抗性湿地松体细胞胚的发育、成熟及萌发  

Development,Maturation and Germination of Resistant Slash Pine Somatic Embryos

作　　者：程方[1,2] 叶建仁 CHENG Fang;YE Jianren(Co-Innovation Center for Sustainable Forestry in Southern China,Nanjing Forest University,Nanjing Jiangsu 210037;Guangdong Xiangtoushan National Nature Reserve Administration,Huizhou Guangdong 516001)

机构地区：[1]南京林业大学南方现代林业协同创新中心,江苏南京210037 [2]广东象头山国家级自然保护区管理局,广东惠州516001

出　　处：《现代农业科技》2024年第3期166-174,共9页Modern Agricultural Science and Technology

基　　金：“十四五”国家重点研发项目(2021YFD1400900)。

摘　　要：为加快抗松针褐斑病湿地松体细胞胚胎发生技术的产业化应用进程,本文研究了肌醇浓度、脱落酸(ABA)及其添加方式、基本培养基、聚乙二醇(PEG-6000)浓度、琼脂糖种类及浓度、活性炭浓度、液体悬浮培养等因素对抗性湿地松体细胞胚的发育、成熟及萌发的影响。结果表明:添加8 g/L肌醇较适宜,过高浓度的肌醇则不利于湿地松体细胞胚发育;ABA可以采用高压灭菌;基本培养基、ABA、PEG-6000互作对湿地松体细胞胚的成熟影响很大,基本培养基以LP为最佳,ABA的最适浓度为10m g/L,PEG-6000以添加25、50 g/L为宜;在湿地松体细胞胚的成熟培养过程中,PEG-6000的浓度不能高于100 g/L;在湿地松体细胞胚的成熟培养基中,添加60 g/L蔗糖最合适;湿地松体细胞胚的成熟培养基中添加0.5 g/L或1 g/L的活性炭较适宜;培养基状态对体细胞胚的高频率诱导具有一定的影响,悬浮培养以100 mL三角瓶装培养液30 mL、摇床转速130 r/min为宜,液体转固体时吸取培养液1 mL为宜;未建立成熟的体细胞胚发生体系前,以采用固体培养为佳。最佳成熟培养基、激素及部分添加物组合为LP+10 mg/L ABA+50 g/L PEG-6000+60 g/L蔗糖+1.0 g/L活性炭。In order to speed up the process of industrialization and application of slash pine somatic embryogenesis technology against pine needle brown spot,this paper studied the influence of the factors on development,maturation and germination of resistant slash pine somatic embryo,such as inositol concentration,abscisic acid(ABA)and its adding method,the basic medium,polyethylene glycol(PEG-6000)concentration,agarose species and concentration,activated carbon concentration,liquid suspension culture.The results showed that it was advisable to add 8 g/L inositol,and too high concentration of inositol was not conducive to the development of slash pine somatic embryos;ABA could be autoclaved;interaction of basic medium,ABA and PEG-6000 had a great influence on maturation of slash pine somatic embryos,LP was the best basic medium,the optimum concentration of ABA was 10 mg/L,the suitable concentrations of PEG-6000 were 25 g/L and 50 g/L;during the maturation culture of slash pine somatic embryos,the concentration of PEG-6000 should not be higher than 100 g/L;adding 60 g/L sucrose was the most suitable in the maturation medium of slash pine somatic embryos;adding 0.5 g/L or 1.0 g/L of activated carbon to the maturation medium of slash pine somatic embryos was suitable;the medium state had a certain effect on the high-frequency induction of somatic embryos,for suspension culture,30 mL culture fluid filled in a 100 mL triangular flask and 130 r/min of shaker speed were appropriate;when liquid was changed to solid,1 mL of culture medium was appropriated;it was better to use solid culture before the establishment of a mature somatic embryogenesis system.The best combination of maturation medium,hormones and additives was LP+10 mg/L ABA+50 g/L PEG-6000+60 g/L sucrose+1.0 g/L activated carbon.

关 键 词：抗性湿地松 体细胞胚胎发生 胚性愈伤组织 发育培养基 成熟培养基 

分 类 号：S791.246[农业科学—林木遗传育种]