【目的】分析镉胁迫下中辽1号杨的转录水平,筛选差异表达基因,挖掘与镉胁迫相关的功能基因,为深入探索中辽1号杨对镉胁迫响应的分子机制提供理论依据。【方法】以中辽1号杨为试验材料,采用盆栽试验的方法将其扦插于土壤Cd含量为20 mg/kg...【目的】分析镉胁迫下中辽1号杨的转录水平,筛选差异表达基因,挖掘与镉胁迫相关的功能基因,为深入探索中辽1号杨对镉胁迫响应的分子机制提供理论依据。【方法】以中辽1号杨为试验材料,采用盆栽试验的方法将其扦插于土壤Cd含量为20 mg/kg(M20)的花盆中,以不加Cd为对照(CK),80 d后采集不同处理中辽1号杨叶片进行转录组测序,使用DESeq2软件筛选CK与M20处理中辽1号杨的差异表达基因,将得到的差异基因在基因本体数据库(gene ontology,GO)、京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)、蛋白相邻类聚簇数据库(clusters of orthologous groups of proteins,COG)中进行注释,分析差异表达基因在不同数据库中的注释信息。随机挑选6个差异表达基因(超氧化物歧化酶(SOD)、丝氨酸/苏氨酸蛋白激酶(STK)、脱落酸(ABA)、T复合蛋白1(TCP1)、MYB基因(MYB)、丝裂原活蛋白激酶12(MAPK12)),利用Primer 5软件设计特异性引物进行实时荧光定量PCR(RT-qPCR)试验,检测这6个基因表达水平的变化,验证转录组结果的准确性。【结果】在CK和M20处理的中辽1号杨中,共发现3812个差异表达基因,其中上调表达的有2209个,下调表达的有1603个。GO分析发现,差异表达基因注释到3大类49个功能组中,其中与代谢过程有关基因最多。转录组KEGG代谢通路分析发现,差异表达基因主要在信号转导途径、代谢途径和生物合成途径富集。COG分析发现,1455个差异基因被注释到22种分类中,其中一般功能预测基因注释最多,其次是信号转导机制基因。结合各数据库分析结果发现,Cd胁迫下中辽1号杨转录组的注释结果中代谢过程和信号转导过程相关基因较多。镉胁迫下差异表达基因最多的家族是ABC、MYB、WRKY、bHLH和NAC基因家族,挖掘出与镉胁迫相关基因WRKY家族基因47(WRKY47)、硝酸盐转运蛋白基因(NRT)、ABC家族转运蛋白基因2(ABC2)、苹果酸脱氢酶(MDH)、谷胱甘肽S-转移酶基因(GSTs)、NAC家族基因2(NAC2)的表达量显著上调,MYB家族基因44(MYB44)、重金属相关异戊二烯化植物蛋白基因39(HIPP39)的表达量显著下调。RT-qPCR结果显示,随机挑选6个差异表达基因表达量的变化趋势与转录组测序结果一致。【结论】镉胁迫后中辽1号杨转录组的差异表达基因主要富集在代谢过程和信号转导过程,挖掘出与镉胁迫相关基因,分别是WRKY47、NRT、ABC2、MDH、GSTs、NAC2、MYB44、HIPP39,为深入探索杨树对镉胁迫响应的分子机制提供了理论依据。展开更多
It is less known whether and how soil metal lead （Pb） impacts the invasion of exotic plants. A greenhouse experiment was conducted to estimate the effects of lead on the growth and mycorrhizae of an invasive species...It is less known whether and how soil metal lead （Pb） impacts the invasion of exotic plants. A greenhouse experiment was conducted to estimate the effects of lead on the growth and mycorrhizae of an invasive species （Solidago canadensis L.） in a microcosm system. Each microcosm unit was separated into HOST and TEST compartments by a replaceable mesh screen that allowed arbuscular mycorrhizal （AM） fungal hyphae rather than plant roots to grow into the TEST compartments. Three Pb levels （control, 300, and 600 mg/kg soil） were used in this study to simulate ambient soil and two pollution sites where S. canadensis grows. Mycorrhizal inoculum comprised five indigenous arbuscular mycorrhizal fungal species （ Glomus mosseae, Glomus versiform, Glomus diaphanum, Glomus geosporum, and Glomus etunicatum）. The ^15N isotope tracer was used to quantify the mycorrhizally mediated nitrogen acquisition of plants. The results showed that S. canadensis was highly dependent on mycorrhizae. The Pb additions significantly decreased biomass and arbuscular mycorrhizal colonization （root length colonized, RLC%） but did not affect spore numbers, N （including total N and ^15N） and P uptake. The facilitating efficiency of mycorrhizae on nutrient acquisition was promoted by Pb treatments. The Pb was mostly sequestered in belowground of plant （root and rhizome）. The results suggest that the high efficiency of mycorrhizae on nutrient uptake might give S. canadensis a great advantage over native species in Pb polluted soils.展开更多
Horseweed （Conyza canadensis）, an invasive alien weed, is one of the main weeds in orchards in China. Although glyphosate has been used for control of horseweed and many other weeds in orchards for more than 25 year...Horseweed （Conyza canadensis）, an invasive alien weed, is one of the main weeds in orchards in China. Although glyphosate has been used for control of horseweed and many other weeds in orchards for more than 25 years in China, a case of glyphosate-resistant horseweed has not been identified in orchard in China so far despite glyphosate-resistant horseweed cases have been reported in some other countries. Seeds of 25 horseweed populations were collected from different orchards with different glyphosate application history. Potted seedlings with 11-13-leaf growth stage were treated with glyphosate at 0.035, 0.07, 0.14, 0.28, 0.56, 1.12, 2.24, 4.48, and 8.96 kg a.i. ha-1. The dosage dependence response curve of each population was constructed with Log-logistic dose response regression equations. The ED50 value of each population was calculated and compared with the susceptible population from China. Different populations had different relative glyphosate-resistant levels which increased with the number of years of glyphosate application. Two populations with the highest resistance levels, 8.28 and 7.95 times, were found in Ningbo, Zhejiang Province, China, where glyphosate was used for weed control in orchards twice each year for 15 yr. The two resistant populations accumulated approximately two to four times less shikimic acid than the two susceptible populations 48 h after glyphosate application.展开更多
文摘【目的】分析镉胁迫下中辽1号杨的转录水平,筛选差异表达基因,挖掘与镉胁迫相关的功能基因,为深入探索中辽1号杨对镉胁迫响应的分子机制提供理论依据。【方法】以中辽1号杨为试验材料,采用盆栽试验的方法将其扦插于土壤Cd含量为20 mg/kg(M20)的花盆中,以不加Cd为对照(CK),80 d后采集不同处理中辽1号杨叶片进行转录组测序,使用DESeq2软件筛选CK与M20处理中辽1号杨的差异表达基因,将得到的差异基因在基因本体数据库(gene ontology,GO)、京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)、蛋白相邻类聚簇数据库(clusters of orthologous groups of proteins,COG)中进行注释,分析差异表达基因在不同数据库中的注释信息。随机挑选6个差异表达基因(超氧化物歧化酶(SOD)、丝氨酸/苏氨酸蛋白激酶(STK)、脱落酸(ABA)、T复合蛋白1(TCP1)、MYB基因(MYB)、丝裂原活蛋白激酶12(MAPK12)),利用Primer 5软件设计特异性引物进行实时荧光定量PCR(RT-qPCR)试验,检测这6个基因表达水平的变化,验证转录组结果的准确性。【结果】在CK和M20处理的中辽1号杨中,共发现3812个差异表达基因,其中上调表达的有2209个,下调表达的有1603个。GO分析发现,差异表达基因注释到3大类49个功能组中,其中与代谢过程有关基因最多。转录组KEGG代谢通路分析发现,差异表达基因主要在信号转导途径、代谢途径和生物合成途径富集。COG分析发现,1455个差异基因被注释到22种分类中,其中一般功能预测基因注释最多,其次是信号转导机制基因。结合各数据库分析结果发现,Cd胁迫下中辽1号杨转录组的注释结果中代谢过程和信号转导过程相关基因较多。镉胁迫下差异表达基因最多的家族是ABC、MYB、WRKY、bHLH和NAC基因家族,挖掘出与镉胁迫相关基因WRKY家族基因47(WRKY47)、硝酸盐转运蛋白基因(NRT)、ABC家族转运蛋白基因2(ABC2)、苹果酸脱氢酶(MDH)、谷胱甘肽S-转移酶基因(GSTs)、NAC家族基因2(NAC2)的表达量显著上调,MYB家族基因44(MYB44)、重金属相关异戊二烯化植物蛋白基因39(HIPP39)的表达量显著下调。RT-qPCR结果显示,随机挑选6个差异表达基因表达量的变化趋势与转录组测序结果一致。【结论】镉胁迫后中辽1号杨转录组的差异表达基因主要富集在代谢过程和信号转导过程,挖掘出与镉胁迫相关基因,分别是WRKY47、NRT、ABC2、MDH、GSTs、NAC2、MYB44、HIPP39,为深入探索杨树对镉胁迫响应的分子机制提供了理论依据。
基金This work was supported by the National Basic Research Program(973)of China(No.2006CB 100206)the Zhejiang Provincial Natural Science Foundation of China(No.R505024,Y307418).
文摘It is less known whether and how soil metal lead （Pb） impacts the invasion of exotic plants. A greenhouse experiment was conducted to estimate the effects of lead on the growth and mycorrhizae of an invasive species （Solidago canadensis L.） in a microcosm system. Each microcosm unit was separated into HOST and TEST compartments by a replaceable mesh screen that allowed arbuscular mycorrhizal （AM） fungal hyphae rather than plant roots to grow into the TEST compartments. Three Pb levels （control, 300, and 600 mg/kg soil） were used in this study to simulate ambient soil and two pollution sites where S. canadensis grows. Mycorrhizal inoculum comprised five indigenous arbuscular mycorrhizal fungal species （ Glomus mosseae, Glomus versiform, Glomus diaphanum, Glomus geosporum, and Glomus etunicatum）. The ^15N isotope tracer was used to quantify the mycorrhizally mediated nitrogen acquisition of plants. The results showed that S. canadensis was highly dependent on mycorrhizae. The Pb additions significantly decreased biomass and arbuscular mycorrhizal colonization （root length colonized, RLC%） but did not affect spore numbers, N （including total N and ^15N） and P uptake. The facilitating efficiency of mycorrhizae on nutrient acquisition was promoted by Pb treatments. The Pb was mostly sequestered in belowground of plant （root and rhizome）. The results suggest that the high efficiency of mycorrhizae on nutrient uptake might give S. canadensis a great advantage over native species in Pb polluted soils.
基金supported by the National Basic Research and Development Program of China (2007CB109202)the National Natural Science Foundation of China (30400059)the National Specific Program on Environmental Protection for Public-welfare Industry of China (200709017)
文摘Horseweed （Conyza canadensis）, an invasive alien weed, is one of the main weeds in orchards in China. Although glyphosate has been used for control of horseweed and many other weeds in orchards for more than 25 years in China, a case of glyphosate-resistant horseweed has not been identified in orchard in China so far despite glyphosate-resistant horseweed cases have been reported in some other countries. Seeds of 25 horseweed populations were collected from different orchards with different glyphosate application history. Potted seedlings with 11-13-leaf growth stage were treated with glyphosate at 0.035, 0.07, 0.14, 0.28, 0.56, 1.12, 2.24, 4.48, and 8.96 kg a.i. ha-1. The dosage dependence response curve of each population was constructed with Log-logistic dose response regression equations. The ED50 value of each population was calculated and compared with the susceptible population from China. Different populations had different relative glyphosate-resistant levels which increased with the number of years of glyphosate application. Two populations with the highest resistance levels, 8.28 and 7.95 times, were found in Ningbo, Zhejiang Province, China, where glyphosate was used for weed control in orchards twice each year for 15 yr. The two resistant populations accumulated approximately two to four times less shikimic acid than the two susceptible populations 48 h after glyphosate application.
