Selection of suitable reference genes for quantitative real-time PCR gene expression analysis in Mulberry (Morus alba L.) under different abiotic stresses
Mulberry ( Morus alba L.) is the sole food source for the mulberry silkworm, Bombyx mori and therefore important for sericulture industry. Different abiotic stress conditions like drought, salt, heat and cold stress adversely affect the productivity and quality of mulberry leaves. Quantitative real...
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| Veröffentlicht in: | Molecular biology reports 2019-04, Vol.46 (2), p.1809-1817 |
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| description | Mulberry (
Morus alba
L.) is the sole food source for the mulberry silkworm,
Bombyx mori
and therefore important for sericulture industry. Different abiotic stress conditions like drought, salt, heat and cold stress adversely affect the productivity and quality of mulberry leaves. Quantitative real time PCR (qPCR) is a reliable and widely used method to identify abiotic stress responsive genes and molecular mechanism in different plant species. Selection of suitable reference genes is important requirement for normalizing the expression of genes through qRT-PCR study. In the present study, we have selected eight candidate reference genes in mulberry for analyzing their expression stability in different abiotic stress treatments including drought, salt, heat and cold stresses. The expression stability of these reference genes was determined using geNorm, NormFinder and RefFinder statistical algorithms. The results showed that Ubiquitin and protein phosphatase 2A regulatory subunit A (PP2A) were the most stable genes across all the treatment samples. However, analysis of individual stresses revealed different expression profiles and stability of reference genes. Actin3 and PP2A were most stable in drought and salt conditions respectively. RPL3 most preferred in heat stress and Ubiquitin was most stable in cold stress. We propose the ubiquitin and PP2A are the preferred reference genes for normalization of gene expression data from abiotic stresses. In addition, Actin3 are preferred for drought stress, PP2A for salt stress, RPL3 for heat stress and Ubiquitin for cold stress studies. |
| doi_str_mv | 10.1007/s11033-019-04631-y |
| format | Article |
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Morus alba
L.) is the sole food source for the mulberry silkworm,
Bombyx mori
and therefore important for sericulture industry. Different abiotic stress conditions like drought, salt, heat and cold stress adversely affect the productivity and quality of mulberry leaves. Quantitative real time PCR (qPCR) is a reliable and widely used method to identify abiotic stress responsive genes and molecular mechanism in different plant species. Selection of suitable reference genes is important requirement for normalizing the expression of genes through qRT-PCR study. In the present study, we have selected eight candidate reference genes in mulberry for analyzing their expression stability in different abiotic stress treatments including drought, salt, heat and cold stresses. The expression stability of these reference genes was determined using geNorm, NormFinder and RefFinder statistical algorithms. The results showed that Ubiquitin and protein phosphatase 2A regulatory subunit A (PP2A) were the most stable genes across all the treatment samples. However, analysis of individual stresses revealed different expression profiles and stability of reference genes. Actin3 and PP2A were most stable in drought and salt conditions respectively. RPL3 most preferred in heat stress and Ubiquitin was most stable in cold stress. We propose the ubiquitin and PP2A are the preferred reference genes for normalization of gene expression data from abiotic stresses. In addition, Actin3 are preferred for drought stress, PP2A for salt stress, RPL3 for heat stress and Ubiquitin for cold stress studies.</description><identifier>ISSN: 0301-4851</identifier><identifier>EISSN: 1573-4978</identifier><identifier>DOI: 10.1007/s11033-019-04631-y</identifier><identifier>PMID: 30694457</identifier><language>eng</language><publisher>Dordrecht: Springer Netherlands</publisher><subject>Abiotic stress ; Animal Anatomy ; Animal Biochemistry ; Biomedical and Life Sciences ; Bombyx mori ; Cold ; Drought ; Droughts ; Food sources ; Gene expression ; Gene Expression - genetics ; Gene Expression Profiling - methods ; Gene Expression Profiling - standards ; Gene Expression Regulation, Plant - genetics ; Genes, Plant - genetics ; Heat ; Heat-Shock Response ; Histology ; Life Sciences ; Morphology ; Morus - genetics ; Morus alba ; Original Article ; Phosphoprotein phosphatase ; Plant Leaves - metabolism ; Plant Proteins - genetics ; Protein phosphatase ; Real-Time Polymerase Chain Reaction - methods ; Reference Standards ; Sericulture ; Stress, Physiological - genetics ; Ubiquitin</subject><ispartof>Molecular biology reports, 2019-04, Vol.46 (2), p.1809-1817</ispartof><rights>Springer Nature B.V. 2019</rights><rights>Molecular Biology Reports is a copyright of Springer, (2019). All Rights Reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c375t-3e510d955272aa41f0c928b250c576b42941b1a9f3c4967c885dff4e80873b853</citedby><cites>FETCH-LOGICAL-c375t-3e510d955272aa41f0c928b250c576b42941b1a9f3c4967c885dff4e80873b853</cites><orcidid>0000-0002-5401-7188</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s11033-019-04631-y$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s11033-019-04631-y$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,27924,27925,41488,42557,51319</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/30694457$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Shukla, Pawan</creatorcontrib><creatorcontrib>Reddy, Ramesha A.</creatorcontrib><creatorcontrib>Ponnuvel, Kangayam M.</creatorcontrib><creatorcontrib>Rohela, Gulab Khan</creatorcontrib><creatorcontrib>Shabnam, Aftab A.</creatorcontrib><creatorcontrib>Ghosh, M. K.</creatorcontrib><creatorcontrib>Mishra, Rakesh Kumar</creatorcontrib><title>Selection of suitable reference genes for quantitative real-time PCR gene expression analysis in Mulberry (Morus alba L.) under different abiotic stresses</title><title>Molecular biology reports</title><addtitle>Mol Biol Rep</addtitle><addtitle>Mol Biol Rep</addtitle><description>Mulberry (
Morus alba
L.) is the sole food source for the mulberry silkworm,
Bombyx mori
and therefore important for sericulture industry. Different abiotic stress conditions like drought, salt, heat and cold stress adversely affect the productivity and quality of mulberry leaves. Quantitative real time PCR (qPCR) is a reliable and widely used method to identify abiotic stress responsive genes and molecular mechanism in different plant species. Selection of suitable reference genes is important requirement for normalizing the expression of genes through qRT-PCR study. In the present study, we have selected eight candidate reference genes in mulberry for analyzing their expression stability in different abiotic stress treatments including drought, salt, heat and cold stresses. The expression stability of these reference genes was determined using geNorm, NormFinder and RefFinder statistical algorithms. The results showed that Ubiquitin and protein phosphatase 2A regulatory subunit A (PP2A) were the most stable genes across all the treatment samples. However, analysis of individual stresses revealed different expression profiles and stability of reference genes. Actin3 and PP2A were most stable in drought and salt conditions respectively. RPL3 most preferred in heat stress and Ubiquitin was most stable in cold stress. We propose the ubiquitin and PP2A are the preferred reference genes for normalization of gene expression data from abiotic stresses. In addition, Actin3 are preferred for drought stress, PP2A for salt stress, RPL3 for heat stress and Ubiquitin for cold stress studies.</description><subject>Abiotic stress</subject><subject>Animal Anatomy</subject><subject>Animal Biochemistry</subject><subject>Biomedical and Life Sciences</subject><subject>Bombyx mori</subject><subject>Cold</subject><subject>Drought</subject><subject>Droughts</subject><subject>Food sources</subject><subject>Gene expression</subject><subject>Gene Expression - genetics</subject><subject>Gene Expression Profiling - methods</subject><subject>Gene Expression Profiling - standards</subject><subject>Gene Expression Regulation, Plant - genetics</subject><subject>Genes, Plant - genetics</subject><subject>Heat</subject><subject>Heat-Shock Response</subject><subject>Histology</subject><subject>Life Sciences</subject><subject>Morphology</subject><subject>Morus - genetics</subject><subject>Morus alba</subject><subject>Original Article</subject><subject>Phosphoprotein phosphatase</subject><subject>Plant Leaves - metabolism</subject><subject>Plant Proteins - genetics</subject><subject>Protein phosphatase</subject><subject>Real-Time Polymerase Chain Reaction - methods</subject><subject>Reference Standards</subject><subject>Sericulture</subject><subject>Stress, Physiological - genetics</subject><subject>Ubiquitin</subject><issn>0301-4851</issn><issn>1573-4978</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNp9kctq3DAUhkVpaSbTvkAXRdBNslAqWZJlL8vQG0xI6GVtZPk4KHjkiY4U6lfp08aeSRvooistzne-g_6fkDeCXwjOzXsUgkvJuKgZV6UUbHpGVkIbyVRtqudkxSUXTFVanJBTxFvOuRJGvyQnkpe1UtqsyO_vMIBLfgx07Clmn2w7AI3QQ4TggN5AAKT9GOldtiHN8-TvF8AOLPkd0OvNtwNE4dc-AuKissEOE3qkPtDLPLQQ40TPLseYkdqhtXR7cU5z6CDSzveHU4na1o_JO4pp0QC-Ii96OyC8fnzX5Oenjz82X9j26vPXzYctc9LoxCRowbta68IU1irRc1cXVVto7rQpW1XUSrTC1r10qi6Nqyrd9b2CildGtpWWa3J29O7jeJcBU7Pz6GAYbIAxY1MIM4dVLlmvybt_0Nsxx_mzB6pQlTRioYoj5eKIOEfZ7KPf2Tg1gjdLc82xuWZurjk010zz0ttHdW530P1d-VPVDMgjgPMo3EB8uv0f7QMg8aVI</recordid><startdate>20190401</startdate><enddate>20190401</enddate><creator>Shukla, Pawan</creator><creator>Reddy, Ramesha A.</creator><creator>Ponnuvel, Kangayam M.</creator><creator>Rohela, Gulab Khan</creator><creator>Shabnam, Aftab A.</creator><creator>Ghosh, M. K.</creator><creator>Mishra, Rakesh Kumar</creator><general>Springer Netherlands</general><general>Springer Nature B.V</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7TK</scope><scope>7TM</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7P</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>Q9U</scope><scope>RC3</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0002-5401-7188</orcidid></search><sort><creationdate>20190401</creationdate><title>Selection of suitable reference genes for quantitative real-time PCR gene expression analysis in Mulberry (Morus alba L.) under different abiotic stresses</title><author>Shukla, Pawan ; Reddy, Ramesha A. ; Ponnuvel, Kangayam M. ; Rohela, Gulab Khan ; Shabnam, Aftab A. ; Ghosh, M. K. ; Mishra, Rakesh Kumar</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c375t-3e510d955272aa41f0c928b250c576b42941b1a9f3c4967c885dff4e80873b853</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2019</creationdate><topic>Abiotic stress</topic><topic>Animal Anatomy</topic><topic>Animal Biochemistry</topic><topic>Biomedical and Life Sciences</topic><topic>Bombyx mori</topic><topic>Cold</topic><topic>Drought</topic><topic>Droughts</topic><topic>Food sources</topic><topic>Gene expression</topic><topic>Gene Expression - genetics</topic><topic>Gene Expression Profiling - methods</topic><topic>Gene Expression Profiling - standards</topic><topic>Gene Expression Regulation, Plant - genetics</topic><topic>Genes, Plant - genetics</topic><topic>Heat</topic><topic>Heat-Shock Response</topic><topic>Histology</topic><topic>Life Sciences</topic><topic>Morphology</topic><topic>Morus - genetics</topic><topic>Morus alba</topic><topic>Original Article</topic><topic>Phosphoprotein phosphatase</topic><topic>Plant Leaves - metabolism</topic><topic>Plant Proteins - genetics</topic><topic>Protein phosphatase</topic><topic>Real-Time Polymerase Chain Reaction - methods</topic><topic>Reference Standards</topic><topic>Sericulture</topic><topic>Stress, Physiological - genetics</topic><topic>Ubiquitin</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Shukla, Pawan</creatorcontrib><creatorcontrib>Reddy, Ramesha A.</creatorcontrib><creatorcontrib>Ponnuvel, Kangayam M.</creatorcontrib><creatorcontrib>Rohela, Gulab Khan</creatorcontrib><creatorcontrib>Shabnam, Aftab A.</creatorcontrib><creatorcontrib>Ghosh, M. 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K.</au><au>Mishra, Rakesh Kumar</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Selection of suitable reference genes for quantitative real-time PCR gene expression analysis in Mulberry (Morus alba L.) under different abiotic stresses</atitle><jtitle>Molecular biology reports</jtitle><stitle>Mol Biol Rep</stitle><addtitle>Mol Biol Rep</addtitle><date>2019-04-01</date><risdate>2019</risdate><volume>46</volume><issue>2</issue><spage>1809</spage><epage>1817</epage><pages>1809-1817</pages><issn>0301-4851</issn><eissn>1573-4978</eissn><abstract>Mulberry (
Morus alba
L.) is the sole food source for the mulberry silkworm,
Bombyx mori
and therefore important for sericulture industry. Different abiotic stress conditions like drought, salt, heat and cold stress adversely affect the productivity and quality of mulberry leaves. Quantitative real time PCR (qPCR) is a reliable and widely used method to identify abiotic stress responsive genes and molecular mechanism in different plant species. Selection of suitable reference genes is important requirement for normalizing the expression of genes through qRT-PCR study. In the present study, we have selected eight candidate reference genes in mulberry for analyzing their expression stability in different abiotic stress treatments including drought, salt, heat and cold stresses. The expression stability of these reference genes was determined using geNorm, NormFinder and RefFinder statistical algorithms. The results showed that Ubiquitin and protein phosphatase 2A regulatory subunit A (PP2A) were the most stable genes across all the treatment samples. However, analysis of individual stresses revealed different expression profiles and stability of reference genes. Actin3 and PP2A were most stable in drought and salt conditions respectively. RPL3 most preferred in heat stress and Ubiquitin was most stable in cold stress. We propose the ubiquitin and PP2A are the preferred reference genes for normalization of gene expression data from abiotic stresses. In addition, Actin3 are preferred for drought stress, PP2A for salt stress, RPL3 for heat stress and Ubiquitin for cold stress studies.</abstract><cop>Dordrecht</cop><pub>Springer Netherlands</pub><pmid>30694457</pmid><doi>10.1007/s11033-019-04631-y</doi><tpages>9</tpages><orcidid>https://orcid.org/0000-0002-5401-7188</orcidid></addata></record> |
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| ispartof | Molecular biology reports, 2019-04, Vol.46 (2), p.1809-1817 |
| issn | 0301-4851 1573-4978 |
| language | eng |
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| source | MEDLINE; SpringerNature Journals |
| subjects | Abiotic stress Animal Anatomy Animal Biochemistry Biomedical and Life Sciences Bombyx mori Cold Drought Droughts Food sources Gene expression Gene Expression - genetics Gene Expression Profiling - methods Gene Expression Profiling - standards Gene Expression Regulation, Plant - genetics Genes, Plant - genetics Heat Heat-Shock Response Histology Life Sciences Morphology Morus - genetics Morus alba Original Article Phosphoprotein phosphatase Plant Leaves - metabolism Plant Proteins - genetics Protein phosphatase Real-Time Polymerase Chain Reaction - methods Reference Standards Sericulture Stress, Physiological - genetics Ubiquitin |
| title | Selection of suitable reference genes for quantitative real-time PCR gene expression analysis in Mulberry (Morus alba L.) under different abiotic stresses |
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