Responses of maize hybrids to water stress conditions at different developmental stages: accumulation of reactive oxygen species, activity of enzymatic antioxidants and degradation in kernel quality traits.

Sustainable maize production under changing climatic conditions, especially heat and water stress conditions is one of the key challenges that need to be addressed immediately. The current field study was designed to evaluate the impact of water stress on morpho-physiological, biochemical, reactive oxygen species, antioxidant activity and kernel quality traits at different plant growth stages in maize hybrids. Four indigenous i.e., YH-5427, YH-5482, YH-5395, JPL-1908, and one multinational maize hybrid i.e., NK-8441 (Syngenta Seeds) were used for the study. Four stress treatments (i) Control (ii) 3-week water stress at pre-flowering stage (iii) 3-week water stress at anthesis stage (iv) 3-week water stress at grain filling/post-anthesis stage. The presence of significant oxidative stress was revealed by the overproduction of reactive oxygen species (ROXs) i.e., H2O2 (1.9 to 5.8 µmole g-1 FW) and malondialdehyde (120.5 to 169.0 nmole g-1 FW) leading to severe negative impacts on kernel yield. Moreover, a severe reduction in photosynthetic ability (50.6%, from 34.0 to 16.8 µmole m-2 s-1), lower transpirational rate (31.3%, from 3.2 to 2.2 mmol m-2 s-1), alterations in plant anatomy, reduced pigments stability, and deterioration of kernel quality was attributed to water stress. Water stress affected all the three studied growth stages, the pre-flowering stage being the most vulnerable while the post-anthesis stage was the least affected stage to drought stress. Antioxidant activity was observed to increase under all stress conditions in all maize hybrids, however, the highest antioxidant activity was recorded at the anthesis stage and in maize hybrids YH-5427 i.e., T-SOD activity was increased by 61.3% from 37.5 U mg-1 pro to 60.5 U mg-1 pro while CAT activity was maximum under water stress conditions 8.3 U mg-1 pro as compared to 10.3 U mg-1 pro under control (19.3%). The overall performance of maize hybrid YH-5427 was much more promising than other hybrids, attributed to its higher photosynthetic activity, and better antioxidant defense mechanism. Therefore, this hybrid could be recommended for cultivation in drought-prone areas.

Biofortification of Cereals and Pulses Using New Breeding Techniques: Current and Future Perspectives.

Cereals and pulses are consumed as a staple food in low-income countries for the fulfillment of daily dietary requirements and as a source of micronutrients. However, they are failing to offer balanced nutrition due to deficiencies of some essential compounds, macronutrients, and micronutrients, i.e., cereals are deficient in iron, zinc, some essential amino acids, and quality proteins. Meanwhile, the pulses are rich in anti-nutrient compounds that restrict the bioavailability of micronutrients. As a result, the population is suffering from malnutrition and resultantly different diseases, i.e., anemia, beriberi, pellagra, night blindness, rickets, and scurvy are common in the society. These facts highlight the need for the biofortification of cereals and pulses for the provision of balanced diets to masses and reduction of malnutrition. Biofortification of crops may be achieved through conventional approaches or new breeding techniques (NBTs). Conventional approaches for biofortification cover mineral fertilization through foliar or soil application, microbe-mediated enhanced uptake of nutrients, and conventional crossing of plants to obtain the desired combination of genes for balanced nutrient uptake and bioavailability. Whereas, NBTs rely on gene silencing, gene editing, overexpression, and gene transfer from other species for the acquisition of balanced nutritional profiles in mutant plants. Thus, we have highlighted the significance of conventional and NBTs for the biofortification of cereals and pulses. Current and future perspectives and opportunities are also discussed. Further, the regulatory aspects of newly developed biofortified transgenic and/or non-transgenic crop varieties via NBTs are also presented.

CRISPR-Based Crop Improvements: A Way Forward to Achieve Zero Hunger.

Zero hunger is one of the sustainable development goals set by the United Nations in 2015 to achieve global food security by 2030. The current harvest of crops is insufficient; feeding the world's population and meeting the goal of zero hunger by 2030 will require larger and more consistent crop production. Clustered regularly interspaced short palindromic repeats-associated protein (CRISPR-Cas) technology is widely used for the plant genome editing. In this review, we consider this technology as a potential tool for achieving zero hunger. We provide a comprehensive overview of CRISPR-Cas technology and its most important applications for food crops' improvement. We also conferred current and potential technological breakthroughs that will help in breeding future crops to end global hunger. The regulatory aspects of deploying this technology in commercial sectors, bioethics, and the production of transgene-free plants are also discussed. We hope that the CRISPR-Cas system will accelerate the breeding of improved crop cultivars compared with conventional breeding and pave the way toward the zero hunger goal.

Harnessing the potential of plant transcription factors in developing climate resilient crops to improve global food security: Current and future perspectives.

Crop plants should be resilient to climatic factors in order to feed ever-increasing populations. Plants have developed stress-responsive mechanisms by changing their metabolic pathways and switching the stress-responsive genes. The discovery of plant transcriptional factors (TFs), as key regulators of different biotic and abiotic stresses, has opened up new horizons for plant scientists. TFs perceive the signal and switch certain stress-responsive genes on and off by binding to different cis-regulatory elements. More than 50 families of plant TFs have been reported in nature. Among them, DREB, bZIP, MYB, NAC, Zinc-finger, HSF, Dof, WRKY, and NF-Y are important with respect to biotic and abiotic stresses, but the potential of many TFs in the improvement of crops is untapped. In this review, we summarize the role of different stress-responsive TFs with respect to biotic and abiotic stresses. Further, challenges and future opportunities linked with TFs for developing climate-resilient crops are also elaborated.

The level of Cry1Ac endotoxin and its efficacy against H. armigera in Bt cotton at large scale in Pakistan.

A biophysical survey was conducted in 15 cotton-growing districts of Pakistan. Four hundred cotton growers were approached and inquired about the production technology of Bt cotton. Further, 25 strip tests using combo strips (Cry1Ac, Cry2Ab, Vip3Aa and Cp4, EPSPS gene) were performed at each farmer's field. Out of 10,000 total-tested samples, farmers claimed 9682 samples as Bt and 318 samples as non-Bt. After performing a strip test, 1009 and 87 samples were found false negative and false positive, respectively. Only 53 samples were found positive for Cry2Ab, 214 for EPSPS and none for Vip3Aa gene. Quantification of Cry endotoxin and bioassay studies were performed by taking leaves from upper, middle, and lower canopies, and fruiting parts at approximately 80 days after sowing from 89 varieties. Expression was highly variable among different canopies and fruiting parts. Moreover, Cry endotoxin expression and insect mortality varied significantly among varieties from 0.26 µg g-1 to 3.54 µg g-1 with mortality ranging from 28 to 97%, respectively. Highest Cry1Ac expression (3.54 µg g-1) and insect mortality (97%) were observed for variety FH-142 from DG Khan. Cry endotoxin expression varied significantly across various plant parts, i.e., IUB-13 variety from upper canopy documented 0.34 µg g-1 expression with 37% insect mortality in Layyah to 3.42 µg g-1 expression and 96% insect mortality from DG Khan. Lethal dose, LD95 (2.20 µg g-1) of Cry1Ac endotoxin was optimized for effective control of H. armigera. Our results provided evidence of practical resistance in H. armigera and way forward.

Role of Genetics, Genomics, and Breeding Approaches to Combat Stripe Rust of Wheat.

Puccinia striiformis (Pst) is a devastating biotrophic fungal pathogen that causes wheat stripe rust. It usually loves cool and moist places and can cause 100% crop yield losses in a single field when ideal conditions for disease incidence prevails. Billions of dollars are lost due to fungicide application to reduce stripe rust damage worldwide. Pst is a macrocyclic, heteroecious fungus that requires primary (wheat or grasses) as well as secondary host (Berberis or Mahonia spp.) for completion of life cycle. In this review, we have summarized the knowledge about pathogen life cycle, genes responsible for stripe rust resistance, and susceptibility in wheat. In the end, we discussed the importance of conventional and modern breeding tools for the development of Pst-resistant wheat varieties. According to our findings, genetic engineering and genome editing are less explored tools for the development of Pst-resistant wheat varieties; hence, we highlighted the putative use of advanced genome-modifying tools, i.e., base editing and prime editing, for the development of Pst-resistant wheat.

Genetic Variability among Different Populations of Root Knot Nematodes Based on Their Encumbrance Response to Pasteuria Isolates Using PCR-RFLP.

A great variable response was observed when PP-3 and PP-J encumbered with 116 populations of root knot nematode (RKN) at two different temperatures (25 ± 2°C and 30 ± 2°C) and concentrations (104 and 105 spores/ml). The PCR reaction amplified intergenic region between cytochrome oxidase subunit II gene (COII) and large subunit of rRNA gene (lrRNA) of the mitochondrial genome of different RKN species. The primer C2F3 and 1108 identified M. incognita with the highest frequency (52.6%) followed by M. javanica (36.8%) and M. arenaria (10.5%). The sizes of PCR products were 1.7 kb for M. incognita and M. javanica populations while populations of M. arenaria produced 1.1 kb fragment. The digestion with Hinf I yielded three different fragment length patterns on 1.5 % agarose gel. From current research it is concluded that intra-Meloidogyne genetic variability exist in RKN populations which have better encumbrance with P. penetrans.