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评价一种用于评估油菜籽根茎腐病(核盘菌)的无伤接种技术。

Evaluation of non-injury inoculation technique for assessing Sclerotinia stem rot (Sclerotinia sclerotiorum) in oilseed Brassica.

机构信息

ICAR-National Institute for Plant Biotechnology, Pusa, New Delhi 110 012, India.

ICAR-Directorate of Rapeseed-Mustard Research, Bharatpur 321 303, Rajasthan, India.

出版信息

J Microbiol Methods. 2020 Aug;175:105983. doi: 10.1016/j.mimet.2020.105983. Epub 2020 Jun 13.

Abstract

The field assessment technique to evaluate the plants with a fungal phytopathogen for their tolerance to the disease is one of the crucial steps in dissecting their genetic control and in developing the resistant crop varieties. The objective behind this study was to develop and evaluate a field-based non-injury method of inoculation technique for Sclerotinia stem rot (SSR) in oilseed Brassica, caused by Sclerotinia sclerotiorum (Lib.) de Bary. The non-injury method of screening technique involves stem inoculation using a five days old mycelial mat on potato dextrose agar (PDA) plug placed on the top of sterile water-soaked cotton pad firmly wrapped over the internodal region with parafilm at the basal portion of the stem (15-20 cm above the ground) in the field. Inoculation without injury substantiates the natural means of infection in the field and the use of moist cotton pad keeps humidity for longer to initiate infection even in case of adverse climatic conditions. Disease development on the inoculated stem was measured by the length and width of the lesion. The symptom appears with water-soaked lesion formation and spreading deeper and wider on the stem in >90% of inoculated plants. During the experiment, about 800 Brassica germplasms including their wild relatives were screened and evaluated for three consecutive years using near-natural (non-injury) method of disease inoculation in the field. The Inoculation severity index (ISI) obtained during these years at Pusa, New Delhi were significantly similar and correlated with the natural infection measured in terms of disease severity index (DSI) on selected germplasm in the sick plot at ICAR-DRMR, Bharatpur. The significant correlations obtained among the used Brassica lines that were earlier not subjected for natural screening suggest the potential of this technique in evaluating the breeding material for SSR before confirmation with natural infection in the field.

摘要

采用田间鉴定技术评估植物对真菌病原体病害的抗性是解析其遗传控制并培育抗性作物品种的关键步骤之一。本研究的目的是开发和评估一种针对由核盘菌(Sclerotinia sclerotiorum (Lib.) de Bary)引起的油菜籽黑胫病(SSR)的基于田间的非损伤接种技术。该非损伤筛选技术包括在田间通过在无菌水浸湿的棉花垫上放置一个在 PDA 平板上生长五天的菌丝垫来进行茎部接种,棉花垫用帕尔膜紧紧包裹住茎的节间区域的基部(离地面 15-20 厘米)。非损伤接种可模拟田间自然感染方式,湿润棉花垫可保持更长时间的湿度,即使在不利的气候条件下也能引发感染。通过接种茎部的病变长度和宽度来测量接种后的发病情况。接种后,症状表现为水渍状病斑的形成,并在茎部更深更广地扩散,超过 90%的接种植物都会出现这种症状。在实验过程中,连续三年采用近自然(非损伤)田间接种方法对包括其野生近缘种在内的约 800 份油菜种质资源进行了筛选和评估。在新德里的 Pusa 获得的接种严重度指数(ISI)在这三年中与在 Bharatpur 的 ICAR-DRMR 的病圃中用选定的种质资源按病害严重度指数(DSI)测量的自然感染显著相似且相关。在之前未进行自然筛选的 Brassica 品系中获得的显著相关性表明,该技术在通过田间自然感染确认之前,具有评估 SSR 育种材料的潜力。

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