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.

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 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 × 10⁵ CFUmL⁻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%–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.

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.

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–sour, sour–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.