A transcriptome analysis uncovers Panax notoginseng resistance to Fusarium solani induced by methyl jasmonate

Diqiu Liu,Qin Zhao,Xiuming Cui,Rui Chen,Xin Li,Bingling Qiu,Feng Ge 한국유전학회 2019 Genes & Genomics Vol.41 No.12

Background Panax notoginseng is a famous Chinese herbal medicine, but the root rot disease mainly caused by Fusarium solani severely reduces the yield and quality of its medicinal materials. Objective The defense priming in P. notoginseng through exogenous application of signaling molecule will supply theoretical support for the exogenous regulation of disease resistance in P. notoginseng. Methods In this study, the exogenous application of methyl jasmonate (MeJA) increased P. notoginseng’s resistance to F. solani. Furthermore, the P. notoginseng transcriptome during F. solani infection was investigated through next-generation sequencing to uncover the resistance mechanism of P. notogingseng induced by MeJA. Results The de novo assembly of transcriptome sequences produced 80,551 unigenes, and 36,771 of these unigenes were annotated by at least one database. A differentially expressed gene analysis revealed that a large number of genes related to terpenoid backbone biosynthesis, phenylalanine metabolism, and plant–pathogen interactions were predominantly upregulated by MeJA. Moreover, jasmonic acid (JA) biosynthesis-related genes and the JA signaling pathway genes, such as linoleate 13S-lipoxygenase, allene oxide cyclase, allene oxide synthase, TIFY, defensin, and pathogenesis-related proteins, showed increased transcriptional levels after inoculation with F. solani. Notably, according to the gene expression analysis, JA and ethylene signaling pathways may act synergistically to positively regulate the defense responses of P. notoginseng to F. solani. Conclusion JA signaling appears to play a vital role in P. notoginseng responses to F. solani infection, which will be helpful in improving the disease resistance of P. notoginseng cultivars as well as in developing an environmentally friendly biological control method for root rot disease.

The PR10 gene family is highly expressed in Lilium regale Wilson during Fusarium oxysporum f. sp. lilii infection

Hua He,Diqiu Liu,Nannan Zhang,Wei Zheng,Qing Han,Bo Ji,Feng Ge,Chaoyin Chen 한국유전학회 2014 Genes & Genomics Vol.36 No.4

Pathogenesis-related (PR) proteins play keyroles in plant responses to pathogens and abiotic stresses. In this study, nine novel PR genes were isolated from Liliumregale Wilson, which is a wild lily species of Chinawith high-level resistance to the soilborne fungal pathogenFusarium oxysporum f. sp. lilii, and homology analysisclassified them into the PR10 family. These novel LrPR10swere clustered together with PR10s from monocotyledonsin a phylogenetic tree, moreover, phylogenetic analysisdivided the nine LrPR10s into two groups. The main-chainconformation and folding patterns of the LrPR10s werehighly conserved with other plant PR10s. The expressionpatterns of the nine LrPR10s in L. regale during normaldevelopment were examined by QRT-PCR, and the transcriptionlevels of the LrPR10s were relatively high inroots. Furthermore, QRT-PCR analysis indicated that theexpression levels of LrPR10-1, LrPR10-2, LrPR10-5,LrPR10-6, and LrPR10-7 in L. regale roots were up-regulatedby two or more stress-related signaling moleculesincluding salicylic acid, jasmonic acid, ethylene, and H2O2,while the other four LrPR10s were repressed by these foursignaling molecules. In addition, five members of theLrPR10 gene family including LrPR10-2, LrPR10-4,LrPR10-5, LrPR10-6, LrPR10-7, and LrPR10-9 werestrongly induced by F. oxysporum in resistant L. regalecompared with the susceptible Lilium Oriental hybrid‘Siberia’. The other four LrPR10s were down-regulated byF. oxysporum infection. In summary, our results indicatethat the members of PR10 gene family are involved inL. regale defense responses against F. oxysporum f. sp. lilii.

A bZIP transcription factor, LrbZIP1, is involved in Lilium regale Wilson defense responses against Fusarium oxysporum f. sp. lilii

Nannan Zhang,Diqiu Liu,Wei Zheng,Hua He,Bo Ji,Qing Han,Feng Ge,Chaoyin Chen 한국유전학회 2014 Genes & Genomics Vol.36 No.6

The basic leucine zipper (bZIP) proteins areubiquitous in plants and play important roles in plantdefense responses. In this study, based on an expressedsequence tag from a suppression subtractive hybridizationcDNA library of Lilium regale Wilson during Fusariumoxysporum f. sp. lilii infection, a novel bZIP transcriptionfactor gene LrbZIP1 was isolated from L. regale root usingthe rapid amplification of cDNA ends method. The predictedprotein of LrbZIP1 with 142 amino acid residuescontains a basic domain signature and a leucine zippermotif. The quantitative reverse transcription-PCR (qRTPCR)analysis showed that the transcription level of Lrb-ZIP1 was higher in roots of L. regale than in young stemsand leaves. Moreover, the expression of LrbZIP1 was upregulatedin the incompatible interaction between L. regaleand F. oxysporum f. sp. lilii as well as after treatments withstress-related signaling molecules. To verify the function ofLrbZIP1, a constitutive expression vector of LrbZIP1 wasconstructed and transferred into tobacco (Nicotiana tabacumL. cv Xanthi). The results of Southern blotting andqRT-PCR analyses demonstrated that the LrbZIP1 wasintegrated into genome of the tobacco transformants andhighly expressed. Under normal conditions, the T1 transgenictobacco lines showed higher antioxidant enzymeactivities and transcription levels of several resistancerelatedgenes than the wild type. Moreover, the T1 transgenictobacco plants showed strong resistance to F. oxysporumf. sp. lilii infection.

Transcriptome analysis reveals putative pathogenesis genes in Alternaria panax during infecting Panax notoginseng leaves

Shah Taif,Liu Diqiu,Cui XiuMing 한국유전학회 2022 Genes & Genomics Vol.44 No.7

Background: Alternaria panax is the causative agent of black spot disease in Panax notoginseng, which causes significant yield loss. However, the molecular mechanisms underlying its pathogenicity remain mostly unknown. Objective: We sequenced the transcriptome of A. panax during infecting P. notoginseng leaves using next-generation RNA-seq to understand the molecular aspects of black spot disease. Methods: In this study, we sequenced the A. panax transcriptome during infecting P. notoginseng leaves through next-generation sequencing to explore the pathogenesis genes that may be responsible for black spot disease on P. notoginseng. Result: The de novo transcriptome assembly of A. panax produced 23,036 unigenes, of which 18,096 genes were functionally annotated by at least one protein database. GO enrichment analysis and KEGG pathways of differentially up-regulated genes suggest that most genes are associated with metabolic processes, catalytic activity, starch, and sucrose metabolism during infection. Many pathogenesis-associated genes, including genes encoding secreted proteins, candidate secreted effectors, cell wall degrading enzymes, transcription factors, and transporters, were up-regulated in A. panax during infection. In addition, the secondary metabolite biosynthesis genes, including cytochrome P450, and nonribosomal peptide synthetases, were also identified in this study. Conclusions: Differential gene expression analysis has confirmed that A. panax infection was mainly present in the middle and final stages. The findings show that these pathogenesis-associated genes in A. panax may be critical for the P. notoginseng black spots disease.

Arabidopsis MiR396 Mediates the Development of Leaves and Flowers in Transgenic Tobacco

Fengxi Yang,Gang Liang,Dongmei Liu,Diqiu Yu 한국식물학회 2009 Journal of Plant Biology Vol.52 No.5

MicroRNAs (miRNAs) are single-stranded, noncoding small RNAs that usually function as posttranscriptional negative regulators by base pairing to target genes. They are pivotal to plant development. MiR396 is conserved among plant species and is predicted to target GRF (growth-regulating factor) genes in Arabidopsis. Here, overexpression of ath-miR396 in tobacco reduced the levels of three NtGRF-like genes containing an miR396 match site. Furthermore, its elevated expression resulted in a small, narrow leaf phenotype similar to that found with the Arabidopsis grf1grf2grf3 triple mutant. We also demonstrated that 35S:MIR396a transgenic plants were defective in the four whorls of floral organs. These results provide a link between the miR396- mediated regulatory pathway of NtGRF-like gene expression and the developmental processes for leaves and flowers in tobacco.

The transcription factor PjERF1 enhances the biosynthesis of triterpenoid saponins in Panax japonicus

Chen Qin,Yu Yilin,Zhang Xiang,Zhao Ren,Zhang Jinyu,Liu Diqiu,Cui Xiuming,Ge Feng 한국식물생명공학회 2021 Plant biotechnology reports Vol.15 No.5

The ERF-type transcription factors (TFs) play vital roles in plant secondary metabolism. ERF TFs simultaneously regulate the expression levels of key enzyme genes involved in the biosynthesis of secondary metabolites due to its “multi-point control” function. In this study, one gene of ERF TFs from Panax japonicus (PjERF1) was cloned. The open reading frame of PjERF1 was 801 bp and encoded 266 amino acids. Phylogenetic analysis showed that PjERF1 belonged to ERF subfamily with a typical conserved domain. Subcellular localization found that PjERF1 protein might be located in eukaryotic cell nucleus. Yeast one-hybrid assay demonstrated that PjERF1 could bind to the promoters of PjβAS, PjCAS, and PjSE specifically and regulate the expression levels of such key enzyme genes involved in the triterpene saponins biosynthesis. Therefore, in the PjERF1 overexpression cell lines, the expression levels of some key enzyme genes involved in the triterpenoid saponins biosynthesis were significantly increased compared with those in non-transgenic cell line. As a result of it, the biosynthesis of chikusetsusaponin IV and IVa, and other ginsenosides (Rd, Rb1, Re, and R0) were also promoted in the PjERF1 over- expression cell lines. This study indicated that PjERF1 could regulate the biosynthesis of saponins in P. japonicus through controlling the expression levels of key enzyme genes related to the biosynthesis of triterpenoid saponins.

Optimization of Production Conditions for Antioxidant Peptides from Walnut Protein Meal Using Solid-state Fermentation

Wanxing Wu,Shenglan Zhao,Chaoyin Chen,Feng Ge,Diqiu Liu,Xiaoming He 한국식품과학회 2014 Food Science and Biotechnology Vol.23 No.6

Solid-state fermentation (SSF) of walnut proteinmeal (WPM) by Bacillus subtilis was optimized for themaximum degree of hydrolysis (DH) and reducing power(RP) using response surface methodology (RSM). Optimumfermentation conditions were fermentation time of 82.01 h,inoculum concentration of 10.40, and water content of 1.50mL/g. Optimized values were 41.80% and 0.78 for DH andRP, respectively. Walnut peptides (WP) were ultrafiltrationmembrane fractionated. The WPs-II (molecular weight <5kDa) fraction showed the highest RP value. WPs-II showedgood DPPH free radical scavenging and Fe2+ chelatingactivities, higher than for GSH (L-glutathione reduced) atthe same concentration. Amino acid composition analysisof WPs-II showed that Asp, Glu, and Arg were the majoramino acids playing important roles in the antioxidantactivity. WPs is an efficient antioxidant suitable for use asa food additive and as a pharmaceutical agent.

Quantitative Indirect ELISA for Determination of Walnut Proteins in Foods

Juan Fang,Dan Chen,Chaoyin Chen,Feng Ge,Diqiu Liu,Benyong Han,Xiangfeng Xiong 한국식품과학회 2015 Food Science and Biotechnology Vol.24 No.6

An indirect enzyme-linked immunosorbent assay (ELISA) capable of detecting walnut proteins in commercial products was developed. Polyclonal antibodies against soluble walnut protein were used. Specific epitopes of walnut protein were analyzed for cross activity with peanut, soybean, sesame, rice, wheat, jujube, and defatted milk protein. Epitopes of walnut proteins had no cross activity. The indirect ELISA was highly specific for soluble walnut proteins. Recovery values from walnut protein solutions ranged from 88.47 to 115%, and low coefficients of variation (<10%) indicated good repeatability. Intra and inter-assay precisions were <6 and <7 %, respectively. The limit of detection was 10 ng/mL of soluble walnut protein. The method was applied to detect some commercial food products in China and the results showed that indirect ELISA could be promisingly used to quantify adulteration of walnut processed foods.