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基于RT-PCR的辣椒种子中辣椒轻斑驳病毒检测及种子传播试验

RT-PCR based detection of Pepper mild mottle virus from capsicum seeds and seed transmission assay.

作者信息

Patel Priyankaben, Kumari Nidhi, Sharma P N

机构信息

Department of Plant Pathology, CSK HPKV, Palampur, Himachal Pradesh 176062 India.

Division of Crop Protection, ICAR-Central Institute for Subtropical Horticulture, Rehmankhera, Lucknow, 226101 India.

出版信息

Virusdisease. 2023 Mar;34(1):50-55. doi: 10.1007/s13337-023-00807-0. Epub 2023 Feb 8.

Abstract

UNLABELLED

Pepper mild mottle virus (PMMoV), a from family, is highly contagious and transmitted by seeds as well as soil in nature. PMMoV has become a greater threat to capsicum cultivation worldwide. To develop an indigenous, rapid, and sensitive protocol for routine detection of PMMoV from seeds, the sensitivity of DAS-ELISA and RT-PCR was compared in the present study. The infected seeds of California Wonder were included in the study. Through DAS-ELISA the virus was successfully detected from 20 mg of seeds. However, using RT-PCR, we were able to detect the virus even from one infected seed with reproducibility. In the present study, vertical seed transmission of the test virus was investigated by employing a grow-out test under greenhouse conditions as well as directly through RT-PCR omitting the grow-out test in three capsicum cultivars. Based on symptoms observations in grow out test, seed transmission was observed in the 3 capsicum cultivars viz., California Wonder (63.04%), Yolo Wonder (33.80%) and Doux des LAndes (33.30%). Through RT-PCR it was estimated to be 55.56% (California Wonder), 28.96% (Yolo Wonder), and 40.64% (Doux des Landes), respectively. Thus, indicating 100% seed-to-seedling PMMoV transmission and reliability of RT-PCR in direct PMMoV detection from seeds. Even a small percentage of infected seed has the potential to greatly increase the PMMoV inoculum in the field and result in 100% plant infection. Therefore, we suggest using the established procedure for PMMoV detection right from the seed.

SUPPLEMENTARY INFORMATION

The online version contains supplementary material available at 10.1007/s13337-023-00807-0.

摘要

未标记

辣椒轻斑驳病毒(PMMoV)属于 科,具有高度传染性,在自然界中可通过种子和土壤传播。PMMoV 已对全球辣椒种植构成更大威胁。为制定一种本土、快速且灵敏的从种子中常规检测 PMMoV 的方法,本研究比较了双抗夹心酶联免疫吸附测定(DAS - ELISA)和逆转录聚合酶链反应(RT - PCR)的灵敏度。研究中纳入了加州奇迹(California Wonder)的感染种子。通过 DAS - ELISA 成功从 20 毫克种子中检测到该病毒。然而,使用 RT - PCR,我们甚至能够从一粒感染种子中检测到病毒,且具有可重复性。在本研究中,通过在温室条件下进行实生苗检测以及直接通过 RT - PCR(省略实生苗检测)在三个辣椒品种中研究了测试病毒的垂直种子传播。基于实生苗检测中的症状观察,在三个辣椒品种即加州奇迹(63.04%)、约洛奇迹(Yolo Wonder,33.80%)和杜克斯德朗德(Doux des LAndes,33.30%)中观察到了种子传播。通过 RT - PCR 估计分别为 55.56%(加州奇迹)、28.96%(约洛奇迹)和 40.64%(杜克斯德朗德)。因此,表明 PMMoV 从种子到幼苗的传播率为 100%,以及 RT - PCR 在直接从种子中检测 PMMoV 方面的可靠性。即使是一小部分感染种子也有可能极大地增加田间的 PMMoV 接种量,并导致 100%的植株感染。因此,我们建议从种子开始就使用既定的 PMMoV 检测程序。

补充信息

在线版本包含可在 10.1007/s13337 - 023 - 00807 - 0 获取的补充材料。

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