The invention provides an ultra low temperature preservation method for the seed of a light bark tree. The following steps are as follows: the seeds of light bark tree are washed with water at room temperature and cultivated with A reagents. After 20 to 50min, the cultivated light bark tree seeds are taken out. In the cryopreservation tube, the B reagents are added to the room temperature for 15 ~ 35min, and then put into the gradient cooling box. In the low temperature refrigerator, the temperature of the gradient cooling box with the seed of the light bark tree is put in the low temperature refrigerator, and the temperature of the cooling box is 35 ~ 60 C, the cooling rate is 1 /min, then the cooling box is taken out, and the cooling box is put in the liquid nitrogen, and the liquid nitrogen is kept at low temperature for 6 years and then the cooling box is from the liquid nitrogen. Take slowly thaw and wash it for seed germination.
【技术实现步骤摘要】
一种光皮树种子超低温保存方法
本专利技术涉及一种光皮树种子超低温保存方法,属于药材种子资源保存
技术介绍
光皮树又叫斑皮抽水树,落叶乔木,高5-18米,稀达40米;树皮灰色至青灰色,块状剥落;幼枝灰绿色,略具4棱,被灰色平贴短柔毛,小枝圆柱形,深绿色,老时棕褐色,无毛,具黄褐色长圆形皮孔。冬芽长圆锥形,长3-6毫米,密被灰白色平贴短柔毛。叶对生,纸质,椭圆形或卵状椭圆形,长6-12厘米,宽2-5.5厘米,先端渐尖或突尖,基部楔形或宽楔形,边缘波状,微反卷,上面深绿色,有散生平贴短柔毛,下面灰绿色,密被白色乳头状突起及平贴短柔毛,主脉在上面稍显明,下面凸出,侧脉3-4对,弓形内弯,在上面稍显明,下面微凸起;叶柄细圆柱形,长0.8-2厘米,幼时密被灰白色短柔毛,老后近于无毛,上面有浅沟,下面圆形。顶生圆锥状聚伞花序,宽6-10厘米,被灰白色疏柔毛;总花梗细圆柱形,长2-3厘米,被平贴短柔毛;花小,白色,直径约7毫米;花萼裂片4,三角形,长约0.4-0.5毫米,长于花盘,外侧被白色短柔毛;花瓣4,长披针形,长约5毫米,上面无毛,下面密被灰白色平贴短柔毛;雄蕊4 ...
【技术保护点】
一种光皮树种子超低温保存方法,包括步骤以下:(1)室温下将光皮树种子用水洗净,用A试剂培育,培育20~50min后,取出培育过的光皮树种子;(2)培育过的光皮树种子放入冻存管中,再加入B试剂进行室温温育15~35min,再将其放入梯度降温盒中;(3)将带有光皮树种子的梯度降温盒放入低温冰箱中,使其降温盒的温度在‐35℃~‐60℃,降温速度为‐1℃/min,然后取出降温盒,马上将此降温盒放入液氮中,进行液氮低温保存备用;(4)经低温保存6年后,再将降温盒从液氮中取,缓慢解冻,洗净后用于种子发芽。
【技术特征摘要】
1.一种光皮树种子超低温保存方法,包括步骤以下:(1)室温下将光皮树种子用水洗净,用A试剂培育,培育20~50min后,取出培育过的光皮树种子;(2)培育过的光皮树种子放入冻存管中,再加入B试剂进行室温温育15~35min,再将其放入梯度降温盒中;(3)将带有光皮树种子的梯度降温盒放入低温冰箱中,使其降温盒的温度在‐35℃~‐60℃,降温速度为‐1℃/min,然后取出降温盒,马上将此降温盒放入液氮中,进行液氮低温保存备用;(4)经低温保存6年后,再将降温盒从液氮中取,缓慢解冻,洗净后用于种子发芽。2.根据权利要求1的方法,其中步骤(1)所述A试剂指B5液体培养液,还包含有1.5~2.5mol/L甘油和1.2~1.6mol/L二甲基亚砜和0.35~0.55mol/L蔗糖。3.根据权利要求1的方法,其中所述A试剂优选110~121℃高压蒸汽灭菌15min以上,...
【专利技术属性】
技术研发人员:罗婵,
申请(专利权)人:长沙爱扬医药科技有限公司,
类型:发明
国别省市:湖南,43
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