2016
DOI: 10.1371/journal.pone.0154818
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Optimization of In Vitro Techniques for Distinguishing between Live and Dead Second Stage Juveniles of Heterodera glycines and Meloidogyne incognita

Abstract: Heterodera glycines (Soybean Cyst nematode, or SCN) and Meloidogyne incognita (Root-Knot nematode, or RKN) are two damaging plant-parasitic nematodes on important field crops. Developing a quick method to distinguish between live and dead SCN and RKN second stage juveniles (J2) is vital for high throughput screening of pesticides or biological compounds against SCN and RKN. The in vitro assays were conducted in 96-well plates to determine the optimum chemical stimulus to distinguish between live and dead SCN a… Show more

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Cited by 25 publications
(24 citation statements)
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“…[Eggs were extracted from roots by placing the root system in a 0.625% NaOCl solution and agitating the roots for 4 min using a rotary shaker at 120 rpm Eggs were rinsed with tap water, collected on a 25-μm-pore sieve, then processed by sucrose centrifugation-flotation at 240 g for 1 minute [60]. M. incognita eggs were placed in a modified Baermann funnel [65] on a slide warmer (Model 77) (Marshall Scientific, Brentwood, NH) and incubated at 31˚C for 5 to 7 days to obtain second stage juveniles (J2) [66]. The J2 were collected on a 25-μm-pore sieve, transferred to 1.5 ml microcentrifuge tubes, centrifuged at 5,000 g for 1 minute, rinsed with sterile distilled water, and centrifuged at 5,000 g for 1 minute.…”
Section: Infection By M Incognita and Analysis Of Resultsmentioning
confidence: 99%
See 1 more Smart Citation
“…[Eggs were extracted from roots by placing the root system in a 0.625% NaOCl solution and agitating the roots for 4 min using a rotary shaker at 120 rpm Eggs were rinsed with tap water, collected on a 25-μm-pore sieve, then processed by sucrose centrifugation-flotation at 240 g for 1 minute [60]. M. incognita eggs were placed in a modified Baermann funnel [65] on a slide warmer (Model 77) (Marshall Scientific, Brentwood, NH) and incubated at 31˚C for 5 to 7 days to obtain second stage juveniles (J2) [66]. The J2 were collected on a 25-μm-pore sieve, transferred to 1.5 ml microcentrifuge tubes, centrifuged at 5,000 g for 1 minute, rinsed with sterile distilled water, and centrifuged at 5,000 g for 1 minute.…”
Section: Infection By M Incognita and Analysis Of Resultsmentioning
confidence: 99%
“…The J2 were collected on a 25-μm-pore sieve, transferred to 1.5 ml microcentrifuge tubes, centrifuged at 5,000 g for 1 minute, rinsed with sterile distilled water, and centrifuged at 5,000 g for 1 minute. The J2 suspension was adjusted to 30 to 40 J2 per 10 μl of water [66]. M. incognita extraction has been performed by gravity screening and centrifugal flotation (sucrose specific gravity = 1.13) [60].…”
Section: Infection By M Incognita and Analysis Of Resultsmentioning
confidence: 99%
“…The mortality rate of J2 at different concentrations (500 μg ml − 1 , 600 μg ml − 1 , and 1000 μg ml − 1 ) of 4-vinylphenol, L-methionine, piperine, and palmitic acid was assessed in vitro as described previously [ 35 ] and calculated as follows: ((live J2 prior to exposure − live J2 at 24 or 48 h post exposure)/live J2 prior to exposure) × 100. All compounds were of analytical grade and purchased from Sigma-Aldrich (St Louis, MO, USA), except for piperine that purchased from Jianglai (Shanghai, China).…”
Section: Methodsmentioning
confidence: 99%
“…Nematoda yang inaktif akan kembali aktif setelah ditetesi NaOH. Nematoda yang masih hidup akan bergerak aktif kembali atau akan menunjukkan bentuk tubuh yang tidak lurus, sedangkan nematoda yang mati akan terdeteksi bentuk tubuhnya lurus (Harada & Yoshiga 2015;Xiang & Lawrence 2016).…”
Section: Konfirmasi Mortalitas Nematodaunclassified