Development of droplet digital PCR for the detection of Tilletia laevis, which causes common bunt of wheat, based on the SCAR marker derived from ISSR and real-time PCR

Abstract Common bunt of wheat caused by Tilletia laevis and/or T. caries (syn. T. tritici), is a major disease in wheat-growing regions worldwide that could lead to 80% or even total loss of production. Even though T. laevis can be distinguished from T. caries on the bases of morphology of teliospor...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Tongshuo Xu, Zhaoqun Yao, Jianjian Liu, Han Zhang, Ghulam Muhae Ud Din, Sifeng Zhao, Wanquan Chen, Taiguo Liu, Li Gao
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/902214a08bf64a749adea366c4eea7f0
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:902214a08bf64a749adea366c4eea7f0
record_format	dspace
spelling	oai:doaj.org-article:902214a08bf64a749adea366c4eea7f02021-12-02T18:51:13ZDevelopment of droplet digital PCR for the detection of Tilletia laevis, which causes common bunt of wheat, based on the SCAR marker derived from ISSR and real-time PCR10.1038/s41598-020-72976-72045-2322https://doaj.org/article/902214a08bf64a749adea366c4eea7f02020-09-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-72976-7https://doaj.org/toc/2045-2322Abstract Common bunt of wheat caused by Tilletia laevis and/or T. caries (syn. T. tritici), is a major disease in wheat-growing regions worldwide that could lead to 80% or even total loss of production. Even though T. laevis can be distinguished from T. caries on the bases of morphology of teliospores using microscopy technique. However, molecular methods could serve as an additional method to quantify the pathogen. To develop a rapid diagnostic and quantify method, we employed the ISSR molecular marker for T. laevis in this study. The primer ISSR857 generated a polymorphic pattern displaying a 1385 bp T. laevis-specific DNA fragment. A pair of specific primers (L57F/L57R) was designed to amplify a sequence-characterized amplified region (SCAR) (763 bp) for the PCR detection assay. The primers amplified the DNA fragment in the tested isolates of T. laevis but failed in the related species, including T. caries. The detection limit of the primer set (L57F/L57R) was 5 ng/µl of DNA extracted from T. laevis teliospores. A SYBR Green I real-time PCR method for detecting T. laevis with a 100 fg/µl detection limit and droplet digital PCR with a high sensitivity (30 fg/µl detection limit) were developed; this technique showed the most sensitive detection compared to the SCAR marker and SYBR Green I real-time PCR. Additionally, this is the first study related the detection of T. laevis with the droplet digital PCR method.Tongshuo XuZhaoqun YaoJianjian LiuHan ZhangGhulam Muhae Ud DinSifeng ZhaoWanquan ChenTaiguo LiuLi GaoNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-10 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Tongshuo Xu Zhaoqun Yao Jianjian Liu Han Zhang Ghulam Muhae Ud Din Sifeng Zhao Wanquan Chen Taiguo Liu Li Gao Development of droplet digital PCR for the detection of Tilletia laevis, which causes common bunt of wheat, based on the SCAR marker derived from ISSR and real-time PCR
description	Abstract Common bunt of wheat caused by Tilletia laevis and/or T. caries (syn. T. tritici), is a major disease in wheat-growing regions worldwide that could lead to 80% or even total loss of production. Even though T. laevis can be distinguished from T. caries on the bases of morphology of teliospores using microscopy technique. However, molecular methods could serve as an additional method to quantify the pathogen. To develop a rapid diagnostic and quantify method, we employed the ISSR molecular marker for T. laevis in this study. The primer ISSR857 generated a polymorphic pattern displaying a 1385 bp T. laevis-specific DNA fragment. A pair of specific primers (L57F/L57R) was designed to amplify a sequence-characterized amplified region (SCAR) (763 bp) for the PCR detection assay. The primers amplified the DNA fragment in the tested isolates of T. laevis but failed in the related species, including T. caries. The detection limit of the primer set (L57F/L57R) was 5 ng/µl of DNA extracted from T. laevis teliospores. A SYBR Green I real-time PCR method for detecting T. laevis with a 100 fg/µl detection limit and droplet digital PCR with a high sensitivity (30 fg/µl detection limit) were developed; this technique showed the most sensitive detection compared to the SCAR marker and SYBR Green I real-time PCR. Additionally, this is the first study related the detection of T. laevis with the droplet digital PCR method.
format	article
author	Tongshuo Xu Zhaoqun Yao Jianjian Liu Han Zhang Ghulam Muhae Ud Din Sifeng Zhao Wanquan Chen Taiguo Liu Li Gao
author_facet	Tongshuo Xu Zhaoqun Yao Jianjian Liu Han Zhang Ghulam Muhae Ud Din Sifeng Zhao Wanquan Chen Taiguo Liu Li Gao
author_sort	Tongshuo Xu
title	Development of droplet digital PCR for the detection of Tilletia laevis, which causes common bunt of wheat, based on the SCAR marker derived from ISSR and real-time PCR
title_short	Development of droplet digital PCR for the detection of Tilletia laevis, which causes common bunt of wheat, based on the SCAR marker derived from ISSR and real-time PCR
title_full	Development of droplet digital PCR for the detection of Tilletia laevis, which causes common bunt of wheat, based on the SCAR marker derived from ISSR and real-time PCR
title_fullStr	Development of droplet digital PCR for the detection of Tilletia laevis, which causes common bunt of wheat, based on the SCAR marker derived from ISSR and real-time PCR
title_full_unstemmed	Development of droplet digital PCR for the detection of Tilletia laevis, which causes common bunt of wheat, based on the SCAR marker derived from ISSR and real-time PCR
title_sort	development of droplet digital pcr for the detection of tilletia laevis, which causes common bunt of wheat, based on the scar marker derived from issr and real-time pcr
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/902214a08bf64a749adea366c4eea7f0
work_keys_str_mv	AT tongshuoxu developmentofdropletdigitalpcrforthedetectionoftilletialaeviswhichcausescommonbuntofwheatbasedonthescarmarkerderivedfromissrandrealtimepcr AT zhaoqunyao developmentofdropletdigitalpcrforthedetectionoftilletialaeviswhichcausescommonbuntofwheatbasedonthescarmarkerderivedfromissrandrealtimepcr AT jianjianliu developmentofdropletdigitalpcrforthedetectionoftilletialaeviswhichcausescommonbuntofwheatbasedonthescarmarkerderivedfromissrandrealtimepcr AT hanzhang developmentofdropletdigitalpcrforthedetectionoftilletialaeviswhichcausescommonbuntofwheatbasedonthescarmarkerderivedfromissrandrealtimepcr AT ghulammuhaeuddin developmentofdropletdigitalpcrforthedetectionoftilletialaeviswhichcausescommonbuntofwheatbasedonthescarmarkerderivedfromissrandrealtimepcr AT sifengzhao developmentofdropletdigitalpcrforthedetectionoftilletialaeviswhichcausescommonbuntofwheatbasedonthescarmarkerderivedfromissrandrealtimepcr AT wanquanchen developmentofdropletdigitalpcrforthedetectionoftilletialaeviswhichcausescommonbuntofwheatbasedonthescarmarkerderivedfromissrandrealtimepcr AT taiguoliu developmentofdropletdigitalpcrforthedetectionoftilletialaeviswhichcausescommonbuntofwheatbasedonthescarmarkerderivedfromissrandrealtimepcr AT ligao developmentofdropletdigitalpcrforthedetectionoftilletialaeviswhichcausescommonbuntofwheatbasedonthescarmarkerderivedfromissrandrealtimepcr
_version_	1718377459920928768

Development of droplet digital PCR for the detection of Tilletia laevis, which causes common bunt of wheat, based on the SCAR marker derived from ISSR and real-time PCR

Ejemplares similares