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Single nucleotide polymorphism (SNP) marker-assisted breeding of Heterorhabditis bacteriophora for improvement of reproductive potential and stress tolerance 单核苷酸多态性(SNP)标记辅助细菌杂环虫育种以提高繁殖潜力和抗逆性
IF 1.2 4区 生物学 Q2 Agricultural and Biological Sciences Pub Date : 2024-02-06 DOI: 10.1163/15685411-bja10307
Christopher Ogaya, Michelle Ann B. Diano, Innocent Hategekimana, Verena Dörfler, Carlos Molina, Ralf-Udo Ehlers
The entomopathogenic nematode (EPN), Heterorhabditis bacteriophora, is an efficient biological control agent against several economically important insect pests. Recent research has assessed the possibility to correlate desired beneficial traits with genotype data to pave a way for marker-assisted breeding approaches. A collection of H. bacteriophora WT inbred lines has been phenotyped in this framework for stress- and virulence-related traits. However, these traits are rarely combined in a single line. Thus, unifying these traits in commercial strains is of high priority. This investigation unified beneficial traits in hybrid pools through marker-assisted breeding using single nucleotide polymorphisms (SNPs) associated with reproductive potential, longevity, virulence and cold tolerance. Recombinant inbred lines (RILs) generated from a cross between a stress tolerant WT inbred line (XX21) and a line high in in vitro reproduction potential (IL3) were genotyped via SeqSNP and screened for SNP markers associated with beneficial traits. Thereafter, a genotypic pool (X21L3) comprising 22 ILs was formed. The X21L3 pool was subsequently evaluated for the target traits in comparison with the cross parents and a commercial strain HB4. An improvement of oxidative stress tolerance at 2°C (cold tolerance) was recorded with X21L3 surviving 1 day longer than the best performing parent (XX21). The hybrid pool also survived 1 day longer than the least performing parent IL3 for the trait longevity at 25°C under oxidative stress conditions. A higher dauer juvenile (DJ) recovery (58%) and DJ yield (209 000 DJ ml−1) than the least performing parent XX21 was recorded for the pool. The storage stability in diatomaceous earth formulation at 2°C and 7.5°C was also improved by 2 and 5 days, respectively, in comparison to the least performing parent XX21. This study depicts the potential of precision marker-assisted breeding for beneficial trait improvement of H. bacteriophora.
昆虫病原线虫(EPN)--细菌异habditis bacteriophora--是一种高效的生物防治剂,可防治多种具有重要经济价值的害虫。最近的研究评估了将所需的有益性状与基因型数据相关联的可能性,从而为标记辅助育种方法铺平道路。在这一框架下,已对一系列菌核病 WT 近交系进行了压力和毒力相关性状的表型分析。然而,这些性状很少结合在一个品系中。因此,在商业品系中统一这些性状是当务之急。这项研究通过使用与繁殖潜力、寿命、毒力和耐寒性相关的单核苷酸多态性(SNPs)进行标记辅助育种,统一了杂交种池中的有益性状。通过 SeqSNP 对抗逆性强的 WT 近交系(XX21)和体外繁殖潜力高的品系(IL3)杂交产生的重组近交系(RIL)进行基因分型,并筛选与有益性状相关的 SNP 标记。之后,形成了一个由 22 个 IL 组成的基因型库(X21L3)。随后,通过与杂交亲本和商业菌株 HB4 的比较,对 X21L3 基因库的目标性状进行了评估。结果表明,X21L3 比表现最好的亲本(XX21)多存活 1 天,从而提高了在 2°C 下的氧化应激耐受性(耐寒性)。在 25°C 的氧化胁迫条件下,杂种池的寿命也比表现最差的亲本 IL3 长 1 天。与表现最差的亲本 XX21 相比,该杂交种池的 Dauer 幼虫(DJ)恢复率(58%)和 DJ 产量(209 000 DJ ml-1)都更高。与性能最差的亲本 XX21 相比,硅藻土制剂在 2°C 和 7.5°C 下的储存稳定性也分别提高了 2 天和 5 天。这项研究展示了精确标记辅助育种在改良菌核病有益性状方面的潜力。
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引用次数: 0
Description of Deladenus tonekabonensis sp. n. from Iran with a key to species 描述来自伊朗的 Deladenus tonekabonensis sp.
IF 1.2 4区 生物学 Q2 Agricultural and Biological Sciences Pub Date : 2024-01-24 DOI: 10.1163/15685411-bja10306
F. Aliramaji, A. Taheri, E. Shokoohi
Deladenus tonekabonensis sp. n. was isolated from dried wood of the Tree of Heaven (Ailanthus altissima) from the Sehezar forests in Tonekabon city in Mazandaran Province. The new species is characterised by a body length of 655 (561-777) and 592 (488-735) μm for mycetophagous females and males, respectively, a short stylet (7.5-8.8 μm) with three knobs backward. Lateral field with ten incisures, and an excretory pore toward the anterior part of the neck, located at 27.5 (22.0-35.5) and 28.0 (21.0-38.0) μm from the anterior end of the mycetophagous females and males, respectively. The distance of the excretory pore to the hemizonid is 66.0 (59.0-71.0) and 61.0 (57.0-67.5) μm for the mycetophagous females and males, respectively. Adults have a long overlapping dorsal pharyngeal gland, lacking a post-uterine sac (PUS), and in some specimens a very short post-uterine sac, 2.4-5.1 μm long, and a conical tail, narrowing towards the tail tip. Males are frequent with slender tylenchoid spicules and a leptoderan bursa reaching the tail tip. Molecular phylogenetic analyses based on 28S rDNA placed the new species close to Deladenus and Anguillonema species in a clade with 0.99 posterior probability support. The measurements, line illustration and light microphotographs are given for the new species.
Deladenus tonekabonensis sp. n.是从马赞达兰省托内卡本市 Sehezar 森林的天堂树(Ailanthus altissima)干燥木材中分离出来的。新物种的特征是:雌性和雄性噬菌体的体长分别为 655 (561-777) μm 和 592 (488-735) μm,花柱短(7.5-8.8 μm),向后有三个节。雌性和雄性噬菌体的侧领域有 10 个切口,颈部前端有一个排泄孔,分别位于距前端 27.5(22.0-35.5)和 28.0(21.0-38.0)微米处。噬菌雌虫和雄虫的排泄孔与半球体的距离分别为 66.0 (59.0-71.0) μm 和 61.0 (57.0-67.5) μm。成虫有一个重叠的长背咽腺体,缺乏子宫后囊(PUS),在某些标本中,子宫后囊非常短,长 2.4-5.1 μm,尾部呈圆锥形,向尾端收窄。雄性标本经常带有细长的泰伦刺囊和到达尾端的钩端囊。基于 28S rDNA 的分子系统学分析表明,该新物种与 Deladenus 和 Anguillonema 接近,同属一个支系,后验概率为 0.99。文中给出了新物种的测量值、线图和光显微照片。
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引用次数: 0
Sensitivity of stubby root nematodes (Trichodorus and Paratrichodorus spp.) to isothiocyanates associated with Brassicaceae in an in vitro assay 茎根线虫(Trichodorus 和 Paratrichodorus spp.)对与十字花科植物有关的异硫氰酸盐的敏感性体外试验
IF 1.2 4区 生物学 Q2 Agricultural and Biological Sciences Pub Date : 2024-01-12 DOI: 10.1163/15685411-bja10302
Nyambura G. Mwangi, Mark Stevens, Alistair Jd Wright, S. Edwards, Martin C. Hare, M. Back
Brassicas contain glucosinolates (GSLs), which are converted into different isothiocyanates (ITCs) that possess biocidal activity. These different ITCs result in a range of toxicities to various target species. Laboratory assays were conducted to evaluate the sensitivity of stubby root nematodes (SRN), Trichodorus and Paratrichodorus spp., to three pure, commercially available ITCs, i.e., 2-phenylethyl (PEITC), allyl (AITC) and sulforaphane (SITC) at different concentrations (1.625, 3.125, 6.25, 12.5, 25 and 50 μg ml−1). The effect on nematode mobility was assessed after 24, 48 and 72 h. Mortality of SRN was assessed after 48 h incubation of the nematodes in distilled water post ITC treatment. Mortality for all ITCs at all tested concentrations was significantly higher than the controls, distilled water and 1% DMSO. Concentration and type of ITC had a significant effect on SRN mobility and mortality, whilst increase in exposure time did not significantly increase the immobility of SRN. The average 24 h ED50 (dose that resulted in 50% immobility) for SRN were 7, 5 and 44 μg ml−1, and the average LD50 (dose that resulted in 50% mortality) after 48 h recovery in distilled water was 7, 11 and 24.3 μg ml−1 for PEITC, AITC and SITC, respectively. SITC was significantly less potent compared to PEITC and AITC, which had LD50 values that were four times and two times lower, respectively. These results indicate the potential use of brassica associated with the tested ITC in the process of biofumigation for SRN suppression.
十字花科植物含有葡萄糖苷酸盐(GSLs),可转化为具有杀菌活性的不同异硫氰酸盐(ITCs)。这些不同的异硫氰酸酯会对不同的目标物种产生不同的毒性。实验室试验评估了粗壮根线虫(SRN)(Trichodorus 和 Paratrichodorus spp.)在不同浓度(1.625、3.125、6.25、12.5、25 和 50 μg ml-1)下对三种市售纯异硫氰酸酯(即 2-苯基乙基(PEITC)、烯丙基(AITC)和磺基(SITC))的敏感性。线虫在 ITC 处理后的蒸馏水中培养 48 小时后,评估 SRN 的死亡率。在所有测试浓度下,所有 ITC 的死亡率都明显高于对照组、蒸馏水和 1%二甲基亚砜。ITC的浓度和类型对SRN的移动性和死亡率有明显的影响,而暴露时间的增加并没有明显增加SRN的不移动性。对于 PEITC、AITC 和 SITC,SRN 在蒸馏水中恢复 48 小时后的平均 ED50(导致 50%不动的剂量)分别为 7、5 和 44 μg ml-1,平均 LD50(导致 50%死亡的剂量)分别为 7、11 和 24.3 μg ml-1。与 PEITC 和 AITC 相比,SITC 的药效明显较低,其半数致死剂量分别低四倍和两倍。这些结果表明,在抑制 SRN 的生物熏蒸过程中,与测试的 ITC 相关的黄铜具有潜在用途。
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引用次数: 0
Steinernema yirgalemense (Steinernematidae) to control Phlyctinus callosus and Phlyctinus xerophilus (Curculionidae) 用 Steinernemema yirgalemense(Steinernematidae)来控制 Phlyctinus callosus 和 Phlyctinus xerophilus(Curculionidae)。
IF 1.2 4区 生物学 Q2 Agricultural and Biological Sciences Pub Date : 2024-01-09 DOI: 10.1163/15685411-bja10305
Steffan P. Hansen, A. Malan, J. Haran, M. Dunn, P. Addison
Considerable progress has been made in the surveying, taxonomy, screening, mass production and formulation of entomopathogenic nematodes (EPN) and their associated symbiotic bacteria in South Africa. Steinernema yirgalemense isolate 157-C is one of the most promising native EPN candidates with regards to virulence, its broad insect-host spectrum, and can be readily mass-produced and formulated into a commercial product. The banded fruit weevils, Phlyctinus callosus sensu stricto and Phlyctinus xerophilus, previously grouped together under the Phlyctinus callosus sensu lato species concept, are native entimine weevils of economic importance to deciduous fruit, grapevine and berries in the Western Cape province of South Africa. This study investigated potential differences in baseline susceptibility of larvae and pupae of the two weevil species to S. yirgalemense in laboratory screenings. The test arena used was 24-well bioassay plates, with an inoculation concentration of 200 infective juveniles (IJ) insect−1 for larvae and 100 IJ insect−1 for pupae. Infection was determined 48 h and 96 h after inoculation. Field-efficacy of S. yirgalemense, applied at a concentration of 60 IJ cm−2, against larvae of the two weevil species was determined in an ecologically relevant semi-field trial. In all cases in vitro mass-produced IJ of S. yirgalemense were used. No baseline differential susceptibility between P. callosus and P. xerophilus larvae was obtained in laboratory screenings. Phlyctinus pupae were approximately twice as susceptible compared to larvae, with significant differences between bioassay batches. Approximately 45% control of P. xerophilus larvae was obtained after 96 h of exposure to S. yirgalemense in the field, differing significantly from the control and P. callosus treatment. Low levels of Phlyctinus larval infection by native EPN (confirmed as Heterorhabditis bacteriophora from one P. xerophilus cadaver) occurred in both control and EPN treatment groups under field conditions.
南非在昆虫病原线虫(EPN)及其相关共生细菌的调查、分类、筛选、大规模生产和配制方面取得了长足的进步。Steinernema yirgalemense 分离物 157-C 是最有前途的本地 EPN 候选者之一,它具有毒性强、昆虫寄主范围广等特点,而且可以很容易地大规模生产并配制成商业产品。带果象鼻虫(Phlyctinus callosus sensu stricto和Phlyctinus xerophilus)以前被归入Phlyctinus callosus sensu lato物种概念下,是对南非西开普省落叶水果、葡萄藤和浆果具有重要经济价值的本地象鼻虫。本研究调查了两种象鼻虫的幼虫和蛹在实验室筛选中对 S. yirgalemense 的基线敏感性的潜在差异。使用的试验场是 24 孔生物测定板,幼虫的接种浓度为 200 感染幼虫 (IJ) 昆虫-1,蛹的接种浓度为 100 感染幼虫 (IJ) 昆虫-1。接种后 48 小时和 96 小时测定感染情况。在与生态相关的半田间试验中,确定了施用浓度为 60 IJ cm-2 的 S. yirgalemense 对两种象鼻虫幼虫的田间药效。在所有情况下,都使用了体外大规模生产的 S. yirgalemense IJ。在实验室筛选中,没有发现 P. callosus 和 P. xerophilus 幼虫之间的基本敏感性差异。与幼虫相比,Phlyctinus 蛹的易感性约为幼虫的两倍,生物测定批次之间存在显著差异。在田间暴露于 S. yirgalemense 96 小时后,P. xerophilus 幼虫的控制率约为 45%,与对照组和 P. callosus 处理有显著差异。在田间条件下,对照组和 EPN 处理组的 Phlyctinus 幼虫都受到了本地 EPN(从一具 P. xerophilus 尸体中确认为细菌异型线虫)的低水平感染。
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引用次数: 0
Nematodes as bioindicators of soil health in different land uses in the São Francisco River Valley, Brazil 线虫是巴西圣弗朗西斯科河谷不同土地用途土壤健康的生物指标
IF 1.2 4区 生物学 Q2 Agricultural and Biological Sciences Pub Date : 2023-12-20 DOI: 10.1163/15685411-bja10303
Larissa de B. Caixeta, J. V. D. L. D. Silva, Nancy Eunice Niño Castañeda, J. M. D. C. Castro, J. Cares
Brazilian Caatinga is one of the most diverse, seasonally dry, tropical forests of the world and is threatened by strong land use pressure and poor protection. One way of understanding changes in the soil conditions caused by anthropogenic activities is using the nematode assemblages as potential bioindicators. This study analysed changes in nematode communities in three different types of land use (natural forest-CA, natural area with accumulation of salts-SS, and agricultural growing areas, such as cover crops-CC for cattle and fruit trees-IC) in the Caatinga, in the sub-median region of the São Francisco River Valley (SFRV), Pernambuco, northeastern Brazil. We recorded a total of 63 262 nematode individuals, representing 42 genera, belonging to 23 families. Nematode abundance, diversity and maturity indices were highest in the CC and lowest in the SS. Among the nematode trophic groups, bacterivores, omnivores + predators and plant parasites were significantly affected by type of land use. Different soil properties were related to the differences in taxonomic composition among the natural and agricultural areas, accounting for 26.6% of the total variation. In general, our results indicate that soil salinisation and desertification in the natural areas, as well as soils subjected to cover crops and fruit tree production, significantly influenced the composition of nematode communities in the São Francisco River Valley.
巴西的 Caatinga 是世界上最多样化、季节性干旱的热带森林之一,受到土地使用压力大和保护不力的威胁。了解人为活动造成的土壤条件变化的一种方法是利用线虫群落作为潜在的生物指标。本研究分析了巴西东北部伯南布哥州圣弗朗西斯科河谷(SFRV)次中间地区卡廷加中三种不同土地利用类型(天然林-CA、盐分积累天然区-SS 和农业种植区,如牛和果树的覆盖作物-CC)中线虫群落的变化。我们共记录了 63 262 个线虫个体,代表 42 个属,隶属 23 个科。线虫丰度、多样性和成熟度指数在 CC 区最高,在 SS 区最低。在线虫营养群中,细菌食性、杂食性+捕食性和植物寄生性受土地利用类型的影响较大。不同的土壤特性与自然区和农业区分类组成的差异有关,占总差异的 26.6%。总之,我们的研究结果表明,自然区的土壤盐碱化和荒漠化以及种植覆盖作物和果树的土壤对圣弗朗西斯科河流域线虫群落的组成有很大影响。
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引用次数: 0
Nematode population dynamics and plant steaming effects on strawberry cultivars under organic field conditions in Florida 在佛罗里达州有机田间条件下,线虫种群动态和植物蒸煮对草莓栽培品种的影响
IF 1.2 4区 生物学 Q2 Agricultural and Biological Sciences Pub Date : 2023-12-20 DOI: 10.1163/15685411-bja10304
Clemen J. Oliveira, Natalia A. Peres, Johan Desaeger
Demand for Florida organic strawberries has significantly increased in recent years, yet this industry is threatened by pests and pathogens that often come with transplants from out-of-state nurseries. Plant steaming has been explored as an alternative to reduce those threats in transplants. This study evaluated the yield performance of seven strawberry cultivars and their impact on nematode densities and plant vigour after steaming was applied to transplants imported from California during two growing seasons (2020-2021 and 2021-2022). The cultivars used were ‘Sensation® Florida127’, ‘Florida Brilliance’, ‘Florida Radiance’, ‘Winterstar™ FL 05-107’, ‘Florida Elyana’, ‘Strawberry Festival’ and ‘Florida Beauty’. Plant steaming reduced plant vigour only 4 weeks after transplanting. Nevertheless, no negative effect was observed in total fruit yield. Differences among cultivars were observed for plant vigour, total fruit yield and abundance of root-knot nematodes. The plant vigour of each cultivar depended on the time of the year, which varied significantly throughout the season. Cultivars ‘Sensation® Florida127’ and ‘Florida Radiance’ attained the highest total yield in both seasons. There was an effect of the cropping system in the nematode community, which increased the bacterivores but drastically reduced omnivores and ring nematodes over time. No effect of cultivar was observed in plant-parasitic nematode populations. The cultivar ‘Winterstar™ FL 05-107’ was an exception since Meloidogyne javanica population densities were significantly higher in ‘Winterstar™ FL 05-107’s experimental plots. Overall, transplant steaming shows the potential to be a component of an integrated pest and disease management programme for the success of sustainable and profitable yields
近年来,对佛罗里达有机草莓的需求大幅增加,但这一产业却受到虫害和病原体的威胁,这些病原体通常来自外州的苗圃。人们一直在探索植物蒸煮作为一种替代方法,以减少移植植物中的这些威胁。本研究评估了七个草莓栽培品种的产量表现,以及在两个生长季节(2020-2021 年和 2021-2022 年)对从加利福尼亚州进口的移栽植物进行蒸煮后对线虫密度和植物活力的影响。使用的栽培品种包括 "Sensation® Florida127"、"Florida Brilliance"、"Florida Radiance"、"Winterstar™ FL 05-107"、"Florida Elyana"、"Strawberry Festival "和 "Florida Beauty"。移栽 4 周后,蒸煮植物会降低植物的活力。不过,在果实总产量方面没有观察到负面影响。不同栽培品种在植株活力、果实总产量和根结线虫数量方面存在差异。各栽培品种的植株活力取决于一年中的时间,在整个季节中差异显著。Sensation® Florida127 "和 "Florida Radiance "两个品种在两个季节的总产量最高。种植系统对线虫群落有影响,随着时间的推移,细菌性线虫增加,但杂食性线虫和环状线虫大幅减少。栽培品种对植物寄生线虫数量没有影响。栽培品种'Winterstar™ FL 05-107'是个例外,因为在'Winterstar™ FL 05-107'的实验地块中,爪哇线虫的种群密度明显较高。总之,移栽蒸腾显示出作为病虫害综合防治计划的一个组成部分的潜力,可成功实现可持续的盈利性产量。
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引用次数: 0
Genotypic markers associated with cold storage survival of the entomopathogenic nematode Heterorhabditis bacteriophora 与昆虫病原线虫冷藏存活率相关的基因型标记
IF 1.2 4区 生物学 Q2 Agricultural and Biological Sciences Pub Date : 2023-12-05 DOI: 10.1163/15685411-bja10295
Christopher Ogaya, Yohannes Moges, Verena Dörfler, Mike Barg, R. Ehlers, Carlos Molina
Limited shelf life is a major constraint to successful commercialisation of entomopathogenic nematodes (EPN), and to extend shelf life, dauer juveniles (DJ) are formulated and stored at low temperatures (4-8°C). We evaluated the cold storage potential of strains of Heterorhabditis bacteriophora formulated in diatomaceous earth at storage temperatures between 5 and 9°C. When assessing DJ decline to reach 75% survival (MT75) in the formulation for the respective temperatures, H. bacteriophora strain HB4 had the highest survival of 25 days at 9°C, while strain D2D6 survived longest at 8°C for 28 days. A set of 22 H. bacteriophora wild type inbred lines was then phenotyped for cold tolerance in water under oxidative stress in 70 mM H2O2 at 2°C. The MT50 (time to 50% survival) ranged from 11 to 23 days. The phenotypic data were correlated with the respective genotypic data, identifying four single nucleotide polymorphic (SNP) markers associated with cold tolerance. The survival of two lines (PT11 and IR11) with opposite extreme cold tolerance pheno- and genotypes was evaluated in diatomaceous earth formulation at 2°C with the cold tolerant IR11 surviving 3 days longer than PT11. Our study yields a set of valuable SNP markers employable in rapid genotyping of cold tolerance and tracking this trait during the breeding process.
有限的保质期是昆虫病原性线虫(EPN)成功商业化的主要制约因素,为了延长保质期,我们配制了dawer幼虫(DJ),并在低温(4-8°C)下储存。我们评估了在硅藻土中配制的异habditis bacteriophora菌株在5 - 9°C的储存温度下的冷藏潜力。在不同温度下,当评估配方中DJ下降达到75%存活率(MT75)时,菌株HB4在9°C下的存活率最高,为25天,而菌株D2D6在8°C下的存活率最长,为28天。在70 mM H2O2和2°C条件下,对22个野生型噬菌体自交系进行了耐冷表型分析。MT50(至50%生存时间)为11 ~ 23天。表型数据与各自的基因型数据相关,鉴定出4个与耐寒性相关的单核苷酸多态性(SNP)标记。在2°C硅藻土配方中,研究了具有相反极端耐寒表型和基因型的两个品系(PT11和IR11)的存活率,耐寒品系IR11比PT11多存活3天。我们的研究产生了一组有价值的SNP标记,可用于快速的耐寒性基因分型和在育种过程中跟踪该性状。
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引用次数: 0
Tylenchodoroides Gagarin, 2001 is a junior synonym of Oostenbrinkia Ali, Suryawanshi & Ahmed, 1971 (Dorylaimida, Aulolaimoididae) Tylenchodoroides Gagarin, 2001 是 Oostenbrinkia Ali, Suryawanshi & Ahmed, 1971 (Dorylaimida, Aulolaimoididae) 的初级异名。
IF 1.2 4区 生物学 Q2 Agricultural and Biological Sciences Pub Date : 2023-12-05 DOI: 10.1163/15685411-bja10298
R. Peña‐Santiago
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引用次数: 0
Automated classification and tracking of microscopic holographic patterns of nematodes using machine learning methods 利用机器学习方法对线虫的显微全息图案进行自动分类和跟踪
IF 1.2 4区 生物学 Q2 Agricultural and Biological Sciences Pub Date : 2023-12-05 DOI: 10.1163/15685411-bja10301
R. D. P. E. S. Ribeiro, A. Sobieranski, Elaine C.D. Gonçalves, Rafael C. Dutra, Aldo von Wangenheim
Analysing nematode behaviour helps estimate biomechanical parameters for applications like cellular biology, pharmacology and cognitive neuroscience. Portable holographic platforms offer cost-effective, high-resolution, high-frame-rate, wide-field imaging compared to conventional microscopy. Holographic methods can reconstruct original shapes using numerical diffraction, although this is computationally expensive. However, video holography remains challenging due to the fast motion and overlapping of holograms when nematodes swim in crowded environments. In this work we address this problem by focusing on automated detection and tracking of nematodes in densely populated environments, using machine learning methods. The main advantage of our approach is to present an automated computational flow to detect and analyse the behaviour of live nematodes in video directly from the raw holographic signals, without the requirement of phase-recovering methods for diffraction. For this purpose, we developed a three-step CNN-based approach consisting of: i) nematode hologram detection; ii) temporal tracking; and iii) behavioural analysis based on mobility parameters. In terms of precision, the obtained results show that using a two-stage detector, the Faster R-CNN architecture with the ResNet18 model as a backbone, presented the best Mean Average Precision (mAP) score with 86% for correct classification. For tracking, the best performing algorithm was IoU with a HOTA, resulting in 62.42% when applied on the individually tagged nematodes, which is comparable to the best current generic multi-tracking approaches available over the literature. Our obtained results show that the use of a convolutional neural network approach associated with a classic tracking algorithm is a very suitable approach for nematode detection and behavioural analysis for biological assays directly from holograms, even in densely populated environments. The proposed approach has been presented as a promising solution for automated inspection of free-living nematode individuals.
分析线虫的行为有助于估计细胞生物学、药理学和认知神经科学等应用的生物力学参数。与传统显微镜相比,便携式全息平台提供成本效益高、高分辨率、高帧率、宽视场成像。全息方法可以使用数值衍射重建原始形状,尽管这在计算上是昂贵的。然而,当线虫在拥挤的环境中游动时,由于全息图的快速运动和重叠,视频全息术仍然具有挑战性。在这项工作中,我们通过使用机器学习方法,专注于在人口密集的环境中自动检测和跟踪线虫来解决这个问题。我们的方法的主要优点是提供了一个自动化的计算流程,直接从原始全息信号中检测和分析视频中活线虫的行为,而不需要衍射的相位恢复方法。为此,我们开发了一种基于cnn的三步方法,包括:i)线虫全息图检测;Ii)时间跟踪;iii)基于流动性参数的行为分析。在精度方面,得到的结果表明,使用两级检测器,以ResNet18模型为骨干的Faster R-CNN架构的平均平均精度(mAP)得分最高,为86%。在跟踪方面,性能最好的算法是带有HOTA的IoU算法,当应用于单个标记的线虫时,其跟踪率为62.42%,与目前文献中可用的最佳通用多跟踪方法相当。我们获得的结果表明,使用卷积神经网络方法与经典跟踪算法相关联是一种非常适合的方法,可以直接从全息图中进行线虫检测和行为分析,即使在人口稠密的环境中也是如此。所提出的方法已被提出作为一个有前途的解决方案,自动检查自由生活的线虫个体。
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引用次数: 0
Effects of cover crops on hatching of and root penetration by Heterodera glycines 覆盖作物对甘蓝杂环虫孵化和根系穿透的影响
IF 1.2 4区 生物学 Q2 Agricultural and Biological Sciences Pub Date : 2023-12-05 DOI: 10.1163/15685411-bja10299
Kamal Neupane, Guiping Yan
The use of non-host cover crops can be a viable strategy for managing soybean cyst nematode (SCN) Heterodera glycines. Cover crops may affect SCN biology and reduce populations by acting as trap crops that promote the hatching and/or penetration by second-stage juveniles (J2) without supporting their development. Two growth chamber experiments were conducted to assess ten cover crops for their impacts on SCN hatching and penetration into the roots. For each experiment, with four replications per treatment, crops were planted in naturally-infested soil in two separate sets to be harvested 15 and 30 days after planting (DAP). SCN susceptible soybean ‘Barnes’ and non-planted natural soil (fallow) were used as controls. Faba bean ‘Petite’, a non-host of SCN, induced the greatest hatching among the cover crops and was statistically similar to soybean. Winter rye ‘ND Dylan’ also induced significant hatching compared to fallow. Root staining revealed that the highest number of J2 penetrated the faba bean roots at 15 DAP, followed by soybean and winter rye. While J2 penetrated all tested crops, they completed their development to become adult females only in soybean and turnip. Soybean cyst nematode development to adult females did not occur in faba bean, and the number of SCN inside the faba bean roots 30 DAP was significantly lower than at 15 DAP and also lower than in soybean. These results suggest that the faba bean affects SCN biology and has the greatest potential to act as a trap crop for managing SCN.
利用非寄主覆盖作物是控制大豆包囊线虫(SCN)的一种可行策略。覆盖作物可能作为诱捕作物,促进第二阶段幼鱼(J2)的孵化和/或渗透,而不支持它们的发育,从而影响SCN生物学并减少种群数量。通过两个生长室试验,评价了10种覆盖作物对SCN孵化和根系渗透的影响。在每个试验中,每个处理4个重复,将作物分两组种植在自然侵染的土壤中,播种后15天和30天收获(DAP)。以对SCN敏感的“巴恩斯”大豆和未种植的自然土壤(休耕)为对照。非SCN寄主的蚕豆‘Petite’在覆盖作物中诱导孵化率最高,与大豆相似。与休耕相比,冬黑麦' ND Dylan '也诱导了显著的孵化。根系染色结果显示,15dap时,J2渗透蚕豆根系的数量最多,其次是大豆和冬黑麦。虽然J2能穿透所有被试作物,但它们只在大豆和萝卜中完成发育成为成虫。大豆包囊线虫在蚕豆中未发育成雌,30 DAP的蚕豆根内SCN数量显著低于15 DAP,也低于大豆。这些结果表明蚕豆影响SCN生物学,并且最有可能作为控制SCN的诱捕作物。
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