Evaluation of Pseudomonas syringae strain ESC-11 for biocontrol of crown rot and anthracnose of banana

Pseudomonas syringae strain ESC-11 and 250 μg/ml each of thiabendazole (TBZ) and imazalil reduced crown rot of banana caused by Fusarium aff. sacchari by 30–36% and 83–86%, respectively, in laboratory experiments. Four field trials performed in Costa Rica varied in treatment combinations. In field trials 1 and 2, 125 and 250 μg/ml each of TBZ and imazalil + 0.5% or 1% alum (aluminum ammonium sulfate) and ESC-11, and 250 μg/ml each of TBZ and imazalil + 1% alum reduced rot and mold. ESC-11 alone or with 0.5% alum significantly reduced rot and mold in field trial 2. In trial 3, 50 and 100 μg/ml of TBZ alone and with ESC-11 reduced mold. In trial 4, 125 μg/ml each of TBZ and imazalil and ESC-11, and 300 μg/ml each of TBZ and imazalil reduced rot, and 50 and 125 μg/ml each of TBZ and imazalil and ESC-11, and 300 μg/ml each of TBZ and imazalil reduced mold. In three field trials, there was no significant difference among treatments for latex staining. In field trial 2 only, combinations of TBZ, imazalil, and alum with or without ESC-11, reduced anthracnose, caused by Colletotrichum musae. The complex of crown rot fungi, order of treatment application, effect of alum and fungicides on ESC-11, concentration of ESC-11, and level of disease may contribute to the variation in crown rot and anthracnose control by ESC-11. Though ESC-11 alone was not effective in reducing disease, further testing in combination with low rates of fungicide should be done.     

Microencapsulating aerial conidia of Trichoderma harzianum through spray drying at elevated temperatures

Conidia of Trichoderma harzianum produced from either solid or liquid fermentation must be dried to prevent spoilage by microbial contamination, and to induce dormancy for formulation development and prolonged self-life. Drying conidia of Trichoderma spp. in large scale production remains the major constraint because conidia lose viability during the drying process at elevated temperatures. Moreover, caking must be avoided during drying because heat generated by milling conidial chunks will kill conidia. It is ideal to dry conidia into a flow-able powder for further formulation development. A method was developed for microencapsulation of Trichoderma conidia with sugar through spray drying. Microencapsulation with sugars, such as sucrose, molasses or glycerol, significantly (P < 0.05) increased the survival percentages of conidia after drying. Microencapsulation of conidia with 2% sucrose solution resulted in the highest survival percentage when compared with other sucrose concentrations and had about 7.5 × 10¹⁰ cfu in each gram of dried conidia, and 3.4 mg of sucrose added to each gram of dried conidia. The optimal inlet/outlet temperature setting was 60/31 °C for spray drying and microencapsulation. The particle size of microencapsulated conidia balls ranged from 10 to 25 μm. The spray dried biomass of T. harzianum was a flow-able powder with over 99% conidia, which could be used in a variety of formulation developments from seed coatings to sprayable formulations.     

Postharvest biocontrol of Monilinia laxa,Monilinia fructicola and Monilinia fructigena on stone fruit by two Aureobasidium pullulans strains

The antagonistic effects of yeasts, L1 and L8, isolated from carposphere of ‘Redhaven’ peaches were tested for the first time in the same experiment against three Monilinia species (Monilinia laxa, Monilinia fructicola and Monilinia fructigena) in in vitro and in vivo trials. The two antagonists were selected after preliminary assays for their ability to reduce brown rot in peaches and nectarines, and both were identified by molecular and morphological tools as Aureobasidium pullulans. In in vivo trials, neither the autoclaved cells, nor the sterile culture filtrates of either antagonist showed any significant reduction of rot incidence produced by inocula of the three Monilinia species, while the washed cells of L1 and L8 completely inhibited M. laxa and M. fructicola rots and reduced M. fructigena infections by 70% and 90%, respectively. In other trials, nectarines treated with antagonist cells and inoculated with the pathogens were stored at 0 °C for 21 days, plus 7 days at 20 °C. The low temperature reduced brown rot development, since all fruit were free from disease symptoms on removal from cold storage. However after 7 d at 20 °C, untreated fruit were rotted over 45% depending on the Monilinia species but the antagonists completely inhibited M. laxa and M. fructicola, while M. fructigena infections were reduced by 89.8% and 91.2% by L1 and L8, respectively. For both strains, 10⁸ CFU ml^?1 was the most active concentration, although L1 showed good activity at a concentration of 10⁷ CFU ml^?1. Isolate L8 at the concentration of 10⁷ CFU ml^?1 was ineffective against M. fructicola and M. fructigena, showing no difference between treated fruit and the control, excepting the case of nectarines inoculated with M. laxa, where L8 at the concentration of 10⁷ CFU ml^?1 reduced the brown rot infections with respect to the control. The increase in population density of A. pullulans strains L1 and L8 in the wounds of nectarines stored at 0° or 20 °C was low but sufficient to control brown rot. In conclusion, the present preliminary study identified two antagonistic strains of A. pullulans as active ingredients for the development of biofungicides for postharvest application against three Monilinia species that are responsible for high economic losses in stone fruit crops.     

Soil microbial biomass influence on growth and biocontrol efficacy of Trichoderma harzianum

Hyphal growth and biocontrol efficacy of Trichoderma harzianum may depend on its interactions with biotic components of the soil environment. Effects of soil microbial biomass on growth and biocontrol efficacy of the green fluorescent protein transformant T. harzianum ThzID1-M3 were investigated using different levels of soil microbial biomass (153, 328, or 517 μg biomass carbon/g of dry soil). Hyphal growth of T. harzianum was significantly inhibited in soil containing 328 or 517 μg biomass carbon/g of dry soil compared with soil containing 153 μg biomass carbon/g. However, when ThzID1-M3 was added to soil as an alginate pellet formulation, recoverable populations of ThzID1-M3 varied, with the highest populations in soil containing 517 μg biomass carbon/g. When sclerotia of Sclerotinia sclerotiorum were added to soils (10 sclerotia per 150 g soil) with ThzID1-M3 (20 pellets per 150 g soil), colonization of sclerotia by ThzID1-M3 was significantly lower in the soil containing the highest level of biomass. Addition of alginate pellets of ThzID1-M3 to soils (10 pellets per 50 g) resulted in increased indigenous microbial populations (total fungi, bacteria, fluorescent Pseudomonas spp., and actinomycetes). Our results suggest that higher levels of microbial soil biomass result in increased interactions between introduced T. harzianum and soil microorganisms, and further that microbial competition in soil favors a shift from hyphal growth to sporulation in T. harzianum, potentially reducing its biocontrol efficacy.     

The seed ecology of an ornamental wattle in South Africa — Why has Acacia elata not invaded a greater area?

Australian Acacia species introduced to South Africa as ornamentals have notably smaller invasive ranges than those introduced for forestry or dune stabilization. We asked whether the relatively small invasive extent of Acacia elata, a species used widely for ornamental purposes, is due to low rates of reproduction. Age at reproductive maturity, seed dispersal, annual seed production, seed bank dynamics and seed germination and viability were assessed at five sites in the Western Cape. Results indicate that A. elata has similar traits to other invasive Australia Acacia species: annual seed input into the leaf litter was high (up to 5000 seeds m^− 2); large seed banks develop (> 20,000 seeds m^− 2) in established stands; seed germinability is high (> 90%); seeds accumulate mostly in the top soil layers but can infiltrate to depths of 40 cm; and seed germination appears to be stimulated by fire. However the age at the onset of reproduction (~ 4 years) is longer than most widespread invaders (~ 3 years) and dispersal is fairly limited (seeds fell up to distances of 6 m from the parent canopy; the highest density of seed rain was found directly under the canopy with over 20% of seeds falling directly under the terminal branches). We suggest that the current limited distribution of invasive A. elata populations is the result of the relatively small size of initial populations (cf. large plantations and widespread plantings for forestry and dune stabilization species), the species' apparent lack of secondary dispersal vectors, and the planting of trees in gardens and urban settings offer limited opportunities for recruitment, proliferation and spread. The species is, however, increasing in abundance and range. We propose methods to improve management of invasions of the species. Management to reduce seed production of this species through classical biological control, as has been achieved for other Australian Acacia species in South Africa, should be prioritised.     

Antimicrobial activity of medicinal plants and induction of defense related compounds in banana fruits cv. Robusta against crown rot pathogens

A total of 72 plant extracts were tested in vitro for their ability to inhibit the mycelial growth of Lasiodiplodia theobromae and Colletotrichum musae the causal agents of crown rot disease of banana. The results showed that the leaf extract of Zimmu (an interspecific hybrid of Allium cepa L. × Allium sativum L.) and tuber extract of Zehneria scabra recorded maximum inhibition of mycelial growth and spore germination of both the test pathogens. The dipping of banana fruits in Zimmu leaf extract at 25% conc. exhibited 100% inhibition of crown rot disease in cold storage (14 °C) up to 35 days and increased the shelf life to 64 days. However, at room storage (28 ± 2 °C), the same treatment exhibited 86% inhibition of crown rot disease up to 12 days. It was found that the treatment of banana fruits with Zimmu leaf extract did not alter the organoleptic properties of banana. The biochemical analysis of banana fruits treated with Zimmu leaf extract showed significant increase in phenylalanine ammonia-lyase (PAL), chitinase and β-1,3-glucanase activities and enhanced accumulation of phenolic compounds compared to other treatments. These findings suggest that the effect of Zimmu leaf extract on crown rot disease may be associated with the direct fungi toxic property against the test pathogens and elicitation of defense related compounds in banana fruits.     

Effect of breed on response to foot rot treatment in mature sheep and lambs

Our objective was to determine whether breed differences existed in response to exposure and treatment of virulent foot rot. Dorset (DS), 1/2 Dorper (DX), 3/4 or greater Dorper (DO), Gulf Coast Native (GC), Katahdin (KA), and St. Croix (SC) mature sheep and lambs were exposed to virulent foot rot in spring 2003. Treatment for foot rot was initiated in 132 lambs and 262 mature sheep in late July. There were eight pasture groups treated, two of which were minimally exposed to foot rot. Treatment included hoof paring, foot bathing with 10% zinc sulfate with surfactant, allowing the zinc sulfate to dry on the foot and moving to a small paddock that had not been exposed to small ruminants for more than 14 d. Foot bathing was repeated every 7 d for a maximum of five treatments. Animals that had not responded (odor or any indication of persistent infection) by then were culled from the flock. As an indication of severity of foot rot for each animal, the number of areas on the foot (interdigital and two digits for each foot), a foot score (0 = no infection found; 1 = infection of digits only; 2 = infection of interdigital area and could include digits), and presence of characteristic odor was recorded. Least squares means for number of areas infected were greater for mature than growing sheep (2.07 ± 0.16 versus 0.88 ± 0.31; P < 0.001), for highly than minimally exposed groups (2.89 ± 0.17 versus 0.05 ± 0.29; P < 0.001), and DX compared with other breed types (P < 0.03). Percentage of sheep with odor was similar between age groups, was greater in the highly exposed groups (11.4 ± 1.9 versus 2.1 ± 3.4; P < 0.02), and greater in DO compared with DS, KA, and SC breeds (P < 0.001). Foot score was similar among breeds and greater in the highly exposed groups (age by group, P < 0.05). Percentage of sheep culled for failure to respond to foot bath treatment was greater for the highly than minimally exposed group (22.9 ± 2.3 versus 0.0 ± 3.9; P < 0.001) and greater for mature sheep compared with lambs (P < 0.001) and similar among breeds. In November, four ewes in a large group and two lambs in a small group were determined to have foot rot and were immediately culled. The two groups containing these ewes were re-treated for 2 weeks and were determined to be free of foot rot (no further signs of lameness). Response to foot rot eradication appeared to be similar among breeds examined.     

Efficient isolation of anthraquinone-derivatives from Trichoderma harzianum ETS 323

Anthraquinone-derivatives, chrysophanol and pachybasin, were purified by a silica column chromatography with two different solvent systems from Trichoderma harzianum ETS 323. The fungus was incubated in sugarcane bagasse solid medium at room temperature without rotation. Structure of chrysophanol was solved by X-ray diffraction and pachybasin by NMR spectra. About 233 ± 13 mg of pure chrysophanol and 773 ± 40 mg of pure pachybasin were recovered per kg of solid cultural medium, with yields 1.7 ± 0.2% and 5.6 ± 0.5%, respectively.     

A new semi-selective medium for Fusarium graminearum,F. proliferatum,F. subglutinans and F. verticillioides in maize seed

Zea mays L., known also as corn and maize, is the most important crop according to the amount of tonnes produced each year. Fungi cause significant destruction of maize in the field as well as during storage rendering the grain unsuitable for human consumption by decreasing its nutritional value and by producing mycotoxins that are detrimental to both human and animal health. Fusarium species are widely distributed and are amongst the most frequently isolated fungal species by plant pathologists. Due to the fact that the Fusarium species involved in maize ear rot vary in fungicide sensitivity, pathogenicity as well as in their capability to produce mycotoxins, accurate quantification and identification is of paramount significance. Currently no method has been developed to test for Fusarium species in maize seed that has been validated and published by the International Seed Testing Association (ISTA). Malachite green agar 2.5 ppm (MGA 2.5) is a potent selective medium for isolation and enumeration of Fusarium spp. In this study, eight different media compositions, potato dextrose agar (PDA), PDA + malachite green oxalate, corn meal agar, 1/2 PDA + malachite green oxalate, 1% malt agar, carnation leaf agar supplemented with potassium chloride (KCLA), malachite green agar (MGA 2.5) and MGA 2.5 + sterile carnation leaf pieces were compared using four Fusarium species (F. graminearum, F. proliferatum, F. subglutinans and F. verticillioides) and five commonly encountered saprophytic fungi (Aspergillus niger, Penicillium crustosum, P. digitatum, Trichoderma harzianum and Rhizopus stolonifer). The maize kernels were surface disinfected using three concentrations of sodium hypochlorite (0.5%, 1% and 1.5% NaOCl) and for different time intervals (1 min, 3 min, 5 min and 10 min). The effect of black-blue light (365 nm) on sporulation of the fungi was also investigated. Surface disinfection of maize seeds with 1% NaOCl for 5 min provided consistent results. PDA, 1/2 PDA, 1% malt agar and KCLA allowed profuse growth of the Fusarium species as well as saprophytes. Media that contained malachite green oxalate was most inhibitory to the radial colony growth of the saprophytes and the Fusarium species. The Fusarium species growing on these media formed underdeveloped morphological structures, thereby obscuring accurate identification. MGA 2.5 showed better hindering of the saprophytes in some instances. MGA 2.5 amended with sterile carnation leaf pieces was the most satisfactory medium in hindering the growth of the saprophytes while allowing adequate sporulation by the four Fusarium species to permit accurate identification. The media also resulted in higher F. verticillioides and lower saprophytic fungal isolation frequency when compared to the other media tested.     

Proximate,mineral, fibre,phytate–phosphate,vitamin E,amino acid and fatty acid composition of Terminalia sericea

Terminalia sericea is widely distributed in the African Savannah bushveld. It is one of the indigenous fruit bearing trees put to multiple uses. Research has focused on the phytochemical composition of its root, bark, and leaf extracts that are used in ethnomedicine neglecting the potential of its seed. This study purposed to determine, by chemical analyses, the nutritive value of T. sericea seed. 78.8% of the seed was found to be crude protein (46.2%) and lipid (32.6%). Ash made up 6.90% of the seed mass. Linoleic and oleic acids constituted 68.63% and 14.05%, respectively, of the seed oil. Phosphorus (1121.75 ± 10.39 mg 100 g^− 1 DM) and glutamic acid (8.07 ± 0.13 g 100^− 1 DM) constituted the most concentrated mineral and amino acid, respectively. T. sericea seed could be utilized as a protein source in feeds and foods and could also be exploited as a non-conventional plant oil source of oleic acid and linoleic acid.     

Biocontrol of major postharvest pathogens on apple using Rhodotorula glutinis and its effects on postharvest quality parameters

The biocontrol activity of Rhodotorula glutinis on gray mold decay and blue mold decay of apple caused by Botrytis cinerea and Penicillium expansum, respectively, was investigated, as well as its effects on postharvest quality of apple fruits. The results show there was a significant negative correlation between concentrations of the yeast cells and the disease incidence of the pathogens. The higher concentration of the R. glutinis, the better effect of the biocontrol capacity. At concentrations of R. glutinis 1 × 10⁸ CFU ml^?1, the amount of gray mold decay was completely inhibited after 5 days incubation at 20 °C, after challenge with B. cinerea spores suspension of 1 × 10⁵ spores ml^?1; While the blue mold decay was completely inhibited at concentrations of 5 × 10⁸ CFU ml^?1, at challenged with P. expansum spores suspension of 5 × 10⁴ spores ml^?1_. These results demonstrated that the efficacy of R. glutinis in controlling of gray mold decay of apples was better than the efficacy of controlling blue mold. R. glutinis within inoculated wounds on apples increased in numbers at 20 °C from an initial level of 9.5 × 10⁵ CFU per wound to 2.24 × 10⁷ CFU at 20 °C after 1 day. The highest population of the yeast was recovered 4 days after inoculation, the yeast population in wounds increased by 56.9 times. After that, the population of the yeast began to decline very slowly. R. glutinis significantly reduced the incidence of natural infections on intact fruit from 75% in the control fruit to 28.3% after 5 days at 20 °C, and from 58.3 to 6.7% after 30 days at 4 °C followed by 4 days at 20 °C. R. glutinis treatment had no deleterious effect on quality parameters after 5 days at 20 °C or after 30 days at 4 °C followed by 4 days at 20 °C.     

Effective control of black Sigatoka disease using a microbial fungicide based on Bacillus subtilis EA-CB0015 culture

Black Sigatoka disease caused by the fungus Mycosphaerella fijiensis Morelet is the most devastating disease of bananas worldwide. Its management is reliant on protectant and systemic fungicides despite their environmental concerns. This study evaluated the effect of a microbial fungicide (MF) based on Bacillus subtilis EA-CB0015 and its metabolites for the control of black Sigatoka disease on banana plants in greenhouse and field conditions. The MF applied at 1.5 L/ha and 3.0 L/ha provided control of the disease comparable to the protectant fungicide chlorothalonil in greenhouse. In the field, the MF applied in solution with water at 0.15 L/ha and 1.5 L/ha every 11 days during 10 weeks reduced black Sigatoka disease severity in 20.2% and 28.1% respectively; reductions comparable to those obtained with the protectant fungicides chlorothalonil (1.5 L/ha) and mancozeb (3.8 L/ha). The MF incorporated into different programs with systemic fungicides reduced disease level up to 42.9% with no significant differences with the conventional program. To determine which component of the MF is responsible for the activity against M. fijiensis, greenhouse and in vitro tests were set up to evaluate individually the spores, vegetative cells and secondary metabolites of B. subtilis EA-CB0015. All components reduced the severity of the disease and the germination of ascospores. For both trials the activity of the metabolites was higher and comparable to the activity obtained with the MF, indicating that the efficacy of the MF depends mainly on the metabolites and in lesser extent to B. subtilis EA-CB0015 cells.     

Temporal changes in wood crystalline cellulose during degradation by brown rot fungi

The degradation of wood by brown rot fungi has been studied intensely for many years in order to facilitate the preservation of in-service wood. In this work we used X-ray diffraction to examine changes in wood cellulose crystallinity caused by the brown rot fungi Gloeophyllum trabeum, Coniophora puteana, and two isolates of Serpula lacrymans. All fungi increased apparent percent crystallinity early in the decay process while decreasing total amounts of both crystalline and amorphous material. Data also showed an apparent decrease of approximately 0.05 Å in the average spacing of the crystal planes in all degraded samples after roughly 20% weight loss, as well as a decrease in the average observed relative peak width at 2θ = 22.2°. These results may indicate a disruption of the outer most semi-crystalline cellulose chains comprising the wood microfibril. X-ray diffraction analysis of wood subjected to biological attack by fungi may provide insight into degradative processes and wood cellulose structure.     

Germination characteristics of the grass weed Digitaria nuda (Schumach.)

The effect of various pre-treatments and their interaction with temperature on cumulative percentage and the rate of germination were evaluated for Digitaria nuda. Stored and fresh seeds were pre-treated with either 0.02 M KNO₃, soaked in water for 24 h (priming), sterilized with 0.5% NaOCl or heat treated at 60 °C. Seeds were germinated at constant temperatures of 25 and 30 °C and fluctuating temperature regimes of 25/10 and 30/15 °C. The effect of pre-chilling on germination of stored and fresh seed was evaluated at 30/15 °C, and seed emergence in two soil types at different burial depths (0, 0.5, 1, 2, 3, 4, 5 and 6 cm) was also determined. The pre-treatment of stored seed with KNO₃ resulted in the highest germination percentage (100%), whereas the pre-treatment of fresh seed with water for 24 h gave the best germination (99%), at constant temperatures of 25 and 30 °C. Pre-chilling of seed increased germination by more than 30%. Emergence from clay loam soil was greater compared with the emergence from sandy loam soil. Total seedling emergence decreased exponentially with increasing burial depths with only 5% of seed germinating from a burial depth of 6 cm. Results from this study showed that germination requirements are species specific and knowledge of factors influencing germination and emergence of grass weed seed can assist in predicting flushes in emergence allowing producers to implement control practices more effectively.     

Effect of lignocellulose degradation products on microbial biomass and lipid production by the oleaginous yeast Cryptococcus curvatus

This work investigated effects of lignocellulose degradation products on cell biomass and lipid production by Cryptococcus curvatus. Furfural was found to have the strongest inhibitory effect. For the three phenolic compounds tested, vanillin was the most toxic, while PHB and syringaldehyde showed comparable inhibitions in the concentration range of 0–1.0 g/L. Generally little significant differences on the relative cell biomass and lipid contents at the same concentrations of tested compounds were observed between glucose and xylose as a sole carbon source. At 1.0 g/L of furfural, the cell biomass and lipid content decreased by 78.4% and 61.0% for glucose as well as 72.0% and 59.3% for xylose, respectively. C. curvatus ceased to grow at concentrations of PHB over 1.0 g/L or vanillin over 1.5 g/L. The strain could survive in the presence of syringaldehyde up to 2.0 g/L for glucose or 1.5 g/L for xylose. The compounds’ negative impact was reduced by an increase in inoculum size and a 10% (v/v) seed was detected to be optimal for cell biomass and lipid production. The results demonstrated C. curvatus could effectively utilize most of the dominant monosaccharides and cellobiose existing in lignocellulosic biomass hydrolysate in the presence of toxic compounds.     

Immobilization of phytase on epoxy-activated Sepabead EC-EP for the hydrolysis of soymilk phytate

In this work, an active phytase concentrated extract from soybean sprout was immobilized on a polymethacrylate-based polymer Sepabead EC-EP which is activated with epoxy groups. The immobilized enzyme exhibited an activity of 0.1 U/g of carrier and activity yield of 64.7%. The optimum temperature and pH for the activity of both free and immobilized enzymes were found as 60 °C and pH 5.0, respectively. The immobilized enzyme was more stable than free enzyme in the range of pH 3.0–8.0 and more than 70% of the original activity was recovered. Both the enzymes completely retained nearly about 84% of their original activity at 65 °C. The K_m and V_max values were measured as 5 mM and 0.63 U/mg for free enzyme and 12.5 mM and 0.71 U/mg for immobilized enzyme, respectively. Free and immobilized soybean sprout phytase enzymes were also used in the biodegradation of soymilk phytate. The immobilized enzyme hydrolysed 92.5% of soymilk phytate in 7 h at 60 °C, as compared with 98% hydrolysis observed for the native enzyme over the same period of time. The immobilization procedure on Sepabead EC-EP is very cheap and also easy to carry out, and the features of the immobilized enzyme are very attractive that the potential for practical application is considerable.     

Density and distribution of seeds in bottom sediments in Zostera marina beds in Ago Bay,central Japan

The density of Zostera marina L. seeds in bottom sediments was examined to study the reproductive patterns of the Z. marina population in Ago Bay, Mie Prefecture, central Japan.Seeds and seed coats were numerous in Tategami, where the annual type of Z. marina grows. In contrast, seeds were scarce in Hamajima, where the perennial type of Z. marina grows. Bottom sediment was sampled with sediment cores at Tategami in November 2004 and March 2005 to examine density and depth distribution of the seeds. Seeds were found as deep as 8 cm, but no deeper. On the other hand, empty seed coats were found as deep as 16 cm in both months. In the upper layers of the sediment to a depth of 8 cm, the average number of seed coats was 7960 ± 2997 m⁻² in November and 16,318 ± 2922 m⁻² in March. Deeper than 8 cm, the number of seed coats gradually decreased owing to decomposition, and none was found below 16 cm. We used the density of reproductive shoots and number of seeds per spadix in Tategami to estimate the fate of seeds and seed coats of the annual type of Z. marina in bottom sediments: out of the 6000 seeds m⁻² produced annually, 72% disappears from the stand and 28% is buried locally. The density and distribution of Z. marina seeds are among the most important factors in maintenance and propagation of the annual population at Tategami.     

Germination response and viability of an endangered tropical conifer Widdringtonia whytei seeds to temperature and light

The tropical conifer Widdringtonia whytei Rendle is an endangered species endemic to Mulanje Mountain in Malawi. A study was conducted for the first time under controlled conditions in order to assess the effects of temperature and light on germination and viability of W. whytei seeds. Seeds incubated at a constant temperature of 20 °C attained the highest cumulative germination percentage (100%) followed by 87% germination under fluctuating temperatures of 15 °C night/25 °C day. No seed germination occurred at temperatures below 15 °C. Seeds that failed to germinate at temperatures below 15 °C showed the highest (> 90%) viability compared to the seeds incubated at 25 °C (60%). Across temperature regimes, germination was significantly higher under light (44.7%) than dark (35.6%) conditions. It is concluded that temperature is one of the critical factors for germination of W. whytei seed. The ability of W. whytei seeds to germinate both in light and darkness implies that the species would unlikely form a persistent soil seed bank, an attribute which is common in species that survive in habitats frequently disturbed by fires.     

Reproductive responses of ram lambs under short-term exposure to endophyte-infected tall fescue seed

Our objective was to determine the influence of short-term exposure to endophyte-infected tall fescue on reproductive function of ram lambs. Rams (214 days of age) were fed a diet free of endophyte-infected fescue seed (EF; n = 8) or endophyte-infected fescue seed (EI; n = 9; 34% of diet; 4.8 μg g⁻¹ ergovaline) for 6 weeks. Feed offered to EF rams, individually fed, was reduced to the average intake of EI lambs from previous day so that intake was similar between treatments and averaged 2.4% BW (DM basis), leading to daily intake of 33.7 μg ergovaline kg⁻¹ BW for the EI fed lambs. Daily high ambient temperature for the trial ranged between 16 and 26 °C. Respiration rate and rectal temperature were measured at 14:00 daily. Blood was collected for serum concentrations of prolactin (weekly) and testosterone (twice weekly). Body weight and body condition scores (BCS; 1 = thin; 5 = fat) were determined every 14 days. Scrotal circumference, scrotal skin temperature, and semen characteristics were determined weekly. Rams were slaughtered after 6 weeks of feeding. Signs of fescue toxicosis in EI fed rams included increased rectal temperature (P < 0.001, R² = 0.11) and respiration rate (day, P < 0.001, R² = 0.25) when high ambient temperature exceeded 22 °C and reduced serum concentrations of prolactin (diet × day, P < 0.001). Body weight of EI fed rams tended to decrease after 36 days of feeding compared with EF fed rams (−3.0 kg versus 0.51 kg; P < 0.07) and BCS was similar between treatment throughout the trial. Serum concentrations of testosterone were greater in EI compared with EF fed rams (diet × day, P < 0.005, R² = 0.08). Scrotal skin temperature, scrotal circumference, semen volume, percent sperm motility, and percent abnormal sperm were similar between treatments. Spermatozoa concentration tended to be greater in EF compared with EI fed rams after 43 days of feeding (P < 0.10; R² = 0.15). Rate of forward movement of spermatozoa tended to increase at a greater rate between Days 8 and 29 in EF compared with EI fed rams (diet × day, P < 0.08). Feeding endophyte-infected fescue seed to ram lambs was associated with potential decreased fertility and increased serum concentrations of testosterone. Short term exposure of endophyte toxins to male ruminants may negatively impact reproductive responses. Feeding for longer periods may further reduce fertility and merits further research.     

UVC-priming mediated modulation of forskolin biosynthesis key genes against Macrophomina root rot of Coleus forskohlii⿿A tissue culture based sustainable approach

Coleus forskohlii is susceptible to root rot disease that reduces yield of root specific metabolite forskolin. In this communication, we first time reported sustainable management of Macrophomina phaseolina root rot in C. forskohlii via Ultraviolet-C (UVC)-hormesis using tissue culture. Compact calli were developed using 2,4-dichlorophenoxy acetic acid (2.0 mg l^⿿1) and tolerance level of UVC exposure was optimized. Calli exhibited 88% viability, 8.4% relative growth in biomass and higher absorbance in 2,3,5-triphenyl tetrazolium chloride analysis subsequent to 10 min of UVC exposure. Pathogenicity of M. phaseolina was reduced in UVC-primed calli-regenerated plants compared with un-primed ones under controlled condition. Plant height, leaf number, shoot and root biomass significantly improved in UVC-primed calli-regenerated plants over un-primed ones when challenged by M. phaseolina in glasshouse. Significant elevation of economically important forskolin via up-regulation of key forskolin biosynthesis genes GGPP synthase, CfTPS2 and CfTPS3 highlighted the potentiality of UVC-priming to boost plant⿿s defence mechanism and protect from drastic economic loss.