Characterization of miRNAs associated with Botrytis cinerea infection of tomato leavesby Weibo Jin, Fangli Wu

BMC Plant Biol


Plant Science


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Society of Automotive Engineers of

Lands of the Thunderbolt

Earl of Ronaldshay

Strategies of social research in Mozambique

by Members of the Centre of African



A o o ed e n ev le

Necrotrophs kill their host cells by secreting toxic com- the genes with complementary sequences. The plant miRJin and Wu BMC Plant Biology (2015) 15:1

DOI 10.1186/s12870-014-0410-4transport inhibitor response 1 (TIR1), auxin signaling

F-Box protein 2 (AFB2), and AFB3, are downregulated.

College of Life Science, Zhejiang Sci-Tech University, Hangzhou, Zhejiang 310018, Chinapounds or lytic enzymes; they also produce an array of pathogenic factors that can subdue host defenses [3,4].

To limit the spread of pathogens, host cells generate signaling molecules to initiate defense mechanisms in the surrounding cells. Abscisic acid and ethylene are plant

NAs are a well-studied class of sRNAs; they are hypersensitive to abiotic or biotic stresses and various physiological processes [9,10]. miR393 participates in bacterial PAMPtriggered immunity (PTI) by repressing auxin signaling [11]. In Arabidopsis plants treated with flg22, miR393 transcription is induced and the mRNAs of miR393 targets, including three F-box auxin receptors, namely* Correspondence: †Equal contributorslosses of 10 to 100 billion US dollars worldwide [2].were differentially expressed in B. cinerea-infected leaves, and 8 of these were further confirmed by quantitative reverse-transcription PCR (qRT-PCR). Moreover, five of these eight differentially expressed miRNAs could hit 10 coding sequences (CDSs) via CleaveLand pipeline and psRNAtarget program. In addition, qRT-PCR revealed that four targets were negatively correlated with their corresponding miRNAs (miR319, miR394, and miRn1).

Conclusion: Results of sRNA high-throughput sequencing revealed that the upregulation of miRNAs may be implicated in the mechanism by which tomato respond to B. cinerea stress. Analysis of the expression profiles of

B. cinerea-responsive miRNAs and their targets strongly suggested that miR319, miR394, and miRn1 may be involved in the tomato leaves’ response to B. cinerea infection.

Keywords: Tomato, High-throughput sequencing, B. cinerea-responsive miRNA, Target expression


Botrytis cinerea, a necrotrophic fungus causing gray mold disease, caused by Botrytis cinerea is considered an important pathogen around throughout the world. It induces decay on in a large number of economically important fruits and vegetables during the growing season and during postharvest storage. It is also a majorcreating serious obstacle problem to in long- distance transport and storage [1]. B. cinerea infection leads to annual hormones that participate in this process [5-7]. Li et al. [8] have found that SlMKK2 and SlMKK4 contribute to the resistance to B. cinerea in tomato. However, despite extensive research efforts, the biochemical and genetic basis of plant resistance to B. cinerea remains poorly understood. sRNAs are non-coding small RNAs (sRNAs), approximately 21–24 nt in length. These RNAs induce gene silencing by binding to Argonaute (AGO) proteins and directing the RNA-induced silencing complex (RISC) toCharacterization of miRN

Botrytis cinerea infection

Weibo Jin*† and Fangli Wu†


Background: Botrytis cinerea Pers. Fr. is an important path extended periods. MicroRNAs (miRNAs) have been report responses and B. cinerea infection in tomato. However, th remains unclear.

Results: The miRNA expression patterns in tomato in respo throughput sequencing. In total, 143 known miRNAs and s expression was detected in mock- and B. cinerea-inoculated© 2015 Jin and Wu; licensee BioMed Central.

Commons Attribution License (http://creativec reproduction in any medium, provided the or

Dedication waiver (http://creativecommons.or unless otherwise stated.Open Access s associated with f tomato leaves gen causing stem rot in tomatoes grown indoors for as gene expression regulators related to several stress function of miRNAs in the resistance to B. cinerea se to B. cinerea stress were investigated by highen novel miRNAs were identified and their corresponding aves. Among those, one novel and 57 known miRNAsThis is an Open Access article distributed under the terms of the Creative, which permits unrestricted use, distribution, and iginal work is properly credited. The Creative Commons Public Domain g/publicdomain/zero/1.0/) applies to the data made available in this article,

Consequently, the resistance to Pseudomonas syringae, a bacterial plant pathogen, is increased [11]. miRNAs are also directly involved in the regulation of disease resistance (R) genes [12-14]. For example, nta-miR6019 and nta-miR6020 are implicated in the regulation of disease resistance inNicotiana benthamiana by controlling the expression of theN gene. This gene encodes a Toll and Interleukin-1 Receptor type of nucleotide we obtained 16,844,708 (representing 6,075,098 unique sequences) and 13,935,908 (representing 4,807,933 unique sequences) clean reads, ranging from 18 nt to 30 nt, from

TC7d and TD7d libraries, respectively (Table 1). Most reads (>86% of redundant reads and >77% of unique reads) had at least 1 perfect match with the tomato genome (Table 1).

The majority of sRNA reads were from 20 nt- to 24

RN )

Jin and Wu BMC Plant Biology (2015) 15:1 Page 2 of 14binding site-leucine-rich-repeat receptor protein that provides resistance to the tobacco mosaic virus [14,15].

The members of different R-gene families in tomato, potato, soybean, andMedicago truncatula are targeted by miR482 and miR2118 miRNAs [12,13]. In addition, pathogen sRNA can also suppress the host immunity by loading into AGO1 and cause enhanced susceptibility to B. cinerea [2].

Tomato (Solanum lycopersicum, 2n = 24), a widespread member of the Solanum species, is an economically important vegetable crop worldwide. Several miRNAs can respond to B. cinerea infection in tomato [16]. To investigate the function of miRNAs in the resistance to this pathogen, we constructed two sRNA libraries from mock- and B. cinerea-inoculated tomato leaves. These libraries were then sequenced using an Illumina Solexa system. This study was conducted to identify and validate B. cinerea-responsive miRNAs from tomato leaves.