拟南芥(Arabidopsis thaliana)ABORTED MICROSPORES(AMS)基因是调控绒毡层和花粉壁发育的关键基因。芦笋(Asparagus officinalis)中与AMS同源的基因尚未报道。本研究通过同源比对、系统进化树构建及基因表达量的综合分析,预测了芦笋的AM...拟南芥(Arabidopsis thaliana)ABORTED MICROSPORES(AMS)基因是调控绒毡层和花粉壁发育的关键基因。芦笋(Asparagus officinalis)中与AMS同源的基因尚未报道。本研究通过同源比对、系统进化树构建及基因表达量的综合分析,预测了芦笋的AMS候选基因并命名AoAMS;进一步以四倍体‘紫色激情’两性株自交后代中两性株花蕾为材料,通过RT-PCR分别扩增并获取AoAMS及与拟南芥DEFECTIVE IN TAPETAL DEVELOPMENT AND FUNCTION 1(TDF 1)同源的基因(AoTDF1)全长CDS序列,开展AoAMS编码蛋白亚细胞定位,与AoTDF1间的互作分析及不同性别的芦笋花发育早期的无性别分化期(T1)、性别分化早期(T2)和晚期(T3)的表达分析。结果显示:AoAMS是具细胞核定位特征且与AoTDF1有互作的蛋白;AoAMS基因在雄花和两性花早期T1、T2、T3期发育的花冠、花药绒毡层、以及花粉中均有表达,但在雌花中相应时期及部位均无表达。研究结果表明AoAMS是专一性地在芦笋雄花或两性花中表达,并且可以与AoTDF1形成互作复合物,通过调控绒毡层及花粉正常发育相关基因的表达而在芦笋性别分化中行使促雄的功能。展开更多
During anther development, the tapetum pro- vides materials and nutrients for pollen development. In Arabidopsis, several transcription factors have been iden- tified to form a genetic pathway （DYT1-TDF1-AMS- MS188-...During anther development, the tapetum pro- vides materials and nutrients for pollen development. In Arabidopsis, several transcription factors have been iden- tified to form a genetic pathway （DYT1-TDF1-AMS- MS188-MS1） for tapetal development and function. DEFECTIVE in TAPETAL DEVELOPMENT and FUNCTION1 （TDF1） is a member of the R2R3 MYB family and is essential for early tapetum development in Arabidopsis. Here, we characterized an ortholog of Ara- bidopsis TDF1 in rice, OsTDF1 （LOC_Os03g18480）. OsTDF1 shares 69 % amino acid sequence identity with AtTDF1 in the putative MYB domain near the N-terminal region. RT-PCR and in situ hybridization show that OsTDF1 is specifically expressed in tapetal cells of rice anthers. The expression of OsTDF1 in Arabidopsis tdfl mutant restores its fertility, suggesting that this homolog can fulfill the normal function of TDF1 in Arabidopsis. The ostdfl knockout mutant exhibits a male-sterile phenotype. Its tapetal cells exhibit a vacuolated and hypertrophic phenotype similar to that of Arabidopsis tdfl mutants. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assays revealed OsTDF1 acts as an essential regulator for tapetum programmed cell death. The qRT-PCR analysis revealed that OsTDF1 acts downstream of UDT1 and upstream of TDR, EAT1, OsMYBI03 and PTCI in rice, suggesting that the genetic pathway for tapetum development is generally conserved between rice and Arabidopsis,展开更多
文摘拟南芥(Arabidopsis thaliana)ABORTED MICROSPORES(AMS)基因是调控绒毡层和花粉壁发育的关键基因。芦笋(Asparagus officinalis)中与AMS同源的基因尚未报道。本研究通过同源比对、系统进化树构建及基因表达量的综合分析,预测了芦笋的AMS候选基因并命名AoAMS;进一步以四倍体‘紫色激情’两性株自交后代中两性株花蕾为材料,通过RT-PCR分别扩增并获取AoAMS及与拟南芥DEFECTIVE IN TAPETAL DEVELOPMENT AND FUNCTION 1(TDF 1)同源的基因(AoTDF1)全长CDS序列,开展AoAMS编码蛋白亚细胞定位,与AoTDF1间的互作分析及不同性别的芦笋花发育早期的无性别分化期(T1)、性别分化早期(T2)和晚期(T3)的表达分析。结果显示:AoAMS是具细胞核定位特征且与AoTDF1有互作的蛋白;AoAMS基因在雄花和两性花早期T1、T2、T3期发育的花冠、花药绒毡层、以及花粉中均有表达,但在雌花中相应时期及部位均无表达。研究结果表明AoAMS是专一性地在芦笋雄花或两性花中表达,并且可以与AoTDF1形成互作复合物,通过调控绒毡层及花粉正常发育相关基因的表达而在芦笋性别分化中行使促雄的功能。
基金This work was supported by the grants from the Ministry of Science and Technology of China （2013CB945100）, the National Natural Science Foundation of China （31100227）.
文摘During anther development, the tapetum pro- vides materials and nutrients for pollen development. In Arabidopsis, several transcription factors have been iden- tified to form a genetic pathway （DYT1-TDF1-AMS- MS188-MS1） for tapetal development and function. DEFECTIVE in TAPETAL DEVELOPMENT and FUNCTION1 （TDF1） is a member of the R2R3 MYB family and is essential for early tapetum development in Arabidopsis. Here, we characterized an ortholog of Ara- bidopsis TDF1 in rice, OsTDF1 （LOC_Os03g18480）. OsTDF1 shares 69 % amino acid sequence identity with AtTDF1 in the putative MYB domain near the N-terminal region. RT-PCR and in situ hybridization show that OsTDF1 is specifically expressed in tapetal cells of rice anthers. The expression of OsTDF1 in Arabidopsis tdfl mutant restores its fertility, suggesting that this homolog can fulfill the normal function of TDF1 in Arabidopsis. The ostdfl knockout mutant exhibits a male-sterile phenotype. Its tapetal cells exhibit a vacuolated and hypertrophic phenotype similar to that of Arabidopsis tdfl mutants. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assays revealed OsTDF1 acts as an essential regulator for tapetum programmed cell death. The qRT-PCR analysis revealed that OsTDF1 acts downstream of UDT1 and upstream of TDR, EAT1, OsMYBI03 and PTCI in rice, suggesting that the genetic pathway for tapetum development is generally conserved between rice and Arabidopsis,
