In this article 1,author name Yanping Wang was incorrectly tagged as corresponding author.Prof.Wang should be the co-first author,instead of the corresponding author.The original article has been corrected.
Abscisic acid(ABA)is a critical regulator of seed development and germination.β-glucosidases(BGs)have been suggested to be contributors to increased ABA content because they catalyze the hydrolysis of ABA-glucose est...Abscisic acid(ABA)is a critical regulator of seed development and germination.β-glucosidases(BGs)have been suggested to be contributors to increased ABA content because they catalyze the hydrolysis of ABA-glucose ester to release free ABA.However,whether BGs are involved in seed development is unclear.In this study,a candidate gene,CiBG1,in watermelon was selected for targeted mutagenesis via the CRISPR/Cas9 system.Seed size and weight were significantly reduced in the Clbg1-mutant watermelon lines,which was mainly attributed to decreased cell number resulting from decreased ABA levels.A transcriptome analysis showed that the expression of 1015 and 1429 unique genes was changed 10 and 18 days after pollination(DAP),respectively.Cytoskeleton-and cell cycle-related genes were enriched in the differentially expressed genes of wild type and Clbg1-mutant lines during seed development.Moreover,the expression of genes in the major signaling pathways of seed size control was also changed.In addition,seed germination was promoted in the Cibg1-mutant lines due to decreased ABA content.These results indicate that ClBG1 may be critical for watermelon seed size regulation and germination mainly through the modulation of ABA content and thereby the transcriptional regulation of cytoskeleton-,cell cycle-and signaling-related genes.Our results lay a foundation for dissecting the molecular mechanisms of controlling watermelon seed size,a key agricultural trait of significant economic importance.展开更多
NAC(NAM,ATAF1/2,and CUC2)transcription factors play important roles in fruit ripening and quality.The watermelon genome encodes 80 NAC genes,and 21 of these NAC genes are highly expressed in both the flesh and vascula...NAC(NAM,ATAF1/2,and CUC2)transcription factors play important roles in fruit ripening and quality.The watermelon genome encodes 80 NAC genes,and 21 of these NAC genes are highly expressed in both the flesh and vascular tissues.Among these genes,CINAC68 expression was signi fi cantly higher in flesh than in rind.However,the intrinsic regulatory mechanism of CINAC68 in fruit ripening and quality is still unknown.In this study,we found that ClNAC68 is a transcriptional repressor and that the repression domain is located in the C-terminus.Knockout of CINAC68 by the CRISPR-Cas9 system decreased the soluble solid content and sucrose accumulation in mutant flesh.Development was delayed,germination was inhibited,and the IAA content was signi ficantly decreased in mutant seeds.Transcriptome analysis showed that the invertase gene CUNV was the only gene involved in sucrose metabolism that was upregulated in mutant flesh,and expression of the indole-3-acetic acid-amido synthetase gene CIGH3.6 in the IAA signaling pathway was also induced in mutant seeds.EMSA and dual-luciferase assays showed that CINAC68 directly bound to the promoters of CUNV and CIGH3.6 to repress their expression.These results indicated that CINAC68 positively regulated sugar and IAA accumulation by repressing CUNV and CIGH3.6.Our findings provide new insights into the regulatory mechanisms by which NAC transcription factors affect fruit quality and seed development.展开更多
Virus-induced gene silencing (VIGS) offers a powerful approach for functional analysis of individual genes by knocking down their expression. We have adopted this approach to dissect gene functions in cotton resista...Virus-induced gene silencing (VIGS) offers a powerful approach for functional analysis of individual genes by knocking down their expression. We have adopted this approach to dissect gene functions in cotton resistant to Verticfllium wilt, one of the most devastating diseases worldwide. We showed here that highly efficient VIGS was obtained in a cotton breeding line (CA4002) with partial resistance to Verticillium wilt, and GhMKK2 and Gh Ve I are required for its resistance to Verticillium wilt. Arabidopsis AtBAK1/SERK3, a central regulator in plant disease resistance, belongs to a subfamily of somatic embryogenesis receptor kinases (SERKs) with five members, AtSERK1 to AtSERK5. Two BAK1 orthologs and one SERK1 ortholog were identified in the cotton genome. Importantly, GhBAK1 is required for CA4002 resistance to Verticillium wilt. Surprisingly, silencing of GhBAK1 is sufficient to trigger cell death accompanied with production of reactive oxygen species in cotton. This result is distinct from Arabidopsis in which AtBAK1 and AtSERK4 play redundant functions in cell death control. Apparently, cotton has only evolved SERK1 and BAK1 whereas AtSERK4/5 are newly evolved genes in Arabidopsis. Our studies indicate the functional importance of BAK1 in Verticillium wilt resistance and suggest the dynamic evolution of SERK family members in different plant species.展开更多
Watermelon(Citrullus lanatus) as non-climacteric fruit is domesticated from the ancestors with inedible fruits. We previously revealed that the abscisic acid(ABA) signaling pathway gene ClSnRK2.3 might infuence waterm...Watermelon(Citrullus lanatus) as non-climacteric fruit is domesticated from the ancestors with inedible fruits. We previously revealed that the abscisic acid(ABA) signaling pathway gene ClSnRK2.3 might infuence watermelon fruit ripening. However,the molecular mechanisms are unclear. Here,we found that the selective variation of ClSnRK2.3 resulted in lower promoter activity and gene expression level in cultivated watermelons than ancestors, which indicated ClSnRK2.3 might be a negative regulator in fruit ripening. Overexpression (OE) of ClSnRK2.3 significantly delayed watermelon fruit ripening and suppressed the accumulation of sucrose, ABA and gibberellin GA4. Furthermore,we determined that the pyrophosphate-dependent phosphofructokinase(ClPFP1) in sugar metabolism pathway and GA biosynthesis enzyme GA20 oxidase(Cl GA20ox) could be phosphorylated by ClSnRK2.3 and thereby resulting in accelerated protein degradation in OE lines and finally led to low levels of sucrose and GA4. Besides that, ClSnRK2.3 phosphorylated homeodomain-leucine zipper protein(ClHAT1) and protected it from degradation to suppress the expression of the ABA biosynthesis gene 9’-cis-epoxycarotenoid dioxygenase 3(Cl NCED3). These results indicated that ClSnRK2.3 negatively regulated watermelon fruit ripening by manipulating the biosynthesis of sucrose, ABA and GA4. Altogether, these findings revealed a novel regulatory mechanism in non-climacteric fruit development and ripening.展开更多
ThThe NAC transcription factor NONRIPENING(NOR)is a master regulator of climacteric fruit ripening.Melon(Cucumis melo L.)has climacteric and nonclimacteric fruit ripening varieties and is an ideal model to study fruit...ThThe NAC transcription factor NONRIPENING(NOR)is a master regulator of climacteric fruit ripening.Melon(Cucumis melo L.)has climacteric and nonclimacteric fruit ripening varieties and is an ideal model to study fruit ripening.Two natural CmNAC-NOR variants,the climacteric haplotype CmNAC-NOR^(S,N) and the non-climacteric haplotype CmNAC-NOR^(A,S),have effects on fruit ripening;however,their regulatory mechanisms have not been elucidated.Here,we report that a natural mutation in the transcriptional activation domain of CmNAC-NORS,Ncontributes to climacteric melon fruit ripening.CmNAC-NOR knockout in the climacteric-type melon cultivar“BYJH”completely inhibited fruit ripening,while ripening was delayed by 5-8 d in heterozygous cmnac-nor mutant fruits.CmN AC-NOR directly activated carotenoid,ethylene,and abscisic acid biosynthetic genes to promote fruit coloration and ripening.Furthermore,CmNAC-NOR mediated the transcription of the“CmNAC-NOR-CmNAC73-CmCWINV2”module to enhance flesh sweetness.The transcriptional activation activity of the climacteric haplotype CmNAC-NORS,Non these target genes was significantly higher than that of the nonclimacteric haplotype CmNAC-NOR^(A,S).Moreover,CmNAC-NORS,Ncomplementation fully rescued the non-ripening phenotype of the tomato(Solanum lycopersicum)cr-nor mutant,while CmNAC-NOR^(A,S) did not.Our results provide insight into the molecular mechanism of climacteric and non-climacteric fruit ripening in melon.展开更多
文摘In this article 1,author name Yanping Wang was incorrectly tagged as corresponding author.Prof.Wang should be the co-first author,instead of the corresponding author.The original article has been corrected.
基金supported by the National Natural Science Foundation of China(NSFC Grant Nos.31701938,31930096,and 1902034)Collaborative Innovation Center of BAAFS(KJCX201907-2)+2 种基金Ministry of Agriculture and Rural Affairs of China(Grant No.CARS-25)Beijing Scholar Program(Grant No.BSP026)Guanxi Bagui Scholar Program(Grant No.2016A11).
文摘Abscisic acid(ABA)is a critical regulator of seed development and germination.β-glucosidases(BGs)have been suggested to be contributors to increased ABA content because they catalyze the hydrolysis of ABA-glucose ester to release free ABA.However,whether BGs are involved in seed development is unclear.In this study,a candidate gene,CiBG1,in watermelon was selected for targeted mutagenesis via the CRISPR/Cas9 system.Seed size and weight were significantly reduced in the Clbg1-mutant watermelon lines,which was mainly attributed to decreased cell number resulting from decreased ABA levels.A transcriptome analysis showed that the expression of 1015 and 1429 unique genes was changed 10 and 18 days after pollination(DAP),respectively.Cytoskeleton-and cell cycle-related genes were enriched in the differentially expressed genes of wild type and Clbg1-mutant lines during seed development.Moreover,the expression of genes in the major signaling pathways of seed size control was also changed.In addition,seed germination was promoted in the Cibg1-mutant lines due to decreased ABA content.These results indicate that ClBG1 may be critical for watermelon seed size regulation and germination mainly through the modulation of ABA content and thereby the transcriptional regulation of cytoskeleton-,cell cycle-and signaling-related genes.Our results lay a foundation for dissecting the molecular mechanisms of controlling watermelon seed size,a key agricultural trait of significant economic importance.
基金This work was financially supported by the Collaborative Innovation Center of BAAFS(KJCX201907-2)Beijing Science&Technology Program(D171100007617001)+3 种基金National Key R&D Program of China(2018YFD0100703)National Natural Science Foun dation of China(31930096 and 1902034)Beijing Natural Science Foundation(6204038 and 6202010)Beijing Scholar Program(BSP026 and YBSP019).
文摘NAC(NAM,ATAF1/2,and CUC2)transcription factors play important roles in fruit ripening and quality.The watermelon genome encodes 80 NAC genes,and 21 of these NAC genes are highly expressed in both the flesh and vascular tissues.Among these genes,CINAC68 expression was signi fi cantly higher in flesh than in rind.However,the intrinsic regulatory mechanism of CINAC68 in fruit ripening and quality is still unknown.In this study,we found that ClNAC68 is a transcriptional repressor and that the repression domain is located in the C-terminus.Knockout of CINAC68 by the CRISPR-Cas9 system decreased the soluble solid content and sucrose accumulation in mutant flesh.Development was delayed,germination was inhibited,and the IAA content was signi ficantly decreased in mutant seeds.Transcriptome analysis showed that the invertase gene CUNV was the only gene involved in sucrose metabolism that was upregulated in mutant flesh,and expression of the indole-3-acetic acid-amido synthetase gene CIGH3.6 in the IAA signaling pathway was also induced in mutant seeds.EMSA and dual-luciferase assays showed that CINAC68 directly bound to the promoters of CUNV and CIGH3.6 to repress their expression.These results indicated that CINAC68 positively regulated sugar and IAA accumulation by repressing CUNV and CIGH3.6.Our findings provide new insights into the regulatory mechanisms by which NAC transcription factors affect fruit quality and seed development.
基金supported by funds from the Texas AgriLife Research Cotton Improvement Program to J.D.,T.W.,P.H., and L.S.the USDA National Institute of Food and Agriculture(Agriculture and Food Research Initiative Competitive Grants Program grant no.2012-67013-19433) to P.H.+1 种基金L.S.A.K.was supported by the NSF REU programF.L.was supported by the China Scholarship Council
文摘Virus-induced gene silencing (VIGS) offers a powerful approach for functional analysis of individual genes by knocking down their expression. We have adopted this approach to dissect gene functions in cotton resistant to Verticfllium wilt, one of the most devastating diseases worldwide. We showed here that highly efficient VIGS was obtained in a cotton breeding line (CA4002) with partial resistance to Verticillium wilt, and GhMKK2 and Gh Ve I are required for its resistance to Verticillium wilt. Arabidopsis AtBAK1/SERK3, a central regulator in plant disease resistance, belongs to a subfamily of somatic embryogenesis receptor kinases (SERKs) with five members, AtSERK1 to AtSERK5. Two BAK1 orthologs and one SERK1 ortholog were identified in the cotton genome. Importantly, GhBAK1 is required for CA4002 resistance to Verticillium wilt. Surprisingly, silencing of GhBAK1 is sufficient to trigger cell death accompanied with production of reactive oxygen species in cotton. This result is distinct from Arabidopsis in which AtBAK1 and AtSERK4 play redundant functions in cell death control. Apparently, cotton has only evolved SERK1 and BAK1 whereas AtSERK4/5 are newly evolved genes in Arabidopsis. Our studies indicate the functional importance of BAK1 in Verticillium wilt resistance and suggest the dynamic evolution of SERK family members in different plant species.
基金financially supported by the National Natural Science Foundation of China (31930096, 31902034, 32172592, 32122077, and 32072601)Foundation of Beijing Academy of Agricultural and Forestry Sciences (QNJJ202032 and QNJJ202206)+4 种基金the Ministry of Agriculture and Rural Affairs of China (CARS-25)the Beijing Scholar Program (YBSP019)Foundation of Xinjiang production and construction corps (2022AB015)The Agriculture Innovation Consortium (BAIC04-2023)The Scientist Training Program of BAAFS (JKZX202211)。
文摘Watermelon(Citrullus lanatus) as non-climacteric fruit is domesticated from the ancestors with inedible fruits. We previously revealed that the abscisic acid(ABA) signaling pathway gene ClSnRK2.3 might infuence watermelon fruit ripening. However,the molecular mechanisms are unclear. Here,we found that the selective variation of ClSnRK2.3 resulted in lower promoter activity and gene expression level in cultivated watermelons than ancestors, which indicated ClSnRK2.3 might be a negative regulator in fruit ripening. Overexpression (OE) of ClSnRK2.3 significantly delayed watermelon fruit ripening and suppressed the accumulation of sucrose, ABA and gibberellin GA4. Furthermore,we determined that the pyrophosphate-dependent phosphofructokinase(ClPFP1) in sugar metabolism pathway and GA biosynthesis enzyme GA20 oxidase(Cl GA20ox) could be phosphorylated by ClSnRK2.3 and thereby resulting in accelerated protein degradation in OE lines and finally led to low levels of sucrose and GA4. Besides that, ClSnRK2.3 phosphorylated homeodomain-leucine zipper protein(ClHAT1) and protected it from degradation to suppress the expression of the ABA biosynthesis gene 9’-cis-epoxycarotenoid dioxygenase 3(Cl NCED3). These results indicated that ClSnRK2.3 negatively regulated watermelon fruit ripening by manipulating the biosynthesis of sucrose, ABA and GA4. Altogether, these findings revealed a novel regulatory mechanism in non-climacteric fruit development and ripening.
基金financially supported by Beijing Academy of Agricultural and Forestry Sciences(KJCX20200204)the National Natural Science Foundation of China(1902034,32122077,and 32172592)+3 种基金the Beijing Scholar Program(YBSP019)the Sci&Tech Innovation of BAAFS(KJCX20200113)the Agriculture Innovation Consortium(BAIC10-2022)the Key Project of Science and Technology of Ningbo(2019B10007)。
文摘ThThe NAC transcription factor NONRIPENING(NOR)is a master regulator of climacteric fruit ripening.Melon(Cucumis melo L.)has climacteric and nonclimacteric fruit ripening varieties and is an ideal model to study fruit ripening.Two natural CmNAC-NOR variants,the climacteric haplotype CmNAC-NOR^(S,N) and the non-climacteric haplotype CmNAC-NOR^(A,S),have effects on fruit ripening;however,their regulatory mechanisms have not been elucidated.Here,we report that a natural mutation in the transcriptional activation domain of CmNAC-NORS,Ncontributes to climacteric melon fruit ripening.CmNAC-NOR knockout in the climacteric-type melon cultivar“BYJH”completely inhibited fruit ripening,while ripening was delayed by 5-8 d in heterozygous cmnac-nor mutant fruits.CmN AC-NOR directly activated carotenoid,ethylene,and abscisic acid biosynthetic genes to promote fruit coloration and ripening.Furthermore,CmNAC-NOR mediated the transcription of the“CmNAC-NOR-CmNAC73-CmCWINV2”module to enhance flesh sweetness.The transcriptional activation activity of the climacteric haplotype CmNAC-NORS,Non these target genes was significantly higher than that of the nonclimacteric haplotype CmNAC-NOR^(A,S).Moreover,CmNAC-NORS,Ncomplementation fully rescued the non-ripening phenotype of the tomato(Solanum lycopersicum)cr-nor mutant,while CmNAC-NOR^(A,S) did not.Our results provide insight into the molecular mechanism of climacteric and non-climacteric fruit ripening in melon.