摘要
玫瑰(Rosa rugosa var.Plena)是一种重要的观赏园艺植物，有关其组培快繁体系的研究报道较少。本试验以玫瑰带芽茎段为外植体，对采样时间、外植体灭菌、遮荫处理控制褐化、诱导芽萌发、不定芽增殖、壮苗和诱导生根等过程中各影响因素进行了研究；并通过组织学观察来阐明不定芽和不定根的起源，从而初步建立起玫瑰的快繁体系，对提高育种效率和进行苗木繁育具有重要意义。
结果表明：利用玫瑰冬季休眠芽的外植体，采用0.5％NaCl+0.1％HgCl_2+0.1％Tween-20处理5min，再用100mg／L庆大霉素浸泡灭菌效果好。
初代培养基中无机盐和激素浓度对诱导芽萌发有着显著的影响，且萌芽率亦受接种材料采集时期的影响，在4～9月的取材范围内，玫瑰最适取材时期是4月和9月。遮荫处理使培养物的褐变百分数降低，而萌芽率显著提高；正常光照条件下多酚氧化酶活性与其他遮荫处理相比并无显著变化，但总酚含量显著高于遮荫处理，二者的相关分析表明，多酚氧化酶活性与组织材料的总酚含量相关性不大。
组培苗在继代增殖培养基MS+0.05 mg／L NAA+1.0 mg／L BA增殖系数最高，达到3.44～6.00。培养基中添加TDZ+NAA+BA，对玫瑰初代培养有显著作用；但将苗子从有TDZ的初代培养基转移到无TDZ的继代培养基上时，增殖系数无明显提高。
壮苗培养以添加0.3mg／L NAA，0.09mg／L BA效果较好。
组培苗在MS+0.05 mg／L NAA的生根培养基上生根率达到41.67％，平均每外植体生根数为2.33。
组织学观察表明，组培苗的不定芽和不定根起源于茎的中柱。
Rose (Rosa rugosa van Plena) is an important ornamental plant but little has been reported about the protocol of its fast propagation. In this research, We used rose stem bearing buds as explants and studied all the factors that affected the fast propagation of rose, including time of collecting explants, sterilization, germination induction, browning controlling through shaded treatments, adventitious buds induction, shoots strengthening and roots induction. Also, we illuminated the origin of adventitious buds and adventitious roots through histological observation. Therefore, we primarily set up a fast propagation protocol of rose, which is important for improving efficiency of breeding and propagating seedlings.
Our results showed that the best sterilization method is treating dormant buds with 0.5% NaCl, 0.1% HgCl2 and 0.1% Tween-20 for 5 minutes, followed by dipping into 100 mg/L gentamycin solution.
In the medium of initial culture, the concentration of inorganic salts and hormone in the medium had significant effect on the germination percentage of buds induced, so did the time of collecting material. And it is optimum to collect material in April and September. Mild shaded treatment decreased the percentage of browning of rose but increased the percentage of germination. As for as controlling browning, the activity of PPO of rose shoot under natural light intensity showed no significant difference from other shaded treatments, whereas the content of phenolic substances was much higher than those treated with shaded illumination. The co-efficiency analysis demonstrated that there was no obvious correlation between the activity of phenol oxidase and content of phenolic substances.
In multiplication MS medium added with 0.05 mg/L NAA and 1.0 mg/L BA, the multiplication rates was best and reached 3.44-6.00. Besides, our study also demonstrated Medium supplemented TDZ+NAA+BA has effect on the germination of rose but had little effect on multiplication of shoots when transferred from initial culture medium containing TDZ into medium without TDZ.
To make shoot grow stronger, the suitable concentration of hormone was 0.3 mg/L NAA, 0.09 mg/LBA.
And the percentage of shoots rooted was 41.67% in the MS medium containing 0.05 mg/L NAA and the average number of roots per shoot was 2.33.
Histological observation displayed the adventitious buds and roots of rose were both originated in stele.
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