Laboratory methods for work with plant and soil nematodes

http://besa.oxfordjournals.org/content/besa/26/4/475.2.full.pdf

Nematodes for Biological Control of Insects

Plant beneficial microbes (PBMs), such as plant growth-promoting bacteria, rhizobia, arbuscular mycorrhizal fungi, and Trichoderma, can reduce the use of agrochemicals and increase plant yield, nutrition, and tolerance to biotic-abiotic stresses. Yet, large-scale applications of PBM have been hampered by the high amounts of inoculum per plant or per cultivation area needed for successful colonization and consequently the economic feasibility. Seed coating, a process that consists in covering seeds with low amounts of exogenous materials, is gaining attention as an efficient delivery system for PBM. Microbial seed coating comprises the use of a binder, in some cases a filler, mixed with inocula, and can be done using simple mixing equipment (e.g., cement mixer) or more specialized/sophisticated apparatus (e.g., fluidized bed). Binders/fillers can be used to extend microbial survival. The most reported types of seed coating are seed dressing, film coating, and pelleting. Tested in more than 50 plant species with seeds of different dimensions, forms, textures, and germination types (e.g., cereals, vegetables, fruits, pulses, and other legumes), seed coating has been studied using various species of plant growth-promoting bacteria, rhizobia, Trichoderma, and to a lesser extent mycorrhizal fungi. Most of the studies regarding PBM applied via seed coating are aimed at promoting crop growth, yield, and crop protection against pathogens. Studies have shown that coating seeds with PBM can assist crops in improving seedling establishment and germination or achieving high yields and food quality, under reduced chemical fertilization. The right combination of biological control agents applied via seed coating can be a powerful tool against a wide number of diseases and pathogens. Less frequently, studies report seed coating being used for adaptation and protection of crops under abiotic stresses. Notwithstanding the promising results, there are still challenges mainly related with the scaling up from the laboratory to the field and proper formulation, including efficient microbial combinations and coating materials that can result in extended shelf-life of both seeds and coated PBM. These limitations need to be addressed and overcome in order to allow a wider use of seed coating as a cost-effective delivery method for PBM in sustainable agricultural systems.

/pdf/seed-coating-a-tool-for-delivering-beneficial-microbes-to-4yr5i5do3t.pdf

Seed Coating: A Tool for Delivering Beneficial Microbes to Agricultural Crops

Root-knot nematode (Meloidogyne incognita) is chief plant parasitic nematode of various crops globally. Meanwhile, the negative side effects on human health and environmental concerns associated with haphazard uses of chemical nematicides. Hence, the search for a safe and effective approach is more relevant. The present study was aimed to evaluate the nematicidal potential of Snef1216 (Penicillium chrysogenum) against M. incognita at different concentrations (5%, 10%, 25%, 50%, 75% and 100%) and with the nutritious medium. The egg hatching inhibition and mortality of second stage juveniles of M. incognita were assessed after 6, 12, 24, 48 and 72 h exposure. Results revealed that egg hatching inhibition and percent mortality of M. incognita increased with increasing concentration and exposure time. The highest mortality of juveniles was recorded at 100% conc. i.e., 24.20%, 36%, 66%, 78% and 97.8% at 6, 12, 24, 48 and 72 h, respectively. The highest ovicidal activity was recorded at 100% concentration with 5.20% of eggs hatching. The outcome suggested that Snef1216 (P. chrysogenum) resulted in the lowest LC50 value was recorded as 3718.496 at 6 h exposure period followed by 10479.87, 11186.682, 14838.58 and 24001.430 at 72, 12, 48 and 24 h respectively via ovicidal assay. Whereas, in the larvicidal assay, the lowest LC50 value demonstrated at 72 h being 17.628% exposure period followed by 28.345, 50.490, 215.710 and 482.595% at 48, 24, 12 and 6 h respectively. It is concluded that Snef1216 has potential being used as a biocontrol agent against M. incognita and can serve as a source of a novel nematicidal agent of fungal origin.

/pdf/in-vitro-evaluation-of-penicillium-chrysogenum-snef1216-4lan7tu78i.pdf

In Vitro Evaluation of Penicillium Chrysogenum Snef1216 Against Meloidogyne Incognita (Root-Knot Nematode)

Entomopathogenic nematodes from the two genera Steinernema and Heterorhabditis are widely used as biological agents against various insect pests and represent a promising alternative to replace pesticides. Efficacy and biocontrol success can be enhanced through improved understanding of their biology and ecology. Many endogenous and environmental factors influence the survival of nematodes following application, as well as their transmission success to the target species. The aim of this paper is to give an overview of the major topics currently considered to affect transmission success of these biological control agents, including interactions with insects, plants and other members of the soil biota including conspecifics.

Transmission Success of Entomopathogenic Nematodes Used in Pest Control

The efficacy of Trichoderma harzianum and Moringa oleifera at various concentrations (S, S/2, S/4) and in combinations was tested on the reproduction, egg hatching and juvenile mortality of Meloidogyne javanica on eggplant. The evaluation was done in vitro as well as in vivo in green house. Both alone and combined application of T. harzianum and M. oleifera caused significant (P = 0.05) reduction in egg hatching and juvenile mortality at all time intervals (24, 48 and 72 h). Reproduction of M. javanica was evaluated on eggplant in green house. Most effective concentration from in vitro experiment was used in further protective and curative experiment. There were three treatments and five replications. The plant growth parameters including shoot length, shoot weight, root length, root weight and number of leaves and nematode parameters including number of galls, egg masses, number of females, number of juveniles per root system and no. of juveniles/100ml soil were recorded after 60 days of growth. Nematode population was significantly reduced when biocontrol agent was applied before the inoculation of nematodes as protective application. So, T. harzianum and M. oleifera might be successfully used in field conditions against M. javanica.

Combined efficacy of Moringa oleifera leaves and a fungus, Trichoderma harzianum against Meloidogyne javanica on eggplant.

Targeted genome editing (GE) technology has brought a significant revolution in fictional genomic research and given hope to plant scientists to develop desirable varieties. This technology involves inducing site-specific DNA perturbations that can be repaired through DNA repair pathways. GE products currently include CRISPR-associated nuclease DNA breaks, prime editors generated DNA flaps, single nucleotide-modifications, transposases, and recombinases. The discovery of double-strand breaks, site-specific nucleases, and repair mechanisms paved the way for targeted genome editing, and the first-generation genome editing tools, ZFNs and TALENs, were successfully utilized in plant genome editing. However, CRISPR-Cas has now become the preferred tool for genome editing due to its speed, reliability, and cost-effectiveness. Plant functional genomics has benefited significantly from the widespread use of CRISPR technology for advancements and developments. This review highlights the progress made in CRISPR technology, including multiplex editing, base editing, and prime editing, as well as the challenges and potential delivery mechanisms. This article is protected by copyright. All rights reserved.

Breakthrough in CRISPR/Cas system: Current and future directions and challenges.

The purpose of this study was to assess the effects of increasing initial population density (Pi) of Meloidogyne incognita on nematode reproduction and growth of susceptible tomato (Lycopersicom esculentum cv. Round-41) at 25°C± 2 in growth chamber. 21-days-old tomato seedlings transplanted in13-cm dia. earthen pot was inoculated with Pi including 250, 500, 1000, 1500 eggs of M. incognita. Nematode reproduction was assessed by determining the number of galls, eggs masses, eggs per root system, and reproduction rate values per root system at 60 days after inoculations. Egg-masses on Phloxine B- stained roots were quantified and root systems were rated for galling and egg mass presence on a 0 to 5 scale where 0 = no gall or egg masses, 1 = 1-2, 2 = 3-10, 3 = 11-30, 4 = 31- 100, and 5 = >100 galls or egg masses per root system. Nematode reproduction was directly proportional to Pi. Reproduction rate (Pf/Pi, where Pf = final number of eggs / initial egg density) for Pi's of 250, 500, 1000 and 1500 eggs/plant was lowest when Pi = 250 (2.23) but similar when Pi = 500, 1000 or 1500. Shoot and root growth were inversely related to Pi. Strong liner relationship exited between root length, root galling index and foliage growth but very poor between root weight and foliage growth. High Pf = final eggs per root system than at all Pi suggests that Round-41 is a good host for M. incognita

The Influence of Meloidogyne incognita Density on Susceptible Tomato

Entomopathogenic nematodes; Steinernema asiaticum (Anis), Steinernema glaseri (Steiner), Heterorhabditis indica (Poinar) and Heterorhabditis bacteriophora (Poinar) were investigated for their effect on Meloidogyne incognita (Kofoid and White) in tomato. They were applied at different levels before and at the same time with the application of root-knot nematodes. Suppression of M. incognita varied with application rates of S. asiaticum, S. glaseri, H. indica and H. bacteriophora in tomato. The high application rates of S. asiaticum, S. glaseri, H. indica and H. bacteriophora applied both at the same time and before reduced M. incognita egg production. The low rates of S. asiaticum, S. glaseri, H. indica and H. bacteriophora were not as effective as high application rates. The findings of this study suggested that entomopathogenic nematodes could be used for the management of root-knot nematodes.

Management of Meloidogyne incognita race 1 through the use of entomopathogenic nematodes in tomato.

Morphological variability of four isolates of Colletotrichum falcatum Went. causing red rot of sugarcane collected from four cultivars: SPF-234, Co-1148, BF-162 and SHF-242 were established. Four red rot isolates were differentiated into two distinct groups based on growth rates. Isolates Co-1148, BF-162 produced medium density colony whereas isolates SHF-242 and SPF-234 had thick density colony. Four isolates also exhibited variability in colony color. The color of colony of CO-1148, BF-162, SHF-242 and SPF-234 Isolates was orange, brown, red and red bottom, respectively. We concluded that C. falcatum isolates are morphologically dissimilar. This investigation demands to substantiate the morphologically

Morphological variability among isolates of Colletotrichum falcatum Went. infecting four cultivars of sugarcane.

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