枇杷属植物茎尖离体培养及褐化抑制研究

Study on shoot tip in vitro culture and inhibit-browning of loquats

【目的】建立野生枇杷成年树茎尖离体培养技术并抑制培养过程中的褐化。【方法】以华南农业大学枇杷属植物种质资源圃所收集保存的14个分类单元的野生枇杷,以及台湾枇杷ב解放钟’枇杷的杂交后代茎尖为培养试材,进行了成年树茎尖的离体培养及培养过程中褐化抑制的研究。研究了不同培养基、植物生长调节剂的配比、抑制褐化物质等对茎尖褐变程度的影响。分析了不同枇杷属植物在离体培养和褐化上差异性。【结果】不同植物生长调节剂及其浓度组合对同种或不同种枇杷的离体培养和抑制褐化作用有很大的差异性。其中,栎叶枇杷老挝变种在MS+ZT0.5 mg·L-1+IBA0.1 mg·L-1条件下获得了100%的萌动率,且无褐化;台湾枇杷ב解放钟’枇杷的杂交后代的茎尖离体培养的萌芽率都达到100%,相对其他枇杷属植物比较容易进行离体培养;而大花枇杷、香花枇杷、窄叶枇杷、细叶枇杷和小叶枇杷褐化程度较高(66.67%~100%),茎尖的萌动率很低(0~26.67%);窄叶枇杷茎尖培养的萌动是最困难的,但在培养基中添加TDZ的条件下,褐化率降至0,萌动率达到100%。试验发现MS+TDZ0.5 mg·L-1+BA1.0 mg·L-1+NAA0.1 mg·L-1的配方组合对所试验的各种野生枇杷茎尖培养都有很强的抑制褐化作用(0~33.33%),并且使茎尖上的芽在1周内就能开始萌动,生长势也最好。【结论】通过不断优化各种培养技术和条件使枇杷属植物的茎尖离体培养和抑制褐化取得了实质性的进展,为以后快速繁殖、种质资源保存甚至转基因奠定一定的技术基础。

【Objective】 Establish wild loquat of adult tree shoot tip in vitro culture basic technique and inhibit browning during the culture. 【Method】 The 14 taxa of wild loquat, as well as Eriobotrya deflexa Nakai × E. japonica Lindl. ‘ Jiefangzhong', picked from State Loquat Germplasm Repository, South China Agricultural University, were used as the materials of shoot tip in vitro culture and inhabit-browning researches. The effects of different media, plant growth regulators and their ratios, types of inhabit-browning substances on shoot tip inhabit-browning are studied. The difference of in vitro culture and browning among various Eriobotrya plants are analyzed. 【Result】 The results showed that: there are great differences among various concentrations of plant growth regulators and their combinations on the same or various kinds of loquats in vitro and inhabit-browning. Among them, E. prinoides var. laotica Rehd. &Wils. cultured in the medium of MS + ZT 0.5 mg·L-1+ IBA 0.1 mg·L-1obtained 100% germination rate without any browning; E. deflexa Nakai × E. japonica Lindl. cv. Jiefangzhong was easier to be cultured than other loquats; E. cavaleriei Rehd, E. fragrans Champ, E. henrjyi Nakai, E. angustissima Hook and E. seguinii Card had higher browning rate which was 66.67% to 100% and their germination rates were low, most of them ranged from 0 to 26.67%; shoot tip in vitro culture of E. henrjyi Nakai was the most difficult, however, when the TDZ was added in the medium with, the browning rate was 0%, germination rate was up to 100%. MS+TDZ 0.5 mg·L-1+BA 1. 0 mg·L-1+NAA 0.1 mg·L-1had a very strong inhibitedbrowning effect on all the wild loquats with the browning rate ranged from 0 to 33.33%, and made the shoot tips sprout in a week. 【Conclusion】 The results could lay certain technical foundation for the rapid propagation, germplasm resource conservation and gene translation in the future.