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利用原位聚合酶链反应和绿色荧光蛋白检测石楠状菌根真菌的定殖情况。

Detecting the colonization of ericoid mycorrhizal fungi in using in situ polymerase chain reaction and green fluorescent protein.

作者信息

Yang Hongyi, Zhao Xingyu, Li Lili, Zhang Jie

机构信息

College of Life Sciences, Northeast Forestry University, Harbin, 150040 China.

Key Laboratory of Saline-alkali Vegetation Ecology Restoration (Northeast Forestry University), Ministry of Education, Harbin, 150040 China.

出版信息

Plant Methods. 2020 Jul 30;16:102. doi: 10.1186/s13007-020-00645-x. eCollection 2020.

DOI:10.1186/s13007-020-00645-x
PMID:32760433
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7391682/
Abstract

BACKGROUND

Ericoid mycorrhizal fungi (EMF) play important roles in mineral cycling and plant nutrient acquisition, and they increase plant survival in nutrient-poor environments. In this study, we detected the colonization of EMF using a green fluorescent protein (GFP) expression method and in situ PCR.

RESULTS

Genetic transformants of and Sordariomycetes sp. expressing GFP were obtained via -mediated transformation. GFP transformants were able to infect , and their fluorescence was visible in the hair roots. Both in situ PCR and the GFP-expressing method indicated that EMF could colonize the hair roots of 2 weeks after inoculation.

CONCLUSIONS

This research represents the first attempt to detect ericoid mycorrhizal colonization using in situ PCR. A GFP-expressing method is an excellent system for detecting the colonization of EMF, but it is dependent on the successful transformation and expression of the gene. In situ PCR and the GFP expression may be developed as new tools to study the interactions of EMF both with ericaceous plants and with the environment.

摘要

背景

石楠状菌根真菌（EMF）在矿物质循环和植物养分获取中发挥着重要作用，并且它们能提高植物在养分贫瘠环境中的存活率。在本研究中，我们使用绿色荧光蛋白（GFP）表达方法和原位PCR检测了EMF的定殖情况。

结果

通过介导转化获得了表达GFP的和炭角菌纲菌的遗传转化体。GFP转化体能够感染，并且在毛根中可见其荧光。原位PCR和GFP表达方法均表明，接种2周后EMF可定殖于的毛根中。

结论

本研究是首次尝试使用原位PCR检测石楠状菌根定殖情况。GFP表达方法是检测EMF定殖的优良系统，但它依赖于基因的成功转化和表达。原位PCR和GFP表达可能会发展成为研究EMF与石楠科植物以及与环境之间相互作用的新工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4313/7391682/571fd1ca7fe7/13007_2020_645_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4313/7391682/24515d53fe51/13007_2020_645_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4313/7391682/f032296e4838/13007_2020_645_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4313/7391682/ea58c56ca6a3/13007_2020_645_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4313/7391682/fe208d3cbb13/13007_2020_645_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4313/7391682/571fd1ca7fe7/13007_2020_645_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4313/7391682/24515d53fe51/13007_2020_645_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4313/7391682/f032296e4838/13007_2020_645_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4313/7391682/ea58c56ca6a3/13007_2020_645_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4313/7391682/fe208d3cbb13/13007_2020_645_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4313/7391682/571fd1ca7fe7/13007_2020_645_Fig5_HTML.jpg

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本文引用的文献

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New Phytol. 2004 Oct;164(1):7-10. doi: 10.1111/j.1469-8137.2004.01180.x.
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Developmental cross talking in the ectomycorrhizal symbiosis: signals and communication genes.外生菌根共生中的发育性相互作用：信号与通讯基因
New Phytol. 2001 Jul;151(1):145-154. doi: 10.1046/j.1469-8137.2001.00169.x.
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Host-Induced Gene Silencing of Arbuscular Mycorrhizal Fungal Genes via Agrobacterium rhizogenes-Mediated Root Transformation in Medicago truncatula.
通过根癌农杆菌介导的毛状根转化在蒺藜苜蓿中进行丛枝菌根真菌基因的宿主诱导基因沉默。
Methods Mol Biol. 2020;2146:239-248. doi: 10.1007/978-1-0716-0603-2_18.
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In situ PCR technique based on pricking microinjection for cDNA cloning in single cells of barley coleoptile and powdery mildew pathogen.基于针刺显微注射的原位PCR技术用于大麦胚芽鞘和白粉病菌单细胞cDNA克隆
Plant Cell Rep. 1997 Jun;16(9):612-618. doi: 10.1007/BF01275501.
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6
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The potential of Dark Septate Endophytes to form root symbioses with ectomycorrhizal and ericoid mycorrhizal middle European forest plants.深色有隔内生真菌与中欧外生菌根和石楠类菌根森林植物形成根系共生关系的潜力。
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