Harnessing the potential of mutation breeding, CRISPR genome editing, and beyond for sustainable agriculture.

By 2050, the global population is projected to exceed 9.5 billion, posing a formidable challenge to ensure food security worldwide. To address this pressing issue, mutation breeding in horticultural crops, utilizing physical or chemical methods, has emerged as a promising biotechnological strategy. However, the efficacy of these mutagens can be influenced by various factors, including biological and environmental variables, as well as targeted plant materials. This review highlights the global challenges related to food security and explores the potential of mutation breeding as an indispensable biotechnological tool in overcoming food insecurity. This review also covers the emergence of CRISPR-Cas9, a breakthrough technology offering precise genome editing for the development of high-yield, stress-tolerant crops. Together, mutation breeding and CRISPR can potentially address future food demands. This review focuses into these biotechnological advancements, emphasizing their combined potential to fortify global food security in the face of a booming population.

A narrative action on the battle against hunger using mushroom, peanut, and soybean-based wastes.

Numerous generations have been affected by hunger, which still affects hundreds of millions of people worldwide. The hunger crisis is worsening although many efforts have been made to minimize it. Besides that, food waste is one of the critical problems faced by most countries worldwide. It has disrupted the food chain system due to inefficient waste management, while negatively impacting the environment. The majority of the waste is from the food production process, resulting in a net zero production for food manufacturers while also harnessing its potential. Most food production wastes are high in nutritional and functional values, yet most of them end up as low-cost animal feed and plant fertilizers. This review identified key emerging wastes from the production line of mushroom, peanut, and soybean (MPS). These wastes (MPS) provide a new source for food conversion due to their high nutritional content, which contributes to a circular economy in the post-pandemic era and ensures food security. In order to achieve carbon neutrality and effective waste management for the production of alternative foods, biotechnological processes such as digestive, fermentative, and enzymatic conversions are essential. The article provides a narrative action on the critical potential application and challenges of MPS as future foods in the battle against hunger.

The Review of Cooking, Drying, and Green Extraction Methods on General Nutritional Properties of Mealworms and Locusts.

The processing of edible insects as an alternative source of nutrition may be a key driver in the development of a sustainable food and feed system. This review will study two industrial types of insects-mealworms and locusts-and summarize evidence related to the impact of processing on their micro- and macronutritional characteristics. The focus will be on their potential use as food for human consumption as opposed to animal feed. Literature has indicated that these two insects have the potential to provide protein and fat qualities comparable to or better than traditional mammalian sources. For example, mealworms-the larval form of the yellow mealworm beetlepossess a higher fat content, while adult locusts are rich in fibers, especially chitin. However, due to the different matrix and nutrient compositions, the processing of mealworms or locusts at a commercial scale needs to be tailored to minimize nutritional loss and maximize cost efficiency. The stages of preprocessing, cooking, drying, and extraction are the most critical control points for nutritional preservation. Thermal cooking applications such as microwave technology have demonstrated promising results, but the generation of heat may contribute to a certain nutritional loss. In an industrial context, drying using freeze dry is the preferred choice due to its uniformity, but it can be costly while increasing lipid peroxidation. During the extraction of nutrients, the use of green emerging technologies such as high hydrostatic pressure, pulsed electric field, and ultrasound may provide an alternative method to enhance nutrient preservation.

Increasing Lovastatin Production by Re-routing the Precursors Flow of Aspergillus terreus via Metabolic Engineering.

Lovastatin is an anti-cholesterol medicine that is commonly prescribed to manage cholesterol levels, and minimise the risk of suffering from heart-related diseases. Aspergillus terreus (ATCC 20542) supplied with carbohydrates or sugar alcohols can produce lovastatin. The present work explored the application of metabolic engineering in A. terreus to re-route the precursor flow towards the lovastatin biosynthetic pathway by simultaneously overexpressing the gene for acetyl-CoA carboxylase (acc) to increase the precursor flux, and eliminate ( +)-geodin biosynthesis (a competing secondary metabolite) by removing the gene for emodin anthrone polyketide synthase (gedC). Alterations to metabolic flux in the double mutant (gedCΔ*accox) strain and the effects of using two different substrate formulations were examined. The gedCΔ*accox strain, when cultivated with a mixture of glycerol and lactose, significantly (p < 0.05) increased the levels of metabolic precursors malonyl-CoA (48%) and acetyl-CoA (420%), completely inhibited the (+)-geodin biosynthesis, and increased the level of lovastatin [152 mg/L; 143% higher than the wild-type (WT) strain]. The present work demonstrated how the manipulation of A. terreus metabolic pathways could increase the efficiency of carbon flux towards lovastatin, thus elevating its overall production and enabling the use of glycerol as a substrate source. As such, the present work also provides a framework model for other medically or industrially important fungi to synthesise valuable compounds using sustainable carbon sources.

Fruiting-body-base flour from an Oyster mushroom waste in the development of antioxidative chicken patty.

In a commercial oyster mushroom farm, from 300 g of the total harvest, only the cap and stem of the fruiting body parts are harvested (200 g) while the unused lower section called fruiting-body-base (FBB) is discarded (50 g). A new antioxidative FBB flour (FBBF) conversion to mixed-ratio chicken patty was recently developed which converts 16.67% of FBB into an edible flour. At the initial stage, pretreatments of FBBF were optimized at particle size (106 µm) and citric acid concentration (0.5 g/100 mL) to improve flour antioxidant responses. Such pretreatments boosted total phenolic content (2.31 ± 0.53 mg GAE/g) and DPPH (51.53 ± 1.51%) of pretreated FBBF. Mixed-ratio chicken patty containing FBBF (10%, 20%, 30%) significantly (P < 0.05) influenced the hardness, cohesiveness, springiness, and chewiness of the patties. However, only the hardness and chewiness increased proportionally with the increase FBBF in concentration. Notably, 60 panellists considered that 10% FBBF-chicken patty sensory attributes, including lightness, redness, and yellowness, is acceptable to consumers. This information could be used to market any type of commercial mushroom farm waste as alternative food products. PRACTICAL APPLICATION: This study shows that unused harvested mushroom waste from a local farm can be used to make an antioxidative chicken patty that is acceptable to consumer panellists. The converted mushroom waste into flour suggests that smaller particles and citric acid pretreatment can increase its nutritional value. This information can be used for waste conversion into new product development from any type of mushroom farm.

Effect of Plant Growth Regulators on Coloured Callus Formation and Accumulation of Azadirachtin, an Essential Biopesticide in Azadirachta indica.

For centuries, Azadirachta indica or neem has been utilized as a primary source of medicine due to its antimicrobial, larvacidal, antimalarial and antifungal properties. Recently, its potential as an effective biopesticide has garnered attention, especially towards efficient and continuous production of its bioactive compounds. The present study investigated the effect of the plant growth regulators (PGRs) thiadiazuron (TDZ) and 2,4-dichlorophenoxyacetic acid (2,4-D) on the induction of colored callus formation and subsequent accumulation of azadirachtin (AZA) in A. indica. An efficient protocol was established for micropropagation and colored callus production of this species, followed by quantification of AZA (a mixture of azadirachtin A and B) and its safety assessment. For induction of the callus, leaf and petiole explants obtained from a young growing neem plant were excised and cultured on Murashige and Skoog (MS) medium supplemented with TDZ (0.2-0.6 mg L-1) and 2,4-D (0.2-0.6 mg L-1), either applied singly or in combination. Callus was successfully induced from both explant types at different rates, where media with 0.6 mg L-1 of TDZ resulted in the highest fresh weight (3.38 ± 0.08 g). In general, media with a single hormone (particularly TDZ) was more effective in producing a high mass of callus compared to combined PGRs. A culture duration of six weeks resulted in the production of green, brown and cream colored callus. The highest callus weight and accumulation of AZA was recorded in green callus (214.53 ± 33.63 mg g-1 dry weight (DW)) induced using TDZ. On the other hand, small amounts of AZA were detected in both brown and cream callus. Further experimentation indicated that the green callus with the highest AZA was found to be non-toxic (LC50 at 4606 µg mL-1) to the zebrafish animal model. These results suggested that the addition of different PGRs during in vitro culture could prominently affect callus and secondary metabolite production and can further be manipulated as a sustainable method for the production of a natural and environmentally friendly pesticide.

The investigation of media components for optimal metabolite production of Aspergillus terreus ATCC 20542.

PURPOSE: This study aimed to assess the effect of nitrogen, salt and pre-culture conditions on the production of lovastatin in A. terreus ATCC 20542. METHODS: Different combinations of nitrogen sources, salts and pre-culture combinations were applied in the fermentation media and lovastatin yield was analysed chromatographically. RESULT: The exclusion of MnSO4 ·5H2O, CuSO4·5H2O and FeCl3·6H2O were shown to significantly improve lovastatin production (282%), while KH2PO4, MgSO4·7H2O, and NaCl and ZnSO4·7H2O were indispensable for good lovastatin production. Simple nitrogen source (ammonia) was unfavourable for morphology, growth and lovastatin production. In contrast, yeast extract (complex nitrogen source) produced the highest lovastatin yield (25.52 mg/L), while powdered soybean favoured the production of co-metabolites ((+)-geodin and sulochrin). Intermediate lactose: yeast extract (5:4) ratio produced the optimal lovastatin yield (12.33 mg/L) during pre-culture, while high (5:2) or low (5:6) lactose to yeast extract ratio produced significantly lower lovastatin yield (7.98 mg/L and 9.12 mg/L, respectively). High spore concentration, up to 107 spores/L was shown to be beneficial for lovastatin, but not for co-metabolite production, while higher spore age was shown to be beneficial for all of its metabolites. CONCLUSION: The findings from these investigations could be used for future cultivation of A. terreus in the production of desired metabolites.

Improved lovastatin production by inhibiting (+)-geodin biosynthesis in Aspergillus terreus.

Lovastatin is widely prescribed to reduce elevated levels of cholesterol and prevent heart-related diseases. Cultivation of Aspergillus terreus (ATCC 20542) with carbohydrates or low-value feedstocks such as glycerol produces lovastatin as a secondary metabolite and (+)-geodin as a by-product. An A. terreus mutant strain was developed (gedCΔ) with a disrupted (+)-geodin biosynthesis pathway. The gedCΔ mutant was created by inserting the antibiotic marker hygromycin B (hyg) within the gedC gene that encodes emodin anthrone polyketide synthase (PKS), a primary gene responsible for initiating (+)-geodin biosynthesis. The effects of emodin anthrone PKS gene disruption on (+)-geodin and lovastatin biosynthesis and the production of the precursors acetyl-CoA and malonyl-CoA were investigated with cultures based on glycerol alone and in combination with lactose. The gedCΔ strain showed improved lovastatin production, particularly when cultivated on the glycerol-lactose mixture, increasing lovastatin production by 80% (113 mg/L) while simultaneously inhibiting (+)-geodin biosynthesis compared to the wild-type strain. This study thus shows that suppression of the (+)-geodin pathway increases lovastatin yield and demonstrates a practical approach of manipulating carbon flux by modulating enzyme activity.