黄牡丹花粉生活力测定方法的比较研究

以黄牡丹的新鲜花粉为试材,利用单因子试验比较了液体培养基中蔗糖浓度、硼、钙、镁、钾对黄牡丹花粉萌发的影响,在此基础上进行了正交试验,比较了蔗糖、H₃BO₃及CaCl₂对黄牡丹花粉萌发的影响;通过对醋酸洋红染色法、I-KI染色法和TTC染色法的比较,寻找快速测定黄牡丹花粉生活力的方法。试验结果表明:蔗糖及H₃BO₃对黄牡丹花粉萌发有极显著影响。在pH值为6.0时,蔗糖150 g·L^-1+H₃BO₃30 mg·L^-1+CaCl₂20 mg·L^-1适宜黄牡丹花粉培养,萌发率为68.7%;纯水培养没有造成花粉原生质体破裂,内含物外流,但萌发率极低,仅为3%;200 g·L^-1以上的高浓度蔗糖溶液和300 mg·L^-1以上的高浓度盐溶液会造成原生质体失水萎缩,质壁分离,这两种情况都抑制花粉萌发;TTC染色法测得的花粉活力率为64.9%,是快速测定黄牡丹花粉生活力的最适染色法。

Comparison of Methods about Paeonia lutea's Pollen Viability Determination

The fresh pollen of Paeonia lutea was employed as experimental material to study the pollen viability. The effects of different concentration of sucrose, boron, calcium, magnesium and potassium on the germination of P. lutea's pollen were compared through simple factorial experiment. Based on these, the orthogonal design was used to compare the effects of sucrose, boron and calcium on the germination of P. lutea's pollen. Carmine acetate staining, I₂-KI staining, and TTC staining were compared in order to seek for a fast determination method of P. lutea's pollen viability. The results showed that the sucrose and boron had great effects on the germination of P. lutea's pollen. Under the optimum pH 6.0 values, the optimum culture solution was sucrose 150 g·L^-1 + H₃BO₃ 30 mg·L^-1 + CaCl₂ 20 mg·L^-1, the germination rate was 68.7%. When cultured in pure water, the protoplasm of the pollen wouldn't split, and the substance inside wouldn't flow out, but its germination rate was very low. When cultured in the solution with high concentration of sucrose or salt, the protoplasm would dehydrate and separate from the cell wall. These two would inhibit the germination of P. lutea's pollen. And TTC staining showed that the viability rate was 64.9% which was the optimum staining for the fast determination of P. lutea's pollen viability.