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作 者 ：吴艳歌, 毕波, 柴明良

期 刊 ：核农学报 2016年 30卷 7期 页码：1288-1295

关键词：沟叶结缕草；细胞分裂素；愈伤组织生长；植株再生；抗氧化酶；

Keywords:Zoysia matrella, cytokinins, callus maintenance, plant regeneration, antioxidant enzymes,

摘 要 ：为探究细胞分裂素对长期继代培养的愈伤组织细胞系分裂、增殖的影响,以及对梢和根的诱导作用,以继代10年的沟叶结缕草愈伤组织为材料,研究0 ~ 0.20 mg·L^-1的6-苄基氨基嘌呤(BA)、激动素(KT)、玉米素(ZT)、反式玉米素核苷(ZR)、噻苯隆(TDZ)及正交设计组合对其生长和再生的影响。结果表明,继代培养4周后,愈伤组织鲜重增长率最高的处理是 0.10 mg·L^-1BA+ 0.20 mg·L^-1KT+ 0.15 mg·L^-1ZT+ 0.10 mg·L^-1TDZ,达到331.3%,略高于0.10 mg·L^-1KT(315.0%),但差异不显著。以1/2MS + 30.0 g·L^-1蔗糖 + 3.0 g·L^-1Phytagel为基本培养基,添加不同浓度的5种细胞分裂素再生培养6周,再生率和再生梢总数最高的是含 0.15 mg·L^-1ZT的培养基,再生率为60.3%,再生梢总数为142.0个,分别是对照的2.41倍和3.67倍。再生过程中过氧化物酶(POD)活性(6.694~14.569 U·g^-1FW·min^-1)显著低于对照(20.802~32.303 U·g^-1FW·min^-1),超氧化物歧化酶(SOD)活性的峰值(382.353 U·g^-1FW·min^-1)显著高于对照(326.760 U·g^-1FW·min^-1),而第3周再生高峰期过氧化氢酶(CAT)活性(0.312 U·g^-1FW·min^-1)显著高于对照(0.141 U·g^-1FW·min^-1)。因此,适宜浓度的细胞分裂素不仅显著促进愈伤组织的生长和分化,也明显影响抗氧化酶的活性。合理调控SOD和POD活性有助于提高胚性愈伤组织的再生能力,为沟叶结缕草现代育种提供基础资料。

Abstract:In order to study the role of cytokinins in cell division, proliferation, shoot and root differentiation, and somatic embryogenesis of long-term maintained callus during in vitro culture, the effects of different cytokinins (BA, KT, ZT, ZR and TDZ, 0 to 0.20 mg·L^-1) and their combinations on maintenance and plant regeneration of the 10-year Zoysia matrella embryonic callus were compared in this study. The highest proliferation rate was observed at 0.10 mg·L^-1 BA + 0.20 mg·L^-1KT + 0.15 mg·L^-1 ZT + 0.10 mg·L^-1 TDZ, reaching 331.3% after subculture for 4 weeks, which was slightly higher than that of 0.10 mg·L^-1 KT (315.0%). Among the plant regeneration media (1/2MS + 30.0 g·L^-1 Sugar + 3.0 g·L^-1 Phytagel) supplemented with five different cytokinins, the highest regeneration rate and shooting differentiation rate were observed in the medium containing 0.15 mg·L^-1 ZT, showing a regeneration rate of 60.3% and a total of 142 differentiated shoots, which were 2.41 times and 3.67 times than those of control respectively. The determination of antioxidant enzyme activities during regeneration indicated that the activities of Peroxide (POD) (6.694 to 14.569 U·g^-1FW·min^-1) were significantly lower than the control (20.802 to 32.303 U·g^-1FW·min^-1) and the peak of Superoxide dismutase (SOD) activity (382.353 U·g^-1FW·min^-1) was significantly higher than the control (326.760 U·g^-1FW·min^-1). In the third week, the activity of Catalase (CAT) was significantly higher than that of the control. The results indicated that cytokinins could promote the growth and differentiation of calli and significantly affect the activities of antioxidant enzymes. Regulation of SOD and POD activity could improve the regeneration of embryogenic callus, and provide experimental material for modern breeding of Z. matrella.

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