To identify and characterize genes involved in reproductive tissue abscission in Brassica oleracea,the transcript data of pollinated pistil was analyzed.A differentially expressed gene,named BoFAZ1(FLOWER ABSCISSION Z...To identify and characterize genes involved in reproductive tissue abscission in Brassica oleracea,the transcript data of pollinated pistil was analyzed.A differentially expressed gene,named BoFAZ1(FLOWER ABSCISSION ZONE1)was identified,which contains one exon and encompass a 139aa.Furthermore,a T-DNA insertion mutant(SALK_302_G01)(faz1 mutant)was obtained from Arabidopsis thaliana mutant library.Floral organ shedding from mutants was delayed and a V-shaped structure in the boundary region between the stalk and torus of the sepal abscission zone was obtained in faz1 mutant.The cell density of this structure was lower than that of the corresponding region in the wild-type control.In the transgenic plants,the normal development of the stalk zone of faz1 was recovered completely by transforming a 1919-bp DNA fragment of BoFAZ1 into the faz1 mutant.In Addition,our data showed that BoFAZ1 was expressed in mature pollen grains,but not in the bracts,roots,stems,leaves,and sepals.Its expression in the filaments,stigma,and pistil exfoliation layer gradually increased after pollination.Subcellular localization experiments showed that BoFAZ1 was located in the cell membrane.A myristoylation site was found at the N-terminus of BoFAZ1.Removal of this site resulted in protein dislocation in the cytoplasm,cell membrane and nucleus.Finally,a yeast two-hybrid test indicated that BoH3.2(histone H3.2),a protein involved in abscission zone development,interacted with BoFAZ1.This interaction was verified by a GST pull-down assay.In summary,our data indicated that BoFAZ1 was involved in the formation of the pistil abscission zone in B.oleracea.展开更多
A MADS-box gene,CiMADS9,was cloned from the male flowers of Carya illinoinensis by rapid amplification of cDNA ends. The gene was 1 077 bp with a 768 bp open reading frame encoding 255 amino acids. Multiple sequence c...A MADS-box gene,CiMADS9,was cloned from the male flowers of Carya illinoinensis by rapid amplification of cDNA ends. The gene was 1 077 bp with a 768 bp open reading frame encoding 255 amino acids. Multiple sequence comparisons revealed that Ci MADS9 is a typical MIKC-type MADS-box gene with a MADS-box domain and a K semi-conserved region. Phylogenetic analysis indicated that CiMADS9 belongs to the AGL15 group of the MADS-box gene family. Quantitative reverse transcription polymerase chain reaction analysis indicated that the expression levels in reproductive organs(i.e.,flowers and young fruits) were considerably higher than in vegetative tissues(i.e.,leaves and branches). The highest expression levels were observed in male flowers. An overexpression vector for CiMADS9 was constructed and the gene was inserted into the Arabidopsis thaliana genome. CiMADS9 expression was confirmed in all transgenic lines. Compared with wild-type plants,transgenic A. thaliana plants overexpressing CiMADS9 exhibited delayed flowering and an increased number of leaves.展开更多
基金This work was supported by the National Natural Science Foundation of China(31572127)A Special Foundation of Central Institution Basic Research(XDJK2017C032).
文摘To identify and characterize genes involved in reproductive tissue abscission in Brassica oleracea,the transcript data of pollinated pistil was analyzed.A differentially expressed gene,named BoFAZ1(FLOWER ABSCISSION ZONE1)was identified,which contains one exon and encompass a 139aa.Furthermore,a T-DNA insertion mutant(SALK_302_G01)(faz1 mutant)was obtained from Arabidopsis thaliana mutant library.Floral organ shedding from mutants was delayed and a V-shaped structure in the boundary region between the stalk and torus of the sepal abscission zone was obtained in faz1 mutant.The cell density of this structure was lower than that of the corresponding region in the wild-type control.In the transgenic plants,the normal development of the stalk zone of faz1 was recovered completely by transforming a 1919-bp DNA fragment of BoFAZ1 into the faz1 mutant.In Addition,our data showed that BoFAZ1 was expressed in mature pollen grains,but not in the bracts,roots,stems,leaves,and sepals.Its expression in the filaments,stigma,and pistil exfoliation layer gradually increased after pollination.Subcellular localization experiments showed that BoFAZ1 was located in the cell membrane.A myristoylation site was found at the N-terminus of BoFAZ1.Removal of this site resulted in protein dislocation in the cytoplasm,cell membrane and nucleus.Finally,a yeast two-hybrid test indicated that BoH3.2(histone H3.2),a protein involved in abscission zone development,interacted with BoFAZ1.This interaction was verified by a GST pull-down assay.In summary,our data indicated that BoFAZ1 was involved in the formation of the pistil abscission zone in B.oleracea.
基金supported by grants from the National Natural Science Foundation of China(31200502,31401854)the Natural Science Foundation of Jiangsu Province(Grant No.BK20140760,BK20150552)
文摘A MADS-box gene,CiMADS9,was cloned from the male flowers of Carya illinoinensis by rapid amplification of cDNA ends. The gene was 1 077 bp with a 768 bp open reading frame encoding 255 amino acids. Multiple sequence comparisons revealed that Ci MADS9 is a typical MIKC-type MADS-box gene with a MADS-box domain and a K semi-conserved region. Phylogenetic analysis indicated that CiMADS9 belongs to the AGL15 group of the MADS-box gene family. Quantitative reverse transcription polymerase chain reaction analysis indicated that the expression levels in reproductive organs(i.e.,flowers and young fruits) were considerably higher than in vegetative tissues(i.e.,leaves and branches). The highest expression levels were observed in male flowers. An overexpression vector for CiMADS9 was constructed and the gene was inserted into the Arabidopsis thaliana genome. CiMADS9 expression was confirmed in all transgenic lines. Compared with wild-type plants,transgenic A. thaliana plants overexpressing CiMADS9 exhibited delayed flowering and an increased number of leaves.