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Sclerotinia sclerotiorum is a necrotrophic fungus causing devastating stem rot and associated yield losses of canola/rapeseed (Brassica napus) worldwide, including in Australia. Developing host resistance against Sclerotinia stem rot is critical if this disease in canola/rapeseed is to be successfully managed, as cultural or chemical control options provide only partial or sporadic control. Three B. napus breeding populations, C2, C5 and C6, including the parents, F1, F2, BC1P1, and BC2P2, were used in a field study with an objective of exploring the inheritance pattern of disease resistance (based on stem lesion length [SLL]) and the genetic relationships of disease with stem diameter (SD) or days to first flowering (DTF), and to compare these new adult plant stem resistances against S. sclerotiorum with those of seedling (cotyledon and leaf) resistances in earlier studies. Heritability (broad sense) of SLL was 0.57 and 0.73 for population C2 at 3 and 5 weeks postinoculation and 0.21 for population C5 at 5 weeks postinoculation. Additive genetic variance was evident within all 3 populations for DTF but not for SD. Narrow-sense heritability for DTF was 0.48 (C2), 0.42 (C5), and 0.32 (C6). SD, DTF, and SLL were all inherited independently, with no significant genetic covariance between traits in bivariate analysis. Genetic variance for SLL in populations C2 and C5 was entirely nonadditive, and there was significant nonadditive genetic covariance of SLL at 3 and 5 weeks postinoculation. Generation means analysis in population C2 supported the conclusion that complex epistatic interactions controlled SLL. Several C2 and C5 progeny showed high adult plant stem resistance, which may be critical in developing enhanced stem resistance in canola/rapeseed. Although population C6 showed no genetic variation for SLL resistance in this study, it showed significant nonadditive genetic variance at the cotyledon and leaf stages in earlier studies. We conclude that host resistance varies across different plant growth stages, and breeding must be targeted for resistance at each growth stage. In populations C2, C5, and C6, resistance to S. sclerotiorum in stem, leaf, and cotyledon was always controlled by nonadditive effects such as complex epistasis or dominance. Overall, our findings in relation to the quantitative inheritance of Sclerotinia stem rot resistance, together with the new high-level resistances identified, will enable breeders to select/develop genotypes with enhanced resistances to S. sclerotiorum.

Background Bactrocera cucurbitae (Coquillett) (Diptera: Tephritidae) is the most destructive pest of bitter gourd. Conventionally, it is controlled by chemical insecticides despite their hazardous impacts. Entomopathogenic fungi are considered as eco-friendly and safer alternative of chemical. This study aimed to evaluate the efficacy of Beauveria bassiana and Metarhizium anisopliae against B. cucurbitae on bitter gourd. Results The study revealed that B. bassiana and M. anisopliae were efficient against B. cucurbitae in controlled as well as field conditions. Both mode of application (i.e., contact and oral) were found to be effective; however, high efficacies of them were observed through contact application (73.43 and 59.72%), respectively. Pathogenicity of both fungi increased with increase in concentration and time intervals. Under field conditions, significantly low fruit infestations and significantly a high population reduction of B. cucurbitae were observed at 30 DAT when both fungi were applied with 108 CFU/ml concentrations. Conclusions Beauveria bassiana and M. anisopliae had the potential to be used against B. cucurbitae; however, the former showed high efficiency comparatively.

In the present study, fifteen cucumber genotypes were screened against Zucchini yellow mosaic virus (ZYMV) and root-knot nematode (Meloidogyne incognita) individually and in combinations. All the cucumber genotypes behaved differently regarding root and shoot lengths and weights, number of galls, eggmasses and ZYMV infection when inoculated with M. incognita and ZYMV alone and in combinations. None of the fifteen cucumber genotypes was found immune, highly or moderately resistant to M. incognita. Two genotypes viz., Alpha Prime and Patio were found resistant to the nematode. Contrarily, Max Pack and Beti-alpha were highly susceptible as evident by maximum galls on their roots and reductions in growth variables. Similarly, the genotypes Best PIC, Songrooh, Northern Pack, C-7 and C-5 appeared as moderately susceptible whereas Shaheen, Jakson, C-1, C-2, C-3 and C-9 genotypes showed susceptible reactions to the nematode alone. Similar trend was noticed when the genotypes were inoculated with both the nematode and the virus. Similarly, on the basis of disease rating scale, two genotypes viz., Alpha Prime and Patio were grouped as moderately resistant to ZYMV. On the other hand, eleven genotypes viz., Max Pack, Shaheen, Songrooh, Northern Pack, Jakson, Beti-alpha, C-1, C-2, C-3, C-5 and C-9 were catalogued as highly resistant while two genotypes Best PIC and C-7 were categorized as susceptible against ZYMV alone. Similarly, all the genotypes behaved in the same way when inoculated with M. incognita. Two genotypes Alpha Prime and Patio were found resistant against both the pathogens inoculated simultaneously and hence are recommended for cultivation. © 2021 Friends Science Publishers

Blue mold caused by Penicillium species is a major fungal disease threatening the viticulture industry in Pakistan, responsible for deteriorating the quality of grapes during handling, transportation, and distribution. Identification-based approaches of Penicillium species provide a better strategy on accurate diagnosis and effective management. In this study, 13 isolates were recovered from symptomatic grape bunches at five main fruit markets of Rawalpindi district, Punjab province. Based on morphological data and multi-loci phylogenetic analysis, the isolates were identified as two distinct species viz. Penicillium expansum (eight isolates) and Penicillium crustosum (five isolates). Meanwhile, the pathogenicity test of Penicillium isolates presented by the inoculation of grape bunches showed various degrees of severity. For improving the fruit quality and eliminating the needs of synthetic fungicides, botanicals such as thyme (Thymus vulgaris L.), fennel (Foeniculum vulgare L.), garlic (Allium sativum L.), ginger (Zingiber officinale L.), and carum (Carum capticum L.) essential oils (EOs) at concentrations of 0.6, 0.9, 1.2, and 1.5 mg/mL were evaluated. In vitro studies indicated that thyme EO showed a highly significant reduction of fungal growth. Furthermore, the experiments related to reducing the decay development and average weight loss percentage of grapes revealed similar findings. Based on the results of this study, it is recommended that thyme EO can be used as an eco-friendly botanical fungicide against Penicillium spp. causing blue mold disease.

Oat predominately cultivated as forage crop and is not explored for its value-added bioactive moieties. This study was aimed to characterize five selected oat (Avena sativa L.) cultivars with respect to organic sugars, antioxidant activities, pesticide residues, mycotoxin and phenolic acids as measured HPLC. Five indigenous oat cultivars (Avon, PD2LV65, S2000, SGD2011, SGD81) were profiled for seven phenolic acids (gallic acid, vanillic acid, caffeic acid, syringic acid, p-coumaric acid, chlorogenic acid and ferulic acid). Highest concentrations of vanillic acid, gallic acid, caffeic acid, syringic acid, ferulic acid, p-coumaric acid and chlorogenic acid were detected in the SGD2011, SGD81, S2000, SGD2011, S2000, SGD2011 and PD2LV65 respectively. TPC in case of all oat cultivars ranged 50.88–101.56 mg/100 g. Among all oat cultivars, SGD-81 exhibited the highest DPPH activity, TPC, TFC and TFoC contents whereas, SGD-2011 showed the highest total anthocyanin content followed by AVON. S2000 showed the highest concentration of mycotoxin M1 (18.51 ppm), while lowest M1 content (10.51 ppm) were exhibited by SGD-81. Abamectin and permethrin pesticide residues were undetected in oat cultivars whereas, Dieldrin was only detected in SGD-81 and chloropyrifos was not detected in S2000 and SGD-81. Mandipropamid, parathion methyl, chlorthalonil, indoxacarb and lufernon were detected in the highest concentrations in SGD2011 and S2000 oat cultivars. Moreover, fructose, glucose and sucrose were found in highest amounts in S2000, PD2LV65 and SGD2011, respectively. Conclusively, it was evident from the results that SGD-81, SGD2011 and SG2000 could be utilized as source of important and promising phytochemicals of nutraceutical significance. Moreover, significant potential of these oat cultivars can be harnessed owing to value addition and exploitation in formulations of functional food ingredients and foodstuffs.

Brinjal (Solanum melongena L.) is the most important vegetable crop in the Asia Pacific region, with Pakistan being the seventh largest producer. There are many biotic and abiotic factors affecting its production in terms of quantity and quality including insect pests as a significant constraint. In Pakistan, the brinjal shoot and fruit borer, Leucinodes orbonalis is the major insect pest of brinjal. However, some other chewing and sap-sucking insects are also essential pests. Bt-brinjal, a transgenic variety, has been introduced in India and Bangladesh for the management of lepidopteran pests. Before the commercialization of transgenic brinjal in Pakistan, it must be assessed for its effects on the environment and non-target organisms. This study is presenting the overview of transgenic brinjal as an option to manage brinjal shoot and fruit borer along with current and future challenges in areas of its commercialization.

Climate warming is impacting the phenology, growth and productivity of diverse cropping systems at local, regional and global levels. Long-term observed chickpea-mungbean system (CMS) phenological changes were used for the determination of the relationship between crop practices, climate warming and phenology for the making strategies for CMS to minimize negative climate change impacts. Observed thermal trend from sowing to maturity was ranging from 0.82 to 1.15 °C decade−1 for chickpea and 0.64 to 0.97 °C decade−1 for mungbean during 1980–2018. Observed chickpea phenology stages was earlier for mean value of 7.04 (sowing; S), 6.76 (emergence; E), 4.31 (anthesis; A), 2.15 (maturity; M) days decade−1, whereas chickpea phases were decreased averagely 2.73 (S–A), 2.16 (A–M), 4.89 (S–M) days decade−1. Mungbean, ‘S’ 6.24, ‘E’ 5.97, ‘A’ 3.76, and ‘M’ 2.01 days decade−1 were occurred earlier. Period of mungbean phenology phases were lessened with averaged 2.45 (S–A), 1.76 (S–M) and 4.23 (A–M) days decade−1, respectively. Phenological stages and phases of both crops chickpea and mungbean correlated negatively with rising temperatures at all sites studied. By using CROPGRO-Chickpea and CROPGRO-Legume models for usual chickpea and mungbean cultivars at the sites for 38 years duration indicated that model predicted phenology stages were accelerated with thermal trend more as compared with observed stages. This showed that, during last decades, growing newly evolved cultivars of pulses having more thermal time requirement have significantly offset the increased temperature induced changes in chickpea (33%) and mungbean (20%) phenology. Therefore, for the mitigation of climate warming influences, newly evolved cultivars for CMS must be familiarized that need greater demand for degree days and having higher tolerance to temperature.

Kinnow orchards grown in different agro-ecological regions of Punjab, Pakistan, namely Sargodha, Toba Tek Singh (TTS) and Vehari districts, were selected to assess the effect of climate variables on fruit-bearing patterns. Experiment was laid out in RCBD while selecting identical features Kinnow plants and labeled twigs at analogous canopy positions in all three sites. Temperature was reported higher in TTS and Vehari areas, while relative humidity in Sargodha accounted for different levels of agrometeorological indices by computing more variations in warm districts. Climate variables influenced fruit-bearing habits and vegetative growth trend in all three flushes while recording heavy fruit-bearing plants during on-year and light fruit-bearing in off-year at Vehari. Similarly, three vegetative flushes were recorded unevenly in all three sites due to different fruit-bearing patterns induced by climate variables. Harvesting pattern of orchards began earlier in Sargodha, where maximum orchards were harvested before new flowering to add evenness to fruiting habits during on & off-years. In warm conditions, fruit ripening arrived in the peak of winter and mostly domestic market-driven harvesting resulted in late start of fruit picking with more erratic fruit-bearing habits. Both physiological and pathological fruit drops have been significantly affected by climate variables with a higher degree of physiological drop in warm regions and pathological effects in the humid conditions of Sargodha on heavy fruit-bearing plants. Fruit yield and grading quality were also affected in both seasons by showing more asymmetrical trend in yield and fruit grading in warm areas of TTS and Vehari due to an irregular fruiting pattern compared to Sargodha. From now on, the climate variables of the three sites directly influenced the fruiting patterns, vegetative flushes, fruit drops, yields and grades of Kinnow mandarin.

Pepper is one of the most important vegetables and spices in the world. Principal pungency is contributed by secondary metabolites called capsaicinoids, mainly synthesized in the placenta of pepper fruit. Various factors, including drought, limit pepper production. Flowering is one of the most sensitive stages affected by drought stress. The current study was conducted to determine the effect of drought on different pepper genotypes at the flowering and pod formation stages. Hot pepper (Pusajuala and Ghotki) and Bell pepper (Green Wonder and PPE-311) genotypes were subjected to drought (35% field capacity) at two different stages (flowering (DF) and pod formation (DP) stage). In comparison, control plants were maintained at 65% field capacity. The data regarding flowering survival rates, antioxidant protein activity, and proline content, were collected. Results indicated that parameters like flower survival percentage, number of fruits per plant, and fruit weight had significant differences among the genotypes in both treatments. A high proline level was observed in Green Wonder at the pod formation stage compared to other genotypes. Capsaicin contents of hot pepper genotypes were affected at the pod formation stage. Antioxidants like GPX were highly active (190 units) in Ghotki at pod formation. Bell pepper genotypes had a high APX activity, highly observed (100 units) in PPE-311 at pod formation, and significantly differ from hot pepper genotypes. In the catalase case, all the genotypes had the highest values in DP compared to control and DF, but Pusajuala (91 units) and Green Wonder (83 units) performed best compared to other genotypes. Overall, the results indicate that drought stress decreased reproductive growth parameters and pungency of pepper fruit as most of the plant energy was consumed in defense molecules (antioxidants). Therefore, water availability at the flowering and pod formation stage is critical to ensure good yield and pepper quality.

Environmental consciousness as well as individual”™s demand for ready to eat food, recently, has changed the trends in food packaging leading to the development of biodegradable and edible packaging. Emulsified edible films have better transparency, superior mechanical properties and provide barriers to water and other atmospheric gases. Edible films if not consumed, biodegrad chemically. In present study, edible films were, initially, prepared using Chitosan and Aloe vera at different concentrations. Films were then subjected to physical and mechanical testing. Films with 20% Aloe vera had low thickness as compared to films with no Aloe vera. These films also had superior mechanical properties and lower water vapor permeability. Films with 20% Aloe vera were, then, selected and beeswax was dispersed in Chitosan-Aloe vera solution at concentration upto 2.0% followed by film preparation through casting technique. Thickness and water vapor permeability were observed to be improved with increase in concentration of beeswax. Tensile strength of edible films was also improved 1.3 times when concentration of beeswax increased from 0.5 to 2.0%. Percentage elongation decreased with increase in beeswax concentration in the emulsified films. No change in particle size was observed with change in concentration of beeswax. Emulsions were also stable at room temperatures. Decrease in transparency of emulsified edible films was observed with increase in beeswax content in the emulsified films. In addition, cost analysis of the films proved them reasonable to be used as an alternate of synthetic packaging materials.

The phenological changes of a long-term observed rice-wheat system (RWS) were used to determine the relationships among management practices, climate change, and crop phenology to devise adaptation strategies for RWS for mitigating the potential impact of climate change. The study comprised of 10 sites of observed and simulated rice-wheat system phenological data for the historical period from 1980 to 2014 in Punjab, Pakistan. The observed climate warming from sowing or/transplanting to maturity ranged from 0.50 to 1.20 °C decade−1 for rice and 0.77 to 1.07 °C decade−1 for wheat. The observed rice phenological stages were advanced by an average of 7.90 (sowing (S)), 6.60 (transplanting (T)), 4.30 (panicle initiation (PI)), 5.00 (anthesis (A)) and 6.40 (maturity (M)) days decade−1, while rice phenological phases were reduced by an average of 1.4 (S-T), 6.40 (T-M), 3.00 (PI-A), 4.70 (PI-M) and 4.10 (A-M) days decade−1. For wheat, sowing (S) and emergence (E) dates were delayed by an average of 9.50 and 1.30 days decade−1, while anthesis (A) (5.30 days decade−1) and maturity (M) (5.40 days decade−1) dates were advanced. The duration of wheat phenological phases was reduced by an average of 5.50 (S-A), 5.70 (S-M) and 4.60 (A-M) days decade−1. The S and E dates were positively correlated with increasing temperature and the A and M dates and phases (S-A, A-M, and S-A) were negatively correlated with increasing temperature for all study locations. Using the CSM-CERES-Rice and CSM-CERES-Wheat models for standard, field-tested cultivars of rice and wheat for all locations for the 35-year period showed that the simulated phenology stages were earlier with climate warming compared to the observed phenology stages. A significant portion of the negative impact of warming on rice (35%) and wheat (21%) was offset by growing new cultivars that had higher thermal time requirements. Thus, to mitigate climate change impacts, new cultivars for RWS should be introduced that require higher growing degree days and have a high temperature tolerance.

Heat shock proteins (Hsps) are stress-responsive molecular chaperones, which uphold proper protein folding in response to external and internal stresses. The Hsp100 gene family plays a substantial role in thermos-tolerance of plants. This study investigated evolutionary relationship and expression of ClpB/Hsp100 gene family in tomato under heat stress. Six SlHsp100 genes were identified using bioinformatics tools. In silico sub-cellular localization indicated that of these 6 ClpB/Hsp100 members, 4 are found in chloroplast, 1 in mitochondria and 1 in the cytoplasm. For evolutionary study, 36 SlHsp100 genes were included in the phylogenetic tree showing a hierarchical clustering shared by the members of the kingdoms Plantae, Archaea, Chromista, Fungi and Bacteria. A total 4 pairs of orthologous and 5 pairs of paralogous genes were identified. Functional divergence between different Hsp100 clusters showed considerable functional homology. Thermo-tolerance measured in terms of cell viability, cell membrane stability and pollen viability indicated that it was paralleled by thermal resistance of Hsps. Reverse transcriptase polymerase chain reaction was used to analyze gene expression in leaves of five-week-old tomato seedlings following exposure to heat stress (45°C) and control (25°C). Chloroplastic LeHSP110/ClpB gene was upregulated in all tomato genotypes after exposure to heat stress highlighting the crucial role of this gene family in acquired thermo-tolerance.

Research to control yield losses from Sclerotinia (Sclerotinia sclerotiorum) in oilseed rape (Brassica napus) has focused on stem resistance. However, resistance to leaf infection against this pathogen would also be beneficial, both in limiting additional plant leaf damage and in reducing inoculum build up within-crop and resultant spread onto stems. Three B. napus breeding populations, C2 (NC-8 × RQ-001-NCA-8 NC2-7), C5 (cv. Charlton × RQ-001-NCA-8 NC2-7) and C6 (cv. Charlton × NC4-5), were screened for leaf resistance (based on mean lesion diameter) under controlled environment conditions. Each population consisted of parents (P1 and P2), F1, F2, BC1P1 and BC2P2, except for C5 that lacked BC1P1. Moderate broad sense heritability for leaf resistance (0.45) to S. sclerotiorum was only found in population C6, where genetic variance was mostly non-additive. Analyses of generation means and variances indicated that both dominance and complex epistatic interactions were present in C6. Bivariate analysis revealed a positive genetic covariance between the non-additive effects for mean leaf lesion and cotyledon lesion diameters, and significant negative covariance of residuals, which supports a common genetic control of cotyledon and leaf resistance to S. sclerotiorum. These results will guide breeders in selection and development of genotypes with both cotyledon and leaf resistance against this important pathogen worldwide.

Sclerotinia rot, caused by the fungal pathogen Sclerotinia sclerotiorum, is a devastating disease on oilseed rape and mustard worldwide. While the focus to effectively control yield losses from Sclerotinia has been on locating stem resistance, resistance to leaf and to early (cotyledon) stage resistance to this pathogen are also important, both not only limiting additional plant damage, but also inoculum build up and spread onto stems. Three Brassica napus breeding populations developed in India, C2 (NC-8 × RQ-001-NCA-8 NC2-7), C5 (cv. Charlton × RQ-001-NCA-8 NC2-7) and C6 (cv. Charlton × NC4-5), were screened for cotyledon resistance (based on lesion diameter) under controlled environmental conditions to investigate the inheritance of disease resistance. Each population consisted of parents (P1 and P2), F1, F2, BC1P1 and BC2P2, except for population C5 which lacked BC1P1. Moderate broad sense heritability of 0.42, 0.31 and 0.49 for cotyledon resistance was found in populations C2, C5 and C6, respectively, and there was mostly non-additive genetic control of resistance. Analyses of generation means and variances indicated that the additive-dominance model was adequate to explain genetics of cotyledon resistance in population C2. There was heterosis for susceptibility (i.e., larger lesion diameter). The dominance × dominance digenic epistasis explained genetic control in population C6, with heterosis towards resistance (i.e., smaller lesion diameter). This information is critical for breeding for resistance to this important pathogen in Australia, India and elsewhere.

Understanding the influence of thermal trends, crop management practices, and genetics on the crop developmental stages and phases is critical to develop adaptation strategies in the face of warming trends. The specific study objectives were to determine the correlation between observed potato phenology with the trends of rising temperature, and to investigate the impacts of thermal trend, crop management practices, and changes in cultivars using a modeling approach. The study was conducted at 12 sites in Punjab, Pakistan from 1980 to 2018 using phenological observations for both the spring and autumn potato crop. For the stages observed during spring, there was an average advance of 6.2 days decade−1 for sowing, 6.0 for emergence, 3.8 for tuber initiation, and 2.0 for maturity. However, for the stages observed during autumn, there was an average delay of 5.2 days decade−1 for sowing, 5.1 for emergence, 3.3 for tuber initiation, and 2.3 for maturity. The average phase duration decreased on average by 2.4 days decade−1 for sowing to tuber initiation, 1.8 days decade−1 for tuber initiation to maturity for spring, and 4.2 days decade−1 for sowing to maturity. The average autumn phase duration decreased on average by 1.9 days decade−1 for sowing to tuber initiation, 1.0 days decade−1 for tuber initiation to maturity, and 2.9 days decade−1 for sowing to maturity. With respect to the local weather observation, the average air temperature had increased 0.8 °C decade−1 for spring and autumn from 1980 to 2018. The differences in spring and autumn phenology had a statistically significant negative correlation with the increase in temperature from 1980 to 2018. When the CSM-SUBSTOR-Potato model was used for a standard variety across locations and years, the predicted phenological stages, on average, occurred earlier due to increase in temperature from 1980 to 2018, while there was less impact on the observed phenological stages. This indicated that during the last four decade, adaptation strategies such as earlier planting for spring potato, and later planting for autumn, as well as the release of new cultivars that require more thermal time compared to the traditional cultivars have been implemented by growers have compensated for some part of temperature induced warming trends of spring and autumn potato phenology.

Understanding the effects of warming trends, genetics, and management on crop phenology is crucial for adaptation against the increasing trend in temperature in the upcoming decades. The goal of this research was to quantify the impact of historic climate warming and management on sunflower phenology (stages and phases). In this project, spring and fall sunflower phenology and daily weather data were obtained for 20 locations in Punjab, Pakistan. We investigated the impact of thermal trend on the phenological stages and phase duration for spring and fall sunflower from 1980 to 2016. The results showed that the mean temperature increased by 0.9 and 0.8 °C decade−1 for spring and fall, respectively. The observed phenological stages such as sowing, emergence, anthesis, and maturity dates were earlier for spring by an average of 6.6, 6.3, 3.8, and 2.2 days decade-1and delayed for fall by an average of 5.7, 5.4, 3.1, and 1.8 days decade-1. The observed sowing to anthesis, anthesis to maturity, and sowing to maturity phases were reduced by an average of 2.8, 1.6 and 4.4 days decade-1for spring and 2.5, 1.2, and 3.8 days decade-1 for fall. The changes in phenology of sunflower for spring and fall were highly correlated with the enhancement in warming trend during 1980–2016. The CSM-CROPGRO-Sunflower model using standard hybrids for all sites and years showed that the simulated phenology had accelerated due to the change in climate and the simulated phenological dates were earlier than the observed dates. These results indicate that earlier sowing dates for spring, delayed sowing dates for fall, and shifts of varieties that require elevated total growing degree days for the duration of 1980 to 2016 have somewhat mitigated the adverse impact of climate warming on spring- and fall-grown sunflower phenology for ensuring sustainable productivity and ensuring food security.

A field experiment with five nitrogen (N) application rates (120, 140, 160, 180, 200 kg hm−2) under peanut/maize intercropping regime was conducted to evaluate cropping patterns and N application rates on yield, soil enzyme activities, and rhizosphere microbial community structure. The field experiment showed that the land equivalent ratio (LER) in maize/peanut intercropping system was not decreased as compared with the monoculture pattern, though the total N supply was approximately reduced by 20%. Terminal restriction fragment length polymorphism (T-RFLP) results showed that the treatments of intercropping (IMP100%, IMP80% and IMP60%) increased the abundance of Rhizobium hainanense, Rhizobium leguminosarum and Frankia which are associated with nitrogen fixation as compared to monoculture pattern (M100% and P100%). The qPCR results revealed that the copy number of nifH was significantly higher in the treatments of intercropping than in monoculture pattern. The result was also consistent with the T-RFLP analysis. In conclusion, peanut and maize under the condition of intercropping can promote the population of microorganisms associated with nitrogen-fixing in the rhizospheric soil. Our results provide a theoretical basis and technical support to soil N utilization, development of low carbon agriculture and protection of farmland ecological environment.

The human serum is a vital component of the innate immunity of the host that acts as the first line of defence against invading pathogens. A key player in serum-mediated innate immune defence is a system of more than 35 proteins, collectively named as the complement system. After exposure of the pathogen, these proteins are activated in a cascade manner, ultimately forming a membrane attack complex (MAC) on the surface of the pathogen that directly lyses the bacterial cell. Formation of the MAC can be demonstrated in vitro by using serum bactericidal assay (SBA) that works in the absence of cellular components of blood after incubating the serum along with bacteria. Here, we describe the age-related differences in the bactericidal activity of human serum against Pseudomonas aeruginosa, an opportunistic human pathogen causing an array of hospital and community-acquired infections. We demonstrate that adult sera were highly effective in the in vitro killing of Pseudomonas aeruginosa as compared to children and the elderly (p < 0.0001). Sera from children were seriously compromised in the killing P. aeruginosa, whereas elderly sera showed a reduced level of killing. Data revealed a positive correlation between age and serum-killing with higher coefficient of determination values of 0.34, 0.27, and 0.58 and p values of < 0.0001, < 0.001, and < 0.0001, respectively, after 60, 90, and 120 minutes of incubation. Hence, our study highlights the age-related difference in the bactericidal activity of human sera. We conclude that sera of children are totally compromised, whereas elderly sera are only partially compromised, in the killing of P. aeruginosa.

Crop production is greatly impacted by growing season duration, which is driven by prevailing environmental conditions (mainly temperature) and agronomic management practices (particularly changes in cultivars and shifts in sowing dates). It is imperative to evaluate the impact of climate change and crop husbandry practices on phenology to devise future management strategies to prepare for climate change. Historical changes in spring and autumn maize phenology were observed in Punjab, Pakistan during 1980–2014. Sowing (S) of spring maize was earlier by an average of 4.6 days decade−1, while autumn maize ‘S’ and emergence (E) were delayed on average 3.0and 1.9 days decade−1. Observed anthesis (A) plus maturity (M) dates were earlier by 7.1 and 9.2 days decade−1 and 2.8 and 4.4 days decade−1for spring and autumn maize, respectively. Similarly, S-A, S-M and A-Mphases were shortened on average by 2.4, 4.6 and 1.9 days decade−1 and 5.5, 7.8 and 2.2 days decade−1 for spring and autumn maize, respectively. The variability in phenological phases of spring and autumn maize had significant correlation,with the increase in temperature during 1980–2014. Employing the CSM-CERES-Maize model using standard hybrid for all locations and years illustrated that model-predicted phenology has accelerated with climate change more than infield-observed phenology. These findings suggest that earlier late sowing and shifts of cultivars requiring high total growing degree day during 1980–2014, have partially mitigated the negative impact of climate change on phenology of both spring and autumn grown maize.

Yield is influenced by the length of the growing season, which is affected by weather conditions and management practices of a crop, including sowing dates and shifting of cultivars. It is necessary to understand the effects of agronomic management practices and weather variables on phenological stages and crop phases in order to develop strategies for adaptation of agricultural systems to changes in climatic conditions. The goal of this study was to determine the impact of warming trends on phenology of canola from 1980 to 2014 for central and southern Punjab, Pakistan. Sowing, emergence, anthesis and physiological maturity dates were delayed by an average of 6.02, 3.14, 3.31 and 1.89 days per decade, respectively. The duration of sowing to anthesis, sowing to physiological maturity and anthesis to physiological maturity phases decreased an average 2.71, 4.13 and 1.42 days per decade, respectively, for all 10 locations that were analysed in this study. The sowing, emergence, anthesis and physiological maturity dates were positively correlated with an increase in temperature by an average 2.71, 1.41, 1.49 and 0.85 days per °C, respectively. However, the phenological phases such as sowing to anthesis, anthesis to maturity and sowing to maturity were negatively correlated with an increase in temperature by an average of 1.22, 0.64 and 1.86 days per °C, respectively, for all 10 locations. Applying a process-based CSM-CROPGRO-Canola model using a standard cultivar (field tested) for all locations and years indicated that the simulated phenological stages occurred earlier due to the warming trend compared to the observed phenological stages. One-quarter of the negative effects of this thermal trend was compensated by growing new cultivars that had higher thermal time requirements. Therefore, new canola genotypes with a higher number of growing degree day requirement and high temperature tolerance should be a priority for evolving new cultivars.

Cereals are grown as foods and consumed by humans. Cereals are vital source of carbohydrates, vitamins, fats, minerals, oils, and protein. Among cereals, wheat (Triticum aestivum L.) is sown globally for its highly nutritive and useful grain. Cultivation of wheat started from ancient times and domesticated. About thousands years it has been a major staple food of many civilizations in Asia, Europe, and Africa. Mostly, wheat crop is grown in temperate region and where average rainfall is between 10 and 70 inches. In order to feed the burgeoning population of the world, advanced wheat production technology should be adopted.

Stresses induced by abiotic and biotic factors badly effect sessile natured plants. Several innate mechanism has been established by plants during evolution. Expression of specialized proteins under harsh conditions to mitigate the effects is one of the conserved mechanisms. Heat shock proteins (hsp) are one of those conserved proteins. Among HSP, Hsp70 holds critical role in stress tolerance as well as development. HSP70 are structurally conserved across the genomes and reported to mitigate stress at cellular level. At the onset of stress, HSP70 releases HSF to translocate and transcribe HSP resulting in increased tolerance. While, abundant HSP70 bind to HSF to down-regulate the expression of HSP. Hsp70 associated with mitochondria and endoplasmic reticulum respond to accumulation of unfolded proteins (unfolded protein response) and repair them. Moreover, they are also associated to control programmed cell death in plants by interacting with BAG proteins.

High temperature has deleterious impacts on tomato growth and development and limits its production. Acetyl salicylic acid (ASA) and 24-epibrassinolide (EBL) have been widely reported as stress-ameliorating agents. The effect of exogenous application of varying levels of EBL (0.75, 1.5, and 3 µM) and ASA (0.25, 0.75, and 1.25 mM) on root activity (RA) in terms of 2,3,5 triphenyl tetrazolium chloride (TTC) reduction and root morphological features was evaluated in four-week-old tomato seedlings (cultivar: Mei Jie Lo) grown under high-temperature stress (46°C/4 h per day) for 21 days. The daily heat stress treatment almost ceased the root growth of chemically untreated seedlings. However, both EBL and ASA significantly attenuated the deleterious impacts of heat stress to different extents regarding root activity, total root length, surface area, volume, and number of nodes and connections. Different concentrations demonstrated signature effects. EBL (3 µM) was over all the best treatment to improve root act...

Rehmannia glutinosa, a widely used Chinese medicinal herb, has been shown to suffer from serious consecutive monoculture problems that cause significant decline in both yield and quality. The objective of this study was to evaluate the response of the soil microbial community and the effect on catabolic diversity to consecutive monoculture regime using three different techniques: substrate-induced respiration (SIR), phospholipid fatty acid (PLFA) and community-level physiological profiles (CLPP) analyses. We found that basal soil respiration (BSR) was significantly higher in the control and newly planted soils than in the second and third year consecutive monoculture soils. However, no significant difference was observed in SIR among the newly planted, second and third year consecutive monoculture soils. The PLFA signatures indicated that the bacterial biomass was larger than the fungal biomass in all four treatments and both enhanced with the increasing years of monoculture and attained the peak in SM. The ratio of cyclopropyl PLFAs to their metabolic precursors (cy/pre), a measure of physiological stress in microbial communities, in the second and third year consecutive monoculture soils was significantly greater than that in the control and newly planted soils. Biolog analysis results revealed that the consumption of carboxylic acids, phenolic acids and amines, especially acid carbon substrates, in the consecutively monocultured soil was significantly greater than that in the newly planted soil. Both PLFA- and CLPP-based principal component analysis (PCA) and cluster analysis revealed the distinct separation between the control, newly planted plots and the second, third year consecutive monoculture plots. Through our PLFA-based and Biolog analysis, together with microbial respiration determination, we were able to reveal characteristic differences in the microbial community composition and activities in the rhizosphere following R. glutinosa monoculture.

Continuous cropping obstacle is prevalent for protected cultivated cucumber in China. Intercropping garlic may effectively relieve this obstacle due to its allelopathic and antimicrobial effects. A two-growing season investigation was carried out during autumn 2009 to spring 2010 in plastic tunnel to determine the effects of intercropped garlic and green garlic on the overall growth of cucumber and soil biological properties. Results showed that green garlic exhibited the inhibitory effect on the growth of cucumber in spring cultivation 2010. Garlic–cucumber intercropping system increased yield of cucumber differently depending on garlic cultivars, with only cv. G005 showing significant increase (13.4%). Intercropping systems were evaluated as a greater net benefit system as compared to monoculture cultivation. Populations of soil bacteria and actinomyces were stimulated, while fungi were inhibited under intercropping system. Activities of soil invertase, urease, and alkaline phosphatase were enc...

The objectives of this study were to evaluate the performance of the Cropping System Model (CSM)-CERES (Crop-Environment Resource Synthesis)-Rice for simulating growth and yield of rice under irrigated conditions for a semiarid environment in Pakistan and to determine the impact of plant density and irrigation regime on grain yield and economic returns. The crop simulation model was evaluated with experimental data collected in 2000 and 2001 in Faisalabad, Punjab, Pakistan. The experiment utilized a randomized complete block design with three replications and included three plant densities (one seedling hill−1, PD1; two seedlings hill−1, PD2; and three seedlings hill−1, PD3) and five irrigation regimes (625 mm, I1; 775 mm, I2; 925 mm, I3; 1075 mm, I4; and 1225 mm, I5). To determine the most appropriate combination of plant densities and irrigation regimes, four plant densities from one seedling hill−1 to four seedlings hill−1 and 17 irrigation regimes ranging from 0 to 1600 mm, for a total of 68 different scenarios, were simulated for 35 years of historical daily weather data. The evaluation of CSM-CERES-Rice showed that the model was able to accurately simulate growth and yield of rice for irrigated semiarid conditions, with an average error of 11% between simulated and observed grain yield. The results of the biophysical analysis showed that the combination of the two seedlings hill−1 plant density and the 1,300 mm irrigation regime produced the highest yield compared to all other scenarios. Furthermore, the economic analysis through the Mean-Gini Dominance (MGD) also showed the superiority of this treatment compared to the other treatment combinations. The mean monetary return ranged from −47 to 1,265 $ ha−1 among all 68 scenarios. However, to be able to furnish the demand of rice grain for local consumption and to increase export, there is a need to expand this technology among the rice growers of other rice producing areas in Pakistan through extension workers.

Molecular chaperones of plasmid pBI121 carrying CaMV35S promoter and a nucleotide sequence of plasmid pBI221 were inserted into plasmid pCAMBIA2300 to construct an intermediate vector: pVBG2307. This novel vector pVBG2307 contains a greatly expanded multiple cloning site with an adjacent imported CaMV35S promoter sequence. This vector allows controlled transformation of DNA in both Escherichia coli and Agrobacterium tumefaciens. Cloned PG, orf456, ipt genes and E8, a fruiting promoter, were amplified by PCR of cDNA libraries of Capsicum annum and Lycopersicon esculentum and were then transferred into vector pVBG2307. The viability of this vector was demonstrated, as it regulated PG, orf456, ipt and E8 genes in E. coli and could be transferred into Agrobacterium strain EHA105-4.

In vitro treatment of tissue cultures of two garlic cultivars with crude culture filtrates from Sclerotium cepivorium was used to select somaclonal variants with enhanced resistance to white rot disease. The crude culture filtrate had a negative effect on callus induction and growth. It also inhibited the differentiation of adventitious buds. The inhibitory effects were greater at higher culture filtrate concentration. With increase in concentration of crude pathogen culture filtrate, rate of callus induction, callus survival, relative growth rate of callus, callus differentiation and number of divided shoots per callus decreased. The inhibitory effect for all the parameters was more visible when crude pathogen culture filtrate was used at 50% concentration or more. However, the performance of disease resistant cultivar Hangzhong Red Skin was better than Gailiang (disease susceptible cultivar) at higher concentrations of crude pathogen culture filtrate. Some somaclonal lines and plantlets (predominantly from Hangzhong Red Skin) that showed resistance to Sclerotium cepivorum were obtained at 50%concentration level of crude pathogen culture filtrate.

The overreliance on and overuse of fungicides is not only a health hazard but also induces natural resistance in plant pathogens, resulting in an economic burden on agricultural producers and a potential threat to natural systems across the globe. It is therefore necessary to identify natural substitutes of fungicides. This study was designed to evaluate the inhibitory effect of the root exudates of two different garlic cultivars, cv. Gailiang (G064) and Cangshan (G025), against Phytophthora capsici, a pepper fungus. All treatments (T1 to T4, i.e., 25%, 50%, 75% and 100% root exudate concentrations) of garlic cultivar G025 showed lower inhibition effects than cultivar G064. An intervarietal inhibition effect comparison at 100% concentration (T4) exhibited a 69.24% decline in hyphal growth for G064 compared with 49.06% for G025. The mycelial growth measured in the control was found to be significantly greater compared with the garlic treatments. The results of high performance liquid chromatography (HPLC) revealed that G064 possessed a large amount of allicin compared with G025. Scanning and transmission electron microscopy showed that, compared with the controls and hyphae treated with G025, the fungal hyphae treated with G064 exudates were misshaped, fragmented and had a smaller diameter, as well as empty cytoplasmic contents in the cell wall. Thus, the root exudates of the G064 cultivar had a significant fungicidal effect on P. capsici.

Bergenin is well-established and potent antioxidant compound with known gastro-protective effects. Main aim of the current investigation was to study the urease inhibitory potential of bergenin both experimentally and computationally. The target compound bergenin was isolated from acetone extract of Mallotus philippinensis bark. Bergenin inhibited the Bacillus pasteurii urease (IC50 value: 21.7 ± 0.2 μM). Thiourea was used as standard inhibitor (IC50 value: 25 ± 0.09 μM). Molecular docking studies revealed the potential binding mode of bergenin, according to which, Bergenin penetrated deeply into the active catalytic site of urease and was found in close interaction with nickel ions and the various important amino acid residues surrounding the catalytic site of B. pasteurii urease. This might be an additional gastro-protective mechanism by which, bergenin inhibits urease released by various urease producing bacterial strains especially Helicobacter pylori.

In July 2016, a severe fruit rot was observed on table grapes (Vitis vinifera) from five different fruit packing houses in the Rawalpindi district (33°38′19.2″N, 73°01′45.0″E) of Punjab province, Pakistan. During the survey, dark gray to black mycelium was observed on the surface of the berries and rapidly expanded to entire bunches. After 3 to 5 days of postharvest storage at ambient temperature, prevalence of fruit rot was 100% in all locations of Rawalpindi district. A total of 200 symptomatic berries were surface disinfested with 1% sodium hypochlorite for 2 min, rinsed three times with sterile distilled water, and then dried on a filter. Excised pieces of sterile tissue (4 mm²) at lesion margins were transferred to potato dextrose agar (PDA) and incubated at 25 ± 2°C for 3 to 5 days. Fungal cultures derived from the isolates gave rise to colonies that were dark olivaceous green with light brown margins, velvety, appressed, and cottony at the center. Conidiophores were septate, bent or sometimes branched, 25.6 to 55.4 μm long and 2.5 to 3.6 μm wide. Conidia were borne singly or in short chains and were obpyriform to obclavate, measuring 9.5 to 55.5 × 6.1 to 25.6 μm with zero to three longitudinal and two to six transverse septa. Morphological characters matched those of Alternaria alternata (Fr.) Keissl (Simmons 2007). For molecular identification, ITS1, 5.8S; ITS2, RNA polymerase II (RPB2); and endopolygalacturonase (endoPG) gene of two representative isolates (Alt 01 and Alt 05) were amplified with primers ITS1/ITS4 (White et al. 1990), fRPB2-7cF/fRpb2-7cR (Liu et al. 1999), and PG3/PG2b (Andrew et al. 2009), respectively. The amplicons were sequenced and deposited in GenBank (accession nos. MG763130, MF785102, MG773211, MG773212, MG786192, and MG786193). A BLAST search of the GenBank database revealed close identity (98 to 100%) with the sequences of various A. alternata isolates (accession nos. KU182490 and MF141012 for ITS gene, KX938348 and KX938349 for RPB2 gene, and KY969535 and KY969535 for endoPG gene) of A. alternata in NCBI. To confirm pathogenicity, 10-µl aliquots of conidial suspension (10⁶ spores/ml) were pipetted onto three nonwounded and four wounded asymptomatic grape berries per isolate. The berries were incubated at 25 ± 2°C in sterile moist chambers, and the experiment was conducted twice. After 3 days, dark gray to black mycelium, similar in appearance to the original isolates, developed on both wounded and nonwounded inoculated berries, whereas control berries inoculated with sterile distilled water remained symptomless. The morphology of the fungus isolated on PDA was identical to that of the original cultures. A. alternata has been reported to cause postharvest fruit rot on grapes in South Africa (Swart and Holz 1991) and Slovakia (Kakalikova et al. 2009). To our knowledge, this is the first report of A. alternata causing fruit rot of grapes in Pakistan, where the disease poses a significant threat to sustainability of grape producers.

Resources are limited, thus improving resource use efficiency is a key objective for cereal-based cropping systems. This field study was carried out to quantify resource use efficiencies in selected C3 and C4 cereals under split nitrogen (N) application regimes. The study included the following treatments: six cereals (three C3: wheat, oat, and barley; and three C4: maize, millet, and sorghum) and four split N application regimes (NS1 = full amount of N at sowing; NS2 = half N at sowing + half N at first irrigation; NS3 = ⅓ N at sowing + ⅓ N at first irrigation + ⅓ N at second irrigation; NS4 = ¼ N at sowing + ¼ N at first irrigation + ¼ N at second irrigation + ¼ N at third irrigation). Results revealed that C4 cereals out-yielded C3 cereals in terms of biomass production, grain yield, and resource use efficiencies (i.e., radiation use efficiency (RUE) and nitrogen use efficiency (NUE)), while splitting N into three applications proved to be a better strategy for all of the selected winter and summer cereals. The results suggest that C4 cereals should be added into existing cereal-based cropping systems and N application done in three installments to boost productivity and higher resource use efficiency to ensure food security for the burgeoning population.

From ancient times, plants are being used in treatment of various diseases. Many of today’s drugs have been derived from plant sources. This research work reveals the indigenous medicinal knowledge of important medicinal plants from district Kotli Azad Jammu and Kashmir (AJK), Pakistan. A total 25 common medicinal plants belonging to the 14 families were reported. Their medicinal and other botanically important uses are described by conducting a meeting and interviews from a total of 137 local inhabitants including 73 males, 47 females and 17 Hakims (herbal specialists). Primary source of indigenous medicines were herbs (56%), shrubs (28%) and trees (16%). Herbal preparations were made by the different plant parts. Most common plant part used to make the herbal preparation was leaf (39%) followed by the root (19%), whole plant (12%), seed (9%), bark (7%), fruit (7%), flower (5%) and tuber (2%). Key words: Ethnobotany, Kotli, Azad Jammu and Kashmir, indigenous knowledge.