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An improved method for RNA extraction from urediniospores of and wheat leaves infected by an obligate fungal pathogen, Puccinia striiformis f. sp. tritici

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Journal of Integrative Agriculture 2016年第6期15卷 1293-1303页

作者： MA Li-Jie QIAO Jia-xing KONG Xin-yu WANG Jun-juan XU Xiang-ming HU Xiao-ping State Key Laboratory of Crop Stress Biology for Arid Areas College of Plant Protection Northwest A&F University Yangling 712100 P.R.China Genetics and Crop Improvement East Mailing Research East Mailing ME19 6B J UK Erdos Ecological Environment of Career Academy Ordos 017010 P.R. China

Stripe rust, caused by Puccinia striiformis f. sp. tritici, is an important wheat disease in China, seriously threatening wheat production. Understanding the winter survival of the fungus is a key for predicting the spring epidemics of the disease, which determines the crop loss. Estimation of P. striiformis f. sp. tritici winter survival requires processing a large number of samples for sensitive detection of the pathogen in wheat leaf tissue using real-time quantitative reverse transcription PCR （qRT-PCR）. A bottleneck for the analysis is the acquisition of a good yield of high quality RNA suitable for qRT-PCR to distinguish dead and alive fungal hyphae inside leaves. Although several methods have been described in the literatures and commercial kits are available for RNA extraction, these methods are mostly too complicated, expensive and inefficient. Thus, we modified three previously reported RNA extraction methods with common and low-cost reagents （LiCI, SDS and NaCI） to solve the problems and selected the best to obtain high quality and quantity RNA for use in qRT-PCR. In the three improved methods, the NaCI method was proven to be the best for extracting RNAfrom urediniospores of and wheat leaves infected by P. striiformis f. sp. tritici, although the modified LiCI and SDS methods also increased yield of RNA compared to the previous methods. The improved NaCI method has the following advantages： 1） Complete transfer of urediniospores of P. striiformis f. sp. tritici from the mortar and pestle can ensure the initial amount of RNA for the qRT-PCR analysis; 2） the use of low-cost NaCI to replace more expensive Trizol can reduce the cost; 3） the yield and quality of RNA can be increased; 4） the improved method is more suitable for a large number and high quantity of samples from fields. Using the improved NaCI method, the amount of RNA was increased three times from urediniospores of P. striiformis f. sp. tritici compared from the extraction kit. Approximately, 1

关键词： Puccinia striiformis f. sp. tritici RNA extraction RT-PCR urediniospore mycelium

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Transcriptomic reprogramming of wheat stripe rusturediniospores and basidiospores during infect wheat and barberry

Transcriptomic reprogramming of wheat stripe rusturediniospo...

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中国植物病理学会2019年学术年会

作者： Zhao Jing Duan Wanlu Wang Long Tian Song Zhuang Hua Kang Zhensheng State Key Laboratory of Crop Stress Biology for Arid Areas and College of Plant Protection Northwest A＆F University

Puccinia striiformis ***（Pst） is the causal agent of wheat stripe rust,which cause severe yield losses to wheat production all over the *** a macrocyclic heteroecious rust,Pst can infect two exclusive and unrelated plant *** infect wheat and cause disease epidemic,while basidiospores infect barbery to complete sexual *** complex life cycle poses interesting questions in the different mechanisms of pathogenesis underlying the infection of two *** the present study,transcriptomes of Pst from leaves from wheat and barberry were *** a result,198 genes specifically expressed in wheat（WEG） were identified,which were far less than 2575 genes specifically expressed in barberry（BEG）.BEGs are mainly enriched in "gene expression","cellular protein metabolic process","nucleic acid metabolic process" and "biosynthesis of antibiotics",while WEGs are related to "carbohydrate catabolic process".Additionally,the percentage of new genes in WEGs was significantly greater than that in BEGs,implying that the original host of Pst is barberry other than wheat or *** patterns of genes encoding cell wall degrading enzymes were compared,and the results demonstrated that urediniospores and basidiospores use different strategies to destroy the host cell walls according to their *** results represent the first analysis of the transcriptional profile of the Pst basidiospores in barberry,and will contribute to a better understanding of evolutionary and pathogenic strategies for different type of rust spores during infecting wheat and barberry.

关键词： wheat stripe rust barberry transcriptome urediniospore basidiospore

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