Kaolinite nanoclay-shielded dsRNA drenching for management of Globodera pallida: An environmentally friendly pest management approach.

Globodera pallida, an obligate sedentary endoparasite, is a major economic pest that causes substantial potato yield losses. This research aimed to study the effects of gene silencing of three FMRFamide-like peptides (FLPs) genes to reduce G. pallida infestation on potato plants by using kaolinite nanoclay as a carrier to deliver dsRNAs via drenching. A dsRNA dosage of 2.0 mg/ml silenced flp-32c by 89.5%, flp-32p by 94.6%, and flp-2 by 94.3%. J2s incubated for 5 and 10 h showed no phenotypic changes. However, J2s of G. pallida efficiently uptake dsRNA of all targeted genes after 15 h of incubation. On the other hand, J2s that had been kept for 24 h had a rigid and straight appearance. Under fluorescence microscopy, all dsRNA-treated nematodes showed fluorescein isothiocyanate (FITC) signals in the mouth, nervous system, and digestive system. The untreated population of J2s did not show any FITC signals and was mobile as usual. The drenching of potato cultivar Kufri Jyoti with the dsRNA-kaolinite formulations induced deformation and premature death of J2s, compared with untreated J2s that entered J3 or J4 stages. This study validates that the nanocarrier-delivered RNAi system could be employed effectively to manage G. pallida infestations.

Identification of genes governing resistance to PCN (Globodera rostochiensis) through transcriptome analysis in Solanum tuberosum.

Potato cyst nematodes (PCNs) are major pests worldwide that affect potato production. The molecular changes happening in the roots upon PCN infection are still unknown. Identification of transcripts and genes governing PCN resistance will help in the development of resistant varieties. Hence, differential gene expression of compatible (Kufri Jyoti) and incompatible (JEX/A-267) potato genotypes was studied before (0 DAI) and after (10 DAI) inoculation of Globodera rostochiensis J2s through RNA sequencing (RNA-Seq). Total sequencing reads generated ranged between 33 and 37 million per sample, with a read mapping of 48-84% to the potato reference genome. In the infected roots of the resistant genotype JEX/A-267, 516 genes were downregulated, and 566 were upregulated. In comparison, in the susceptible genotype Kufri Jyoti, 316 and 554 genes were downregulated and upregulated, respectively. Genes encoding cell wall proteins, zinc finger protein, WRKY transcription factors, MYB transcription factors, disease resistance proteins, and pathogenesis-related proteins were found to be majorly involved in the incompatible reaction after PCN infection in the resistant genotype, JEX/A-267. Furthermore, RNA-Seq results were validated through quantitative real-time PCR (qRT-PCR), and it was observed that ATP, FLAVO, CYTO, and GP genes were upregulated at 5 DAI, which was subsequently downregulated at 10 DAI. The genes encoding ATP, FLAVO, LBR, and GP were present in > 1.5 fold before infection in JEX-A/267 and upregulated 7.9- to 27.6-fold after 5 DAI; subsequently, most of these genes were downregulated to 0.9- to 2.8-fold, except LBR, which was again upregulated to 44.4-fold at 10 DAI.

Development and evaluation of loop-mediated isothermal amplification assay for rapid and sensitive detection of potato cyst nematode, Globodera pallida from soil.

Potato cyst nematodes, Globodera pallida and G. rostochiensis, are economically important and difficult to manage pests of the potato crop. The cyst of both the species looks similar and it is difficult to differentiate once it turns brown upon maturity. Early detection of the PCN at the species level is crucial to avoid its further spread and for adopting the appropriate management strategies. Therefore, in the present study, highly specific and sensitive loop-mediated isothermal amplification (LAMP) assay was developed to amplify mitochondrial-Sequence Characterized Amplified Region (SCAR) sequence of potato cyst nematode, G. pallida. The LAMP assay was completed within a shorter incubation period of 60 min at 60 °C followed by the reaction termination at 80 °C for 5 min. The developed LAMP assay exhibited high specificity for G. pallida and did not detect any other species including its sibling species, G. rostochiensis. In sensitivity tests, the assay detected G. pallida at 1000 times less DNA concentration (10 fg/µl) as compared to conventional PCR (10 pg/µl). In addition to this, the developed LAMP assay was tested for the detection of G. pallida directly from the soil samples, and even a single cyst mixed with soil was successfully detected by the developed assay. Moreover, the utility of low-cost instruments like hot water bath was also demonstrated for the detection of G. pallida from the soil. The developed LAMP is a rapid, highly specific, sensitive, and cost-effective technique for the species-specific detection of G. pallida. The developed assay will facilitate the rapid detection of G. pallida at quarantine stations as well as from the fields which will help to stop its further spread in new areas and also to devise effective management strategies for sustainable potato production. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-023-03542-x.

Chaetomium globosum KPC3: An Antagonistic Fungus Against the Potato Cyst Nematode, Globodera rostochiensis.

The potato cyst nematode (Globodera rostochiensis) is one of the most economically important pests of potato (Solanum tuberosum L.), causing significant economic losses worldwide. The identification of biocontrol agents for the sustainable management of G. rostochiensis is crucial. In this study, a potential biocontrol agent, Chaetomium globosum KPC3, was identified based on sequence analysis of the DNA internal transcribed spacer (ITS) region, the translation elongation factor 1-alpha (TEF1-α) gene, and the second largest subunit of the RNA polymerase II (RPB2) gene. The pathogenicity test of C. globosum KPC3 against cysts and second-stage juveniles (J2s) revealed that fungus mycelium fully parasitized the cyst after 72 h of incubation. The fungus was also capable of parasitizing the eggs inside the cysts. The culture filtrate of C. globosum KPC3 caused 98.75% mortality in J2s of G. rostochiensis after 72 h of incubation. The pot experiments showed that the combined application of C. globosum KPC3 as a tuber treatment at a rate of 1 lit kg-1 of tubers and a soil application at a rate of 500 ml kg-1 of farm yard manure (FYM) resulted in significantly lesser reproduction of G. rostochiensis compared to the rest of the treatments. Altogether, C. globosum KPC3 has the potential to be used as a biocontrol agent against G. rostochiensis and can be successfully implemented in integrated pest management programs.

Application of ultrasonication as pre-treatment for freeze drying: An innovative approach for the retention of nutraceutical quality in foods.

Freeze drying (FD) is an important and highly effective technology in food industry for retaining the quality in final dried product. This drying technique is performed at lower temperatures, restricting the damage suffered by thermally sensitive ingredients. However, FD consumes large amount of energy and required more time than conventional drying methods. The utilization of ultrasonic technology (US) as pre-treatment before FD represents a promising alternative in accelerating the drying process, decreases energy consumption and maintaining quality as compared to the non pre-treated sample. This review summarizes research progress and current studies in ultrasonic as pre-treatment for freeze drying (US + FD) technique. The impact of US + FD on phytochemical, color, texture and micro-structure of food are well summarized. The review also suggests that the optimised US treatment parameters are required to improve heat and mass transfer in food samples which help in speed up the drying process and reduction of drying time.

Management of potato cyst nematodes with special focus on biological control and trap cropping strategies.

Potato cyst nematodes (PCNs; Globodera spp.) are one of the most difficult pests of potato to manage worldwide. Indiscriminate use of pesticides and their hazardous effects discourage the use of many chemicals for the management of PCNs. As a result, biological control agents and trap crops have received more attention from growers as safer ways to manage PCNs. The biological control agents such as Pochonia chlamydosporia, Purpureocillium lilacinum, Trichoderma spp., Pseudomonas fluorescens, Bacillus spp., Pasteuria spp., and others are recognized as potential candidates for the management of PCNs. Moreover recently, the use of trap crop Solanum sisymbriifolium also showed promise by drastically reducing soil populations of PCNs. Integration of these management strategies along with other practices including identification, conservation, and multiplication of native antagonists, will facilitate efficient management of the PCNs in potato cropping system. Some of the promising research approaches that are being used against PCNs are addressed in this review. © 2022 Society of Chemical Industry.

RNA-Seq of Cyst Nematode Infestation of Potato (Solanum tuberosum L.): A Comparative Transcriptome Analysis of Resistant and Susceptible Cultivars.

Potato (Solanum tuberosum L.) is an important food crop worldwide, and potato cyst nematodes (PCNs) are among the most serious pests. The identification of disease resistance genes and molecular markers for PCN infestation can aid in crop improvement research programs against PCN infestation. In the present study, we used high-throughput RNA sequencing to investigate the comprehensive resistance mechanisms induced by PCN infestation in the resistant cultivar Kufri Swarna and the susceptible cultivar Kufri Jyoti. PCN infestation induced 791 differentially expressed genes in resistant cultivar Kufri Swarna, comprising 438 upregulated and 353 downregulated genes. In susceptible cultivar Kufri Jyoti, 2225 differentially expressed genes were induced, comprising 1247 upregulated and 978 downregulated genes. We identified several disease resistance genes (KIN) and transcription factors (WRKY, HMG, and MYB) that were upregulated in resistant Kufri Swarna. The differentially expressed genes from several enriched KEGG pathways, including MAPK signaling, contributed to the disease resistance in Kufri Swarna. Functional network analysis showed that several cell wall biogenesis genes were induced in Kufri Swarna in response to infestation. This is the first study to identify underlying resistance mechanisms against PCN and host interaction in Indian potato varieties.

Biocontrol potential of entomopathogenic nematodes for the sustainable management of Spodoptera frugiperda (Lepidoptera: Noctuidae) in maize.

BACKGROUND: The occurrence of Spodoptera frugiperda (J.E. Smith) in Asia was reported for the first time from Karnataka in 2018. This pest is widely distributed in India, causing significant damage to maize. Management of this recent invasive pest in maize-growing regions of India relies on chemical control. Resistance is the greatest obstacle to the successful use of chemical insecticides to control this pest. Indiscriminate use of chemical insecticides destroys beneficial natural enemies, therefore effective and sustainable alternative control strategies are needed. In this case, the use of biological control agents is the alternative option to mitigate this pest. Thus, this study aimed to select virulent entomopathogenic nematodes (EPNs) isolates based on the laboratory assay and further to test the efficacy of virulent isolates in the field conditions along with commonly used chemical insecticide emamectin benzoate against S. frugiperda. RESULTS: Laboratory results revealed that both Heterorhabditis indica 1 NBAIIH38 and Steinernema carpocapsae NBAIRS59 caused 100% mortality in third- and fourth-instar larvae of S. frugiperda, while these two species caused 85% and 72% mortality in pupae, respectively. When pupae of S. frugiperda were exposed to EPNs, pupae died after metamorphosis to malformed adults. All the nematode species were able to penetrate and reproduce within S. frugiperda larvae, but the reproduction rate for Heterorhabditids was higher than that of Steinernematids. Field trial results showed that H. indica 1 NBAIIH38 significantly reduced the number of larvae and leaf damage scores compared to S. carpocapsae NBAIRS59. Emamectin benzoate was more effective in reducing the larval population compared to EPN species. The cob yield was significantly higher in EPN- and emamectin benzoate-treated plots than in untreated control plots. CONCLUSION: Overall, these experiments suggest H. indica 1 NBAIIH38 is a promising biocontrol agent against S. frugiperda in maize production. © 2022 Society of Chemical Industry.

Delineation of mechanistic approaches of rhizosphere microorganisms facilitated plant health and resilience under challenging conditions.

Sustainable agriculture demands the balanced use of inorganic, organic, and microbial biofertilizers for enhanced plant productivity and soil fertility. Plant growth-enhancing rhizospheric bacteria can be an excellent biotechnological tool to augment plant productivity in different agricultural setups. We present an overview of microbial mechanisms which directly or indirectly contribute to plant growth, health, and development under highly variable environmental conditions. The rhizosphere microbiomes promote plant growth, suppress pathogens and nematodes, prime plants immunity, and alleviate abiotic stress. The prospective of beneficial rhizobacteria to facilitate plant growth is of primary importance, particularly under abiotic and biotic stresses. Such microbe can promote plant health, tolerate stress, even remediate soil pollutants, and suppress phytopathogens. Providing extra facts and a superior understanding of microbial traits underlying plant growth promotion can stir the development of microbial-based innovative solutions for the betterment of agriculture. Furthermore, the application of novel scientific approaches for facilitating the design of crop-specific microbial biofertilizers is discussed. In this context, we have highlighted the exercise of "multi-omics" methods for assessing the microbiome's impact on plant growth, health, and overall fitness via analyzing biochemical, physiological, and molecular facets. Furthermore, the role of clustered regularly interspaced short palindromic repeats (CRISPR) based genome alteration and nanotechnology for improving the agronomic performance and rhizosphere microbiome is also briefed. In a nutshell, the paper summarizes the recent vital molecular processes that underlie the different beneficial plant-microbe interactions imperative for enhancing plant fitness and resilience under-challenged agriculture.