Targeted Genome Editing for Cotton Improvement

Conventional tools induce mutations randomly throughout the cotton genome—making breeding difficult and challenging. During the last decade, progress has been made to edit the gene of interest in a very precise manner. Targeted genome engineering with engineered nucleases (ENs) specifically zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regularly interspaced short palindromic repeat (CRISPR) RNA-guided nucleases (e.g., Cas9) has been described as a “game-changing technology” for diverse fields as human genetics and plant biotechnology. In eukaryotic systems, ENs create double-strand breaks (DSBs) at the targeted DNA sequence which are repaired by nonhomologous end joining (NHEJ) or homology-directed recombination (HDR) mechanisms. ENs have been used successfully for targeted mutagenesis, gene knockout, and multisite genome editing (GenEd) in model plants and crop plants such as cotton, rice, and wheat. Recently, cotton genome has also been edited for targeted mutagenesis through CRISPR/Cas for improved lateral root formation. In addition, an efficient and fast method has been developed to evaluate guide RNAs transiently in cotton. The targeted disruption of undesirable genes or metabolic pathway can be achieved to increase quality of cotton. Undesirable metabolites like gossypol in cottonseed can be targeted efficiently using ENs for seed-specific low-gossypol cotton. Moreover, ENs are also helpful in gene stacking for herbicide resistance, insect resistance, and abiotic stress tolerance

Comparative Expression Studies of Fiber Related Genes in Cotton Spp.

Cotton fibers are the seed trichomes that are developed around the seed and are used to make clothes and yarn for the textile industry. Expression profiling of cotton fiber genes is very important to estimate the differential gene expression level at different fiber developmental stages. Expression analysis of fiber developing genes are very important to enhance the fiber length of cotton. The expression profiling of three gene families in five stages (0, 5, 10, 15 and 20 DPA) of cotton fiber tissues was carried out through real-time PCR. Expression analysis revealed that transcripts of GA-20 Oxidase, XTH, and PEPc were elevated from 5 to 20 days post-anthesis (DPA) fibers. Total RNA was extracted from various stages of cotton fiber development and was reverse transcribed to cDNA for PCR amplification. For data normalization, 18s rRNA was used as an internal control. The objective of this study was to explore the expression level of fiber developing genes at specific stages of fiber development. The results showed that most of the genes were expressed during the elongation phase in between 5 DPA to 15 DPA. Results obtained from this study may be helpful for the further identification of fiber genes and the improvement of fiber characteristics in cotton. PEPc and XTH genes that are expressed with a high rate during the fiber development may be used in breeding programmes for the improvement of fiber quality and quantity

Climate Smart Crops for Food Security

After ensuring the food security for over 50 years, the green revolution is eventually reaching its biological limits which are very much reflected by the ongoing stagnancy in yield increased over the past few decades. Meeting the increasing food demands due to increasing population is the greatest challenge for today’s plant scientists. Changing climatic conditions are posing additional threats to crop growth, productivity and yield. After successfully deploying gene editing to modify simple traits, scientists are now embarked on more ambitious adventures in genomics to combat challenges of food security in the wake of increasing population and climate change adversaries. The chapter outlines use of new technologies in tailoring crops beyond simple traits aiming to harvest the desired diversity lost during domestication and manipulating complex traits, which evolved over evolutionary timescale with special emphasis on the development of climate smart crops

Drought Susceptibility Index; a Preferred Criterion in Screening for Tolerance in Soybean

Soybean (Glycine max L.) yield and yield related traits are constrained by drought. Adaptation of soybean to changing environment could be improved by exploitation and introgression of diverse germplasm in breeding program. In present study, the response to drought conditions, especially at flowering stage, was evaluated to determine the potential soybean germplasm for future soybean breeding programs in Pakistan. Field experiment was conducted under two water regimes i.e. well-water and water-limited, to assess the effect of drought in seed yield and yield related traits. Although, drought led to overall reduction of ~15 % in thousand seed weight but still some soybean genotypes performed relatively better under water-limited conditions. These genotypes were also tolerant to drought, with a drought susceptibility index of \u3c 0.5. PCA also explained the pattern of variation existing in soybean germplasm grown under given water regimes i.e. well-water and water-limited conditions. The identified soybean genotypes could be a favorable resource to introduce high yielding soybean in local environment

Chemo-Modulatory Potential of Flaxseed Oil as Natural Anticancer Therapeutic

Background: Cancer is a disease which is characterised by uncontrolled cell proliferation and development. Surgery, chemotherapy, radiotherapy, and photodynamic therapy are the most frequent cancer treatments. On the other hand, there are many negative health impacts of radiation and chemotherapy that limit the efficient use of these therapies.Methods: This scenario needs natural treatments that are cost-effective and has no adverse effects. Flaxseed oil can be used as a nutraceuticals for the management of cancer. The current research was aimed on the exploration of the flaxseed (Linum usitatissimum) oil for in vitro anticancer activity as a natural therapy for the management of cancer.Result: The flaxseed powder contained 36.6±0.04% oil contents with an average yield of 36.6±0.03% by using hexane as solvent for extraction. Moreover, the oil contained polyunsaturated fatty acid with omega 3 fatty acid (alpha-linolenic acid) as a dominant content of the oil. In-vitro anticancer activity of flaxseed oil was observed by Cytotoxic (3T3 cell line) and Prostate Cancer (PC3 cell line) indicating that the oil possessed anticancer activity which was dose-dependent.Conclusion: On the basis of results, it was concluded that the flaxseed or its oil can be used for the management of cancer as a natural therapy by using optimized dose levels for a different types of cancer

Comparative Expression Studies of Fiber Related Genes in Cotton Spp.

Cotton fibers are the seed trichomes that are developed around the seed and are used to make clothes and yarn for the textile industry. Expression profiling of cotton fiber genes is very important to estimate the differential gene expression level at different fiber developmental stages. Expression analysis of fiber developing genes are very important to enhance the fiber length of cotton. The expression profiling of three gene families in five stages (0, 5, 10, 15 and 20 DPA) of cotton fiber tissues was carried out through real-time PCR. Expression analysis revealed that transcripts of GA-20 Oxidase, XTH, and PEPc were elevated from 5 to 20 days post-anthesis (DPA) fibers. Total RNA was extracted from various stages of cotton fiber development and was reverse transcribed to cDNA for PCR amplification. For data normalization, 18s rRNA was used as an internal control. The objective of this study was to explore the expression level of fiber developing genes at specific stages of fiber development. The results showed that most of the genes were expressed during the elongation phase in between 5 DPA to 15 DPA. Results obtained from this study may be helpful for the further identification of fiber genes and the improvement of fiber characteristics in cotton. PEPc and XTH genes that are expressed with a high rate during the fiber development may be used in breeding programmes for the improvement of fiber quality and quantity

Determination of Crop Coefficient of Hybrid Wheat under Arid Climate: A Pot Study

Climate change increases vulnerabilities for crop productivity in Pakistan. Water crises are increasing with an increase in temperature and change in precipitation patterns due to climate change which ultimately imposed a threat to the food security of the country. Water is indispensable for all plants to complete life cycle as the unavailability of water at critical growth stages drastically affects the development of the plant. The present pot study was conducted for the estimation of crop coefficient of hybrid wheat for irrigation scheduling at Muhammad Nawaz Shareef University of Agriculture, Multan during two growing seasons 2018-19 and 2019-20. In this experiment, three wheat varieties were used were Hybrid-1 (R26-3-1/DH-16), Hybrid-2(AR 7-5 / ZWB-14), and Galaxy-2013 as treatment. The soil moisture content was maintained between 50 to 100 % available water content (AWC) during both growing seasons. The crop coefficient (Kc) and actual evapotranspiration (Eta) were maximum in galaxy-13 and minimum in hybrid wheat. The grain yield for Hybrid-1, Hybrid-2, and galaxy-13 was 1, 1.5, and 0.6 g plant-1, respectively while the straw output was 4.8, 4.3, and 3 g plant-1, respectively. The harvest index for Hybrid-1, Hybrid-2, and galaxy-13 were 20, 34, and 20% respectively. The water use efficiency (WUE) for Hybrid-1, Hybrid-2, and galaxy-13 was 0.2. 0.3 and 0.1 g plant-1mm-1, respectively. The Hybrid-1 and Hybrid-2 produced more grain yield, straw yield, more spikes, and more grains per spikes and showed more water use efficiency with short plant height as compared to galaxy-13. The results of the study revealed that Hybrid-2 is more water-efficient with low water requirement and it was followed by Hybrid-1. The growing of Hybrid-2 will enhance the wheat yield to meet the food requirements of the increasing population under the climate change scenario with less water

Improving Growth and Yield of Sunflower with Integrated Use of Compost and PGPR (Variovorax Paradoxus) with Different Levels of N-Chemical Fertilizer

Plant growth-promoting rhizobacteria (PGPRs) stimulate plant growth through their ability, to increasing the root length and growth, by asymbiotic nitrogen fixation, by producing siderophores, solubilization of mineral phosphates and mineralization of other nutrients. Organic waste material of fruits and vegetables was collected and composted in a locally fabricated composting unit. A pot trial was conducted to study the effectiveness of compost and PGPR (Variovorax paradoxus) with recommended rate of PK and with different rates of N fertilizer i.e. 50 %, 75 % and 100 %, on growth and yield of sunflower. Results showed that the integrated use of 75 % N of recommended dose in combination with PGPR inoculation and compost caused a significant increase in grain yield and yield-contributing parameters compared with control. PGPR isolate Variovorax paradoxus in combination with 75 % N of recommended dose and compost gave maximum total achene’s yield 73 % more than control where recommended NPK was applied. Similarly, in case of Plant height, root length, head diameter, head weight, 100 – Achene’s weight and nitrogen content in straw were also increased by PGPR isolate Variovorax paradoxus in combination with 75 % N of the recommended dose and compost up to 11, 109, 27, 26, 17, and 54.0 % respectively over control

Effect of Drought on Trichome Density and Length in Cotton (Gossypium Hirsutum)

Cotton is a major cash crop and backbone of the textile industry in Pakistan which is badly affected by sucking insects. Drought is an important abiotic factor in trichome development. The objective of the study was to determine the effects of drought on trichome density and length. Trichome density was measured in two ways, one through the scaling method and the other through counting the trichome density manually. The scaling method is qualitative grading while quantitative grading includes trichomecount in a card of optimized length. Three scales were finalized to classify leaves on the basis of trichomes which were counted in a specific area (0.25cm2) on abaxial side of the leaf. In drought stress, trichomes density and length were measured and compared to that in normal conditions. Trichome density varies from 12 to 56 in 0.25cm2 under drought stress. On the basis of correlation of trichome density with stomatal conductance, photosynthetic rate, PAR and transpiration ratio under drought and normal conditions, it was concluded that trichome density increased as a result of drought stress

Evolutionary Integrated Heuristic with Gudermannian Neural Networks for Second Kind of Lane–Emden Nonlinear Singular Models

In this work, a new heuristic computing design is presented with an artificial intelligence approach to exploit the models with feed-forward (FF) Gudermannian neural networks (GNN) accomplished with global search capability of genetic algorithms (GA) combined with local convergence aptitude of active-set method (ASM), i.e., FF-GNN-GAASM to solve the second kind of Lane–Emden nonlinear singular models (LE-NSM). The proposed method based on the computing intelligent Gudermannian kernel is incorporated with the hidden layer configuration of FF-GNN models of differential operatives of the LE-NSM, which are arbitrarily associated with presenting an error-based objective function that is used to optimize by the hybrid heuristics of GAASM. Three LE-NSM-based examples are numerically solved to authenticate the effectiveness, accurateness, and efficiency of the suggested FF-GNN-GAASM. The reliability of the scheme via statistical valuations is verified in order to authenticate the stability, accuracy, and convergence