湿地松体细胞胚胎发生和植株再生的方法

本发明涉及一种湿地松体细胞胚胎发生和植株再生的方法,包括(1)采集湿地松的球果,分离出未成熟合子胚;(2)诱导产生胚性愈伤组织;(3)胚性愈伤组织的增殖;(4)熟化培养获得具有萌发潜力的子叶胚;(5)萌发培养获得萌发苗和(6)炼苗及移植的步骤,所述诱导培养基为高浓度硝态氮的培养基，硝酸根离子的总浓度为13.8-14.6mM:所述诱导培养基中的激素组分包括:2.5-3μM6-苄氨基嘌呤、10.5-11μM萘乙酸、2.5-3μM激动素、18.7-19.2μM脱落酸和0.8-1.2μM芸苔素内酯。该方法具有较高的胚性愈伤组织的诱导率、体胚成熟率以及萌发率，并且移殖后的植株成活率高。

This invention relates to a method for somatic embryogenesis and plant regeneration of Pinus elliottii. The method comprises the following steps: (1) Collecting pine cones of Pinus elliottii and isolating immature zygotic embryos; (2) Inducing embryogenic callus; (3) Proliferating embryogenic callus; (4) Carrying out maturation culture to obtain cotyledonary embryos with germination potential; (5) Conducting germination culture to obtain germinated seedlings; and (6) Hardening-off and transplanting the seedlings. The induction medium is a high-concentration nitrate nitrogen medium, with a total nitrate ion concentration of 13.8–14.6 mM. The hormone components in the induction medium are as follows: 2.5–3 μM 6-benzylaminopurine, 10.5–11 μM 1-naphthaleneacetic acid, 2.5–3 μM kinetin, 18.7–19.2 μM abscisic acid, and 0.8–1.2 μM brassinolide. This method features high induction rate of embryogenic callus, somatic embryo maturation rate and germination rate, as well as high survival rate of transplanted plants.