Water scarcity poses a significant challenge for farmers in dryland regions. The availability of water in these dryland areas greatly impacts crop cultivation and directly affects the livelihoods of farmers. As a result, preserving water resources is crucial for achieving successful crop cultivation. Scientists emphasize that in-situ water conservation is more effective than other methods. However, previous research has indicated that farmers have been reluctant to adopt in-situ water conservation (ISWC) practices Therefore, this study was conducted to analyse dryland farmers’ attitude towards ISWC practices in the Pudukkottai district. The data was collected from 120 participants during September October 2023. Statistical analysis techniques, such as the correlation, regression, and path analysis, were used to analyze the data. The results indicate that a majority of the farmers (53.33%) displayed a highly favourable attitude towards in-situ water conservation. Furthermore, variables such as cropping intensity, innovativeness, information seeking behaviour, cosmopoliteness, risk orientation, scientific orientation, awareness of soil erosion problems and accessibility of farm implements were found to have a positive association with the attitude towards in-situ water conservation. The policy implications include providing incentives to farmers who adopt water conservation measures and implementing effective training programs.

The solitary species of the genus Cocos is the coconut palm (Cocos nucifera L.). The coconut palm is affectionately known as "Kalpa Vriksha," which translates to "the tree of heaven. The causal organism of grey blight was identified based on the morphological characteristics i.e., five-celled conidia had three middle cells that ranged in colour from light brown to dark brown further confirmed through PCR analysis as Pestalotiopsis palmarum. In cultural characteristics, maximum radial growth occurred in PDA, with V-8 juice agar showing the least growth (65.00 mm). The optimal pH for pathogen growth was 6 (338.11 mg). Fungicide evaluation revealed ziram as highly efficient among non-systemic fungicides, while carbendazim and thiophonate methyl were most effective among systemic ones. Trifloxystrobin 25% + tebuconazole 50% EC emerged as the most effective combi product, inhibiting pathogen growth by 88.02%. Among bio-agents, T. viride-2 displayed the highest mycelial inhibition (71.30%). Turmeric exhibited the highest botanical efficacy (48.03%), whereas lantana had the least impact (3.37%).

The present study was conducted in the Sathyamanagalam block of Erode district during 2021 with the objective of studying the Extent of Awareness level of Sugarcane Farmers on Crop Residue Management Innovations. 60 respondents were selected from three villages. The results of the study shows that 100 per cent of respondents were aware that crop residue burning could create health risks. Crop residue management inventions such as trash mulching, trash shredders, and Harvest mounted cane trash shredder and collection systems were known by 100 per cent of the respondents. Only 10 per cent of those polled had no idea what bio-decomposers were. The Majority of the respondents (93.33%), regularly used trash mulching of sugarcane leaves to manage crop residue beneficially. Only 6.66 per cent of respondents used trash shredders regularly. Even though the subsidy given by the Government was available, due to timely unavailability of machinery led to the irregular use of trash shredders. Even though many of the respondents are aware of the negative aspects of residue burning and innovations available to manage sugarcane trash, they continue to burn due to time and cost constraints. With regard to constraints in adopting crop residue management technologies cent (100.00%) of respondents reported that lack of awareness on subsidies and involves more time and cost as the major constraints. Continuous monitoring of fields by the officials and demonstrations should be conducted to enhance the knowledge of the sugarcane growers on trash management.

De novo shoot apical meristem (SAM) organogenesis during regeneration in tissue culture has been investigated for several decades, but the precise mechanisms governing early-stage cell fate specification remain elusive. In contrast to SAM establishment during embryogenesis, in vitro SAM formation occurs without positional cues and is characterized by autonomous initiation of cellular patterning. Here, we report on the initial stages of SAM organogenesis and on the molecular mechanisms that orchestrate gene patterning to establish SAM homeostasis. We found that SAM organogenesis in tobacco calli starts with protuberance formation followed by the formation of an intact L1 layer covering the nascent protuberance. We also exposed a complex interdependent relationship between L1 and WUS expression and revealed that any disruption in this interplay compromises shoot formation. Silencing WUS in nascent protuberances prevented L1 formation and caused the disorganization of the outer cell layers exhibiting both anticlinal and periclinal divisions, suggesting WUS plays a critical role in the proper establishment and organization of L1 during SAM organogenesis. We further discovered that silencing TONNEAU1 prevents the exclusive occurrence of anticlinal divisions in the outermost layer of the protuberances and suppresses the acquisition of L1 cellular identity and L1 formation, ultimately impeding SAM formation and regeneration. This study provides a novel molecular framework for the characterization of a WUS/L1 interplay that mediates SAM formation during regeneration.

Background: Rice being staple food has qualities beyond its nutritive value with high digestibility and least allergic properties compared to other cereal grains. Research on exploring the nutritional value of traditional rice varieties with its inherent medicinal values and productivity has poor documentation and hence should be encouraged and supported. Field experiment was conducted with the objective to study the productivity of medicinal rice varieties as influenced by nutrient sources (inorganic and organic) and soil conditions (reduced and oxidized) under direct seeded rice. Methods: A field experiment was conducted in the wetland during Rabi season (October-February) at Tamil Nadu Agricultural University, Coimbatore with an objective to study the effect of crop establishment of medicinal rice varieties as influenced with nutrient sources and soil conditions under direct seeded rice. The experiment was laid out in factorial randomized block design with three replications. The treatments consisted of soil conditions (reduced, oxidized) as one factor, sources of nutrients (inorganic, organic) as the second factor and four medicinal rice varieties (Black Kavuni, Red Kavuni, Local Kavuni and Njavara) as the third factor. Result: The reduced condition and inorganic source of nutrient recorded higher plant height, maximum tiller population dry matter production, leaf area index, panicle length, higher number of spikelet panicle-1, 1000-grain weight and filled grains. Among the medicinal rice varieties, Red Kavuni recorded higher growth parameters and yield attributes. From the above results, it could be concluded that Red Kavuni medicinal rice under a reduced condition with inorganic nutrients resulted in higher growth parameters and yield attributes.

Symbiotic interactions play a vital role in maintaining the phosphate (Pi) nutrient status of host plants and providing resilience during biotic and abiotic stresses. Serendipita indica, a mycorrhiza-like fungus, supports plant growth by transporting Pi to the plant. Despite the competitive behaviour of arsenate (AsV) with Pi, the association with S. indica promotes plant growth under arsenic (As) stress by reducing As bioavailability through adsorption, accumulation, and precipitation within the fungus. However, the capacity of S. indica to enhance Pi accumulation and utilization under As stress remains unexplored. Axenic studies revealed that As supply significantly reduces intracellular ACPase activity in S. indica, while extracellular ACPase remains unaffected. Further investigations using Native PAGE and gene expression studies confirmed that intracellular ACPase (isoform2) is sensitive to As, whereas extracellular ACPase (isoform1) is As-insensitive. Biochemical analysis showed that ACPase (isoform1) has a Km of 0.5977 µM and Vmax of 0.1945 Unit/min. In hydroponically cultured tomato seedlings, simultaneous inoculation of S. indica with As on the 14thday after seed germination led to hyper-colonization, increased root/shoot length, biomass, and induction of ACPase expression and secretion under As stress. Arsenic-treated S. indica colonized groups (13.33 µM As+Si and 26.67 µM As+Si) exhibited 8.28–19.14 and 1.71–3.45-fold activation of ACPase in both rhizospheric media and root samples, respectively, thereby enhancing Pi availability in the surrounding medium under As stress. Moreover, S. indica (13.33 µM As+Si and 26.67 µM As+Si) significantly improved Pi accumulation in roots by 7.26 and 9.46 times and in shoots by 4.36 and 8.85 times compared to the control. Additionally, S. indica induced the expression of SiPT under As stress, further improving Pi mobilization. Notably, fungal colonization also restricted As mobilization from the hydroponic medium to the shoot, with a higher amount of As (191.01 ppm As in the 26.67 µM As+Si group) accumulating in the plant's roots. The study demonstrates the performance of S. indica under As stress in enhancing Pi mobilization while limiting As uptake in the host plant. These findings provide the first evidence of the As-Pi interaction in the AM-like fungus S. indica, indicating reduced As uptake and regulation of PHO genes (ACPase and SiPT genes) to increase Pi acquisition. These data also lay the foundation for the rational use of S. indica in agricultural practices.

As climate change induces habitat shifts, bioclimatic variables are used in suitable habitat modeling to predict species migration. Begonia aborensis Dunn of sect. Sphenanthera (Hassk.) Warb. (Begoniaceae), a native species to Abor hills was categorized as ‘Rare’ in India. The study aimed to generate ecological niche modeling of B. aborensis, identify the influential bioclimatic factors, and trait analysis to detect the informative morphological traits of the species by ancestral reconstruction. Maxent version 3.3.3 was employed for ENM at 30 arc-second which showed an AUC value < 1, and highly suitable habitats in parts of Arunachal Pradesh, Meghalaya, and Nagaland. The annual precipitation, precipitation of the driest month, elevation, and annual mean temperature were found as the most influential bioclimatic predictors. The Representative Concentration Pathway (RCP) scenarios of greenhouse gas emission for 2050 and 2070 showed suitability potential as 3.62

Abiotic stresses, including drought, salinity, cold, heat, and heavy metals, extensively reducing global agricultural production. Traditional breeding approaches and transgenic technology have been widely used to mitigate the risks of these environmental stresses. The discovery of engineered nucleases as genetic scissors to carry out precise manipulation in crop stress-responsive genes and associated molecular network has paved the way for sustainable management of abiotic stress conditions. In this context, the clustered regularly interspaced short palindromic repeat-Cas (CRISPR/Cas)-based gene-editing tool has revolutionized due to its simplicity, accessibility, adaptability, flexibility, and wide applicability. This system has great potential to build up crop varieties with enhanced tolerance against abiotic stresses. In this review, we summarize the latest findings on understanding the mechanism of abiotic stress response in plants and the application of CRISPR/Cas-mediated gene-editing system towards enhanced tolerance to a multitude of stresses including drought, salinity, cold, heat, and heavy metals. We provide mechanistic insights on the CRISPR/Cas9-based genome editing technology. We also discuss applications of evolving genome editing techniques such as prime editing and base editing, mutant library production, transgene free and multiplexing to rapidly deliver modern crop cultivars adapted to abiotic stress conditions.

In pulse crops, synchronized flowering altered the source-sink relationship due to the rapid translocation of nutrients from leaves to the developing pods. Additional nutrition through foliar feeding plays a vital role in pulse production by stimulating root development, nodulation, energy transformation, various metabolic processes and increasing pod setting, thereby increasing the yield. Many researchers are trying to reduce transpiration losses, flower shedding and maximizing productivity, foliar application of nutrient formulations and growth regulators in pulses. Thus, the foliar application of macro and micronutrients and growth regulators is considered an efficient and economical method of supplementing part of the nutrient requirements and moisture stress tolerance at critical stages. The PPFM (Pink pigmented facultative methylobacteria), when used as a foliar spray, it releases osmoprotectants (sugars and alcohols) on the surface of the plants and it increases chlorophyll content, thereby increasing the photosynthetic efficiency and makes drought tolerance ability of plants. This matrix helped to protect the plants from desiccation and high temperatures. Whereas potassium as spray also enhances drought tolerance in plants by mitigating harmful effects by increasing translocation, maintaining water balance and increasing pod filling. Further, Salicylic acid is an endogenous growth regulator of phenolic nature, which regulates physiological processes to mitigate stress, acts as a chelate for phosphorous uptake, increases pod setting, flowering and grain yield. Pulse wonder decreases flower shedding, increases yield by up to 20% and offers moisture stress tolerance.

Plant cells are endowed with a plastic nature that confers them the capacity to regenerate most plant organs or even an entire plant body from somatic cells. Plant tissue culture procedures take advantage of this asset to induce specific morphogenic responses using hormonal treatments. Still many plants, for example, peppers ( Capsicum annuum ), are considered recalcitrant because they respond poorly to exogenous hormone applications, which strongly limits transformation and genetic manipulations in this species. In this study, we exposed pepper cotyledon and hypocotyl explants to various cytokinins-containing media and characterized the morphological and gene-expression events that took place during adventitious shoot formation. Cellular organization events were observed and led to the formation of small protrusions, which in turn developed and expanded to form abundant rosette-leaf-like organs. Shoot formation was also obtained, although in rarer cases, and promoted the growth of elongated plantlets that at last rooted well. Moreover, gene-expression analyses of pepper cotyledons exposed to cytokinins revealed the induction of shoot morphogenesis-related genes, reflecting the occurrence of events of shoot apical meristem formation. In addition, we identified an increase in the expression of genes related to carbon and primary metabolism, suggesting the need for energy in this process. Here, we developed a protocol for shoot regeneration in pepper and our results provide new insights into adventitious shoot formation in pepper plants.

An experiment consisting of fifteen testers, three lines and their forty five crosses was conducted at crop research farm of Birsa Agriculture University, Kanke, Ranchi, Jharkhand during kharif season. Each parent and F1s were sown in five rows plot of 5.0×1.0 m2 spaced at 20×15 cm2 between rows and plants respectively. The experiment was laid out in Randomized Complete Block Design replicated thrice. All the recommended packages of practices were adopted to raise a good crop except irrigations as crop was irrigated through rains itself. The observations were recorded on five randomly selected plants from each of parents and F1 crosses on fifteen yield and yield attributing traits. Heterosis over mid parent for yield and its components was calculated as usual procedure. Cross combinations namely; BAU-274-92×IR-36 gave significantly positive heterosis to the tune of 31.39% followed by BR-8×IR-36 (27.57%) and BR-8×BD-202 (27%) and BAU-211-90×IR-36 (27.44%), BAU-211×BD- 202 (25.83%) and BAU-269-92×IR-36 (25.97%) for grain yield plant-1. The gca status of the parents involved revealed high×high, high×low and low×low combinations means involvement of both additive and non additive gene effects. The combinations can be further improved through simple selection procedures (additive×additive) or after advancing the generations through transgression effects.

The review of the literature demonstrated that the diverse properties of the organotin (IV) attributed to the various moieties contained inside the molecule account for the functions and utility of the organotin (IV) complexes. Furthermore, the capacity of organometallic compounds to stabilise complexes with unique stereochemistry is well documented. Due to their robust coordination chemistry, consistency, and varied molecular structures, these complexes exhibit a wide spectrum of biological activity. This article provides an overview of complexes' arrangement and geometry, spectroscopic research, and physical, chemical, and biological properties. This review also focuses on recent developments in conventional chemistry, practical synthesis methods, and the diverse functions of organotin (IV) complexes.

Arsenic contamination in rice is a major health concern that affects millions of people worldwide. Several strategies are recommended to reduce arsenic in rice, with AMF being one of the most important and cost-effective. This work is a pot study under environmental condition on rice plants under arsenic (AsV) treatment to demonstrate the ameliorative property of axenically cultivable AMF Serendipita indica (S. indica) in arsenic stress and accumulation in colonised plants. Colonisation of S. indica is 12 times higher under arsenic stress which is correlated to improved growth, biomass, and yield in fungal-colonised plants compared to arsenic-treated plants. Fungal colonisation results in a 1.9 times reduction in H2O2 content with the induction of the antioxidative enzyme system, proline and glutathione levels (up to 1.5 times). The fungal colonisation results in the recovery of pigment content (Chl a, Chl b and carotenoids) with improved photosynthesis, transpiration, and water use efficiency under arsenic stress. S. indica colonisation in plant roots resulted in a massive accumulation of arsenic (31.5 ppm) in the root leading to reduced arsenic mobilisation from the root to shoot, husk and grains. The restriction of arsenic in the root by the fungus was due to the modulation in iron (Fe) partitioning, resulting in the protection of the plant from arsenic stress and reduced level of arsenic in the grain. We show that using axenically cultivable fungus S. indica in highly arsenic-contaminated agricultural soil may reduce arsenic stress and accumulation in rice plants through modulation in Fe homeostasis.

Fruit diseases brought on by fungus infestation leads to postharvest losses of fresh fruit. Approximately 30% of harvested fruits do not reach consumers’ plates due to postharvest losses. Fungal pathogens play a substantial part in those losses, as they cause the majority of fruit rots and consumer complaints. Understanding fungal pathogenic processes and control measures is crucial for developing disease prevention and treatment strategies. In this review, we covered the presented pathogen entry, environmental conditions for pathogenesis, fruit’s response to pathogen attack, molecular mechanisms by which fungi infect fruits in the postharvest phase, production of mycotoxin, virulence factors, fungal genes involved in pathogenesis, and recent strategies for protecting fruit from fungal attack. Then, in order to investigate new avenues for ensuring fruit production, existing fungal management strategies were then assessed based on their mechanisms for altering the infection process. The goal of this review is to bridge the knowledge gap between the mechanisms of fungal disease progression and numerous disease control strategies being developed for fruit farming.

KEY MESSAGE:An optimal RNAi configuration that could restrict gene expression most efficiently was determined. This approach was also used to target PTGS and yielded higher rates of gene-editing events. Although it was characterized long ago, transgene silencing still strongly impairs transgene overexpression, and thus is a major barrier to plant crop gene-editing. The development of strategies that could prevent transgene silencing is therefore essential to the success of gene editing assays. Transgene silencing occurs via the RNA silencing process, which regulates the expression of essential genes and protects the plant from viral infections. The RNA silencing machinery thereby controls central biological processes such as growth, development, genome integrity, and stress resistance. RNA silencing is typically induced by aberrant RNA, that may lack 5' or 3' processing, or may consist in double-stranded or hairpin RNA, and involves DICER and ARGONAUTE family proteins. In this study, RNAi inducing constructs were designed in eleven different configurations and were evaluated for their capacity to induce silencing in Nicotiana spp. using transient and stable transformation assays. Using reporter genes, it was found that the overexpression of a hairpin consisting of a forward tandem inverted repeat that started with an ATG and that was not followed downstream by a transcription terminator, could downregulate gene expression most potently. Furthermore, using this method, the downregulation of the NtSGS3 gene caused a significant increase in transgene expression both in transient and stable transformation assays. This SGS3 silencing approach was also employed in gene-editing assays and caused higher rates of gene-editing events. Taken together, these findings suggested the optimal genetic configuration to cause RNA silencing and showed that this strategy may be used to restrict PTGS during gene-editing experiments.

Symbiotic interactions play a crucial role in the phosphate (Pi) nutrient status of the host plant and offer resilience during biotic and abiotic stresses. Despite a competitive behavior of arsenic (AsV) with Pi, Serendipita indica association promotes plant growth by reducing arsenic bioavailability in the rhizosphere. Reduced arsenic availability is due to the adsorption, accumulation, and precipitation of arsenic in the fungus. The present investigation focused on the fitness and performance of Pi acquisition and utilization in S. indica for growth and metabolism under arsenic stress. The fungus accumulates a massive amount of arsenic up to 2459.3 ppm at a tolerable limit of arsenic supply (1 mM) with a bioaccumulation factor (BAF) 32. Arsenic induces Pi transporter expression to stimulate the arsenic acquisition in the fungus. At the same time, Pi accumulation was also enhanced by 112.2 times higher than the control with an increase in poly-P (polyphosphate) content (6.69 times) of the cell. This result suggests arsenic does not hamper poly-P storage in the cell but shows a marked delocalization of stored poly-P from the vacuoles. Furthermore, an enhanced exopolyphosphatase activity and poly-P storage during arsenic stress suggest induction of cellular machinery for the utilization of Pi is required to deal with arsenic toxicity and competition. However, at high arsenic supply (2.5 and 5 mM), 14.55 and 22.07 times reduced Pi utilization, respectively, was observed during the Pi uptake by the fungus. The reduction of Pi uptake reduces the cell growth and biomass due to competition between arsenic and phosphate. The study suggests no negative impact of arsenic on the Pi acquisition, storage, and metabolism in symbiotic fungus, S. indica, under environmental arsenic contamination.

Arsenic (As) is a toxic metalloid that is present in natural surroundings in many forms with severe consequences to sustainable agriculture and human health. Plant growth-promoting Rhizobia have been found involved in the induction of plant tolerance under various biotic and abiotic stresses. An endofungal Rhizobium species associated with arbuscular mycorrhizal fungi (AMF) Serendipita indica deploy beneficial role in the promotion of plant growth and tolerance against various biotic and abiotic stresses. In the current study, we have determined the role of endofungal Rhizobium species in protection of host plant growth under As stress. We observed that endofungal Rhizobium species strain Si001 tolerate AsV up to 25 mM and its inoculation enhances tomato seed germination and seedling growth. A hyper-colonization of Rhizobium species Si001 in tomato roots was observed under As stress and results in modulation of GSH and proline content with reduced ROS. Rhizobium species Si001 colonization in host plant recovered pigment contents (chlorophyll-a and chlorophyll-b up to 189.5% and 192%, respectively), photosynthesis (157%), and water use efficiency (166%) compared to As-treated plants. Interestingly, bacterial colonization results in 40% increased As accumulation in the root, while a reduction in As translocation from root to shoot up to 89% was observed as compared to As treated plants. In conclusion, endofungal Rhizobium species Si001 association with the host plant may improve plant health and tolerance against As stress with reduced As accumulation in the crop produce.

Salinity-induced ethylene production and reactive oxygen species (ROS) inhibit agricultural productivity. The plant synthesizes ethylene directly from aminocyclopropane-1-carboxylic acid (ACC). By using ACC as a nitrogen source, bacteria with ACC deaminase (ACCD) inhibit the overproduction of ethylene, thereby maintaining the ROS. The present study investigated the ACCD activity of previously identified rhizobacterial strains in Dworkin and Foster (DF) minimal salt media supplemented with 5 mM ACC (as N-source). Bacterial isolates GKP KS2_7 (Pseudomonas aeruginosa) and MBD 133 (Bacillus subtilis) could degrade ACC into α-ketobutyrate, exhibiting ACCD activity producing more than ~257 nmol of α-ketobutyrate mg protein−1 h−1, and were evaluated for other plant growth-promoting (PGP) traits including indole acetic acid production (>63 µg/mL), phosphate solubilization (>86 µg mL−1), siderophore (>20%) ammonia and exopolysaccharide production. Furthermore, Fourier Transform Infrared analysis also demonstrated α-ketobutyrate liberation from ACC deamination in DF minimal salt media, thereby confirming the ACCD activity. These isolates also showed enhanced tolerance to salinity stress of 3% w/v NaCl in vitro, in addition to facilitating multifarious PGP activities. Seed bacterization by these ACCD-producing bacterial isolates (GKP KS2_7 and MBD 133) revealed a significant decline in stress-stimulated ethylene levels and its associated growth inhibition during seedling germination. They also mitigated the negative effects of salt stress and increased the root-shoot length, fresh and dry weight of root and shoot, root-shoot biomass, total sugar, protein, reducing sugar, chlorophyll content, and antioxidants enzymes in Pisum sativum. As a result, these strains (GKP KS2_7 and MBD 133) might be applied as biofertilizers to counteract the negative effects of soil salinity.

Climate change has devastating effects on plant growth and yield. During ontogenesis, plants are subjected to a variety of abiotic stresses, including drought and salinity, affecting the crop loss (20–50%) and making them vulnerable in terms of survival. These stresses lead to the excessive production of reactive oxygen species (ROS) that damage nucleic acid, proteins, and lipids. Plant growth-promoting bacteria (PGPB) have remarkable capabilities in combating drought and salinity stress and improving plant growth, which enhances the crop productivity and contributes to food security. PGPB inoculation under abiotic stresses promotes plant growth through several modes of actions, such as the production of phytohormones, 1-aminocyclopropane-1-carboxylic acid deaminase, exopolysaccharide, siderophore, hydrogen cyanide, extracellular polymeric substances, volatile organic compounds, modulate antioxidants defense machinery, and abscisic acid, thereby preventing oxidative stress. These bacteria also provide osmotic balance; maintain ion homeostasis; and induce drought and salt-responsive genes, metabolic reprogramming, provide transcriptional changes in ion transporter genes, etc. Therefore, in this review, we summarize the effects of PGPB on drought and salinity stress to mitigate its detrimental effects. Furthermore, we also discuss the mechanistic insights of PGPB towards drought and salinity stress tolerance for sustainable agriculture.

MAIN CONCLUSION:An efficient method of DNA-free gene-editing in potato protoplasts was developed using linearized DNA fragments, UBIQUITIN10 promoters of several plant species, kanamycin selection, and transient overexpression of the BABYBOOM transcription factor. Plant protoplasts represent a reliable experimental system for the genetic manipulation of desired traits using gene editing. Nevertheless, the selection and regeneration of mutated protoplasts are challenging and subsequent recovery of successfully edited plants is a significant bottleneck in advanced plant breeding technologies. In an effort to alleviate the obstacles related to protoplasts' transgene expression and protoplasts' regeneration, a new method was developed. In so doing, it was shown that linearized DNA could efficiently transfect potato protoplasts and that UBIQUITIN10 promoters from various plants could direct transgene expression in an effective manner. Also, the inhibitory concentration of kanamycin was standardized for transfected protoplasts, and the NEOMYCIN PHOSPHOTRANSFERASE2 (NPT2) gene could be used as a potent selection marker for the enrichment of transfected protoplasts. Furthermore, transient expression of the BABYBOOM (BBM) transcription factor promoted the regeneration of protoplast-derived calli. Together, these methods significantly increased the selection for protoplasts that displayed high transgene expression, and thereby significantly increased the rate of gene editing events in protoplast-derived calli to 95%. The method developed in this study facilitated gene-editing in tetraploid potato plants and opened the way to sophisticated genetic manipulation in polyploid organisms.