Lily is a well-known ornamental plant with a diversity of fragrant types.Basic information on lily floral scent compounds has been obtained for only a few accessions,and little is known about Lilium aroma types,the te...Lily is a well-known ornamental plant with a diversity of fragrant types.Basic information on lily floral scent compounds has been obtained for only a few accessions,and little is known about Lilium aroma types,the terpene synthase genes that may play roles in the production of key volatiles,or the range of monoterpenes that these genes produce.In this study,41 cultivars were analyzed for volatile emissions,and a total of 46 individual volatile compounds were identified,16 for the first time in lilies.Lily accessions were classified into six groups according to the composition of major scent components:faint-scented,cool,fruity,musky,fruity-honey,and lily.Monoterpenes were one of the main groups of volatiles identified,and attention was focused on terpene synthase(TPS)genes,which encode enzymes that catalyze the last steps in monoterpene synthesis.Thirty-two candidate monoterpene synthase cDNAs were obtained from 66 lily cultivars,and 64 SNPs were identified.Two InDels were also shown to result from variable splicing,and sequence analysis suggested that different transcripts arose from the same gene.All identified nucleotide substitution sites were highly correlated with the amounts of myrcene emitted,and InDel site 230 was highly correlated with the emission of all major monoterpenoid components,especially(E)-β-ocimene.Heterologous expression of five cDNAs cloned from faint-scented and strong-scented lilies showed that their corresponding enzymes could convert geranyl diphosphate to(E)-β-ocimene,α-pinene,and limonene.The findings from this study provide a major resource for the assessment of lily scent volatiles and will be helpful in breeding of improved volatile components.展开更多
目的探讨RNA干扰半乳糖凝集素-3(galectin-3,Gal-3)表达对胰腺癌细胞增殖和凋亡的影响及其机制.方法将培养的Panc-1细胞随机分为对照组(未处理)、NC组(转染control-si RNA)和Gal-3干扰组(转染Gal-3-si RNA),以小干扰RNA(Small interferi...目的探讨RNA干扰半乳糖凝集素-3(galectin-3,Gal-3)表达对胰腺癌细胞增殖和凋亡的影响及其机制.方法将培养的Panc-1细胞随机分为对照组(未处理)、NC组(转染control-si RNA)和Gal-3干扰组(转染Gal-3-si RNA),以小干扰RNA(Small interfering RNA,si RNA)技术干扰Panc-1细胞中Gal-3表达后,RT-PCR和Western blot检测干扰效果,CCK-8法检测细胞增殖,流式细胞仪检测细胞凋亡,Western blot检测细胞中ki67、细胞周期蛋白D1(Cyclin D1)、活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved caspase-3)抗体和β-连环蛋白(β-catenin)蛋白表达.结果与对照组相比,NC组中Gal-3 m RNA(0.99±0.08vs 1.01±0.06)和蛋白(0.36±0.03 vs 0.34±0.05)的表达差异不显著(P>0.05),而Gal-3干扰组中Gal-3m RNA(0.38±0.02 vs 1.01±0.06)和蛋白(0.10±0.01vs 0.34±0.05)的表达均显著降低(P<0.05);Gal-3干扰组细胞增殖能力减弱(24 h:0.55±0.03 vs 0.71±0.05;48 h:0.76±0.05 vs 0.97±0.06;72 h:1.08±0.06 vs1.32±0.09),G0/G1期细胞百分比升高(79.48±1.32 vs71.52±1.15),S期(14.26±1.08 vs 18.24±1.06)和G2/M期(6.21±0.78 vs 10.19±1.52)降低,凋亡能力增强(13.26±2.28 vs 5.82±0.35),与对照组相比,差异均有统计学意义(P<0.05);与对照组相比,Gal-3干扰组中ki67(0.24±0.02 vs 0.96±0.07)、CyclinD 1(0.26±0.03vs 0.88±0.09)和β-catenin(0.42±0.05 vs 0.87±0.05)蛋白的表达水平均明显降低,而Cleaved Caspase-3(0.70±0.06 vs 0.32±0.03)蛋白表达水平明显升高,差异有统计学意义(P<0.05);NC组与对照组间各指标差异均无统计学意义(P>0.05).结论 RNA干扰Panc-1细胞中Gal-3表达,可抑制细胞增殖并促进细胞凋亡,其机制可能与抑制Wnt/β-catenin信号通路有关.展开更多
Joubert syndrome is characterized by unique malformation of the cerebellar vermis.More than thirty Joubert syndrome genes have been identified,including ARL13 B.However,its role in cerebellar development remains unexp...Joubert syndrome is characterized by unique malformation of the cerebellar vermis.More than thirty Joubert syndrome genes have been identified,including ARL13 B.However,its role in cerebellar development remains unexplored.We found that knockdown or knockout of arl13b impaired balance and locomotion in zebrafish larvae.Granule cells were selectively reduced in the corpus cerebelli,a structure homologous to the mammalian vermis.Purkinje cell progenitors were also selectively disturbed dorsomedially.The expression of atoh1 and ptf1,proneural genes of granule and Purkinje cells,respectively,were selectively down-regulated along the dorsal midline of the cerebellum.Moreover,wnt1,which is transiently expressed early in cerebellar development,was selectively reduced.Intriguingly,activating Wnt signaling partially rescued the granule cell defects in arl13b mutants.These findings suggested that Arl13 b is necessary for the early development of cerebellar granule and Purkinje cells.The arl13b-deficient zebrafish can serve as a model organism for studying Joubert syndrome.展开更多
基金sponsored by the Natural Science Foundation of Shanxi Province(201601D011077)Shanxi Key Project in Agriculture(201803D221011-2).
文摘Lily is a well-known ornamental plant with a diversity of fragrant types.Basic information on lily floral scent compounds has been obtained for only a few accessions,and little is known about Lilium aroma types,the terpene synthase genes that may play roles in the production of key volatiles,or the range of monoterpenes that these genes produce.In this study,41 cultivars were analyzed for volatile emissions,and a total of 46 individual volatile compounds were identified,16 for the first time in lilies.Lily accessions were classified into six groups according to the composition of major scent components:faint-scented,cool,fruity,musky,fruity-honey,and lily.Monoterpenes were one of the main groups of volatiles identified,and attention was focused on terpene synthase(TPS)genes,which encode enzymes that catalyze the last steps in monoterpene synthesis.Thirty-two candidate monoterpene synthase cDNAs were obtained from 66 lily cultivars,and 64 SNPs were identified.Two InDels were also shown to result from variable splicing,and sequence analysis suggested that different transcripts arose from the same gene.All identified nucleotide substitution sites were highly correlated with the amounts of myrcene emitted,and InDel site 230 was highly correlated with the emission of all major monoterpenoid components,especially(E)-β-ocimene.Heterologous expression of five cDNAs cloned from faint-scented and strong-scented lilies showed that their corresponding enzymes could convert geranyl diphosphate to(E)-β-ocimene,α-pinene,and limonene.The findings from this study provide a major resource for the assessment of lily scent volatiles and will be helpful in breeding of improved volatile components.
文摘目的探讨RNA干扰半乳糖凝集素-3(galectin-3,Gal-3)表达对胰腺癌细胞增殖和凋亡的影响及其机制.方法将培养的Panc-1细胞随机分为对照组(未处理)、NC组(转染control-si RNA)和Gal-3干扰组(转染Gal-3-si RNA),以小干扰RNA(Small interfering RNA,si RNA)技术干扰Panc-1细胞中Gal-3表达后,RT-PCR和Western blot检测干扰效果,CCK-8法检测细胞增殖,流式细胞仪检测细胞凋亡,Western blot检测细胞中ki67、细胞周期蛋白D1(Cyclin D1)、活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved caspase-3)抗体和β-连环蛋白(β-catenin)蛋白表达.结果与对照组相比,NC组中Gal-3 m RNA(0.99±0.08vs 1.01±0.06)和蛋白(0.36±0.03 vs 0.34±0.05)的表达差异不显著(P>0.05),而Gal-3干扰组中Gal-3m RNA(0.38±0.02 vs 1.01±0.06)和蛋白(0.10±0.01vs 0.34±0.05)的表达均显著降低(P<0.05);Gal-3干扰组细胞增殖能力减弱(24 h:0.55±0.03 vs 0.71±0.05;48 h:0.76±0.05 vs 0.97±0.06;72 h:1.08±0.06 vs1.32±0.09),G0/G1期细胞百分比升高(79.48±1.32 vs71.52±1.15),S期(14.26±1.08 vs 18.24±1.06)和G2/M期(6.21±0.78 vs 10.19±1.52)降低,凋亡能力增强(13.26±2.28 vs 5.82±0.35),与对照组相比,差异均有统计学意义(P<0.05);与对照组相比,Gal-3干扰组中ki67(0.24±0.02 vs 0.96±0.07)、CyclinD 1(0.26±0.03vs 0.88±0.09)和β-catenin(0.42±0.05 vs 0.87±0.05)蛋白的表达水平均明显降低,而Cleaved Caspase-3(0.70±0.06 vs 0.32±0.03)蛋白表达水平明显升高,差异有统计学意义(P<0.05);NC组与对照组间各指标差异均无统计学意义(P>0.05).结论 RNA干扰Panc-1细胞中Gal-3表达,可抑制细胞增殖并促进细胞凋亡,其机制可能与抑制Wnt/β-catenin信号通路有关.
基金supported by grants from the National Natural Science Foundation of China (31171044,81160144, and 81760216)the Young Scientist Project of Jiangxi Province, China (20122BCB23007)。
文摘Joubert syndrome is characterized by unique malformation of the cerebellar vermis.More than thirty Joubert syndrome genes have been identified,including ARL13 B.However,its role in cerebellar development remains unexplored.We found that knockdown or knockout of arl13b impaired balance and locomotion in zebrafish larvae.Granule cells were selectively reduced in the corpus cerebelli,a structure homologous to the mammalian vermis.Purkinje cell progenitors were also selectively disturbed dorsomedially.The expression of atoh1 and ptf1,proneural genes of granule and Purkinje cells,respectively,were selectively down-regulated along the dorsal midline of the cerebellum.Moreover,wnt1,which is transiently expressed early in cerebellar development,was selectively reduced.Intriguingly,activating Wnt signaling partially rescued the granule cell defects in arl13b mutants.These findings suggested that Arl13 b is necessary for the early development of cerebellar granule and Purkinje cells.The arl13b-deficient zebrafish can serve as a model organism for studying Joubert syndrome.