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A simple and effective method for total rna isolation of appressoria in Magnaporthe oryzae

Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2008年第10期9卷 811-817页

作者： Tong-bao LIU Jian-ping LU Xiao-hong LIU Hang MIN Fu-cheng LIN State Key Laboratory for Riee Biology Biotechnology Institute Zhejiang University Hangzhou 310029 China College of Life Sciences Zhejiang University Hangzhou 310058 China

Appressorium formation is an important event in establishing a successful interaction between the rice blast fungus, Magnaporthe oryzae, and its host plant, rice. An understanding of molecular events occurring in appressorium differentiation will give new strategies to control rice blast. A quick and reliable method to extract total rna from appressorium is essential for studying gene expression during appressorium formation and its mechanism. We found that duplicate film is an efficient substratum for appressorium formation, even when inoculated with high density conidia. When inoculated with conidia at 1 × 106 ml^-1, the percentages of conidium germination and appressorium formation were （97.98±0.67）% and （97.88±0.45）%, respectively. We applied Trizol before appressorium collection for total rna isolation, and as much as 113.6 lag total rna was isolated from the mature appressoria at 24 h after inoculation. Functional analysis of two genes, MNH6 and MgATG1, isolated from the cDNA subtractive library, revealed that the quantity of rna was good enough to construct a cDNA （complementary DNA） library or a cDNA subtractive library. This method may be also applicable for the appressorium rna isolation of other pathogenic fungi in which conidia differentiate into appressoria in the early stages of host infection.

关键词： Appressorium Magnaporthe oryzae rna isolation

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A modified one-step procedure for rapid rna isolation from insect

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Journal of Forestry Research 2006年第2期17卷 129-131页

作者：景天忠王志英刘宽余齐凤慧东北林业大学

A modified guanidinium isothiocyanate method was used to extract total rna from two forest insect species Clostera anastomosis and Saperda populnea. The integrity of rna was demonstrated by the methods of gel electrophoresis and cDNA analysis. Typical A260/ A280 absorbance ratio of the total rna was in range of 1.8 to 2.0. The size of double strand cDNAs obtained by RT-PCR was more than 2 kb, which indicated that intact mrna was obtained. The fragments of β-actin and chitinase gene from the rna of C. anastomosis were obtained by RT-PCR, which indicated that the rna could be used for other molecular operation. By this procedure, rnas could be extracted and analyzed by electrophoresis from at least 8 samples within 4 hours. These results showed that this method was time- and cost-saving and effective.

关键词： rna isolation β-actin gene Clostera anastomosis Saperda populnea

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isolation of Functional rna from Heavily Infested, Wilted and Necrotic Leaf Tissues of Tea with High Polyphenol Content

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Journal of Agricultural Science and Technology(B) 2012年第1期2卷 121-127页

作者： Tirthankar Bandyopadhyay Raju Bharalee Bomali Gohain Sushmita Gupta Niraj Agarwala H. RanjitSingh Setu Chakrabarty Priyadarshini Bhorali Mohan C. Kalita Sudripta Das Department of Biotechnology Tea Research Association Tocklai Jorhat 785008 Assam India Department of Biotechnology Gauhati University Guwahati 781014 Assam India

A procedure is described to isolate intact rna from tissues not previously undertaken-highly infested and wilted apical buds and leaves of tea [Camellia sinensis （L.） O. Ktze.]. The protocol uses a final concentration of 450 mM β mercaptoethanol （βME） and 10% Polyvinylpyrrolidone （PVP） to circumvent problems associated with large amounts of polyphenols, polysaccharides, pigments and other secondary metabolites not easily removed by conventional procedures. Furthermore, the proposed protocol is applicable to normal tissues and other plant tissues with similar stresses, containing compounds that interfere with rna extractions. Total rna could be used for downstream applications such as mrna isolation, reverse transcription, quantitative Polymerase Chain Reaction （PCR）, cDNA library construction and Rapid amplification of cDNA ends （RACE）.

关键词： rna isolation Camellia sinensis Helopeltis theivora infestation necrosis polyphenols.

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Efficient isolation of high quality rna from tropical palms for rna-seq analysis

Efficient isolation of high quality RNA from tropical palms ...

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第十四届全国植物基因组学大会

作者： Lixia Zhou Yong Xiao Yaodong Yang Hongxing Cao Haikuo Fan Zilong Ma Xintao Lei Annaliese S.Mason Zhihui Xia Xi Huang Coconuts Research Institute Chinese Academy of Tropical Agricultural Sciences

Currently,rna-seq as a high throughput technology is being widely applied to various species to elucidate the complexity of the transcriptome and to discover large number of novel ***,the technology has had poor succe... 详细信息

关键词： rna isolation rna-seq Cocos nucifera Palmaceace rrna

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A Simple and High Quality Method for isolation and Extraction of Total rna of Pholiota adipose

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Asian Agricultural Research 2018年第1期10卷 40-44页

作者： Zhimin WEI Shunguo LI Meng LIU Xueyan XIA Yu ZHAO Hanzhang ZHOU Chengang GUO Institute of Millet Crops Hebei Academy of Agricultural & Forestry Sciences Shijiazhuang 050035 China Dezhou University Dezhou 253023 China

Analyzing functional values of rna using rna-seq technology is a hot spot of researches. In order to study the medicinal value of Pholiota adipose from the transcriptome level,it is necessary to extract and isolate rna samples of high purity and high quality P. adipose. In this study,liquid nitrogen grinding Trizol one-step method was used to extract the total rna of P. adipose. Quality test and statistical comparative analysis were carried out for rna extract of liquid nitrogen grinding treated and untreated P. adipose. The results showed that the concentration of rna in the samples treated with liquid nitrogen was much higher than that of the samples without grinding treatment. The OD260/280 of both was about 2,indicating that the purity of rna was very high. Besides,the ratio of fluorescence intensity of 25 S and 18 S subunit strips of three replicate samples was 1. 8,1. 9,and 1. 9,close to 2,indicating rna integrity is good. RIN test results of Agilent 2100 were 9. 1,8. 7,and 9. 3,higher than the standard value 6. 8,further proving the integrity. In sum,liquid nitrogen grinding Trizol one-step method is a very simple and efficient method for extracting high quality total rna of P. adipose.

关键词： rna isolation Pholiota adipose rna-seq Antineoplastic

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Endocarpon pusillum(地衣型Ascomycota)的mycobiont cDNA全长度文库的结构(英文)

Endocarpon pusillum(地衣型Ascomycota)的mycobiont cDNA全长度...

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2010全国地衣与苔藓系统学研讨会

作者：王燕燕张涛周启明魏江春 College of Life Sciences Shangdong Agriculture University Systematic Mycology and Lichenology Laboratory Institute of MicrobiologyChinese Academy of Sciences

As biological weathering agents in the rock-weathering and pedogenic process lichens are pioneer of the organisms in the ***,as terrestrial organisms,can be distributed almost everywhere,including in extreme conditions,such as Cold desert,Hot desert and Mid-latitude *** experiments showed that the mycobiont of Endocarpon pusillum can survive for 7 months under the starvation and desiccation ***-mentioned information showed that lichens,as the life-support system in fungal-algal symbiosis,were able to regulate the expression of genes in response to the drought and cold stresses.

关键词： Desert lichen Full-length cDNA library rna isolation

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