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Soil salinization, especially in arid and semi-arid regions, is one of the major abiotic stresses that affect plant growth. To mediate and boost plant tolerance against this abiotic stress, arbuscular mycorrhizal fungi (AMF) symbiosis is commonly thought to be an effective tool. So, the main purpose of this study was to estimate the role of AMF (applied as a consortium of Claroideoglomus etunicatum, Funneliformis mosseae, Rhizophagus fasciculatum, and R. intraradices species) symbiosis in mitigating deleterious salt stress effects on the growth parameters (shoot length (SL), root length (RL), shoot dry weight (SDW), root dry weight (RDW), root surface area (RSA), total root length (TRL), root volume (RV), root diameter (RD), number of nodes and leaves) of Cenchrus ciliaris L. plants through improved accumulations of photosynthetic pigments (chlorophyll a, chlorophyll b, total chlorophyll), proline and phenolic compounds. The results of this experiment revealed that the roots of C. ciliaris plants were colonized by AMF under all the applied salinity levels (0, 75, 150, 225, and 300 mM NaCl). However, the rate of colonization was negatively affected by increasing salinity as depicted by the varied colonization structures (mycelium, vesicles, arbuscules and spores) which were highest under non-saline conditions. This association of AMF induced an increase in the growth parameters of the plant which were reduced by salinity stress. The improved shoot/root indices are likely due to enhanced photosynthetic activities as the AMF-treated plants showed increased accumulation of pigments (chlorophyll a, chlorophyll b and total chlorophyll), under saline as well as non-saline conditions, compared to non-AMF (N-AMF) plants. Furthermore, the AMF-treated plants also exhibited enhanced accumulation of proline and phenolic compounds. These accumulated metabolites act as protective measures under salinity stress, hence explaining the improved photosynthetic and growth parameters of the plants. These results suggest that AMF could be a good tool for the restoration of salt-affected habitats. However, more research is needed to check the true efficacy of different AMF inoculants under field conditions.

A greenhouse experiment was conducted to determine the potential of arbuscular mycorrhizal fungi (AMF) to mitigate the allelopathic effects of invasive Nicotiana glauca on the growth of Acacia gerrardii seedlings. Aqueous extracts from N. glauca leaves and flowers (at concentrations of 20, 60, and 100 g/L) and AMF treatment (±AMF) were applied to A. gerrardii seedlings arranged in a factorial experiment with four replicates. After 6 months, vegetative (plant fresh and dry weight, height, leaf number area), physiological (photosynthesis, stomatal conductance, transpiration rate), chlorophyll a and b, proline, and root (length, surface area, diameter, volume, tip number) parameters as well as root distribution in five root diameter classes (0–5 mm, in 1 mm increments) were examined. The presence of AMF increased the seedling growth parameters. The 20 g/L treatment stimulated most of the vegetative growth, root, chlorophyll, and photosynthesis parameters at both inoculant levels for all extract types and concentrations, and the 100 g/L treatment inhibited seedling growth whether inoculated with AMF or not. This decline was alleviated in AMF-treated seedlings. For both extract sources, root length decreased as extract concentration increased, and roots N. glauca on A. gerrardii seedling growth.

The world's population is increasing and is anticipated to spread 10 billion by 2050, and the issue of food security is becoming a global concern. To maintain global food security, it is essential to increase crop productivity under changing climatic conditions. Conventional agricultural practices frequently use artificial/chemical fertilizers to enhance crop productivity, but these have numerous negative effects on the environment and people's health. To address these issues, researchers have been concentrating on substitute crop fertilization methods for many years, and biofertilizers as a crucial part of agricultural practices are quickly gaining popularity all over the globe. Biofertilizers are living formulations made of indigenous plant growth-promoting rhizobacteria (PGPR) which are substantial, environment-friendly, and economical biofertilizers for amassing crop productivity by enhancing plant development either directly or indirectly, and are the renewable source of plant nutrients and sustainable agronomy. The review aims to provide a comprehensive overview of the current knowledge on microbial inoculants as biofertilizers, including their types, mechanisms of action, effects on crop productivity, challenges, and limitations associated with the use of microbial inoculants. In this review, we focused on the application of biofertilizers to agricultural fields in plant growth development by performing several activities like nitrogen fixation, siderophore production, phytohormone production, nutrient solubilization, and facilitating easy uptake by crop plants. Further, we discussed the indirect mechanism of PGPRs, in developing induced system resistance against pest and diseases, and as a biocontrol agent for phytopathogens. This review article presents a brief outline of the ideas and uses of microbial inoculants in improving crop productivity as well as a discussion of the challenges and limitations to use microbial inoculants.

Nicotian glauca is a noxious invasive shrub in Saudi Arabia, and it is well known for its harmful impact on biodiversity, competing with native plants using various strategies. Among their strategies is their allelochemical activity, i.e., the use of chemicals to dominate and compete. The present study aims to evaluate the allelopathic potential of N. glauca aqueous extracts from leaves, flowers, and twigs on Acacia gerrardi seed germination and seedling growth. Petri dishes containing extracts from N. glauca parts with different concentrations were incubated in a growth chamber. The results indicated that extracts and concentrations negatively impact seed germination and hypocotyl. Relative to the control, the leaf extract with concentrations of 60 and 100 g/L reduced the seed germination rate by 81.11% and 93.33%, respectively. Flower extracts of similar concentrations decreased germination by 81.91 and 92.55%, while the twig extracts declined germination by 79.56 and 95.69%, respectively. The leaf, flower, and twig extracts (100 g/L) decreased hypocotyl radicle by 76.54, 78.05, and 65.75%, respectively. Overall, the concentrations of 20, 60, and 100 g/L showed the lowest growth indices. Generally, aqueous extracts, particularly 100 g/L, impacted the germination and protrusion of A. gerrardii hypocotyl. In conclusion, our study draws attention to the harmful impact of N. glauca on A. gerrardii regeneration.

Abstract Context Withania coagulans (Stocks) Dunal fruits are used in the therapeutics of several ailments due to possessing of potent phytoconstituents which is also used traditionally for curing the diabetes. Objective The present study was assessing the amelioration potential of the phytochemicals of an ethanol fruit extract of W. coagulans (Stocks) Dunal in the HOMA (Homeostatic model assessment) indices and pancreatic endocrinal tissues by inhibition of DPP-4 and antioxidants activities. Material and methods The identification of phytoconstituents of the test extract was performed by LCMS. Further, assessments of in-vitro, in-vivo and in-silico were achieved by following standard methods. In-vivo studies were conducted on type-2 diabetic rats. Results The chosen extract inhibited DPP-4 activity by 63.2% in an in vitro assay as well as significantly inhibit serum DPP-4 levels. Accordingly, the administration of the ethanol fruit extract resulted in a significant (P ≤ 0.001) alterations in the lipid profile, antioxidant levels, and HOMA indices. Moreover, pancreatic endocrinal tissues (islet of Langerhans) appeared to have the restoration of normal histoarchitecture as evidenced by increased cellular mass. Molecular docking (Protein-ligands) of identified phytoconstituents with DPP-4 (target enzyme) shown incredibly low binding energy (Kcal/mol) as required for ideal interactions. ADMET analysis of the pharmacokinetics of the identified phytoconstituents indicated an ideal profile as per Lipinski laws. Conclusion It can be concluded that the phytoconstituents of an ethanol fruit extract of W. coagulans have the potential to inhibit DPP-4 which result in improved glucose homeostasis and restoration of pancreatic endocrinal tissues in type-2 diabetic rats.

Brassinosteroids (BRs) are well recognized for their defensive role in plants under abiotic stress conditions, but 28-homobrassinolide (HBR)-induced tolerance to drought stress has not been reported in soybean (Glycine max L.). The present study investigated the effect of HBR on soybean seedlings under drought stress. Drought stress suppressed growth and photosynthetic systems while increased the proline, glycine betaine (GB), anthocyanin, total phenolic (TP), and total flavonoid (TF) levels in soybean seedlings. HBR restricted reactive oxygen species (ROS) accumulation and decreased the hydrogen peroxide (H2O2) and malondialdehyde (MDA) content by triggering the antioxidant systems. HBR acts as a shield in soybean, protecting the plant against the harmful effects of methylglyoxal (MG) effects by upregulating the enzymes glyoxalase I, (Gly I;15%) and glyoxalase II (Gly II;29.1%) compared to the levels in drought stressed seedlings. Overall, HBR improved drought tolerance in soybean seedlings by modulating osmolytes, the AsA–GSH cycle, and enzyme activities.

By assessing plant species composition and distribution in biodiversity hotspots influenced by environmental gradients, we greatly advance our understanding of the local plant community and how environmental factors are affecting these communities. This is a proxy for determining how climate change influences plant communities in mountainous regions ("space-for-time" substitution). We evaluated plant species composition and distribution, and how and which environmental variables drive the plant communities in moist temperate zone of Manoor valley of Northwestern Himalaya, Pakistan. During four consecutive years (2015–2018), we sampled 30 sampling sites, measuring 21 environmental variables, and recording all plant species present in an altitudinal variable range of 1932–3168 m.a.s.l. We used different multivariate analyses to identify potential plant communities, and to evaluate the relative importance of each environmental variable in the species composition and distribution. Finally, we also evaluated diversity patterns, by comparing diversity indices and beta diversity processes. We found that (i) the moist temperate zone in this region can be divided in four different major plant communities; (ii) each plant community has a specific set of environmental drivers; (iii) there is a significant variation in plant species composition between communities, in which six species contributed most to the plant composition dissimilarity; (iv) there is a significant difference of the four diversity indices between communities; and (v) community structure is twice more influenced by the spatial turnover of species than by the species loss. Overall, we showed that altitudinal gradients offer an important range of different environmental variables, highlighting the existence of micro-climates that drive the structure and composition of plant species in each micro-region. Each plant community along the altitudinal gradient is influenced by a set of environmental variables, which lead to the presence of indicator species in each micro-region.

Philodendron bipinnatifidum Schott ex Endl. is a popular ornamental plant that is normally propagated by tissue culture methods. However, the growth and acclimatization of micropropagated plants are tarrying processes. Therefore, in the present study we examined the effect of arbuscular mycorrhizal fungi (AMF) Gigaspora albida and G. marginata on the success in the establishment, growth, and development of P. bipinnatifidum plantlets during the acclimatization phase. AMF plants had significantly more leaves (10.67 per plant), leaf area (75.63 cm2), plant height (14.17 cm), shoot fresh weight (3.30 g) and shoot dry weight (0.31 g), according to an analysis of growth characteristics. In comparison, non-AMF plants had lower values for these metrics. In addition, AMF plants had significantly longer main roots (23 cm), total length roots per plantlet (485.73 cm), average root diameter (4.58 mm) per plantlet, number of root tips (236) per plant, total root surface area (697.76 cm2), total root volume (79.98 cm3), roots fresh weight (1.51 g), roots dry weight (0.16 g) than non-AMF plants. AMF-treated plants showed better performance in leaf gas exchange, chlorophyll, and carotenoid content. These results emphasize the need for mycorrhization of micropropagated plants to promote vegetative growth, especially during the acclimatization stage.

In order to evaluate the impact of Bacillus sonorensis on propagation of Abelmoschus esculentus and its antimicrobial activity was investigated. In the present study, A. esculentus was cultivated in B. sonorensis inoculated soil and also assessed the morphological as well as biochemical parameters of crops. The culture inoculum of B. sonorensis influenced growth and yield of treated plant. The root (39.5 cm), shoot length (20 cm), chlorophyll (1.06 mg), carotenoid (0.445 mg), protein and total soluble sugar content, NR & starch activity were higher in bioinoculated treated plant than control. The fresh biomass and dry weight were drastically increased in bioinoculated plants when compared to control. Gradual increase in composition of pivotal nutrients (N, P, K) and minor nutrients were observed in the bioinoculated plants. The culture filtrate possessed phytopathogenic activity against different phytopathogens. Among the three different phytopathogens, the maximum zone of inhibition (21 mm) was noticed in Aspergillus colletotrichum infected plants. The fermentation study was carried out in pilot scale fermentor and the synthesis of plant growth promoting substance was found to be 30.00 mg/l IAA.

The presence of secondary metabolite biosynthetic gene clusters (BGCs) makes actinobacteria well-known producers of diverse metabolites. These ubiquitous microbes are extensively exploited for their ability to synthesize diverse secondary metabolites. The extent of their ability to synthesize various molecules is yet to be evaluated. Current advancements in genome sequencing, metabolomics, and bioinformatics have provided a plethora of information about the mechanism of synthesis of these bioactive molecules. Accessing the biosynthetic gene cluster responsible for the production of metabolites has always been a challenging assignment. The genomic approach developments have opened a new gateway for examining and manipulating novel antibiotic gene clusters. These advancements have now developed a better understanding of actinobacterial physiology and their genetic regulation for the prolific production of natural products. These new approaches provide a unique opportunity to discover novel bioactive compounds that might replenish antibiotics’ exhausted stock and counter the microbes’ resistance crisis.

Cadmium and drought are the most destructive of the abiotic stresses with negative consequences in terms of impaired metabolism, restricted nutrient use efficiency and disruptive photosynthesis of plants. The present study investigated the mitigation strategy of both aforementioned stresses by the application of iron oxide (IONPs) and hydrogel nanoparticles (HGNPs) simultaneously probably for the first time. IONPs were biofabricated by using a locally identified Bacillus strain RNT1, while HGNPs were produced chemically followed by the confirmation and characterization of both NPs through nanomaterials characterization techniques. Results of FTIR and XRD showed the capping of NPs by different functional groups together with their crystalline structure, respectively. SEM and TEM analysis showed the spherical shape along with the particle size ranging from 18 to 94 nm of both NPs, while EDS analysis confirmed the elemental purity of NPs. The results revealed that IONPs-treated rice plants increased biomass, antioxidant enzyme contents, photosynthesis efficiency, nutrient acquisition together with the decrease in reactive oxygen species and acropetal Cd translocation under normal and drought stress conditions as compared with control plants. Furthermore, the expression of the Cd transporter genes, OsHMA2, OsHMA3 and OsLCT1 were curtailed in NPs-treated rice plants under normal and drought stress conditions. The overall significance of the study lies in devising the NPs-based solutions of increasing heavy metal pollution and water availability challenges being faced the farmers around the world.

Salt stress is one of the most significant abiotic stresses that substantially negatively impact plant growth and productivity. However, a slew of research thus far has investigated the ameliorating properties of arbuscular mycorrhizal (AM) symbiosis and their potential to improve plant tolerance to salt stress. The present study aimed to evaluate and compare the role of mycorrhizal inocula obtained from Sabkha (S-AMF) and non-Sabkha (NS-AMF) habitats of Saudi Arabia on the morphological, physiological, and biochemical behaviors of the Lasiurus scindicus plant. For this reason, arbuscular mycorrhizal fungi (AMF) isolated from Sabkha and non-Sabkha soils were treated with salinity-exposed L. scindicus. The results revealed that the AMF-treated plants had higher growth metrics and increased synthesis of photosynthetic pigments, which were reduced by salt stress. Furthermore, the application of AM symbiosis induced an increase in the activities of the antioxidant system, which resulted in a reduction of the plant oxidative damage. It was also found that the increased accumulation of proline and phenols acted as a protective measure. Moreover, plants inoculated with S-AMF had the highest ameliorating responses on all the studied parameters compared to NS-AMF. This could be attributed to the presence of habitat-specific AMF, which may have induced adaptive plasticity in plants to tolerate or resist extreme salinity. However, further study in exploring the S-AMF diversity is needed to make it an ecofriendly choice for the restoration of salinity-affected ecosystems.

Upon publication of this article [1], it was brought to our attention that Figs. 3, 4 and 5 are incorrectly presented in the original version of the article. The figures were inadvertently swapped in the original submission and published. Figure 3 should be treated as Fig. 5; Fig. 4 should be 3 and Fig. 5 should be Fig. 4.

Abstract Background Actinobacteria from freshwater habitats have been explored less than from other habitats in the search for compounds of pharmaceutical value. This study highlighted the abundance of actinobacteria from freshwater sediments of two rivers and one lake, and the isolates were studied for their ability to produce antimicrobial bioactive compounds. Results 16S rRNA gene sequencing led to the identification of 84 actinobacterial isolates separated into a common genus (Streptomyces) and eight rare genera (Nocardiopsis, Saccharopolyspora, Rhodococcus, Prauserella, Amycolatopsis, Promicromonospora, Kocuria and Micrococcus). All strains that showed significant inhibition potentials were found against Gram-positive, Gram-negative and yeast pathogens. Further, three biosynthetic genes, polyketide synthases type II (PKS II), nonribosomal peptide synthetases (NRPS) and aminodeoxyisochorismate synthase (phzE), were detected in 38, 71 and 29% of the strains, respectively. Six isolates based on their antimicrobial potentials were selected for the detection and quantification of standard antibiotics using ultra performance liquid chromatography (UPLC–ESI–MS/MS) and volatile organic compounds (VOCs) using gas chromatography mass spectrometry (GC/MS). Four antibiotics (fluconazole, trimethoprim, ketoconazole and rifampicin) and 35 VOCs were quantified and determined from the methanolic crude extract of six selected Streptomyces strains. Conclusion Infectious diseases still remain one of the leading causes of death globally and bacterial infections caused millions of deaths annually. Culturable actinobacteria associated with freshwater lake and river sediments has the prospects for the production of bioactive secondary metabolites.

Plant growth-promoting endophytic bacteria can stimulate the growth, nutrient acquisition, symbiotic performance and stress tolerance of chickpea plants under saline soil conditions. The aim of this study was to investigate the stress-adaptive mechanisms of chickpea plants mediated by Bacillus subtilis (BERA 71) under saline conditions. Inoculation with BERA 71 enhanced plant biomass and the synthesis of photosynthetic pigments and reduced the levels of reactive oxygen species (ROS) and lipid peroxidation in plants under conditions of stress. Furthermore, the activities of ROS-scavenging antioxidant enzymes (superoxide dismutase, peroxidase, catalase and glutathione reductase), the levels of non-enzymatic antioxidants (ascorbic acid and glutathione) and the total phenol content were increased in stressed plants during bacterial association. The bacteria decreased sodium accumulation and enhanced the nitrogen, potassium, calcium and magnesium content in the plants. The suppression of ROS generation and of lipid peroxidation and the accumulation of proline in BERA-71-inoculated plants enhanced the membrane stability under salinity stress and non-stress conditions.

Plant species are distributed in different types of habitats, forming different communities driven by different sets of environmental variables. Here, we assessed potential plant communities along an altitudinal gradient and their associations with different environmental drivers in the unexplored Manoor Valley (Lesser Himalaya), Pakistan. We have implemented various ecological techniques and evaluated phytosociological attributes in three randomly selected 50 m-transects within each stand (a total of 133) during different seasons for four years (2015–2018). This phytosociological exploration reported 354 plant species representing 93 different families. The results revealed that the Therophytic life form class dominated the flora, whereas Nanophyll dominated the leaf size spectra. There were a total of twelve plant communities identified, ranging from the lowest elevations to the alpine meadows and cold deserts. The maximum number of species were found in Cedrus–Pinus–Parrotiopsis community (197 species), in the middle altitudinal ranges (2292–3168 m). Our results showed that at high altitudes, species richness was reduced, whereas an increase in soil nutrients was linked to progression in vegetation indicators. We also found different clusters of species with similar habitats. Our study clearly shows how altitudinal variables can cluster different plant communities according to different microclimates. Studies such as ours are paramount to better understanding how environmental factors influence ecological and evolutionary aspects.

The ecotonal zones support populations that are acclimated to changing, fluctuating, and unstable conditions, and as a result, these populations are better equipped to adjust to expected change. In this context, a hypothesis was tested that there must be vegetation dominated by unique indicator plant species under the influence of ecological gradients in the ecotonal zone of Manoor Valley (northwestern Himalaya), Pakistan. Keeping the aforementioned hypothesis in mind, detailed field studies were conducted during different seasons in 2015-18. Line transect sampling and phytosociological characteristics (density, frequency, cover, and their relative values and Importance Value) were implemented as ecological methods. This investigation documented 97 plant species recorded from seven sampling sites. The community distribution modelling revealed that the ecological variables separate the seven sampling sites into two major plant communities (Indigofera-Parrotiopsis-Bistorta and Ziziphus-Leptopus-Quercus) recognized by TWINSPAN. The IBP communities showed a positive and significant correlation with altitude (1789.6–1896.3 m), sandy soil texture with a slightly acidic pH (6.4–6.5), and higher phosphorous (9–13 mg kg−1). In contrast with this, the ZLQ community was recognized on the southern slope under the strong influence of high electrical conductivity (2.82–5.4 dsm−1), organic matter (1.08–1.25%), calcium carbonate (5.8–7.6 mg kg−1), potassium (202–220 mg kg−1), and temperature (28.8–31.8 °C). Hence, both communities were found on opposite axes with clear differences based on the ecological gradients. NMDS clustered different species with similar habitats and different stands with common species, showing that plant species and stands were in a linear combination with ecological gradients. The IPB community has the maximum number of plant species (87 species), Shannon value (H’ = 4), Simpson value (0.98), and Pielou’s evenness value (0.96). Thus, the multivariate approaches revealed unique vegetation with sharp boundaries between communities which might be due to abrupt environmental changes.

The nomadic pastoral indigenous communities of the Ladakhi people share roots with Tibetan culture in terms of food, clothing, religion, festivals, and habits, and rely widely on plant resources for survival and livelihood. This survey was conducted during 2019–2021 to document the indigenous knowledge about plant resources of the Balti, Beda, and Brokpa communities of the Ladakh region, trans-Himalayas. Open- and close-ended semi-structured interviews (N = 184) and group discussions (N = 17) were used to collect the data. Quantitative data was further analyzed using various statistical tools. A total of 105 plant species belonging to 82 genera and 39 families were used as medicine, fuel wood, fragrance, oil, food, flavor, fodder, decoration, and dye. Among these, medicinal use was most prevalent, with 70% of use reports, followed by fodder and fuel wood. Leaves (27%) were the most preferred plant part used, followed by roots and flowers. The principal component analysis revealed five clusters of ethnobotanical usage, i.e., food, medicine, fuel wood, fodder, and fragrance, oil, dye, and flavor. The maximum number of plant species used was reported by the Brokpa, while the Beda reported the minimum number of plant species uses. Delphinium brunonianum, Waldheimia tomentosa, and Juniperus indica played a significant role in the cultural and religious ritual aspects, whereas Allium przewalskianum, Waldheimia tomentosa, Juniperus indica, and Hippophae rhamnoides were commonly used as a livelihood source among Ladakhi communities. The local people collected most plants (65%) for self-consumption, while the rest (35%) were sold in markets as a source of income. The sustainable utilization and management of plant resources by local people is a strategy to boost livelihoods and food security and alleviate poverty.

This research was performed to explore the vital role of seed priming with a 0.01 µM concentration of brassinosteroids (EBL) to alleviate the adverse effects of Cr (100 µM) in two different rice cultivars. Seed priming with EBL significantly enhanced the germination attributes (germination percentage, germination energy, germination index, and vigor index, etc.), photosynthetic rate as well as plant growth (shoot and root length including the fresh and dry weight) under Cr toxicity as compared to the plants primed with water. Cr toxicity induced antioxidant enzyme activities (SOD, POD, CAT, and APX) and ROS level (MDA and H2O2 contents) in both rice cultivars; however, a larger increment was observed in YLY-689 (tolerant) than CY-927 (sensitive) cultivar. EBL application stimulatingly increased antioxidant enzyme activities to scavenge ROS production under Cr stress. The gene expression of SOD and POD in EBL-primed rice plants followed a similar increasing trend as observed in the case of enzymatic activities of SOD and POD compared to water-primed rice plants. Simultaneously, Cr uptake was observed to be significantly higher in the water-primed control compared to plants primed with EBL. Moreover, Cr uptake was significant in YLY-689 compared to CY-927. In ultra-structure studies, it was observed that EBL priming relieved the rice plants from sub-cellular damage. Conclusively, our research indicated that seed priming with EBL could be adopted as a promising strategy to enhance rice growth by copping the venomous effect of Cr.

Abstract: Plant secondary metabolites play vital role in plant stress response. In this study we investigated whether root colonization of tomato (Solanum lycopersicum) infected by Trichoderma harzianum leads to alterations in the biosynthesis of secondary plant metabolites including phytohormones and osmolyte proline under drought stress. Exposure of tomato to drought caused a drastic decline in plant growth and physiological parameters. Tomato inoculated with T. harzianum showed increased root and shoot growth and chlorophyll pigments as compared to uninoculated controls as well as drought stressed plants. Proline and total soluble protein content was increased in plants inoculated with T. harzianum under both normal as well as drought conditions. An obvious increase in phenol and flavonoid content was observed due to T. harzianum. In addition, T. harzianum inoculated plants maintained higher levels of growth regulators indole acetic acid, indole butyric acid, and gibberellic acid under drought stress. Improved secondary metabolites which play an important role in plant stress tolerance by T. harzianum may have coordinately worked for bringing the growth regulation by protecting membranes from reactive oxygen species (ROS) and enhance plant growth through accessing more nutrients by root system.

In this study, the effects of calcium (Ca2+) application on acquired systemic tolerance mechanism to cadmium (Cd) stress in sesame (Sesamum indicum L.) were studied. The Cd stress reduced the root and shoot growth of sesame, and plant contents of photosynthetic pigments; however, the application of Ca2+ improved these parameters under Cd stress condition. The hydrogen peroxide, malondialdehyde and soluble sugar contents were higher under Cd stress, and were reduced by Ca2+ treatment. The antioxidant enzyme activities in the leaves of sesame, superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), ascorbate peroxidase (APX) and glutathione reductase (GR) were higher under Cd stress, whereas reduced concentration was observed in Ca2+-treated plants. Cd stress increased the contents of diacylglycerol and sterol ester; however Ca2+ treatment resulted in a significant increase in phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol and phosphatidylserine. Our results indicated that application of calcium enables sesame plants to withstand the deleterious impact of cadmium through upregulating acquired systemic tolerance system as lipid fractions (galactolipids, phospholipids, neutral lipids), antioxidant enzymes (SOD, POD, CAT, APX, GR) hence protect membrane functions.

This study was aimed to elucidate the mitigation mechanism of an endophytic bacterium, Bacillus subtilis (BERA 71) against Macrophomina phaseolina (Tassi) Goid disease in mung bean. M. phaseolina reduced the plant growth by inducing disease, hydrogen peroxide (H2O2) and lipid peroxidation. The inoculation of B. subtilis to diseased plants increased chlorophyll, ascorbic acids, and superoxide dismutase, catalase, peroxidase, ascorbate peroxidase and glutathione reductase activities, and while inhibited H2O2 and lipid peroxidation for enhancing plant growth. In addition, B. subtilis association in plants mitigated the M. phaseolina infection due to increase of indole acetic acids and indole butyric acid, and also a decrease of abscisic acid. However, the nutrients (N, K, Ca, Mg, Zn, Cu, Mn and Fe) were increased, except Na in M. phaseolina diseased plants treated with B. subtilis. The result of this study suggests that B. subtilis interaction with plants can modulate the metabolism of pigments, hormones, antioxidants and nutrients against M. phaseolina to induce disease resistance in mung bean.

Resistant Starch (RS), plays a crucial role in human health and nutrition by controlling glucose metabolism. RS or dietary fibre content in rice is low because it goes through a variety of process before it is ready for cooking and consumption. Hence, this study was carried out to develop a rice mutant with increased RS. The rice mutant (γ278) with increased RS was developed by utilizing gamma (γ) rays as a mutagen. Mutant γ278 was characterized for mutations in the starch biosynthetic genes viz., GBSSI, SSI, SSIIa, SSIIIa, SBEIa, and SBEIIb to reveal the functional mutations/variations led to high RS content in rice. A total of 31 sequence variants/mutations in six genes were identified. We report the discovery of three deleterious mutation/variants each in GBSSI, SSIIa, and SSIIIa with the potential to increase RS content in rice. Further, wild × mutant crosses were made to develop an F2 population to study the effect of combination of deleterious mutations. The SNP (GBSSI:ssIIa:ssIIIa) combination responsible for high RS content in F2 population was identified and recorded highest amylose content (AC) (26.18%) and RS (8.68%) content. In conclusion, this marker combination will be highly useful to develop a rice variety with increased RS.

Ajmalicine is naturally present in the root bark of Catharanthus roseus L. and Rauvolfia serpentina (L.) Benth ex.Kurz. It has been extensively utilized in the treatment of hypertension across the world. The increased demand, overconsumption, and low content of the alkaloid in the plants have raised the issue of the depletion of natural sources. The metabolic engineering approach has not been successful in improving the content of the ajmalicine because the metabolic regulation of this metabolite is not known. The regulation of a metabolite in the metabolic pathway requires a tool that can carry out real-time measurement of the flux of the metabolite in living system. Given this, the present study was conducted to develop a genetically encoded FRET-based nanosensor by engineering human Cytochrome P450-2D6, an ajmalicine binding protein. The Cytochrome P450-2D6 was sandwiched between two FRET fluorophores. The design of the nanosensor brings two fluorescent proteins in conjunction with the ajmalicine binding protein, such that it undergoes FRET (Fluorescence Resonance Energy Transfer) upon binding of the ligand. The nanosensor, named as FLIP-Ajn (Fluorescence Indicator Protein for Ajmalicine), was pH stable and ajmalicine specific. The affinity of the FLIP-Ajn was 582 μM. The FLIP-Ajn successfully performed real-time measurement of ajmalicine in prokaryotic (bacteria) and eukaryotic systems (yeast, animal cell line, and plant suspension culture), thereby, establishing its biocompatibility in monitoring of ajmalicine in living cells. Besides, several affinity mutants of the nanosensor were generated through mutations in the ajmalicine binding protein to increase the detection range of the nanosensor at varying physiological scales. The non-invasiveness and high spatial and temporal resolution of the tool holds a great significance in the bio-imaging of a highly compartmentalized metabolic pathway. The flux study of ajmalicine will help in identifying the regulatory steps involved in the synthesis of the alkaloids and, hence, will improve the production rate of ajmalicine from its natural sources.

Fusarium graminearum is a leading plant pathogen that causes Fusarium head blight, stalk rot, and Gibberella ear rot diseases in cereals and posing the immense threat to the microbiological safety of the food. Herein, we report the green synthesis of zinc oxide nanoparticles from Syzygium aromaticum (SaZnO NPs) flower bud extract by combustion method and investigated their application for controlling of growth and mycotoxins of F. graminearum. Formation of SaZnO NPs was confirmed by spectroscopic methods. The electron microscopic (SEM and TEM) analysis revealed the formation of triangular and hexagonal shaped SaZnO NPs with size range 30–40 nm. The synthesized SaZnO NPs reduced the growth and production of deoxynivalenol and zearalenone of F. graminearum in broth culture. Further analysis revealed that treatment of mycelia with SaZnO NPs resulted in the accumulation of ROS in the dose-dependent manner. Also, SaZnO NPs treatment enhanced lipid peroxidation, depleted ergosterol content, and caused detrimental damage to the membrane integrity of fungi. Moreover, SEM observations revealed that the presence of diverged micro-morphology (wrinkled, rough and shrank surface) in the macroconidia treated with SaZnO NPs. Taken together, SaZnO NPs may find a potential application in agriculture and food industries due to their potent antifungal activity.

The present work reports the assessment of the effectiveness of a foliar-spray of salicylic acid (SA) on growth attributes, biochemical characteristics, antioxidant activities and osmolytes accumulation in wheat grown under control (100% field capacity) and water stressed (60% field capacity) conditions. The total available water (TAW), calculated for a rooting depth of 1.65 m was 8.45 inches and readily available water (RAW), considering a depletion factor of 0.55, was 4.65 inches. The water contents corresponding to 100 and 60% field capacity were 5.70 and 1.66 inches, respectively. For this purpose, seeds of two wheat cultivars (Fsd-2008 and S-24) were grown in pots subjected to water stress. Water stress at 60% field capacity markedly reduced the growth attributes, photosynthetic pigments, total soluble proteins (TSP) and total phenolic contents (TPC) compared with control. However, cv. Fsd-2008 was recorded as strongly drought-tolerant and performed better compared to cv. S-24, which was moderately drought tolerant. However, water stress enhanced the contents of malondialdehyde (MDA), hydrogen peroxide (H2O2) and membrane electrolyte leakage (EL) and modulated the activities of antioxidant enzymes (superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), as well as accumulation of ascorbic acid (AsA), proline (Pro) and glycine betaine (GB) contents. Foliar-spray with salicylic acid (SA; 0, 3 mM and 6 mM) effectively mitigated the adverse effects of water stress on both cultivars. SA application at 6 mM enhanced the shoot and root length, as well as their fresh and dry weights, and improved photosynthetic pigments. SA foliage application further enhanced the activities of antioxidant enzymes (SOD, POD, and CAT) and nonenzymatic antioxidants such as ascorbic acid and phenolics contents. However, foliar-spray of SA reduced MDA, H2O2 and membrane permeability in both cultivars under stress conditions. The results of the present study suggest that foliar-spray of salicylic acid was effective in increasing the tolerance of wheat plants under drought stress in terms of growth attributes, antioxidant defense mechanisms, accumulation of osmolytes, and by reducing membrane lipid peroxidation.

This research work reports the magnetic adsorption of fluoride from drinking water through silica-coated Fe3O4 nanoparticles. Chemical precipitation and wet impregnation methods were employed to synthesize the magnetic nanomaterials. Moreover, the synthesized nanomaterials were characterized for physicochemical properties through scanning electron microscopy, Fourier-transform infrared spectroscopy, and X-ray powder diffraction. Screening studies were conducted to select the best iron oxide loading (0.0–1.5 wt%) and calcination temperature (300–500 °C). The best selected nanomaterial (0.5Fe-Si-500) showed a homogenous FeO distribution with a 23.79 nm crystallite size. Moreover, the optimized reaction parameters were: 10 min of contact time, 0.03 g L−1 adsorbent dose, and 10 mg L−1 fluoride (F−) concentration. Adsorption data were fitted to the Langmuir and Freundlich isotherm models. The Qm and KF (the maximum adsorption capacities) values were 5.5991 mg g−1 and 1.869 L g−1 respectively. Furthermore, accelerated adsorption with shorter contact times and high adsorption capacity at working pH was among the outcomes of this research work.

Protected areas are a critical tool to conserve biodiversity in the face of the global crisis of species extinction. Here, we present the first ever management effectiveness assessment of Pakistan’s Protected Areas (PAs). We link these assessments to the delivery of conservation outcomes focusing on the threatened Western Tragopan (Tragopan melanocephalus) endemic to Pakistan and India. We used two approaches, first mapping the spatial distribution of potential habitat coverage using machine learning ensemble models and second, an assessment of the management effectiveness of protected areas. Our results show that only Machiara National Park scored just above 40% (indicating relatively weak management), 22 of the PAs fell within the 25–50% quantile (indicating weak management), and 3 scored below 25% (indicating poor management). PAs within the species distributional range covered 92,387 ha which is only 2% of the total potential habitat of the Tragopan. Scoring of Planning element was insufficient both in term of the site and species. Likewise, inputs (e.g., research and monitoring program, staff numbers, staff training, current budget, security of budget, and management after process) were also inadequate. Finally, we recommend the establishment of more protected areas within the species potential habitat and inclusion of species-specific plans in Pakistan’s PAs management.

Symbiotic arbuscular mycorrhizal fungi (AMF) play a major role in plant development, growth, and relationships with the environment through a change in the accumulation of secondary metabolites; hence, we planned to investigate AMF’s influence on sennoside A and B accumulation in Senna alexandrina (SA) and Senna italica (SI). Seeds of SA (S. alexandrina free of mycorrhizae) and SI (S. italica free of mycorrhizae) were planted in two types of soils: +mycorrhiza and—mycorrhiza. The plant leaves of SA, SI, S. alexandrina with mycorrhizae (SAM) and S. italica with mycorrhizae (SIM) were collected and extracted (with 85% methanol), and sennoside A and B content was evaluated by the HPLC–UV method. The antioxidant activity of SA, SI, SAM and SIM was evaluated by using 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) methods, while antimicrobial properties were evaluated by the minimum inhibitory concentration method (MIC). The AMF colonization was 85.66% and 85%, respectively, in the roots of SA and SI. The HPLC analysis showed a significant increase in (%) the content of sennoside A/sennoside B by 71.11/88.21, respectively, in SAM and 6.76/36.37 in SIM, which clearly indicated positive AMF effects. The DPPH/ABTS [The half maximal inhibitory concentration (IC50): 235.9/321.5 µg/mL] scavenging activity of SAM was comparatively higher and it also exhibited strong antibacterial action (MIC: 156.25 µg/mL), which supported the increase in sennoside content. This finding may be useful for further investigations of the symbiotic relation of mycorrhizal fungi with other plant species.

Reactive oxygen species (ROS) and other free radicals cause oxidative damage in cells under biotic and abiotic stress. Endophytic microorganisms reside in the internal tissues of plants and contribute to the mitigation of such stresses by the production of antioxidant enzymes and compounds. We hypothesized that the endophytic actinobacterium Streptomyces sp. strain DBT34, which was previously demonstrated to have plant growth-promoting (PGP) and antimicrobial properties, may also have a role in protecting plants against several stresses through the production of antioxidants. The present study was designed to characterize catalase and superoxide dismutase (SOD), two enzymes involved in the detoxification of ROS, in methanolic extracts derived from six endophytic actinobacterial isolates obtained from the traditional medicinal plant Mirabilis jalapa. The results of a preliminary screen indicated that Streptomyces sp. strain DBT34 was the best overall strain and was therefore used in subsequent detailed analyses. A methanolic extract of DBT34 exhibited significant antioxidant potential in 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2′-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) assays. The cytotoxicity of DBT34 against liver hepatocellular cells (HepG2) was also determined. Results indicated that methanolic extract of Streptomyces sp. strain DBT34 exhibited significant catalase and SOD-like activity with 158.21 U resulting in a 55.15% reduction in ROS. The IC50 values of a crude methanolic extract of strain DBT34 on DPPH radical scavenging and ABTS radical cation decolorization were 41.5 µg/mL and 47.8 µg/mL, respectively. Volatile compounds (VOC) were also detected in the methanolic extract of Streptomyces sp. strain DBT34 using GC-MS analysis to correlate their presence with bioactive potential. Treatments of rats with DBT34 extract and sitagliptin resulted in a significant (p ≤ 0.001) reduction in total cholesterol, LDL-cholesterol, and VLDL-cholesterol, relative to the vehicle control and a standard diabetic medicine. The pancreatic histoarchitecture of vehicle control rats exhibited a compact volume of isolated clusters of Langerhans cells surrounded by acinies with proper vaculation. An in-vivo study of Streptomyces sp. strain DBT34 on chickpea seedlings revealed an enhancement in its antioxidant potential as denoted by lower IC50 values for DPPH and ABTS radical scavenging activity under greenhouse conditions in relative comparison to control plants. Results of the study indicate that strain DBT34 provides a defense mechanism to its host through the production of antioxidant therapeutic agents that mitigate ROS in hosts subjected to biotic and abiotic stresses.

Isoleucine is one of the branched chain amino acids that plays a major role in the energy metabolism of human beings and animals. However, detailed investigation of specific receptors for isoleucine has not been carried out because of the non-availability of a tool that can monitor the metabolic flux of this amino acid in live cells. This study presents a novel genetically-encoded nanosensor for real-time monitoring of isoleucine in living cells. This nanosensor was developed by sandwiching a periplasmic binding protein (LivJ) of E. coli between a fluorescent protein pair, ECFP (Enhanced Cyan Fluorescent Protein), and Venus. The sensor, named GEII (Genetically Encoded Isoleucine Indicator), was pH stable, isoleucine-specific, and had a binding affinity (Kd) of 63 ± 6 μM. The GEII successfully performed real-time monitoring of isoleucine in bacterial and yeast cells, thereby, establishing its bio-compatibility in monitoring isoleucine in living cells. As a further enhancement, in silico random mutagenesis was carried out to identify a set of viable mutations, which were subsequently experimentally verified to create a library of affinity mutants with a significantly expanded operating range (96 nM−1493 μM). In addition to its applicability in understanding the underlying functions of receptors of isoleucine in metabolic regulation, the GEII can also be used for metabolic engineering of bacteria for enhanced production of isoleucine in animal feed industries.

Phosphate (Pi) deficiency is a critical environmental constraint that affects the growth and development of several legume crops that are usually cultivated in semi-arid regions and marginal areas. Pi deficiency is known to be a significant limitation for symbiotic nitrogen (N2) fixation (SNF), and variability in SNF is strongly interlinked with the concentrations of Pi in the nodules. To deal with Pi deficiency, plants trigger various adaptive responses, including the induction and secretion of acid phosphatases, maintenance of Pi homeostasis in nodules and other organs, and improvement of oxygen (O2) consumption per unit of nodule mass. These molecular and physiological responses can be observed in terms of changes in growth, photosynthesis, and respiration. In this mini review, we provide a brief introduction to the problem of Pi deficiency in legume crops. We then summarize the current understanding of how Pi deficiency is regulated in legumes by changes in the transcriptomes and metabolomes found in different plant organs. Finally, we will provide perspectives on future directions for research in this field.

Cancer is a major cause of death worldwide, with an increasing number of cases being reported annually. The elevated rate of mortality necessitates a global challenge to explore newer sources of anticancer drugs. Recent advancements in cancer treatment involve the discovery and development of new and improved chemotherapeutics derived from natural or synthetic sources. Natural sources offer the potential of finding new structural classes with unique bioactivities for cancer therapy. Endophytic fungi represent a rich source of bioactive metabolites that can be manipulated to produce desirable novel analogs for chemotherapy. This review offers a current and integrative account of clinically used anticancer drugs such as taxol, podophyllotoxin, camptothecin, and vinca alkaloids in terms of their mechanism of action, isolation from endophytic fungi and their characterization, yield obtained, and fungal strain improvement strategies. It also covers recent literature on endophytic fungal metabolites from terrestrial, mangrove, and marine sources as potential anticancer agents and emphasizes the findings for cytotoxic bioactive compounds tested against specific cancer cell lines.

Water deficit and phosphate (Pi) deficiency adversely affect growth and biological nitrogen fixation (BNF) of legume crops. In this study, we examined the impact of interaction between soil water conditions and available soil-Pi levels on growth, nodule development and BNF potential of nine cowpea varieties grown on dry savanna soils. In our experimental design, soils with different available soil-Pi levels, i.e., low, moderate, and high soil-Pi levels, collected from various farming fields were used to grow nine cowpea varieties under well-watered and water-deficit conditions. Significant and severe water deficit-damaging effects on BNF, nodulation, growth, levels of plant-nitrogen (N) and -phosphorus (P), as well as shoot relative water content and chlorophyll content of cowpea plants were observed. Under well-watered and high available soil-Pi conditions, cowpea varieties IT07K-304-9 and Dan'Ila exhibited significantly higher BNF potential and dry biomass, as well as plant-N and -P contents compared with other tested ones. Significant genotypic variations among the cowpeas were recorded under low available soil-Pi and water-deficit conditions in terms of the BNF potential. Principal component (PC) analysis revealed that varieties IT04K-339-1, IT07K-188-49, IT07K-304-9, and IT04K-405-5 were associated with PC1, which was better explained by performance for nodulation, plant biomass, plant-N, plant-P, and BNF potential under the combined stress of water deficit and Pi deficiency, thereby offering prospects for development of varieties with high growth and BNF traits that are adaptive to such stress conditions in the region. On another hand, variety Dan'Ila was significantly related to PC2 that was highly explained by the plant shoot/root ratio and chlorophyll content, suggesting the existence of physiological and morphological adjustments to cope with water deficit and Pi deficiency for this particular variety. Additionally, increases in soil-Pi availability led to significant reductions of water-deficit damage on dry biomass, plant-N and -P contents, and BNF potential of cowpea varieties. This finding suggests that integrated nutrient management strategies that allow farmers to access to Pi-based fertilizers may help reduce the damage of adverse water deficit and Pi deficiency caused to cowpea crop in the regions, where soils are predominantly Pi-deficient and drought-prone.

Plants are subjected to various abiotic stresses, such as drought, extreme temperature, salinity, and heavy metals. Abiotic stresses have negative impact on the physiology and morphology of plants through defects in the genetic regulation of cellular pathways. Plants employ several tolerance mechanisms and pathways to avert the effects of stresses that are triggered whenever alterations in metabolism are encountered. Phytohormones are among the most important growth regulators; they are known for having a prominent impact on plant metabolism, and additionally, they play a vital role in the stimulation of plant defense response mechanisms against stresses. Exogenous phytohormone supplementation has been adopted to improve growth and metabolism under stress conditions. Recent investigations have shown that phytohormones produced by root-associated microbes may prove to be important metabolic engineering targets for inducing host tolerance to abiotic stresses. Phytohormone biosynthetic pathways have been identified using several genetic and biochemical methods, and numerous reviews are currently available on this topic. Here, we review current knowledge on the function of phytohormones involved in the improvement of abiotic stress tolerance and defense response in plants exposed to different stressors. We focus on recent successes in identifying the roles of microbial phytohormones that induce stress tolerance, especially in crop plants. In doing so, this review highlights important plant morpho-physiological traits that can be exploited to identify the positive effects of phytohormones on stress tolerance. This review will therefore be helpful to plant physiologists and agricultural microbiologists in designing strategies and tools for the development of broad spectrum microbial inoculants supporting sustainable crop production under hostile environments.

Plants have evolved several metabolic pathways as a response to environmental stressors such as low temperatures. In this perspective, it is paramount to highlight physiological mechanisms of plant responses to altitudinal gradients as a proxy to evaluate changing environments. Here, we aimed to determine the impact of elevation on the physiological attributes of two plant species along an altitudinal gradient. Our hypothesis was that the altitudinal gradient influences proline, protein, and sugar contents, as well as abscisic acid (ABA) and indole acetic acid (IAA) concentrations. We studied these physiological components in leaves collected from four different altitudinal ranges in Himalayan region of Pakistan from two native herbs, namely Bistorta affinis and Sibbaldia procumbens. Leaves were collected at the initial blooming phase from each altitudinal range, viz. 2850 m, 3250 m, 3750 m and 4250 m. We observed that most abiotic factors decrease with altitude which induces cold acclimation. A significant increase in the concentration of physiological components was observed as altitude increased, except for IAA, which decreased. Furthermore, we did not find variations in proline, ABA and IAA concentrations between species; only sugar and protein, with higher values for B. affinis. We conclude that altitudinal gradients significantly affect the physiological components of B. affinis and S. procumbens in Himalayan region. This result contributes to the understanding of how plants adapt to environmental pressures, acting as a proxy for the evaluation of impacts caused by climate changes.

The present study demonstrates an economical and eco-friendly method for the synthesis of silver nanoparticles (AgNPs) using the wild mushroom Ganoderma sessiliforme. The synthesis of AgNPs was confirmed and the products characterized by UV-visible spectroscopy, dynamic light scattering spectroscopy and X-ray diffraction analysis. Furthermore, Fourier transform infrared spectroscopy (ATR-FTIR) analysis was performed to identify the viable biomolecules involved in the capping and active stabilization of AgNPs. Moreover, the average sizes and morphologies of AgNPs were analyzed by field emission scanning electron microscopy (FE-SEM). The potential impacts of AgNPs on food safety and control were evaluated by the antimicrobial activity of the synthesized AgNPs against common food-borne bacteria, namely, Escherichia coli, Bacillus subtilis, Streptococcus faecalis, Listeria innocua and Micrococcus luteus. The results of this study revealed that the synthesized AgNPs can be used to control the growth of food-borne pathogens and have potential application in the food packaging industry. Moreover, the AgNPs were evaluated for antioxidant activity (DPPH), for biocompatibility (L-929, normal fibroblast cells), and for cytotoxic effects on human breast adenosarcoma cells (MCF-7 & MDA-MB231) to highlight their potential for use in a variety of bio-applications.

By assessing plant species composition and distribution in biodiversity hotspots influenced by environmental gradients, we greatly advance our understanding of the local plant community and how environmental factors are affecting these communities. This is a proxy for determining how climate change influences plant communities in mountainous regions ("space-for-time" substitution). We evaluated plant species composition and distribution, and how and which environmental variables drive the plant communities in moist temperate zone of Manoor valley of Northwestern Himalaya, Pakistan. During four consecutive years (2015-2018), we sampled 30 sampling sites, measuring 21 environmental variables, and recording all plant species present in an altitudinal variable range of 1932-3168 m.a.s.l. We used different multivariate analyses to identify potential plant communities, and to evaluate the relative importance of each environmental variable in the species composition and distribution. Finally, we also evaluated diversity patterns, by comparing diversity indices and beta diversity processes. We found that (i) the moist temperate zone in this region can be divided in four different major plant communities; (ii) each plant community has a specific set of environmental drivers; (iii) there is a significant variation in plant species composition between communities, in which six species contributed most to the plant composition dissimilarity; (iv) there is a significant difference of the four diversity indices between communities; and (v) community structure is twice more influenced by the spatial turnover of species than by the species loss. Overall, we showed that altitudinal gradients offer an important range of different environmental variables, highlighting the existence of micro-climates that drive the structure and composition of plant species in each micro-region. Each plant community along the altitudinal gradient is influenced by a set of environmental variables, which lead to the presence of indicator species in each micro-region.

Plant defensins possess diverse biological functions that include antifungal and antibacterial activities and α-amylase and trypsin inhibitory properties. Two mutations, G9R and V39R, were confirmed to increase the antifungal activity of Raphanus sativus antifungal protein 2 (RsAFP2). Accelerated Molecular Dynamics (aMD) were carried out to examine the conformational changes present in these RsAFP2 mutants, and its two closest homologs compared to the wild-type protein. Specifically, the root mean square fluctuation values for the eight cysteine amino acids involved in the four disulfide bonds were low in the V39R mutant compared to the wild-type. Additionally, analysis of the free energy change revealed that G9R and V39R mutations exert a neutral and stabilizing effect on RsAFP2 conformation, and this is supported by the observed lower total energy of mutants compared to the wild-type, suggesting that enhanced stability of the mutants. However, MD simulations to a longer time scale would aid in capturing more conformational state of the wild-type and mutants defensin protein. Furthermore, the aMD simulations on fungal mimic membranes with RsAFP2 and its mutants and homologs showed that the mutant proteins caused higher deformation and water diffusion than the native RsAFP2, especially the V39R mutant. The mutant variants seem to interact by specifically targeting the POPC and POPI lipids amongst others. This work highlights the stabilizing effect of mutations at the 9th and 39th positions of RsAFP2 and their increased membrane deformation activity.

Wheat cultivars are genetically crossed to improve end-use quality for traits as per demands of baking industry and broad consumer preferences. The processing and baking qualities of bread wheat are influenced by a variety of genetic make-ups, environmental factors and their interactions. Two wheat cultivars, WL711 and C306, derived recombinant inbred lines (RILs) with a population of 206, were used for phenotyping of quality-related traits. The genetic analysis of quality traits showed considerable variation for measurable quality traits, with normal distribution and transgressive segregation across the years. From the 206 RILs, few RILs were found to be superior to those of the parental cultivars for key quality traits, indicating their potential use for the improvement of end-use quality and suggesting the probability of finding new alleles and allelic combinations from the RIL population. Mapping analysis identified 38 putative QTLs for 13 quality-related traits, with QTLs explaining 7.9-16.8% phenotypic variation spanning over 14 chromosomes, i.e., 1A, 1B, 1D, 2A, 2D, 3B, 3D, 4A, 4B, 4D, 5D, 6A, 7A and 7B. In-silico analysis based on homology to the annotated wheat genes present in database, identified six putative candidate genes within QTL for total grain protein content, qGPC.1B.1 region. Major QTL regions for other quality traits such as TKW have been identified on 1B, 2A, and 7A chromosomes in the studied RIL population. This study revealed the importance of the combination of stable QTLs with region-specific QTLs for better phenotyping, and the QTLs presented in our study will be useful for the improvement of wheat grain and bread-making quality.