Molecular phylogeny and pathotyping of Fusarium solani: a causal agent of Dalbergia sissoo decline

Sissoo (Dalbergia sissoo), commonly known as shisham, is amongst the finest woods of South Asia, but ‘wilt’ disease has caused a rapid decline in this species. The cause of the disease remains uncertain. The aim of this study is to identify the causal agent of the disease and characterize isolates made from diseased trees, based on genomic data and variations in virulence. Samples of infected roots, stems and the ooze exuded from infected trees were obtained from plants showing symptoms in different geographical regions of India for the isolation of microorganisms. Isolates were used to inoculate healthy plants. Based on the morphological characteristics, genus‐ and species‐specific PCR, and in silico analysis of 5.8S rDNA‐ITS regions, of the 38 fungal isolates, 24 and 14 were identified as Fusarium solani and Fusarium sp., respectively. In a pathotyping study, eighteen F. solani isolates, isolated from roots and stem parts of symptomatic plants, induced typical wilt symptoms when inoculated through soil and roots on D. sissoo seedlings of 1–15 months in age. The population of F. solani was the highest in infected roots and the lowest in parts of stems, gradually decreasing with height, and was isolated constantly up to approximately 40% height of the seedling. F. solani isolates used in inoculations were successfully re‐isolated from the rhizosphere, infected roots and wilted stems, as confirmed using isolate‐specific DNA fingerprints. Molecular phylogenies based on rDNA‐ITS sequences showed that the 38 isolates fell into 2 groups. Group I comprised of F. solani isolates from D. sissoo and F. solani sequences in the NCBI GenBank database, whereas group II included Fusarium isolates other than F. solani. These results are helpful in developing integrated control measures for this highly variable pathogen and to establish a base for future population studies.     

Effect of vitamins,probiotics and protein level on semen traits and seminal plasma biochemical parameters of post-moult male broiler breeders

1. A study was designed to investigate the comparative effect of supplementary vitamin E, vitamin C, probiotics and dietary crude protein concentration on semen traits and seminal plasma biochemical parameters in male broiler breeders after Zn-induced moulting.

2. A total of 180 male broiler breeders were induced to moult at 65 weeks of age by mixing ZnO in diet at the rate 3000?mg/kg of feed. After moulting, the males were divided into six groups that were given feed supplemented with: vitamin C (500?IU/kg), vitamin E (100?IU/kg), protein (140?g CP/kg), probiotics (50?mg/kg) and the combination of these components, while one group was kept as a control. Semen samples were collected weekly and semen volume, spermatozoa concentration, motility and dead spermatozoa percentage were determined. Seminal plasma was separated to determine the concentration of total antioxidant capacity (TAC), homocysteine, paraoxonase (PON1), arylesterase, ceruloplasmin, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities.

3. Semen volume was significantly higher in the vitamin E and C groups compared to the control. Spermatozoa motility was higher in the vitamin E group and dead spermatozoa percentage was significantly lower in the vitamin C group compared with the control group.

4. Seminal plasma TAC was higher in the vitamin E group, homocysteine was lower in the vitamin C and E groups. PON1 was higher in the combination group. Arylesterase increased significantly in the vitamin C and combination groups over time. Seminal plasma AST was significantly lower in the vitamin C and E supplemented groups whereas ALT decreased significantly only in the vitamin E group compared with the control. Higher concentrations of ceruloplasmin were observed in the combination group compared with the other treatments.

5. It was concluded that additional vitamin E and C or their combination was the most potent nutrient treatment for improving the semen traits and seminal plasma biochemical characteristics in male broiler breeders after Zn-induced moulting.     

Development and Characterization of Calcium-Alginate Beads of Apigenin: In Vitro Antitumor,Antibacterial, and Antioxidant Activities

The objective of this work was to develop sustained-release Ca-alginate beads of apigenin using sodium alginate, a natural polysaccharide. Six batches were prepared by applying the ionotropic gelation technique, wherein calcium chloride was used as a crosslinking agent. The beads were evaluated for particle size, drug loading, percentage yield, and in vitro drug release. Particle size was found to decrease, and drug entrapment efficiency was enhanced with an increase in the polymer concentration. The dissolution study showed sustained drug release from the apigenin-loaded alginate beads with an increase in the polymer proportion. Based on the dissolution profiles, BD6 formulation was optimized and characterized for FTIR, DSC, XRD, and SEM, results of which indicated successful development of apigenin-loaded Ca alginate beads. MTT assay demonstrated a potential anticancer effect against the breast cancer MCF-7 cell lines. The antimicrobial activity exhibited effective inhibition in the bacterial and fungal growth rate. The DPPH measurement revealed that the formulation had substantial antioxidant activity, with EC50 value slightly lowered compared to pure apigenin. A stability study demonstrated that the BD6 was stable with similar (f2) drug release profiles in harsh condition. In conclusion, alginate-based beads could be used for sustaining the drug release of poorly water-soluble apigenin while also improving in vitro antitumor, antimicrobial, and antioxidant activity.     

Closing the nitrogen use efficiency gap and reducing the environmental impact of wheat-maize cropping on smallholder farms in the Guanzhong Plain,Northwest China

A high crop yield with the minimum possible cost to the environment is generally desirable. However, the complicated relationships among crop production, nitrogen (N) use efficiency and environmental impacts must be clearly assessed. We conducted a series of on-farm N application rate experiments to establish the linkage between crop yield and N₂O emissions in the Guanzhong Plain in Northwest China. We also examined crop yield, partial factor productivity of applied N (PFPN) and reactive N (Nr) losses through a survey of 1 529 and 1 497 smallholder farms that grow wheat and maize, respectively, in the region. The optimum N rates were 175 and 214 kg ha^–1 for winter wheat and summer maize, respectively, thereby achieving the yields of 6 799 and 7 518 kg ha^–1, correspondingly, with low N₂O emissions based on on-farm N rate experiments. Among the smallholder farms, the average N application rates were 215 and 294 kg ha^–1 season^–1, thus producing 6 490 and 6 220 kg ha^–1 of wheat and maize, respectively. The corresponding PFPN values for the two crops were 36.8 and 21.2 kg N kg^–1, and the total N₂O emissions were 1.50 and 3.88 kg ha^–1, respectively. High N balance, large Nr losses and elevated N₂O emissions could be explained by the overdoses of N application and low grain yields under the current farming practice. The crop yields, N application rates, PFPN and total N₂O for wheat and maize were 18 and 24% higher, 42 and 37% less, 75 and 116% higher, and 42 and 47% less, correspondingly, in the high-yield and high-PFPN group than in the average smallholder farms. In conclusion, closing the PFPN gap between the current average and the value for the high-yield and high-PFPN group would increase crop production and reduce Nr losses or the total N₂O emissions for the investigated cropping system in Northwest China.     

Influence of nitrogen application on dry biomass allocation and translocation in two maize varieties under short pre-anthesis and prolonged bracketing flowering periods of drought

Plants have evolved different mechanisms to survive under stress conditions. This field study was conducted to evaluate the influence of nitrogen (N) application on dry biomass allocation and translocation in two maize varieties under short pre-anthesis and prolonged bracketing flowering period of drought. Two maize varieties, ‘Pioneer 30B80? and ‘Suwan 4452? receiving N at 0 (control), 160 (optimal) and 320 (supra-optimal) kg ha^?1 were subjected to short pre-anthesis and prolonged bracketing flowering periods of drought. Prolonged bracketing flowering period of drought had more suppressive effect on anthesis-silking interval, dry matter allocation and translocation, leaf greenness, contribution of current assimilates to grain (CCAG), kernel number, kernel weight and kernel yield of two maize varieties than a short pre-anthesis drought. Nitrogen application at optimal level was the best for all traits, except CCAG. The maize variety ‘Pioneer 30B80? performed better under both drought types due to more root xylem vessels of large size and more accumulation of dry matter in leaves and roots than the variety ‘Suwan 4452?. Therefore, the variety ‘Pioneer 30B80? may be planted in drought prone environments and may be used in breeding program aimed at developing drought-tolerant cultivars.     

Floristic composition and ecological gradient analyses of the Liakot Forests in the Kalam region of District Swat,Pakistan

Kalam Kohistan is a dry,temperate hilly region in the Hindu Raj Series of the Greater Hindu Kush Mountains with diverse forests.As plant distribution and composition is expression of a range of various environmental variables,ecological and floristic attributes of vegetation in the Laikot Forests were therefore evaluated via consecutive explorative trips.These forests have not been evaluated ecologically before due to harsh climatic conditions and issues of accessibility and the study in hand is one of the first in its nature.Quantitative ecological techniques were used to sample forest vegetation and identify species in quadrats of 2 m×2 m for herbs,5 m×5 m for shrubs and 10 m×10 m for trees.All the data of 195 plant species and environmental factors were analyzed via Two-way Cluster Analysis and Canonical Correspondence Analysis using PCORD and CANOCO software.Among 195 plant species from 63 families were 27 species of Asteraceae,the most-speciose family,followed by Poaceae(20 spp.).The dominant life form was therophyte(86 spp.;44%share),followed by phanerophyte(41 spp.;21%share).For the leaf size spectrum,most prevalent was nanophyll(73 spp.;44%)followed by microphyll(66 spp.,34%)and mesophyll(44 spp.;23%).Calcium carbonate,pH,potassium,sand,silt,organic matter,slope aspect and grazing significantly influenced species composition,distribution and habitat.The floristic variation in the region was diverse due to elevational and aspect gradients and thus devided into three zones.This vegetation zonation can be utilized for forest management,species and habitat conservation.Both in situ and ex situ conservation of threatened plant species may improve their conservation status in future if address properly.     

Chemical and organic immobilization treatments for reducing phytoavailability of heavy metals in copper‐mine tailings

This study was conducted to evaluate the efficiency of diammonium phosphate (DAP), agricultural limestone (lime), and green‐waste compost mixed with 30% treated sewage sludge (GCS) applied alone or in combination as chemical immobilization treatment using tomato as a test crop. Mine waste was collected from an abandoned copper‐mine tailing site at Mynydd Parys, Anglesey (UK). Lime was applied at the rate of CaCO₃ equivalent (CCE, pH = 7), DAP at the rate of 23 g P per kg substrate, and 10% by weight, GCS as sole application. Half rate of each amendment was also tested as a combined treatment and an untreated substrate (control). Plant‐available metals (Cd, Cu, Fe, Pb, and Zn) were measured in substrate with conventional diethylenetriaminepentaacetic acid (DTPA) and sequential Ca(NO₃)₂ extraction. Plant–dry biomass yield was significantly (p < 0.001) increased by the combined application of all the three amendments while sole application of DAP reduced yield by 4‐fold compared to unamended soil probably due to P toxicity. Addition of lime reduced the DTPA‐extractable Cu, Fe, and Zn by 75%, 81%, and 85%, respectively, while Pb availability was reduced by 88% in combined DAP + lime + GCS treatment compared to control. The extraction capacity of DTPA was higher than that of Ca(NO₃)₂ by 3‐fold for Cu and Fe, 8‐fold for Pb, and 2‐fold for Zn. The leaf‐tissue concentrations of Cu and Fe were reduced by 77% and 83% in the lime + GCS amendment, respectively, while both Pb and Zn were reduced by 89% and 33%, respectively, in substrate treated with the combined application of all three amendments. These results suggest that alkaline amendments (both lime and GCS) were effective in reducing the phytoavailability of Cu, Fe, and Zn while DAP mixed with either GCS or lime was effective in reducing Pb availability.     

Regulation of DNA synthesis in Leydig cells obtained from neonatal pig testes.

Three distinct waves of Leydig cell development are found in the pig testes, which occur during fetal, perinatal, and prepubertal periods. Proliferation of Leydig cells is primarily regulated by luteinizing hormone (LH); however, effects of LH on proliferation of immature rat Leydig cells are mediated by specific growth factors and cytokines such as transforming growth factor-alpha (TGFalpha), insulin-like growth factor-1 (IGF-1), interleukin-1beta (IL-1beta), steroidogenesis-inducing protein (SIP), and TGFbeta. The objective of the present study was to identify growth factors that could possibly be involved in the proliferation of Leydig cells in the neonatal pig testis. Leydig cells were isolated from 3- to 5-d-old pig testes, cultured for 48 hr in serum-free media, washed, and treated with hCG and/or IGF-1, epidermal growth factor (EGF), IL-1beta, SIP, and TGFbeta for 18 hr. Tritiated thymidine incorporation into DNA was assessed over a subsequent 4-hr period. Incorporation of [3H]-thymidine was stimulated by hCG treatment with a 2.3-fold increase over control cultures. SIP also induced a significant increase (P < 0.0001) in the incorporation of [3H]thymidine into Leydig cell DNA. Similarly, EGF and IGF-1 also increased DNA synthesis in neonatal porcine Leydig cells, whereas IL-1beta had no effect. TGFbeta had very little, if any, effect on DNA synthesis when added alone, but inhibited the stimulatory effects of other mitogens (SIP, hCG, EGF/TGFalpha, and IGF-1). Our results indicate that these growth factors may play a role in the LH/hCG-dependent proliferation of Leydig cells during the perinatal period of development.     

Straw mulching with fertilizer nitrogen: An approach for improving crop yield,soil nutrients and enzyme activities

Field experiments were conducted to study soil properties, soil enzymes activities, water use efficiency (WUE) and crop productivity after six years of soya bean straw mulching in the semi‐arid conditions of China. The experiment included four treatments: CK (Control), N (240 kg N ha^‐1), H (soya bean straw mulching at half rate 700 kg ha^‐1 with 240 kg N ha^‐1) and F (soya bean straw mulching at full rate 1,400 kg ha^‐1 with 240 kg N ha^‐1). Soil organic carbon (SOC), soil labile organic carbon (LOC), soil available N (AN), available P (AP) and enzyme activities were analysed after wheat harvesting in 2016 and 2017. Results show that straw amounts had positive effects on the soil fertility indices being higher for treatment F. The SOC, LOC, AN, AP and enzyme activities (i.e. saccharase, urease and alkaline phosphatase) were in the order of F > H > N > CK. High wheat grain yield and WUE were observed for F treatment. A total of six years mulching along with 240 kg ha^‐1 nitrogen fertilizer application is sufficient for wheat yield stability and improving soil properties except urease activities in the semi‐arid condition of China. However, the straw mulching amount should be further studied with minimum nitrogen fertilizer for an environment‐friendly and effective approach for improving the soil biological properties with adequate crop production on a sustainable basis in the semi‐arid region of China.     

Spatial and temporal variation in otolith chemistry and its relationship with water chemistry: Stock discrimination of Sperata aor

Long‐whiskered catfish, Sperata aor (Hamilton 1822), is commercially important in food, ornamental and sport fisheries. The fish is mainly caught from the wild populations because its aquaculture practices are not commercialised. Inland fishery in the Ganga basin is mostly unorganised; hence, no published report is available on the trend of S. aor production from the selected habitat. In India, S. aor has been categorised vulnerable mainly due to natural and anthropogenic threats. Otolith chemistry shows variation with changing physico‐chemical conditions of the fish habitat. Therefore, the present study was conducted with the objective to analyse spatio‐temporal variations in water chemistry in relation to environmental factors; relationship between water and otolith chemistry; and spatio‐temporal variations in otolith chemistry to discriminate the stocks of S. aor inhabiting the River Ganga. Most of the element: Ca ratios in water samples did not show significant correlations with environmental factors, viz. temperature and conductivity. Only few element: Ca concentrations in otoliths were positively correlated to their corresponding ratios in the ambient water. In the selected study area, the S. aor populations were discriminated into four stocks possibly because of heterogeneous water chemistry at the sampling sites, and physical barriers. In the present study, otolith chemistry showed relatively low temporal variability as compared to spatial variability; thus, the classification accuracy of individuals to their original populations remained consistent over the selected time period. The findings could be useful in devising scientifically sound management strategies and/or any conservation plans for the vulnerable S. aor populations inhabiting the River Ganga.