三种培养方式对橡胶树花药胚性悬浮细胞团再生体系构建的影响  

Efects of three culture methods on the establishment of anther embryogenic suspension cell masses regeneration system of Hevea brasiliensis

作　　者：李淑雅 陈健妙[1,2] 张骐飞 党彤 LI Shuya;CHEN Jianmiao;ZHANG Qifeil;DANG Tong(College of Tropical Crops,Hainan University,Haikou 570228,China;Hainan Key Laboratory for Sustainable Utilization of Tropical Bioresource(State Key Laboratory Breeding Base),Haikou 570228,China)

机构地区：[1]海南大学热带作物学院,海口570228 [2]海南省热带生物资源可持续利用重点实验室(省部共建国家重点实验室培育基地),海口570228

出　　处：《植物生理学报》2022年第7期1236-1244,共9页Plant Physiology Journal

基　　金：国家自然科学基金地区基金项目(31360188);海南省高等学校教育教学改革研究项目(Hnjg2019-3)。

摘　　要：本研究以橡胶树(Hevea brasiliensis)花药胚性悬浮细胞团为材料,比较了液体悬浮、间歇浸没、固体培养三种培养方式对其增殖、体细胞胚发生、成熟以及萌发成苗的影响。结果表明:液体悬浮培养方式有利于胚性悬浮细胞团增殖,其增幅最大,细胞团鲜重增加了9.371倍;体细胞胚早期分化阶段,固体培养方式下体细胞胚分化率为58.6%,显著高于其他两种培养方式;体细胞胚成熟和萌发阶段,液体悬浮培养未见有体胚成熟,而以自动间歇浸没式生物反应器(RITA)进行的间歇浸没式培养下体胚成熟率和体胚萌发成苗率均显著高于固体培养,且间歇浸没式培养大幅缩短了体胚萌发成苗的周期。综上,悬浮胚性细胞团在继代增殖阶段较适宜以液体悬浮方式培养;固体培养方式在体胚早期分化阶段具备优势,然而在体胚成熟萌发阶段,间歇浸没式培养可更好促进体胚发育成熟和萌发成苗,提高植株再生频率。In this study, the embryogenic suspension cell masses of Hevea brasiliensis were cultivated under regeneration conditions in liquid medium, in recipient for automated temporary immersion(RITA) bioreactor, and on gelled medium, respectively. The embryogenic callus proliferation, somatic embryogenesis,embryo development, embryo maturation and germination in different culture methods were investigated.Suspension culture was beneficial to the proliferation of embryonic suspension cell masses. This culture method produced the greatest accumulation of biomass and the embryogenic callus biomass increased by 9.371 times. At the early stage of somatic embryo differentiation, the differentiation rate of somatic embryos was 58.6% under solid culture conditions, which was significantly higher than those of the other two culture methods. At the stage of somatic embryo maturation and germination, there was no mature somatic embryos observed under liquid suspension culture method. The optimal culture method for somatic embryo maturation and germination was temporary immersion culture in RITA bioreactor. Compared with the gelled medium culture, temporary immersion culture in RITA containers significantly increased the maturation rate and germination rate of somatic embryos and greatly shortened the period of somatic embryos germinating to form seedlings. In conclusion, for suspension cell masses, suspension culture in liquid medium is suitable for proliferation stage, and solid culture on gelled medium has advantages in the early somatic embryogenesis stage, but in the somatic embryo maturation and germination stage, the temporary immersion culture in RITA bioreactor is more helpful to promote somatic embryos maturation and germination, thereby effectively increases the frequency of plant regeneration.

关 键 词：橡胶树 胚性悬浮细胞 培养方式 间歇浸没式培养 体细胞胚成熟和萌发 

分 类 号：S794.1[农业科学—林木遗传育种]