热带作物学报2024，Vol.45Issue(7)：1385-1392,8.DOI:10.3969/j.issn.1000-2561.2024.07.011
橡胶树热垦525胚性悬浮细胞系的建立及其胚性能力的维持
Establishment of Embryogenic Cell Suspensions of Hevea brasiliensis Clone Reken 525 and Maintenance of Its Embryogenic Competence
摘要
Abstract
Both friable embryogenic callus and suspension cells are ideal materials for genetic and cell engineering.However,the induction of friable embryogenic callus of Hevea brasiliensis is low frequency and time-consuming,and its embryogenic competence usually decreases gradually or even completely loses with the increase of subculture times.Therefore,to quickly obtain friable embryogenic callus,establish embryogenic cell suspensions with high efficiency of somatic embryogenesis,and maintain the embryogenic competence as long as possible,are important research contents for genetic and cell engineering of H.brasiliensis.In this study,the immature anthers of H.brasiliensis clone Reken 525 was used as the explants to induce callus and establish embryogenic cell suspensions,and the effects of two long-term subculture methods-solid and liquid on the maintenance of embryogenic competence were compared and analyzed.The results showed that the yellow and compact primary callus tissue induced from anther explants on callus induction me-dium had a low ability for somatic embryogenesis,and was not suitable for suspension culture due to poor dispersion.After 70-80 d of culture on the medium for embryogenic callus induction,the formation of embryogenic structures and early somatic embryogenesis were observed,and bright yellow small granular friable embryogenic calli grew around them.The type Ⅰ embryogenic suspension cell line established through routine methods possessed typical characteris-tics of embryogenic cells,and the frequency of somatic embryogenesis was significantly higher than that of embryo-genic callus tissue,but proned to callus formation during somatic embryogenesis;the type Ⅱ embryogenic suspension cell line established by screening specific embryogenic tissue and initiating with low density suspension culture was in an orderly embryonic structure observed by microscope,and its frequency of somatic embryogenesis could reach up to 100％.After two years of continuous subculture in liquid culture medium containing 2 mg/L 2,4-D,the cells were obvi-ously aging with lower proliferation coefficient,and the ability for somatic embryogenesis was almost lost.However,the ability for somatic embryogenesis still remained at a high level after two years of continuous subculture on solid culture medium without 2,4-D but supplemented with abscisic acid(0.1 mg/L)and hydrolyzed casein(0.5 g/L).The type Ⅱ embryogenic suspension cell line could provide high-quality,sufficient and relatively stable material sources for genetic transformation and protoplast culture of Hevea brasiliensis in the long term.关键词
橡胶树/易碎胚性愈伤组织/胚性悬浮细胞系/体胚发生/胚性维持Key words
Hevea brasiliensis/friable embryogenic callus/embryogenic cell suspensions/somatic embryogenesis/maintenance of embryogenic competence分类
农业科技引用本文复制引用
戴雪梅,彭素娜,成镜,顾晓川,黄天带..橡胶树热垦525胚性悬浮细胞系的建立及其胚性能力的维持[J].热带作物学报,2024,45(7):1385-1392,8.基金项目
海南省重点研发计划国际科技合作研发项目(No.GHYF2022013) （No.GHYF2022013）
海南省重点研发计划项目(No.ZDYF2022XDNY-204) （No.ZDYF2022XDNY-204）
海南省国际科技合作人才与交流项目(No.G20241024008E). （No.G20241024008E）
