黑皮油松外植体的抗褐化处理与诱导培养

Anti-browning Treatments and Induced Culture on the Explants of Pinus tabuliformis var.mukdensis

黑皮油松在组织培养过程中存在明显的褐化现象,严重影响离体培养的诱导效果。为有效地抑制黑皮油松组培中外植体的褐化,本试验以黑皮油松休眠叶芽为外植体,研究聚乙烯吡咯烷酮(polyvinyl pyrrolidone, PVP)、活性炭(active carbon, AC)、维生素C (vitamin C, VC)三种不同抗褐化剂对黑皮油松休眠芽外植体褐化与萌发的影响。结果表明,添加3种抗褐化剂对黑皮油松外植体褐化有不同程度的抑制效果,抗褐化剂处理的外植体褐化程度相比对照明显减轻,在培养8 d时抗褐化剂处理开始表现出抑制外植体褐化的作用,大部分处理外植体褐化率在培养至24 d时达到峰值,随后趋于稳定。整体来看,PVP处理的防褐化效果优于AC和VC处理,添加1 500 mg/L PVP抑制黑皮油松外植体褐化的效果最佳,褐化率仅为25.56%,比对照低51.11%,该处理的休眠芽萌发率也最高(37.78%),说明添加PVP不仅有效抑制黑皮油松外植体的褐化,也明显促进黑皮油松休眠芽的萌发启动。本研究为黑皮油松组织培养过程中褐变这一瓶颈问题的解决提供理论依据和技术支撑。

There is obvious browning during the tissue culture of Pinus tabulaeformis var. mukdensis, which has been seriously affecting the induction effectiveness of in vitro culture. In order to effectively inhibit the browning of explants in the tissue culture of P. tabulaeformis, in this study, the effects of three different anti-browning agents,including polyvinyl pyrrolidone(PVP), activated carbon(AC) and vitamin C(VC), on the browning and germination of the explants, dormant leaf buds of P. tabulaeformis were explored. The results showed that the addition of the three anti-browning agents have inhibited the browning of P. tabulaeformis explants to a different extent with the browning degree of explants treated with the anti-browning agents being significantly reduced compared with the control. The anti-browning agents started to inhibit the browning of the explants after 8 days of culture, for most treatments the browning rates of the explants peaked at 24 d and then stabilized. Overall, the anti-browning effect of PVP treatment was better than AC treatment and VC treatment, and the addition of 1 500 mg/L PVP gained the best treatment effect in preventing the browning of P. tabulaeformis explants having a browning rate of only25.56%, which was 51.11% lower than that of the control, and the germination rate of dormant buds was also the highest(37.78%), which indicated that the addition of PVP not only effectively inhibited the browning of P. tabulaeformis explants, but also obviously promoted the germination of the dormant buds. This study provides theoretical basis and technical support for solving the browning bottleneck problems in P. tabulaeformis tissue culture.