Loquat(Eriobotrya japonica Lindl.)is a subtropical evergreen fruit tree that produces fruits with abundant nutrients and medicinal components.Confirming suitable reference genes for a set of loquat samples before qRT-...Loquat(Eriobotrya japonica Lindl.)is a subtropical evergreen fruit tree that produces fruits with abundant nutrients and medicinal components.Confirming suitable reference genes for a set of loquat samples before qRT-PCR experiments is essential for the accurate quantification of gene expression.In this study,eight candidate reference genes were selected from our previously published RNA-seq data,and primers for each candidate reference gene were designed and evaluated.The Cq values of the candidate reference genes were calculated by RT-qPCR in 31 different loquat samples,including 12 subgroups of developing or abiotic-stressed tissues.Different combinations of stable reference genes were screened according to a comprehensive rank,which was synthesized from the results of four algorithms,including the geNorm,NormFinder,BestKeeper andΔCt methods.The screened reference genes were verified by normalizing EjLGA1 in each subgroup.The obtained suitable combinations of reference genes for accurate normalization were GAPDH,EF1αand ACT for floral development;GAPDH,UBCE and ACT for fruit setting;EF1α,GAPDH and eIF2B for fruit ripening;ACT,EF1αand UBCE for leaves under heat stress;eIF2B,UBCE and EF1αfor leaves under freezing stress;EF1α,TUA and UBCE for leaves under salt stress;ACT,EF1αand eIF2B for immature pulp under freezing stress;ACT,UBCE and eIF2B for immature seeds under freezing stress;EF1α,eIF2B and UBCE for both immature pulp and seeds under freezing stress;UBCE,TUB and TUA for red-fleshed fruits under cold-storage stress;eIF2B,RPS3 and TUB for white-fleshed fruits under coldstorage stress;and eIF2B,UBCE and RPS3 for both red-and white-fleshed fruits under cold-storage stress.This study obtained different combinations of stable reference genes for accurate normalization in twelve subgroups of developing or abiotic-stressed tissues in loquat.To our knowledge,this is the first report to obtain stable reference genes for normalizing gene expression of abiotic-stressed tissues in E.japonica.The use of the three most stable reference genes could increase the reliability of future quantification experiments.展开更多
The Gibberellic Acid-stimulated Arabidopsis(GASA)gene family is involved in the regulation of gene expression and plant growth,development,and stress responses.To investigate the function of loquat GASA genes in the g...The Gibberellic Acid-stimulated Arabidopsis(GASA)gene family is involved in the regulation of gene expression and plant growth,development,and stress responses.To investigate the function of loquat GASA genes in the growth and developmental regulation of plants,a loquat EjGASA6 gene homologous to Arabidopsis AtGASA6 was cloned.EjGASA6 expression was induced by gibberellin,and ectopic transgenic plants containing this gene exhibited earlier bloom and longer primary roots since these phenotypic characteristics are related to higher gibberellin content.Transcriptome analysis and qRT-PCR results showed that the expression levels of GA3ox1 and GA20ox1,which encode key enzymes in gibberellin biosynthesis,were significantly increased.Furthermore,we confirmed that EjGASA6 could promote the expression of GA20ox1 via the luciferase reporter system.Overall,our results suggest that EjGASA6 promotes blooming and main-root elongation by positively regulating gibberellin biosynthesis.These findings broaden our understanding of the role of GASAs in plant development and growth,and lay the groundwork for future research into the functions of EjGASA6 in regulating loquat growth and development.展开更多
基金funded by the Natural Science Foundation of Fujian Province(2021J05240)Fujian Provincial Science and Technology Project(2021N5014,2022N5006)+1 种基金Research Project of Putian Science and Technology Bureau(2021ZP08,2021ZP09,2021ZP10,2021ZP11)Scientific Research Project of Putian University(2018064).
文摘Loquat(Eriobotrya japonica Lindl.)is a subtropical evergreen fruit tree that produces fruits with abundant nutrients and medicinal components.Confirming suitable reference genes for a set of loquat samples before qRT-PCR experiments is essential for the accurate quantification of gene expression.In this study,eight candidate reference genes were selected from our previously published RNA-seq data,and primers for each candidate reference gene were designed and evaluated.The Cq values of the candidate reference genes were calculated by RT-qPCR in 31 different loquat samples,including 12 subgroups of developing or abiotic-stressed tissues.Different combinations of stable reference genes were screened according to a comprehensive rank,which was synthesized from the results of four algorithms,including the geNorm,NormFinder,BestKeeper andΔCt methods.The screened reference genes were verified by normalizing EjLGA1 in each subgroup.The obtained suitable combinations of reference genes for accurate normalization were GAPDH,EF1αand ACT for floral development;GAPDH,UBCE and ACT for fruit setting;EF1α,GAPDH and eIF2B for fruit ripening;ACT,EF1αand UBCE for leaves under heat stress;eIF2B,UBCE and EF1αfor leaves under freezing stress;EF1α,TUA and UBCE for leaves under salt stress;ACT,EF1αand eIF2B for immature pulp under freezing stress;ACT,UBCE and eIF2B for immature seeds under freezing stress;EF1α,eIF2B and UBCE for both immature pulp and seeds under freezing stress;UBCE,TUB and TUA for red-fleshed fruits under cold-storage stress;eIF2B,RPS3 and TUB for white-fleshed fruits under coldstorage stress;and eIF2B,UBCE and RPS3 for both red-and white-fleshed fruits under cold-storage stress.This study obtained different combinations of stable reference genes for accurate normalization in twelve subgroups of developing or abiotic-stressed tissues in loquat.To our knowledge,this is the first report to obtain stable reference genes for normalizing gene expression of abiotic-stressed tissues in E.japonica.The use of the three most stable reference genes could increase the reliability of future quantification experiments.
基金financially supported by the National Key R&D Program of China (2023YFD1600800)the National Nature Science Foundation of China (32102321)+4 种基金the Chongqing Science and Technology Commission, China (cstc2024ycjh-bgzxm0202, cstc2021jscx-gksbX0010 and cstc2021jcyj-msxmX1156)the Chongqing Forestry Administration, China (YuLinKeYan2022-14)the Innovation Research Group Funds for Chongqing Universities, China (CXQT19005)the Characteristic Fruit Industry and Technology System Innovation Team of Chongqing Agriculture and Rural Affairs Commission, China [(2022)164 and 2020(3)01]the Chongqing Postgraduate Research and Innovation Programme, China (CYB23128)
文摘The Gibberellic Acid-stimulated Arabidopsis(GASA)gene family is involved in the regulation of gene expression and plant growth,development,and stress responses.To investigate the function of loquat GASA genes in the growth and developmental regulation of plants,a loquat EjGASA6 gene homologous to Arabidopsis AtGASA6 was cloned.EjGASA6 expression was induced by gibberellin,and ectopic transgenic plants containing this gene exhibited earlier bloom and longer primary roots since these phenotypic characteristics are related to higher gibberellin content.Transcriptome analysis and qRT-PCR results showed that the expression levels of GA3ox1 and GA20ox1,which encode key enzymes in gibberellin biosynthesis,were significantly increased.Furthermore,we confirmed that EjGASA6 could promote the expression of GA20ox1 via the luciferase reporter system.Overall,our results suggest that EjGASA6 promotes blooming and main-root elongation by positively regulating gibberellin biosynthesis.These findings broaden our understanding of the role of GASAs in plant development and growth,and lay the groundwork for future research into the functions of EjGASA6 in regulating loquat growth and development.
