Watermelon is a highly cultivated fruit crop renowned for its quality properties of fruit flesh.Among various quality factors,fruit flesh firmness is a crucial quality parameter influencing the fruit texture,shelf lif...Watermelon is a highly cultivated fruit crop renowned for its quality properties of fruit flesh.Among various quality factors,fruit flesh firmness is a crucial quality parameter influencing the fruit texture,shelf life and its commercial value.The auxin/indole-3-acetic acid(Aux/IAA)plays a significant role in fruit development and ripening of non-climacteric fruits.However,the regulatory mechanism of Aux/IAA in controlling fruit flesh firmness and ripening in watermelon remains unknown.In this study,we employed an integrative approach combining genome-wide association study(GWAS)and bulked segregant RNA-Seq analysis(BSR-Seq)to identify an overlapping candidate region between 12776310 and 12968331 bp on chromosome 6,underlying an auxin-responsive gene(Aux/IAA)associated with flesh firmness in watermelon.Transcriptome analysis,followed by real-time quantitative reverse transcription PCR(qRT-PCR),confirmed that the expression of Aux/IAA was consistently higher in fruits with high flesh firmness.The sequence alignment revealed a single base mutation in the coding region of Aux/IAA.Furthermore,the concomitant Kompetitive/Competitive allele-specific PCR(KASP)genotyping data sets for F2 population and germplasm accessions identified Aux/IAA as a strong candidate gene associated with flesh firmness.Aux/IAA was enriched in the plant hormone signal transduction pathway,involving cell enlargement and leading to low flesh firmness.We determined the higher accumulation of abscisic acid(ABA)in fruits with low flesh firmness than hard flesh.Moreover,overexpression of Aux/IAA induced higher flesh firmness with an increased number of fruit flesh cells while reducing ABA content and flesh cell sizes.Additionally,the allelic variation in Aux/IAA for soft flesh firmness was found to exist in Citrullus mucosospermus and gradually fixed into Citrullus lanatus during domestication,indicating that soft flesh firmness was a domesticated trait.These findings significantly enhanced our understanding of watermelon fruit flesh firmness and consequently the watermelon fruit quality.展开更多
Watermelon(Citrullus lanatus)holds global significance as a fruit with high economic and nutritional value.Exploring the regulatory network of watermelon male reproductive development is crucial for developing male st...Watermelon(Citrullus lanatus)holds global significance as a fruit with high economic and nutritional value.Exploring the regulatory network of watermelon male reproductive development is crucial for developing male sterile materials and facilitating cross-breeding.Despite its importance,there is a lack of research on the regulation mechanism of male reproductive development in watermelon.In this study,we identified that ClESR2,a VIIIb subclass member in the APETALA2/Ethylene Responsive Factor(AP2/ERF)superfamily,was a key factor in pollen development.RNA insitu hybridization confirmed significantClESR2 expression in the tapetum and pollen during the later stage of anther development.The pollens of transgenic plants showed major defects in morphology and vitality at the late development stage.The RNA-seq and protein interaction assay confirmed that ClESR2 regulates pollen morphology and fertility by interacting with key genes involved in pollen development at both transcriptional and protein levels.These suggest that Enhancer of Shoot Regeneration 2(ESR2)plays an important role in pollen maturation and vitality.This study helps understand the male reproductive development of watermelon,providing a theoretical foundation for developing male sterile materials.展开更多
基金the Agricultural Science and Technology Innovation Program(Grant No.CAAS-ASTIP-2021-ZFRI)China Agriculture Research System of MOF and MARA(Grant No.CARS-25-03)+3 种基金National Nature Science Foundation of China(Grant Nos.31672178 and 31471893)the Natural Science Foundation of Henan Province(Grant No.212300410312)the scientific and technological research in Henan Province(Grant No.202102110398)the key project of the Action of“Rejuvenating Mongolia with Science and Technology”(Grant No.NMKJXM202114).
文摘Watermelon is a highly cultivated fruit crop renowned for its quality properties of fruit flesh.Among various quality factors,fruit flesh firmness is a crucial quality parameter influencing the fruit texture,shelf life and its commercial value.The auxin/indole-3-acetic acid(Aux/IAA)plays a significant role in fruit development and ripening of non-climacteric fruits.However,the regulatory mechanism of Aux/IAA in controlling fruit flesh firmness and ripening in watermelon remains unknown.In this study,we employed an integrative approach combining genome-wide association study(GWAS)and bulked segregant RNA-Seq analysis(BSR-Seq)to identify an overlapping candidate region between 12776310 and 12968331 bp on chromosome 6,underlying an auxin-responsive gene(Aux/IAA)associated with flesh firmness in watermelon.Transcriptome analysis,followed by real-time quantitative reverse transcription PCR(qRT-PCR),confirmed that the expression of Aux/IAA was consistently higher in fruits with high flesh firmness.The sequence alignment revealed a single base mutation in the coding region of Aux/IAA.Furthermore,the concomitant Kompetitive/Competitive allele-specific PCR(KASP)genotyping data sets for F2 population and germplasm accessions identified Aux/IAA as a strong candidate gene associated with flesh firmness.Aux/IAA was enriched in the plant hormone signal transduction pathway,involving cell enlargement and leading to low flesh firmness.We determined the higher accumulation of abscisic acid(ABA)in fruits with low flesh firmness than hard flesh.Moreover,overexpression of Aux/IAA induced higher flesh firmness with an increased number of fruit flesh cells while reducing ABA content and flesh cell sizes.Additionally,the allelic variation in Aux/IAA for soft flesh firmness was found to exist in Citrullus mucosospermus and gradually fixed into Citrullus lanatus during domestication,indicating that soft flesh firmness was a domesticated trait.These findings significantly enhanced our understanding of watermelon fruit flesh firmness and consequently the watermelon fruit quality.
基金support from the National Key Research and Development Program of China(2022YFD1602000)the National Natural Science Foundation of China(32202514,U22A20498 and 32072596)+2 种基金the Joint Fund of Henan Province Science and Technology Research and Development Plan,China(222103810009)the Science and Technology Innovation Team of Shaanxi,China(2021TD-32)the China Postdoctoral Science Foundation(2022M711064 and 2023M741062).
文摘Watermelon(Citrullus lanatus)holds global significance as a fruit with high economic and nutritional value.Exploring the regulatory network of watermelon male reproductive development is crucial for developing male sterile materials and facilitating cross-breeding.Despite its importance,there is a lack of research on the regulation mechanism of male reproductive development in watermelon.In this study,we identified that ClESR2,a VIIIb subclass member in the APETALA2/Ethylene Responsive Factor(AP2/ERF)superfamily,was a key factor in pollen development.RNA insitu hybridization confirmed significantClESR2 expression in the tapetum and pollen during the later stage of anther development.The pollens of transgenic plants showed major defects in morphology and vitality at the late development stage.The RNA-seq and protein interaction assay confirmed that ClESR2 regulates pollen morphology and fertility by interacting with key genes involved in pollen development at both transcriptional and protein levels.These suggest that Enhancer of Shoot Regeneration 2(ESR2)plays an important role in pollen maturation and vitality.This study helps understand the male reproductive development of watermelon,providing a theoretical foundation for developing male sterile materials.
