Quantitative real-time PCR(qPCR)is an effective and widely used method to analyze expression patterns of target genes.Selection of stable reference genes is a prerequisite for accurate normalization of target gene exp...Quantitative real-time PCR(qPCR)is an effective and widely used method to analyze expression patterns of target genes.Selection of stable reference genes is a prerequisite for accurate normalization of target gene expression by qRT-PCR.In Iris germanica L.,no studies have yet been published regarding the evaluation of potential reference genes.In this study,nine candidate reference genes were assessed at different flower developmental stages and in different tissues by four different algorithms(GeNorm,NormFinder,BestKeeper,and RefFinder).The results revealed that ACT11(Actin 11)and EF1α(Elongation factor 1 alpha)were the most stable reference genes in different tissues,whereas TUA(Tubulin alpha)and UBC9(Ubiquitin-protein ligase 9)were the most stable ones in different flower developmental stages.UBC9 and ACT11 were the most stable reference genes in all of the tested samples,while the SAMDC(S-Adenosylmethionine decarboxylase)showed the least stability.Finally,to validate the suitability of the selected reference genes,the relative expression level of IgTPS(beta-caryophyllene synthase)was assessed and highlighted the importance of suitable reference gene selection.This work constitutes the first systematic evaluation of potential reference genes in I.germanica and provides guidelines for future research on gene function and molecular mechanisms on I.germanica and related species.展开更多
Alzheimer＇s disease（AD） is associated with oxidative stress, and ultimately results in cognitive deficit. Despite existing literature on the pathophysiology of AD, there is currently no cure for AD. The present stu...Alzheimer＇s disease（AD） is associated with oxidative stress, and ultimately results in cognitive deficit. Despite existing literature on the pathophysiology of AD, there is currently no cure for AD. The present study investigated the effects of kaempferol（Kmp） isolated from the extract of Mespilus germanica L.（medlar） leaves on cognitive impairment, hippocampal antioxidants, apoptosis, lipid peroxidation and neuro-inflammation markers in ovariectomized（OVX） rat models of sporadic AD. Kaempferol, as the main flavonoid of medlar extract has been previously known for anti-oxidative, anti-inflammatory and anti-neurotoxic effects. Thirty-two female Wistar rats were ovariectomized, and randomly divided into four groups： sham, OVX ＋ saline, OVX ＋ streptozotocin（STZ） ＋ saline, OVX ＋ STZ ＋ Kmp. Animals received intracerebroventricular injection of STZ（3 mg/kg, twice with one day interval） to establish models of sporadic AD. Intraperitoneal injection of Kmp（10 mg/kg） for 21 days was performed in the OVX ＋ STZ ＋ Kmp group. Spatial learning and memory of rats were evaluated using a Morris water maze. Finally, brain homogenates were used for biochemical analysis by enzyme-linked immunosorbent assay. The results showed a significant improvement in spatial learning and memory as evidenced by shortened escape latency and searching distance in Morris water maze in the OVX ＋ STZ ＋ Kmp group compared with the OVX ＋ STZ group. Kmp also exhibited significant elevations in brain levels of antioxidant enzymes of superoxide dismutase and glutathione, while reduction in tumor necrosis factor-α and malondialdehyde. Our results demonstrate that Kmp is capable of alleviating STZ-induced memory impairment in OVX rats, probably by elevating endogenous hippocampal antioxidants of superoxide dismutase and glutathione, and reducing neuroinflammation. This study suggests that Kmp may be a potential neuroprotective agent against cognitive deficit in AD.展开更多
以白花、短梦、法国回声、金娃娃、金皇后、紫花共6个德国鸢尾(Iris germanica L.)品种的花器官为外植体,进行不定芽的诱导和快速繁殖技术研究。结果表明:外植体采用0.1%HgCl2消毒10min效果最好,污染率为26.67%,死亡率为6.70%;不同外植...以白花、短梦、法国回声、金娃娃、金皇后、紫花共6个德国鸢尾(Iris germanica L.)品种的花器官为外植体,进行不定芽的诱导和快速繁殖技术研究。结果表明:外植体采用0.1%HgCl2消毒10min效果最好,污染率为26.67%,死亡率为6.70%;不同外植体部位愈伤组织诱导与分化能力不同,花茎的不定芽诱导率最高,达59.00%;花茎外植体诱导愈伤组织和分化成苗的能力与花苞大小及植物生长调节剂等因素有关,适宜的花苞长度为4～8cm,最适培养基为MS+6-BA1.0mg·L-1+IBA0.5mg·L-1;将不定芽丛切割至4～6株的株丛转接到MS+6-BA2.0mg·L-1+IBA0.5mg·L-1继代培养基中增殖较好,增殖系数达5.5,玻璃化程度较低,为16.4%;最佳生根培养基为1/2MS+NAA0.5mg·L-1,生根率达92.67%;移栽基质采用珍珠岩最好,成活率达88.20%。展开更多
基金The work was supported by the National Natural Science Foundation of China(31801901)the Natural Science Foundation of Jiangsu(BK20180314)+1 种基金the Foundation of Key Laboratory of Landscaping(KF201901),Ministry of Agriculture and Rural Affairs,Chinathe Jiangsu Key Laboratory for the Research and Utilization of Plant Resources(JSPKLB201814).
文摘Quantitative real-time PCR(qPCR)is an effective and widely used method to analyze expression patterns of target genes.Selection of stable reference genes is a prerequisite for accurate normalization of target gene expression by qRT-PCR.In Iris germanica L.,no studies have yet been published regarding the evaluation of potential reference genes.In this study,nine candidate reference genes were assessed at different flower developmental stages and in different tissues by four different algorithms(GeNorm,NormFinder,BestKeeper,and RefFinder).The results revealed that ACT11(Actin 11)and EF1α(Elongation factor 1 alpha)were the most stable reference genes in different tissues,whereas TUA(Tubulin alpha)and UBC9(Ubiquitin-protein ligase 9)were the most stable ones in different flower developmental stages.UBC9 and ACT11 were the most stable reference genes in all of the tested samples,while the SAMDC(S-Adenosylmethionine decarboxylase)showed the least stability.Finally,to validate the suitability of the selected reference genes,the relative expression level of IgTPS(beta-caryophyllene synthase)was assessed and highlighted the importance of suitable reference gene selection.This work constitutes the first systematic evaluation of potential reference genes in I.germanica and provides guidelines for future research on gene function and molecular mechanisms on I.germanica and related species.
基金supported by a grant from Research and Technology Chancellor of Guilan University of Medical Sciences,Iran(No.IR.GUMS.REC.1936.51)
文摘Alzheimer＇s disease（AD） is associated with oxidative stress, and ultimately results in cognitive deficit. Despite existing literature on the pathophysiology of AD, there is currently no cure for AD. The present study investigated the effects of kaempferol（Kmp） isolated from the extract of Mespilus germanica L.（medlar） leaves on cognitive impairment, hippocampal antioxidants, apoptosis, lipid peroxidation and neuro-inflammation markers in ovariectomized（OVX） rat models of sporadic AD. Kaempferol, as the main flavonoid of medlar extract has been previously known for anti-oxidative, anti-inflammatory and anti-neurotoxic effects. Thirty-two female Wistar rats were ovariectomized, and randomly divided into four groups： sham, OVX ＋ saline, OVX ＋ streptozotocin（STZ） ＋ saline, OVX ＋ STZ ＋ Kmp. Animals received intracerebroventricular injection of STZ（3 mg/kg, twice with one day interval） to establish models of sporadic AD. Intraperitoneal injection of Kmp（10 mg/kg） for 21 days was performed in the OVX ＋ STZ ＋ Kmp group. Spatial learning and memory of rats were evaluated using a Morris water maze. Finally, brain homogenates were used for biochemical analysis by enzyme-linked immunosorbent assay. The results showed a significant improvement in spatial learning and memory as evidenced by shortened escape latency and searching distance in Morris water maze in the OVX ＋ STZ ＋ Kmp group compared with the OVX ＋ STZ group. Kmp also exhibited significant elevations in brain levels of antioxidant enzymes of superoxide dismutase and glutathione, while reduction in tumor necrosis factor-α and malondialdehyde. Our results demonstrate that Kmp is capable of alleviating STZ-induced memory impairment in OVX rats, probably by elevating endogenous hippocampal antioxidants of superoxide dismutase and glutathione, and reducing neuroinflammation. This study suggests that Kmp may be a potential neuroprotective agent against cognitive deficit in AD.
文摘以白花、短梦、法国回声、金娃娃、金皇后、紫花共6个德国鸢尾(Iris germanica L.)品种的花器官为外植体,进行不定芽的诱导和快速繁殖技术研究。结果表明:外植体采用0.1%HgCl2消毒10min效果最好,污染率为26.67%,死亡率为6.70%;不同外植体部位愈伤组织诱导与分化能力不同,花茎的不定芽诱导率最高,达59.00%;花茎外植体诱导愈伤组织和分化成苗的能力与花苞大小及植物生长调节剂等因素有关,适宜的花苞长度为4～8cm,最适培养基为MS+6-BA1.0mg·L-1+IBA0.5mg·L-1;将不定芽丛切割至4～6株的株丛转接到MS+6-BA2.0mg·L-1+IBA0.5mg·L-1继代培养基中增殖较好,增殖系数达5.5,玻璃化程度较低,为16.4%;最佳生根培养基为1/2MS+NAA0.5mg·L-1,生根率达92.67%;移栽基质采用珍珠岩最好,成活率达88.20%。
