal total and specific activities for both the enzymes during 72 h germination. Se supplementation as Na₂SeO₃ up to 0.75 ppm was found to be beneficial to growth and revealed selective enhancement of β-galactosidase activity by 40% at 0.5 ppm Se. The activities of both the enzymes drastically decreased at 1.0 ppm level of Se supplementation. On the contrary, addition of Na₂SeO₃ in vitro up to 1 ppm to the enzyme extracts did not influence these activities. Hydrolytic rates of β-glucosidase in both control and Se-supplemented groups were enhanced by 20% with 0.05M glycerol in the medium and 30% at 0.1M glycerol. The rates were marginally higher in Se-supplemented seedlings than the controls, irrespective of added glycerol in the medium. In contrast, hydrolysis by β-galactosidase showed a trend of decrease in Se-supplemented seedlings compared to the control, when glycerol was present in the medium. Addition of Se in vitro in the assay medium showed no difference in the hydrolytic rate by β-galactosidase when compared to control, while the activity of β-glucosidase declined by 50%. Se-grown seedlings showed an enhancement of transglucosidation rate by 40% in the presence of 0.1M glycerol. The study reveals a differential response to Se among the β-galactosidase and β-glucosidase ofT. foenumgraecum with increase in the levels of β-galactosidase activity.