【目的】建立一种快速、灵敏、特异的检测甘蔗条点病毒(sugarcane striate virus,SStrV)的SYBR Green I荧光定量PCR方法。【方法】从SStrV基因组保守区域序列设计特异性扩增引物,构建含有SStrV基因组序列的重组质粒pMD19-T-SStrV-qN作...【目的】建立一种快速、灵敏、特异的检测甘蔗条点病毒(sugarcane striate virus,SStrV)的SYBR Green I荧光定量PCR方法。【方法】从SStrV基因组保守区域序列设计特异性扩增引物,构建含有SStrV基因组序列的重组质粒pMD19-T-SStrV-qN作为阳性质粒标准品,以其为模板建立SStrV荧光定量PCR检测方法,并对该方法的灵敏性、特异性、稳定性进行了测试,随后用该方法对甘蔗不同组织部位中SStrV载量进行检测。【结果】将含有SStrV基因组序列的重组质粒按10倍比稀释成标准品,将其作为模板进行荧光定量PCR,获得标准曲线y=-3.337 x+38.197,相关系数r2=0.999,说明Cq值与标准品浓度拷贝数的对数呈线性关系;建立的荧光定量PCR最低可以检测到13拷贝重组质粒/μL,是普通PCR灵敏度的100倍。该方法能特异的检测SStrV,特异性高,组内和组间的变异系数在0.13%-0.94%之间,表明该方法重复性良好。SStrV的载量在甘蔗的不同组织部位中差异较大,+4叶中SStrV的载量最高,与其他组织部位达到了显著差异。【结论】建立了能灵敏特异检测SStrV的SYBR Green I荧光定量PCR方法,明确了+4叶是甘蔗中SStrV检测的最佳采样部位。展开更多
AP2/ERF转录因子在调控植物的生长发育、抵抗生物和非生物胁迫方面发挥重要作用。克隆ShERF3,并分析其功能,为甘蔗抗逆遗传改良提供基因资源。基于转录组数据从‘新台糖22号’甘蔗中克隆ShERF3,并通过Real time PCR、生物信息学分析、...AP2/ERF转录因子在调控植物的生长发育、抵抗生物和非生物胁迫方面发挥重要作用。克隆ShERF3,并分析其功能,为甘蔗抗逆遗传改良提供基因资源。基于转录组数据从‘新台糖22号’甘蔗中克隆ShERF3,并通过Real time PCR、生物信息学分析、水稻原生质体亚细胞定位等技术对ShERF3编码的蛋白特性、亚细胞定位及基因表达模式进行分析。结果显示,克隆获得1 142 bp ShERF3的cDNA序列,包含1个1 053 bp的完整开放阅读框,编码350个氨基酸;ShERF3蛋白含有一个AP2结构域,属于AP2/ERF家族ERF亚家族成员,与高粱和柳枝稷ERF3、谷子和哈氏黍ERF118蛋白同源性较高;ShERF3蛋白是一种不稳定的疏水性蛋白,亚细胞定位结果显示其定位于细胞核;ShERF3主要在甘蔗成熟茎节中表达,在叶片和根系中相对表达较低;在PEG处理下,ShERF3表达先下调后上调,在NaCl处理下,随着胁迫时间延长ShERF3表达量降低。甘蔗ShERF3积极响应干旱和盐逆境胁迫,可能在甘蔗茎秆发育、干旱以及盐胁迫应答方面发挥关键作用。展开更多
Soybean yield has traditionally been increased through high planting density,but investigating plant height and petiole traits to select for compact architecture,lodging resistance,and high yield varieties is an under...Soybean yield has traditionally been increased through high planting density,but investigating plant height and petiole traits to select for compact architecture,lodging resistance,and high yield varieties is an underexplored option for further improving yield.We compared the relationships between yield-related traits,lodging resistance,and petioleassociated phenotypes in the short petiole germplasm M657 with three control accessions during 2017–2018 in four locations in the Huang–Huai region,China.The results showed that M657 exhibited stable and high tolerance to high planting density and resistance to lodging,especially at the highest density(8×105 plants ha–1).The regression analysis indicated that a shorter petiole length was significantly associated with increased lodging resistance.The yield analysis showed that M657 achieved higher yields under higher densities,especially in the northern part of the Huang–Huai region.Among the varieties,there were markedly different responses to intra-and inter-row spacing designs with respect to both lodging and yield that were related to location and density.Lodging was positively correlated with planting density,plant height,petiole length,and number of effective branches,but negatively correlated with stem diameter,seed number per plant,and seed weight per plant.The yield of soybean was increased by appropriately increasing the planting density on the basis of the current soybean varieties in the Huang–Huai region.This study provides a valuable new germplasm resource for the introgression of compact architecture traits that are amenable to providing a high yield in high density planting systems,and it establishes a high-yield model of soybean in the Huang–Huai region.展开更多
Phytophthora root and stem rot of soybean caused by Phytophthora sojae(P.sojae)is a devastating disease that affects soybean[Glycine max(L.)Merr.]all over the world.S-phase kinase-associated protein 1(SKP1)proteins ar...Phytophthora root and stem rot of soybean caused by Phytophthora sojae(P.sojae)is a devastating disease that affects soybean[Glycine max(L.)Merr.]all over the world.S-phase kinase-associated protein 1(SKP1)proteins are key members of the SKP1/Cullin/F-box protein(SCF)ubiquitin ligase complex and play diverse roles in plant biology.However,the role of SKP1 in soybean against the phytopathogenic oomycete P.sojae remains unclear.In this study,a novel member of the soybean SKP1 gene family,GmSKP1 which was significantly induced by P.sojae,was reported.The expression of GmSKP1 was simultaneously induced by methyl jasmonate(MeJA),salicylic acid(SA)and ethylene(ET),which might suggest an important role for GmSKP1 of plant in responses to hormone treatments.Functional analysis using GmSKP1 overexpression lines showed that GmSKP1 enhanced resistance to P.sojae in transgenic soybean plants.Further analyses showed that GmSKP1 interacted with a homeodomain-leucine zipper protein transcription factor(GmHDL56)and a WRKY transcription factor(GmWRKY31),which could positively regulate responses to P.sojae in soybean.Importantly,several pathogenesis-related(PR)genes were constitutively activated,including GmPR1a,GmPR2,GmPR3,GmPR4,GmPR5a and GmPR10,in GmSKP1-OE soybean plants.Taken together,these results suggested that GmSKP1 enhanced resistance to P.sojae in soybean,possibly by activating the defense-related PR genes.展开更多
文摘【目的】建立一种快速、灵敏、特异的检测甘蔗条点病毒(sugarcane striate virus,SStrV)的SYBR Green I荧光定量PCR方法。【方法】从SStrV基因组保守区域序列设计特异性扩增引物,构建含有SStrV基因组序列的重组质粒pMD19-T-SStrV-qN作为阳性质粒标准品,以其为模板建立SStrV荧光定量PCR检测方法,并对该方法的灵敏性、特异性、稳定性进行了测试,随后用该方法对甘蔗不同组织部位中SStrV载量进行检测。【结果】将含有SStrV基因组序列的重组质粒按10倍比稀释成标准品,将其作为模板进行荧光定量PCR,获得标准曲线y=-3.337 x+38.197,相关系数r2=0.999,说明Cq值与标准品浓度拷贝数的对数呈线性关系;建立的荧光定量PCR最低可以检测到13拷贝重组质粒/μL,是普通PCR灵敏度的100倍。该方法能特异的检测SStrV,特异性高,组内和组间的变异系数在0.13%-0.94%之间,表明该方法重复性良好。SStrV的载量在甘蔗的不同组织部位中差异较大,+4叶中SStrV的载量最高,与其他组织部位达到了显著差异。【结论】建立了能灵敏特异检测SStrV的SYBR Green I荧光定量PCR方法,明确了+4叶是甘蔗中SStrV检测的最佳采样部位。
文摘AP2/ERF转录因子在调控植物的生长发育、抵抗生物和非生物胁迫方面发挥重要作用。克隆ShERF3,并分析其功能,为甘蔗抗逆遗传改良提供基因资源。基于转录组数据从‘新台糖22号’甘蔗中克隆ShERF3,并通过Real time PCR、生物信息学分析、水稻原生质体亚细胞定位等技术对ShERF3编码的蛋白特性、亚细胞定位及基因表达模式进行分析。结果显示,克隆获得1 142 bp ShERF3的cDNA序列,包含1个1 053 bp的完整开放阅读框,编码350个氨基酸;ShERF3蛋白含有一个AP2结构域,属于AP2/ERF家族ERF亚家族成员,与高粱和柳枝稷ERF3、谷子和哈氏黍ERF118蛋白同源性较高;ShERF3蛋白是一种不稳定的疏水性蛋白,亚细胞定位结果显示其定位于细胞核;ShERF3主要在甘蔗成熟茎节中表达,在叶片和根系中相对表达较低;在PEG处理下,ShERF3表达先下调后上调,在NaCl处理下,随着胁迫时间延长ShERF3表达量降低。甘蔗ShERF3积极响应干旱和盐逆境胁迫,可能在甘蔗茎秆发育、干旱以及盐胁迫应答方面发挥关键作用。
基金funded by the National Natural Science Foundation of China (31271753)the Central Publicinterest Scientific Institution Basal Research Fund, China (S2021ZD02)the Agricultural Science and Technology Innovation Program (ASTIP) of the Chinese Academy of Agricultural Sciences (CAAS-ZDRW202003-1)。
文摘Soybean yield has traditionally been increased through high planting density,but investigating plant height and petiole traits to select for compact architecture,lodging resistance,and high yield varieties is an underexplored option for further improving yield.We compared the relationships between yield-related traits,lodging resistance,and petioleassociated phenotypes in the short petiole germplasm M657 with three control accessions during 2017–2018 in four locations in the Huang–Huai region,China.The results showed that M657 exhibited stable and high tolerance to high planting density and resistance to lodging,especially at the highest density(8×105 plants ha–1).The regression analysis indicated that a shorter petiole length was significantly associated with increased lodging resistance.The yield analysis showed that M657 achieved higher yields under higher densities,especially in the northern part of the Huang–Huai region.Among the varieties,there were markedly different responses to intra-and inter-row spacing designs with respect to both lodging and yield that were related to location and density.Lodging was positively correlated with planting density,plant height,petiole length,and number of effective branches,but negatively correlated with stem diameter,seed number per plant,and seed weight per plant.The yield of soybean was increased by appropriately increasing the planting density on the basis of the current soybean varieties in the Huang–Huai region.This study provides a valuable new germplasm resource for the introgression of compact architecture traits that are amenable to providing a high yield in high density planting systems,and it establishes a high-yield model of soybean in the Huang–Huai region.
基金Supported by the NSFC Projects(31971972)the Natural Science Foundation of Heilongjiang Province(ZD2019C001)the Outstanding Talents and Innovative Team of Agricultural Scientific Research。
文摘Phytophthora root and stem rot of soybean caused by Phytophthora sojae(P.sojae)is a devastating disease that affects soybean[Glycine max(L.)Merr.]all over the world.S-phase kinase-associated protein 1(SKP1)proteins are key members of the SKP1/Cullin/F-box protein(SCF)ubiquitin ligase complex and play diverse roles in plant biology.However,the role of SKP1 in soybean against the phytopathogenic oomycete P.sojae remains unclear.In this study,a novel member of the soybean SKP1 gene family,GmSKP1 which was significantly induced by P.sojae,was reported.The expression of GmSKP1 was simultaneously induced by methyl jasmonate(MeJA),salicylic acid(SA)and ethylene(ET),which might suggest an important role for GmSKP1 of plant in responses to hormone treatments.Functional analysis using GmSKP1 overexpression lines showed that GmSKP1 enhanced resistance to P.sojae in transgenic soybean plants.Further analyses showed that GmSKP1 interacted with a homeodomain-leucine zipper protein transcription factor(GmHDL56)and a WRKY transcription factor(GmWRKY31),which could positively regulate responses to P.sojae in soybean.Importantly,several pathogenesis-related(PR)genes were constitutively activated,including GmPR1a,GmPR2,GmPR3,GmPR4,GmPR5a and GmPR10,in GmSKP1-OE soybean plants.Taken together,these results suggested that GmSKP1 enhanced resistance to P.sojae in soybean,possibly by activating the defense-related PR genes.
