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Bacillus species synthesize antifungal lipopeptides (LPs) making them a sustainable and eco-friendly management option to combat Fusarium wilt of chickpea.In this study, 18 endophytic Bacillus strains were assessed for their antifungal activity against Fusarium oxysporum f. sp. ciceris (FOC) associated with Fusarium wilt of chickpea. Among them, 13 strains produced significant inhibition zones in a direct antifungal assay while five strains failed to produce the inhibition of FOC. Bacillus thuringiensis CHGP12 exhibited the highest inhibition 3.45 cm of FOC. The LPs extracted from CHGP12 showed significant inhibition of the pathogen. Liquid chromatography-mass spectrometry (LC-MS) analysis confirmed that CHGP12 possessed the ability to produce fengycin, surfactin, iturin, bacillaene, bacillibactin, plantazolicin, and bacilysin. In an in vitro qualitative assay CHGP12 exhibited the ability to produce lipase, amylase, cellulase, protease, siderophores, and indole 3-acetic acid (IAA). IAA and gibberellic acid (GA) were quantified using ultra-performance liquid chromatography (UPLC) with 370 and 770 ng mLIn conclusion, our findings suggest that B. thuringiensis CHGP12 is a promising strain with high antagonistic and growth-promoting potential against Fusarium wilt of chickpea. © 2022 Society of Chemical Industry.

Plant disease complexes are caused by variety of interactions between fungal plant pathogens and plant parasitic nematodes but the most significant and frequent association is Fusarium oxysporum and Root-knot nematodes (Meloidogyne spp). Nematode-fungal interactions are frequently categorized as either synergistic or additive. Different plant-parasitic nematodes caused damage to the host plants depending on their life cycle and feeding habit. Nematode feeding sites are areas with intense metabolic activity, such as syncytia or giant cells, and have a large number of mitochondria and golgi bodies. In addition, cytoplasm is dense and packed with ribosomes. Injuries of host plants are vulnerable to infection by fungi that need assistance to enter their host. Nematode-instigated physiological changes can be fundamental but they still have a lot of detrimental effects. Some nematodes make plant tissues more favorable for pathogen that negatively affects the host by causing them to grow and reproduce more rapidly. Moreover, they alter the root exudates of host plant, promoting the various phases of fungal life cycle like germination, development and reproduction. The importance of nematode-fungus synergistic complex in the development of distinct crop species that exhibit resistance to various biotic or abiotic stresses is only infrequently, if ever described, although numerous studies reveal that resistance breaks down during concomitant infection.

Strigolactone (GR24) is a phytohormone, involved in reconfiguration of plant development pattern in response to salinity stress. Current experiment was carried out in vitro conditions to study the role of GR24 on sunflower (Helianthus annuus L. cv. OLIVER-P) under saline and non-saline growth mediums. Two salinity levels [0 mM (control) and 150 mM NaCl] were maintained in growth mediums for callus induction along with four GR24 levels (0, 0.001, 0.01, 0.1 mg L−1) in Somatic Cell Genetic Lab, Centre of Agricultural Biochemistry and Biotechnology (CABB), University of Agriculture Faisalabad, Pakistan. Results of this study showed that among GR24 levels, 0.01 mg L−1 proved to be ideal dose as it improved callus fresh weight (30.59%), dry weight (45.26%), free protein (55.87%), Ca2+ (13.04%) and K+ (11.76%) contents under non-saline, while superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), free proline and glycine betaine (GB) contents by 38.1%, 67.87%, 124.01%, 63.38% and 76.12%, respectively, under salinity stress mediums. However, water potential (Ψw), osmotic potential (Ψs), turgor potential (Ψp), hydrogen peroxide (H2O2) and malondialdehyde (MDA) were reduced up to 32.65%, 39.80%, 40.74%, 65.78% and 30.76%, respectively, due to different levels of GR24. Overall, high saline conditions caused a significant reduction in concentration of callus biomass, Ψw, Ψs, soluble protein, Ca2+ and K+ ions than non-saline mediums. These results reflected that GR24 application could be an effective approach to mitigate hazardous impacts of salinity in sunflower production. Further, the molecular techniques of processes in which GR24 may play a key role in different commercially vital crops is extreme important and may carry new leads for future experimentation in this exciting area of plant hormones.

Many phytochemicals can affect the growth and development of plants and insects which can be used as biological control agents. In this study, different concentrations of crude, hexane, chloroform, butanol, and aqueous extracts of Euphorbia nivulia Buch.-Ham., an endemic plant of the Cholistan desert in South Punjab of Pakistan, were analysed for their chemical constituents. Their various concentrations were also tested for their phytotoxic and insecticidal potential against duckweed, Lemna minor L., and the dusky cotton bug, Oxycarenus hyalinipennis Costa. various polyphenols, i.e., quercetin, gallic acid, caffeic acid, syringic acid, coumaric acid, ferulic acid, and cinnamic acid were detected in different concentrations with different solvents during the phytochemical screening of E. nivulia. In the phytotoxicity test, except for 100 μg/mL of the butanol extract gave 4.5% growth regulation, no phytotoxic lethality could be found at 10 and 100 μg/mL of all the extracts. The highest concentration, 1000 μg/mL, of the chloroform, crude, and butanol extracts showed 100, 63.1, and 27.1% of growth inhibition in duckweed, respectively. In the insecticidal bioassay, the highest O. hyalinipennis mortalities (87 and 75%) were recorded at 15% concentration of the chloroform and butanol extracts of E. nivulia. In contrast, the lower concentrations of the E. nivulia extracts caused the lower mortalities. Altogether, these findings revealed that E. nivulia chloroform extracts showed significant phytotoxicity while all the extracts showed insecticidal potential. This potential can be, further, refined to be developed for bio-control agents.

Keeping in view the yield losses instigated by heat stress in several crops, we carried out an experiment to explore the curative effect of exogenous applications of proline on the morpho-physiological, biochemical, and water-related attributes of okra genotypes under high-temperature stress (controlled conditions). Four contrasting genotypes C1, C2, C3, and C4 heat tolerant and heat sensitive genotypes were selected from a diverse panel of okra genotypes (n = 100) to examine plant responses to high-temperature stress and exogenous application of proline. Four-week-old seedlings were subjected to heat stress by gradually increasing the temperature of a growth chamber from 28/22 °C to 45/35 °C (day/night) and sprayed with an optimized proline concentration 2.5 mM. The experiment consisted of a factorial arrangement of treatments in a completely randomized design. The results showed that there were maximum increases in shoot length (32.7%), root length (58.9%), and shoot fresh (85.7%). The quantities of leaves per plant were increased by 52.9%, 123.6%, 82.5%, and 62.2% in C1, C2, C3, and C4 after proline application. On the other hand, only root fresh weight decreased in all genotypes after proline application by 23.1%, 20%, 266.7%, and 280.8% (C1, C2, C3, C4). A lower leaf temperature of 27.72 °C, minimum transpiration of 3.29 mmol m−2 s−1, maximum photosynthesis of 3.91 μmol m−2 s−1, and a maximum water use efficiency of 1.20 μmol CO2 mmol H2O were recorded in the genotypes C2, C1, C3, and C4, respectively. The highest enzymatic activity of superoxide dismutase, peroxidase and catalase were 14.88, 0.31, and 0.15 U mg-protein in C2, C1, and C3, respectively. Maximum leaf proline, glycinebetaine, total free amino acids, and chlorophyll content 3.46 mg g−1, 4.02 mg g−1, 3.46 mg g−1, and 46.89 (in C2), respectively, due to foliar applications of proline. Another important finding was that heat tolerance in okra was highly linked highly linked to genotypes’ genetic potential, having more water use efficiency, enzymatic activities, and physio-biochemical attributes under the foliar applications of proline.

The root-knot nematode is one of the most damaging plant-parasitic nematodes worldwide, and the ecofriendly alternative approach of biological control has been used to suppress nematode populations. Here the nematicidal activity of Aspergillus japonicus ZW1 fermentation filtrate against Meloidogyne incognita was evaluated in vitro and in greenhouse, and the effects of A. japonicus ZW1 fermentation filtrate on seed germination and the active compound of A. japonicus ZW1 fermentation filtrate were determined. The 2-week fermentation filtrate (2-WF) of A. japonicus ZW1 exhibited markedly inhibitory effects on egg hatching, and 5% 2-WF showed potential nematicidal activities on second-stage juveniles (J2s); the mortality of J2s was 100% after 24 h exposure. The internal contents of nematodes were degraded and remarkable protruded wrinkles were present on the body surface of J2s. The nematicidal activity of the fermentation was stable after boiling and was not affected by storage time. A germination assay revealed that 2-WF did not have a negative effect on the viability and germination of corn, wheat, rice, cowpeas, cucumbers, soybeans, or tomato seeds. The pot-grown study confirmed that a 20% fermentation broth solution significantly reduced root galls and egg numbers on tomatoes, and decreased galls and eggs by 47.3% and 51.8% respectively, over Czapek medium and water controls. The active compound from the A. japonicus ZW1 fermentation filtrate was isolated and identified as 1,5-Dimethyl Citrate hydrochloride ester on the basis of nuclear magnetic resonance (NMR) and LC-MS (liquid chromatograph-mass spectrometer) techniques. Thus, fermentation of A. japonicus ZW1 could be considered a potential new biological nematicide for the control of M. incognita.

Grey mold is one of the most serious and catastrophic diseases, causing significant yield losses in fruits and vegetables worldwide. Iprodione is a broad spectrum agrochemical used as a foliar application as well as a seed protectant against many fungal and nematode diseases of fruits and vegetables from the last thirty years. The extensive use of agrochemicals produces resistance in plant pathogens and is the most devastating issue in food and agriculture. However, the molecular mechanism (whole transcriptomic analysis) of a resistant mutant of B. cinerea against iprodione is still unknown. In the present study, mycelial growth, sporulation, virulence, osmotic potential, cell membrane permeability, enzymatic activity, and whole transcriptomic analysis of UV (ultraviolet) mutagenic mutant and its wild type were performed to compare the fitness. The EC50 (half maximal effective concentration that inhibits the growth of mycelium) value of iprodione for 112 isolates of B. cinerea ranged from 0.07 to 0.87 &micro, g/mL with an average (0.47 &micro, g/mL) collected from tomato field of Guangxi Province China. Results also revealed that, among iprodione sensitive strains, only B67 strain induced two mutants, M0 and M1 after UV application. The EC50 of these induced mutants were 1025.74 &mu, g/mL and 674.48 &mu, g/mL, respectively, as compared to its wild type 1.12 &mu, g/mL. Furthermore, mutant M0 showed higher mycelial growth sclerotia formation, virulence, and enzymatic activity than wild type W0 and M1 on potato dextrose agar (PDA) medium. The bctubA gene in the mutant M0 replaced TTC and GAT codon at position 593 and 599 by TTA and GAA, resulting in replacement of phenyl alanine into leucine (transversion C/A) and aspartic acid into glutamic acid (transversion T/C) respectively. In contrast, in bctubB gene, GAT codon at position 646 is replaced by AAT and aspartic acid converted into asparagine (transition G/A). RNA sequencing of the mutant and its wild type was performed without (M0, W0) and with iprodione treatment (M-ipro, W-ipro). The differential gene expression (DEG) identified 720 unigenes in mutant M-ipro than W-ipro after iprodione treatment (FDR &le, 0.05 and log2FC &ge, 1). Seven DEGs were randomly selected for quantitative real time polymerase chain reaction to validate the RNA sequencing genes expression (log fold 2 value). The gene ontology (GO) enrichment and Kyoto encyclopedia genes and genomes (KEGG) pathway functional analyses indicated that DEG&rsquo, s mainly associated with lysophopholipase, carbohydrate metabolism, amino acid metabolism, catalytic activity, multifunctional genes (MFO), glutathione-S transferase (GST), drug sensitivity, and cytochrome P450 related genes are upregulated in mutant type (M0, M-ipro) as compared to its wild type (W0, W-ipro), may be related to induce resistant in mutants of B. cinerea against iprodione.

Pomegranate adaptation to abiotic stress conditions has led to its wide dispersion in Pakistan and to the appearance of new, local genotypes. These genotypes are important to characterize for breeding programs aimed towards the production of new cultivars suitable to arid, semi-arid, and moderate weather regions. In this study, eleven pomegranate accessions were investigated for fruit morphological and biochemical traits, and root anatomic adaptation under diverse climates. The commercial accession (Sava) had the maximum fruit weight (373.97 g) compared to accessions KK-I (60.94 g) and KK-II (71.63 g), which were old plantations established since United India. Most of the accessions were juicy with a wood portion index (WPI) ranging from 0.93 to 1.06%, however, the non-commercial accession of Khushab Kanhatti (KK-I) showed the highest WPI (4.38%). A high variation occurred among all accessions for total soluble solids, total sugars, and total ascorbic acid contents with a maximum in accession Sava (16.46 &deg, Brix), TK-II (53.16%), and QW-I (0.68%), respectively. Root anatomical studies in accessions depicted significantly high variation, and accession Sava of semi-arid climate produced maximum root epidermis (97.43 &micro, m2), phloem (2730.78 &micro, m2), and metaxylem (717.77 &micro, m2) area. In contrast, high cortex thickness (104.74 &micro, m), cortex area (462.74 &micro, m2), and vascular area (114,382.10 &micro, m2) was measured in accessions KK-II and KK-III of Khushab district with an arid climate. The germplasm of diverse regions showed a strong association and clustered into two main classes based on fruit morpho-chemical and root anatomical characteristics. These traits are likely to provide clues towards plants adaptation to various growing conditions and can be exploited successfully in crop improvement programs.

The soil-borne fungus Fusarium oxysporum (Fo) and the nematode Meloidogyne incognita (Mi) are destructive pathogens that cause substantial yield losses to tomato (Solanum lycopersicum L.) crops worldwide. The present study sought to elucidate the physiological, biochemical, and cytological responses of tomato cultivars (Gailing maofen 802 and Zhongza 09) by root invasion of Fo (1 &times, 105 CFUmL&minus, 1) and Mi (1500 second-stage juveniles (J2) alone and in combination after 14 days. Results revealed that combined inoculation of Fo and Mi significantly increased disease intensity, electrolyte leakage, and hydrogen peroxide and malondialdehyde contents, and decreased photosynthetic capacity and enzyme activity in both cultivars as compared to their solo inoculation. Increasing the disease intensity reduced the maximum morphological traits, such as shoot length, total dry weight, and total chlorophyll contents, in G. maofen 802 (by 32%, 54.2%, and 52.3%, respectively) and Zhongza 09 (by 18%, 32%, and 21%, respectively) as compared to the control. Others factors were also reduced in G. maofen 802 and Zhongza 09, such as photosynthetic capacity (by 70% and 57%, respectively), stomatal conductance (by 86% and 70%, respectively), photochemical quantum yield of photosystem II (YII) (by 36.6% and 29%, respectively), and electron transport rate (by 17.7% and 10%, respectively), after combined inoculation of Fo and Mi. Furthermore, the combined infestation of Fo and Mi resulted in reduced activity of plant-defense-related antioxidants in G. maofen 802 compared with their single application or control. However, these antioxidants were highly up-regulated in Zhongza 09 (by 59%&ndash, 93%), revealing the induction of tolerance against studied pathogens. The transmission electron microscopy (TEM) results further demonstrated that root cells of Zhongza 09 had unique tetrahedral crystal-like structures in the membrane close to mitochondria under all treatments except control. Therefore, it is concluded that Mi caused severe root damage, suppressed plant growth, depleted antioxidants, and caused high generation of ROS in the presence of Fo as compared to its solo inoculation. Tolerant cultivars adopted different mechanistic strategies at the structural and cellular levels to tolerate the Mi and Fo stresses.

Wheat is one of the best-domesticated cereal crops and one of the vital sources of nutrition for humans. An investigation was undertaken to reveal the potential of novel bio-inoculants enriching micronutrients in shoot and grains of wheat crop to eliminate the hazards of malnutrition. Sole as well as consortia inoculation of bio-inoculants significantly enhanced mineral nutrients including zinc (Zn) and iron (Fe) concentrations in shoot and grains of wheat. Various treatments of bio-inoculants increase Zn and Fe content up to 1–15% and 3–13%, respectively. Sole inoculation of Bacillus aryabhattai (S10) impressively improves the nutritious of wheat. However, the maximum increase in minerals contents of wheat was recorded by consortia inoculation of Paenibacillus polymyxa ZM27, Bacillus subtilis ZM63 and Bacillus aryabhattai S10. This treatment also showed a maximum bacterial population (18 × 104 cfu mL-1) in the rhizosphere. The consortium application of these strains showed up to a 17% increase in yield. It is evident from the results that the consortium application was more effective than sole and co-inoculation. A healthy positive correlation was found between growth, yield, and the accessibility of micronutrients to wheat crops at the harvesting stage. The present investigations revealed the significance of novel bacterial strains in improving the nutritional status of wheat crops. These strains could be used as bio-inoculants for the biofortification of wheat to combat hidden hunger in developing countries.

Genome editing is a relevant, versatile, and preferred tool for crop improvement, as well as for functional genomics. In this review, we summarize the advances in gene-editing techniques, such as zinc-finger nucleases (ZFNs), transcription activator-like (TAL) effector nucleases (TALENs), and clustered regularly interspaced short palindromic repeats (CRISPR) associated with the Cas9 and Cpf1 proteins. These tools support great opportunities for the future development of plant science and rapid remodeling of crops. Furthermore, we discuss the brief history of each tool and provide their comparison and different applications. Among the various genome-editing tools, CRISPR has become the most popular; hence, it is discussed in the greatest detail. CRISPR has helped clarify the genomic structure and its role in plants: For example, the transcriptional control of Cas9 and Cpf1, genetic locus monitoring, the mechanism and control of promoter activity, and the alteration and detection of epigenetic behavior between single-nucleotide polymorphisms (SNPs) investigated based on genetic traits and related genome-wide studies. The present review describes how CRISPR/Cas9 systems can play a valuable role in the characterization of the genomic rearrangement and plant gene functions, as well as the improvement of the important traits of field crops with the greatest precision. In addition, the speed editing strategy of gene-family members was introduced to accelerate the applications of gene-editing systems to crop improvement. For this, the CRISPR technology has a valuable advantage that particularly holds the scientist’s mind, as it allows genome editing in multiple biological systems.

Pomegranate is famous for its health benefiting chemical and biochemical constituent compounds. The present study was undertaken to characterize pomegranate germplasm for its various fruit traits, acids, and sugar profiling through high performance liquid chromatography (HPLC) analysis. Among 11 detected acids and 8 sugars, citric acid and fructose were predominant in 18 domestic and 5 wild genotypes, respectively. Fruit weight, aril weight and wood portion index (WPI) were ranged from 15.82% to 24.42%, 10.99% to 113.78%, and 2.39% to 17.25%, respectively. Genotypes were grouped as sweet, sweet&ndash, sour, sour&ndash, sweet, and sour based on citric acid contents. Lactic acid and pyruvic acid showed the highest correlation (r = 0.92), however, sour and sweet genotypes had strong association for acids and sugars, respectively. Straddling of dendrogram showed the flow of genetic material in a cultivated location with wild and cultivated pomegranates grouped in different classes, however, wild and sour landraces grouped in the same class with 71% similarity of traits. Based on the observations of the current study, it was concluded that selected wild and arid zones (Multan, Bahawalpur) genotypes are poor in nutrients (acid and sugars) quality, however, genotypes of Rahim-Yar-Khan, Muzafar Garh, and Khyber Pakhtunkhwa have a better composition of sugars and acids.