通过逆转录聚合酶链式反应（RT-PCR）结合c DNA末端快速扩增（RACE）技术,从杉木Cunninghamia lanceolata中克隆到2个全长为3 235 bp和3 876 bp的纤维素合成酶基因c DNA序列,被分别命名为Cl Ces A1和Cl Ces A2,Gen Bank登录号分别为JQ84...通过逆转录聚合酶链式反应（RT-PCR）结合c DNA末端快速扩增（RACE）技术,从杉木Cunninghamia lanceolata中克隆到2个全长为3 235 bp和3 876 bp的纤维素合成酶基因c DNA序列,被分别命名为Cl Ces A1和Cl Ces A2,Gen Bank登录号分别为JQ844574和JQ844575。相应编码蛋白分别包含992和1 092个氨基酸残基,推测分子量为111 845.3 D和123 105.7 D,等电点为6.04和6.65。氨基酸序列分析发现,它们具有植物纤维素合成酶基因相应的结构特征——锌指结构,2个易变区CSRI和CSRII,2个保守区CRI和CRII,纤维素合成酶底物结合结构域＂D,D,D,QVLRW＂,以及8个跨膜区。荧光定量PCR分析表明：Cl Ces A1和Cl Ces A2基因在茎中表达丰度均为最高,且成熟木质部中表达量均高于皮层中的相应数值;2个基因在根和针叶中的表达量相对较低。以具特殊材质性状的矮生杉木和正常杉木木质部为材料的进一步表达分析发现：Cl Ces A1和Cl Ces A2在正常杉木木质部中的表达量大约是矮生杉木表达量的2~12倍。2个杉木Ces A基因可能在杉木木材形成过程中具有重要作用。展开更多
克隆得到了一个白桦纤维素合成酶基因(CESA7)Gen Ban K登录号(EU591531)启动子序列,通过序列分析发现该启动子含有多个不同功能的顺式作用元件,包括光响应元件、激素响应元件、叶片形态发育元件等,推测该启动子在白桦生长发育过程中具...克隆得到了一个白桦纤维素合成酶基因(CESA7)Gen Ban K登录号(EU591531)启动子序列,通过序列分析发现该启动子含有多个不同功能的顺式作用元件,包括光响应元件、激素响应元件、叶片形态发育元件等,推测该启动子在白桦生长发育过程中具有关键作用。将Bp CESA7启动子克隆至带有GUS报告基因的植物表达载体,命名为pro Bp CESA7-121-GUS,并利用农杆菌介导方法侵染白桦和拟南芥,然后通过GUS组织化学染色观察Bp CESA7基因启动子的组织表达特性。结果在白桦的根、茎、叶和拟南芥的根,叶,萼片、雌蕊中检测到了GUS活性,说明Bp CESA7基因启动子具有启动子活性,并且在白桦的根和叶中染色最深,表明Bp CESA7基因在白桦根和叶中表达量较高,并且其存在组织表达特异性。展开更多
Arabidopsis (Arabidopsis thaliana) WRKY33 is a key transcription factor in pathogen-induced defense signaling, but its function in abiotic stresses remains largely unclear. In this study, we report on the use of a rev...Arabidopsis (Arabidopsis thaliana) WRKY33 is a key transcription factor in pathogen-induced defense signaling, but its function in abiotic stresses remains largely unclear. In this study, we report on the use of a reverse-genetic approach, as well as a yeast (Saccharomyces cerevisiae) expression system, to determine the role of WRKY33 in drought. A T-DNA insertion deletion mutant of WRKY33 is more sensitive to dehydration. Through genome-wide screening the target genes of WRKY33 in yeast, we identified 23 candidate genes including a drought tolerance gene CesA8. Further results revealed that WRKY33 repressed CesA8 expression through binding to the W-box elements of CesA8 distal promoter region and probably interacting with the transcriptional activator of CesA8, MYB46. These findings revealed the primary molecular mechanism underlying the function of WRKY33 in response to展开更多
Cotton is widely distributed worldwide,and improving the quality of its fiber is one of the most important tasks in cotton breeding.Cotton fibers are primarily composed of cellulose,which is synthesized by CesA comple...Cotton is widely distributed worldwide,and improving the quality of its fiber is one of the most important tasks in cotton breeding.Cotton fibers are primarily composed of cellulose,which is synthesized by CesA complexes(CSCs).However,the functions of CesA genes in cotton fiber development have not been comprehensively analysed.In this study,the cotton transcriptome and metabolome were used to investigate the function of CesA genes in fiber development.Finally,321 metabolites were obtained,84 of which were associated with the corresponding genes.Interestingly,a target gene named Gh_A08G144300,one of the CesA gene family members,was closely correlated with the development of cotton fibers.The target CesA gene Gh_A08G144300 was analysed to determine its specific function in cotton fiber development.High-level gene expression of Gh_A08G144300 was found at different fiber development stages by RNA-seq analysis,and the silencing of Gh_A08G144300 visibly inhibited the growth of cotton fibers,showing that it is critical for their growth.This study provides an important reference for research on the gene function of Gh_A08G144300 and the regulatory mechanism of fiber development in cotton.展开更多
文摘通过逆转录聚合酶链式反应（RT-PCR）结合c DNA末端快速扩增（RACE）技术,从杉木Cunninghamia lanceolata中克隆到2个全长为3 235 bp和3 876 bp的纤维素合成酶基因c DNA序列,被分别命名为Cl Ces A1和Cl Ces A2,Gen Bank登录号分别为JQ844574和JQ844575。相应编码蛋白分别包含992和1 092个氨基酸残基,推测分子量为111 845.3 D和123 105.7 D,等电点为6.04和6.65。氨基酸序列分析发现,它们具有植物纤维素合成酶基因相应的结构特征——锌指结构,2个易变区CSRI和CSRII,2个保守区CRI和CRII,纤维素合成酶底物结合结构域＂D,D,D,QVLRW＂,以及8个跨膜区。荧光定量PCR分析表明：Cl Ces A1和Cl Ces A2基因在茎中表达丰度均为最高,且成熟木质部中表达量均高于皮层中的相应数值;2个基因在根和针叶中的表达量相对较低。以具特殊材质性状的矮生杉木和正常杉木木质部为材料的进一步表达分析发现：Cl Ces A1和Cl Ces A2在正常杉木木质部中的表达量大约是矮生杉木表达量的2~12倍。2个杉木Ces A基因可能在杉木木材形成过程中具有重要作用。
文摘克隆得到了一个白桦纤维素合成酶基因(CESA7)Gen Ban K登录号(EU591531)启动子序列,通过序列分析发现该启动子含有多个不同功能的顺式作用元件,包括光响应元件、激素响应元件、叶片形态发育元件等,推测该启动子在白桦生长发育过程中具有关键作用。将Bp CESA7启动子克隆至带有GUS报告基因的植物表达载体,命名为pro Bp CESA7-121-GUS,并利用农杆菌介导方法侵染白桦和拟南芥,然后通过GUS组织化学染色观察Bp CESA7基因启动子的组织表达特性。结果在白桦的根、茎、叶和拟南芥的根,叶,萼片、雌蕊中检测到了GUS活性,说明Bp CESA7基因启动子具有启动子活性,并且在白桦的根和叶中染色最深,表明Bp CESA7基因在白桦根和叶中表达量较高,并且其存在组织表达特异性。
文摘Arabidopsis (Arabidopsis thaliana) WRKY33 is a key transcription factor in pathogen-induced defense signaling, but its function in abiotic stresses remains largely unclear. In this study, we report on the use of a reverse-genetic approach, as well as a yeast (Saccharomyces cerevisiae) expression system, to determine the role of WRKY33 in drought. A T-DNA insertion deletion mutant of WRKY33 is more sensitive to dehydration. Through genome-wide screening the target genes of WRKY33 in yeast, we identified 23 candidate genes including a drought tolerance gene CesA8. Further results revealed that WRKY33 repressed CesA8 expression through binding to the W-box elements of CesA8 distal promoter region and probably interacting with the transcriptional activator of CesA8, MYB46. These findings revealed the primary molecular mechanism underlying the function of WRKY33 in response to
基金This project was supported by the Special Fund for Modern Agriculture of Jiangxi Province(JXARS-22)Science and Technology Research Project of Education Department of Jiangxi Province(181366).
文摘Cotton is widely distributed worldwide,and improving the quality of its fiber is one of the most important tasks in cotton breeding.Cotton fibers are primarily composed of cellulose,which is synthesized by CesA complexes(CSCs).However,the functions of CesA genes in cotton fiber development have not been comprehensively analysed.In this study,the cotton transcriptome and metabolome were used to investigate the function of CesA genes in fiber development.Finally,321 metabolites were obtained,84 of which were associated with the corresponding genes.Interestingly,a target gene named Gh_A08G144300,one of the CesA gene family members,was closely correlated with the development of cotton fibers.The target CesA gene Gh_A08G144300 was analysed to determine its specific function in cotton fiber development.High-level gene expression of Gh_A08G144300 was found at different fiber development stages by RNA-seq analysis,and the silencing of Gh_A08G144300 visibly inhibited the growth of cotton fibers,showing that it is critical for their growth.This study provides an important reference for research on the gene function of Gh_A08G144300 and the regulatory mechanism of fiber development in cotton.
