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Transcriptome Analysis of Pleurotus ostreatus at Different Growth Stages in Response to IAA

CUI Xiao, ZHANG Yuting, KONG Weili, HU Sujuan, LIU Qin, WANG Yanpo, WU Jie, SHI Ziwen

Journal of Henan Agricultural Sciences 2022, 51 (12): 97-109. DOI: 10.15933/j.cnki.1004-3268.2022.12.012

Abstract （112）

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To investigate the mechanism of Pleurotus ostreatus at different growth stages in response to IAA，transcriptome sequencing of mycelium（7 d，15 d）and fruiting body stage（25 d，27 d，31 d）under two concentrations of IAA（high concentration：1×10 ^‑3 mol/L；low concentration：1×10 ^‑8 mol/L） was performed to screen the differentially expressed genes（DEGs），with no treatment（0 mol/L IAA）as the control.Through Venn analysis，cluster analysis，functional enrichment analysis，and the construction of co‑expression network，the molecular mechanism of P.ostreatus at different growth stages in response to IAA concentration was revealed.Results indicated that 217 DEGs，6 735 co‑expression pairs at the mycelium stage and 10 DEGs，32 co‑expression pairs at the fruiting body stage were screened under the high concentration of IAA，while 81 DEGs and 87 DEGs，and 766 co‑expression pairs and 1 565 co‑expression pairs were dug out under the low concentration of IAA at the mycelium and fruiting body stage，respectively.The GO and KEGG results showed that the low concentration of IAA could regulate ribosome synthesis of endogenous plant hormones through TRINITY_DN2732_c0_g1，while promote the metabolism of the fruiting body stage through TRINITY_DN530_c0_g1 and TRINITY_DN3832_c0_g1.High concentration of IAA could regulate glycerophospholipid metabolism through TRINITY_DN41591_c0_g1，regulate chitin metabolism through TRINITY_DN14069_c0_g1，and regulate oxidoreductase activity through TRINITY_DN35150_c0_g1，TRINITY_DN5314_c0_g1 and TRINITY_DN43009_c0_g1，while at the fruiting body stage，high concentration of IAA regulated ATP synthesis via TRINITY_DN21430_c0_g1.The above results indicate that low concentration of IAA affects the phytohormone synthesis in the mycelium stage，and the metabolic process in the fruit body stage of P.ostreatus through DEGs；high concentration of IAA affects the oxidative stress mechanism in the mycelium stage，and the ATP synthesis in the fruit body stage of P.ostreatus through DEGs.

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Analysis of Differentially Expressed Genes in Two Different Nitrogen Efficiency Maize Varieties in Response to Nitrogen Reduction

LI Chuan, ZHANG Panpan, ZHANG Meiwei, NIU Jun, ZHAO Xia, HE Guanhua, QIAO Jiangfang

Journal of Henan Agricultural Sciences 2022, 51 (9): 10-24. DOI: 10.15933/j.cnki.1004-3268.2022.09.002

Abstract （2020）

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In order to explore the differentially expressed genes（DEGs）in Weike 518（WK518）with high nitrogen efficiency and Nongda108（ND108） with low nitrogen efficiency under nitrogen reduction condition and their functions，the ear⁃leaf samples were collected from WK518 and ND108 at mid filling stage under normal nitrogen fertilizer application rate（225 kg/ha nitrogen,HN）and low nitrogen fertilizer application rate（0 kg/hanitrogen,LN）,and used for high⁃throughput RNA sequencing. Then the GO term and KEGG metabolic pathway of DEGs were analyzed from different compared groups，and the differentially expressed transcription factor families were analyzed. The results showed that 2 065 up⁃regulated DEGs and 2 319 down⁃regulated DEGs were detected between WK518 and ND108 under LN condition,2 368 up⁃regulated DEGs and 3 780 down⁃regulated DEGs were detected under HN condition;1 009 up⁃regulated DEGs and 2 268 down⁃regulated DEGs were detected in WK518 under different nitrogen application rates,and 364 up⁃regulated DEGs and 510 down⁃regulated DEGs were detected in ND108 under different nitrogen application rates.Under LN condition,DEGs between WK518 and ND108 were mainly enriched in uroporphyrin⁃Ⅲ C⁃methyltransferase activity,mannose⁃6⁃phosphate isomerase activity,oxidation⁃reduction process,mitochondrion organization,nuclear chromatin and other GO terms,and amino sugar and nucleotide sugar metabolism,phenylalaninemetabolism,monoterpenoid biosynthesis,glycine/serine and threonine metabolism,base excision repair and other KEGG pathways. Under HN condition，DEGs between WK518 and ND108 were mainly enriched in stomatal closure,transmembrane receptor protein serine/threonine kinase activity,chloroplast stroma，thylakoid,chloroplast envelope and other GO terms,and carbon fixation in photosynthetic organisms，amino sugar and nucleotide sugar metabolism,glycolysis/gluconeogenesis,β⁃alanine metabolism,photosynthesis⁃antenna proteins and other KEGG pathways.DEGs in WK518 were mainly enriched in response to chitin,protein phosphorylation，membrane，indole glucosinolate metabolic process，galactinol⁃sucrose galactosyltransferase and other GO terms,and carbon fixation in photosynthetic origination,plant hormone signal transduction,protein processing in endoplasmic reticulum,plant⁃pathogen interaction,MAPK signaling pathway⁃plant and other KEGG pathways under different nitrogen application rates.DEGs in ND108 were mainly enriched in response to water deprivation,toxin catabolic process,chitinase activity,trehalose biosynthetic process,trehalose⁃phosphatase activity and other GO terms,and glyoxylate and dicarboxylate metabolism,MAPK signaling pathway⁃plant，prodigiosin biosynthesis,zeatin biosynthesis,biotin metabolism and other KEGG pathways under different nitrogen application rates.Fifty⁃eight differentially expressed transcription factor families were detected in WK518 and ND108 under different nitrogen application rates,including GRAS,bHLH,MYB⁃related,NAC,C3H,ERF,C2H2,WRKY,FAR1 transcription factor families and so on,which were very important in plants growth,development and response to biotic and abiotic stresses.

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Transcriptome Analysis of Nanocellulose‑Fe Chelate Correcting Iron‑deficiency Chlorosis of Pear

GUO Xianping, BIAN Yiwei, WANG Dongsheng, WU Zhongying, WANG Hezhong, LIAN Xiaodong, GUO Peng

Journal of Henan Agricultural Sciences 2021, 50 (11): 117-129. DOI: 10.15933/j.cnki.1004-3268.2021.11.014

Abstract （211）

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In order to explore the molecular mechanism of nanocellulose‑Fe chelate correctingiron‑deficiency chlorosis of pear，Pyrus betulifolia leaves with iron‑deficiency chlorosis obtained by hydroponics was treated with 4 mmol/L of FeSO₄（T1）and nanocellulose‑Fe chelate in which nanocellulose and FeSO₄ were chelated at a charge ratio of 1∶3 000（T2）by spraying method. Deionized water was served as control（CK）.The active iron content，SPAD and net photosynthetic rate of the leaves were determined after 72 h of treatment，and the leaves transcriptome was sequenced and analyzed.The results showed that the active iron contents of T1 and T2 were significantly increased by 110.7% and 235.8% compared with the control，the SPAD values were significantly increased by 26.1% and 61.7% respectively，the net photosynthetic rates were significantly increased by 70.1% and 98.5%，respectively，and there was a significant difference between T1 and T2. The transcriptome sequencing results showed that T1 vs CK and T2 vs CK had 1 033 and 1 943 differentially expressed genes，respectively.GO functional enrichment analysis showed that the number of GO items enriched in T2 was all greater than T1 in molecular function，cell composition and biological process，mainly involving metal ion fixation，oxidation‑reduction process，chloroplast，photosynthetic light harvesting. KEGG enrichment analysis found that only 55 differentially expressed genes of T1 were annotated into 4 pathways，and 712 differentially expressed genes of T2 were annotated into 18 pathways compared with the control. These differentially expressed genes were mainly involved in photosynthesis‑antenna proteins，photosynthesis，carbon fixation in photosynthetic organisms,and metabolic pathways.According to the analysis of gene expression level of Pyrus betulifolia ferritin gene family，four ferritin genes of T2 were all highly expressed than those of T1. In the pathways of photosynthesis-antenna protein and photosynthesis,the expression levels of 58 genes in T1 and T2 were up-regulated compared with the control，and the expression levels of 56 genes in T2 were higher than in T1.The results of the expression patterns of eight selected differentially expressed genes verified by qRT-PCR were all consistent with the results of RNA-Seq analysis. Comprehensive analysis showed that spraying nanocellulose-Fe chelate could mobilize more genes and metabolic pathways，enhancing its recovery ability from iron-deficiency chlorosis of pear.

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Transcriptome and Metabolome Analysis of Mechanisms Responding to High Temperature Stress during Anthesis Stage in Zhengdan 309

LI Chuan, HUANG Lu, QIAO Jiangfang, ZHANG Meiwei, ZHANG Panpan, NIU Jun, LIU Jingbao, WANG Shufeng

Journal of Henan Agricultural Sciences 2021, 50 (2): 19-31. DOI: 10.15933/j.cnki.1004-3268.2021.02.003

Abstract （1197）

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The ear-leaf samples collected from Zhengdan 309 under normal condition(CK) and high temperature stress for 7 d and 14 d were used for RNA-sequencing using Illumina HiSeqTM2500 high-throughput sequencing technology and metabolome profiling analysis using liquid-chromatographymass spectrometry-based metabolomics. Differentially expressed genes (DEGs) and metabolites were explored to clarify the mechanism of maize responding to high temperature stress at the anthesis stage.The transcriptome sequencing results showed that 515 DEGs were detected in Zhengdan 309 under high temperature stress for 7 d compared with the control under normal growth condition, of which 75 genes were up-regulated,and 440 genes were down-regulated.There were 506 DEGs detected in Zhengdan 309 under high temperature stress for 14 d compared with the control under normal growth condition,including 114 up-regulated genes and 392 down-regulated genes. There were 2 050 DEGs detected in Zhengdan 309 under high temperature stress for 7 d compared with that under high temperature stress for 14 d, of which 790 genes were up-regulated,and 1 260 genes were down-regulated. The DEGs in Zhengdan 309 under high temperature stress for 7 d compared with that under the normal condition were concentrated in extracelluar region, molecular function regulator and other GO classification.The DEGs in Zhengdan 309 under high temperature stress for 14 d compared with that under the normal condition were mainly enriched in rhythmic process, extracelluar region and other GO classification.The DEGs in Zhengdan 309 under high temperature stress for 7 d compared with that under high temperature stress for 14 d were mainly enriched in cell part, single-organism process and other GO classification.The DEGs in Zhengdan 309 under high temperature stress for 7 d compared with that under the normal condition,Zhengdan 309 under high temperature stress for 14 d compared with that under the normal condition and Zhengdan 309 under high temperature stress for 7 d compared with that under high temperature stress for 14 d were mainly distributed in 5, 7 and 15 major KOG/COG classifications, respectively, and were mainly annotated into 6,7,20 KEGG metabolic pathways. There were 654 metabolites detected in Zhengdan 309,40 differentially expressed metabolites(DEMs) were detected in Zhengdan 309 under high temperature stress for 7 d compared with that under the normal condition, of which 8 metablites were up-regulated,and 32 metablites were down-regulated.There were also 40 DEMs detected in Zhengdan 309 under high temperature stress for 14 d compared with that under the normal condition, including 4 up-regulated metablites and 36 down-regulated metablites.There were 46 DEMs detected in Zhengdan 309 under high temperature stress for 7 d compared with that under high temperature stress for 14 d, of which 17 metablites were up-regulated,and 29 metablites were down-regulated.The 40 DEMs in Zhengdan 309 under high temperature stress for 7 d compared with that under the normal condition were concentrated in biosynthesis of secondary metabolites, arginine and proline metabolism and other KEGG pathways.The 40 DEMs in Zhengdan 309 under high temperature stress for 14 d compared with that under the normal condition were concentrated in biosynthesis of secondary metabolites, steroid biosynthesis and other KEGG pathways.The 46 DEMs in Zhengdan 309 under high temperature stress for 7 d compared with that under high temperature stress for 14 d were mainly enriched in aminoacyl-tRNA biosynthesis, alanine,aspartate and glutamate metabolism and other KEGG pathways.

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