Somatic embryogenesis(SE) plays a vital role in genetic transformation and massive propagation of important agronomical and economical crops.Here,we conducted a systematic assessment of the morphological,cytochemical,...Somatic embryogenesis(SE) plays a vital role in genetic transformation and massive propagation of important agronomical and economical crops.Here,we conducted a systematic assessment of the morphological,cytochemical,and cytogenetical characteristics of six culture strains with various embryogenic/regenerative potential during SE process in cotton.Results indicated that the six cell culture strains had stable ploidy levels,and did not reveal any relationship between the cytogenetic state and their morphogenetic potential.Moreover,the six culture strains were compared via double staining with Evans blue and Acetocarmine to efficiently distinguish embryogenic and non-embryogenic cells and determine the embryogenic nature of the calli.In addition,the kind of auxins added in medium affected not only growth property,color,size of cell clumps but also ploidy level and regeneration ability.By combining analysis of morphological,cytochemical,and cytogenetical characteristics of the cell cultures,we are able to obtain and maintain homogeneous cell population with high morphogenic and regeneration ability and establish efficient somatic embryogenesis and regeneration system from short-term cell cultures in upland cotton,which highlight the application of biotechnological approaches in crop breeding,and above all,to better understand totipotency of cells in higher plants.展开更多
Programmed cel death （PCD） plays a critical role in the development of plant pigment glands, while H2O2, which is a kind of reactive oxygen species （ROS） produced by the aerobic metabolism of cels, acts as an impo...Programmed cel death （PCD） plays a critical role in the development of plant pigment glands, while H2O2, which is a kind of reactive oxygen species （ROS） produced by the aerobic metabolism of cels, acts as an important signal in this process. Here, we investigated the temporal and spatial dynamics of accumulated H2O2 in pigment glands ofGossypium hirsutum L. with 3,3-diaminobenzidine （DAB） staining, 2’,7’-dichlorodihydrolfuorescein diacetate （DCFH2）-DA lfuorescent labeling and CeCl3 cytochemical localization techniques. The results showed that thepigment glandsofG. hirsutum could generate H2O2, and the amount and localization of H2O2 variedat different developmental stages. At the early developmental stage, a smal amount of H2O2 accumulated in the vacuole membrane of pigment gland cels. At the intermediate stage, a large number of H2O2 appeared in the vacuole membrane, while cel wals started to accumulate a smal amount of H2O2. When pigment gland cel degraded, H2O2 mainly accumulated on the chloroplast envelope membrane of inner sheath cels. With the degradation of the sheath cels, H2O2was detected in cel wal and the membrane of secretory vesicles which contains the preliminary contents of pigment gland. With the pigment glands completely maturation, H2O2 would disappeared. The accumulation sites of H2O2are consistent with the process of PCD of individual gland cels, which started from the degra-dation of intracelular membrane and ended with the degradation of cel wals. Thus H2O2 probably plays an important role in the development of pigment glands. In addition, the development of pigment glands and the generation of H2O2 are not associated with the light, and no H2O2 was detected in the secretions of pigment glands.展开更多
基金supported by the National Key R&D Program of China (2016YFD0100306)the National Natural Science Fundation of China (31401428)+1 种基金the Fok Ying-Tong Education Foundation of China (151024)the Taishan Scholar Talent Project from China (tsqn20161018)
文摘Somatic embryogenesis(SE) plays a vital role in genetic transformation and massive propagation of important agronomical and economical crops.Here,we conducted a systematic assessment of the morphological,cytochemical,and cytogenetical characteristics of six culture strains with various embryogenic/regenerative potential during SE process in cotton.Results indicated that the six cell culture strains had stable ploidy levels,and did not reveal any relationship between the cytogenetic state and their morphogenetic potential.Moreover,the six culture strains were compared via double staining with Evans blue and Acetocarmine to efficiently distinguish embryogenic and non-embryogenic cells and determine the embryogenic nature of the calli.In addition,the kind of auxins added in medium affected not only growth property,color,size of cell clumps but also ploidy level and regeneration ability.By combining analysis of morphological,cytochemical,and cytogenetical characteristics of the cell cultures,we are able to obtain and maintain homogeneous cell population with high morphogenic and regeneration ability and establish efficient somatic embryogenesis and regeneration system from short-term cell cultures in upland cotton,which highlight the application of biotechnological approaches in crop breeding,and above all,to better understand totipotency of cells in higher plants.
基金supported by the National Natural Science Foundation of China (31270428)
文摘Programmed cel death （PCD） plays a critical role in the development of plant pigment glands, while H2O2, which is a kind of reactive oxygen species （ROS） produced by the aerobic metabolism of cels, acts as an important signal in this process. Here, we investigated the temporal and spatial dynamics of accumulated H2O2 in pigment glands ofGossypium hirsutum L. with 3,3-diaminobenzidine （DAB） staining, 2’,7’-dichlorodihydrolfuorescein diacetate （DCFH2）-DA lfuorescent labeling and CeCl3 cytochemical localization techniques. The results showed that thepigment glandsofG. hirsutum could generate H2O2, and the amount and localization of H2O2 variedat different developmental stages. At the early developmental stage, a smal amount of H2O2 accumulated in the vacuole membrane of pigment gland cels. At the intermediate stage, a large number of H2O2 appeared in the vacuole membrane, while cel wals started to accumulate a smal amount of H2O2. When pigment gland cel degraded, H2O2 mainly accumulated on the chloroplast envelope membrane of inner sheath cels. With the degradation of the sheath cels, H2O2was detected in cel wal and the membrane of secretory vesicles which contains the preliminary contents of pigment gland. With the pigment glands completely maturation, H2O2 would disappeared. The accumulation sites of H2O2are consistent with the process of PCD of individual gland cels, which started from the degra-dation of intracelular membrane and ended with the degradation of cel wals. Thus H2O2 probably plays an important role in the development of pigment glands. In addition, the development of pigment glands and the generation of H2O2 are not associated with the light, and no H2O2 was detected in the secretions of pigment glands.
