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
刘义存;岑伟仪;黄天启;林玉霜;张志珂;高用顺;林顺权;
摘要(Abstract):
【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。
关键词(KeyWords): 枇杷;茎尖;离体培养;植物生长调节剂;褐化抑制
基金项目(Foundation): 公益性行业(农业)科研专项(201003073);; 广东省自然科学基金重点项目(07118121);; 广东省高校华南农业大学园艺植物种质资源创新与利用重点实验室项目(KLB2011018)
作者(Author): 刘义存;岑伟仪;黄天启;林玉霜;张志珂;高用顺;林顺权;
Email:
DOI: 10.13925/j.cnki.gsxb.2014.03.024
参考文献(References):
- [1]LIN Shun-quan,YANG Xiang-hui,LIU Cheng-ming,HU You-li,HE Ye-hua,HU Gui-bing,ZHANG Hai-lan,HE Xiao-long,LIU Yue-xue,LIU Zong-li,Natural geographical distribution of genus eriobotrya plants in China[J].Acta Horticulturae Sinica,2004,31(5):101-104.林顺权,杨向晖,刘成明,胡又厘,何业华,胡桂兵,张海岚,何小龙,刘月学,刘宗莉.中国枇杷属植物的自然地理分布[J].园艺学报,2004,31(5):101-104.
- [2]DING Zhong-kui,CHEN Qi-feng,SUN Tian-lin,XIA Qi-zhou.The germplasm resources and improved varieties of Chinese loquat[J].Crop genetic resources,1993(4):8-11.丁长奎,陈其峰,孙田林,夏起洲.中国枇杷种质资源与枇杷的品种改良[J].作物品种资源,1993(4):8-11.
- [3]WANG Yun-sheng,HU You-li,HE Xiao-long,YANG Xiang-hui,TIAN Zhong-wei,LIN Shun-quan.Investigation report on wild and semi-wild germplasm of loquatin China[J].Fujian Fruits,2012(4):11-15.王云生,胡又厘,何小龙,杨向晖,田宗伟,林顺权.中国部分野生普通枇杷资源调查分析[J].福建果树,2012(4):11-15.
- [4]JIANG Ji-mou,CHEN Xiu-ping,JIANG Fan,HUANG Ai-ping,DENG Chao-jun,HU Wen-shun,XU Qi-zhi,ZHENG Shao-quan.Study on earring characteristic of loquat and its evaluating index innutrition periods[J].Journal of Fruit Science,2013,30(6):938-943.蒋际谋,陈秀萍,姜帆,黄爱萍,邓朝军,胡文舜,许奇志,郑少泉.枇杷种质资源抽穗特性与幼树期评价指标探讨[J].果树学报,2013,30(6):938-943.
- [5]YANG Yong-qing,CHEN Guang-lu,TANG Dao-yi.A study on tissue culture and inducing proliferation from shoot apexes of loquat tree[J].Acta Horticulturae Sinica,1983,10(2):79-82.杨永青,陈光禄,唐道一.枇杷茎尖培养与增殖的研究[J].园艺学报,1983,10(2):79-82.
- [6]HE Wei-qi.In vitro culture of immature embryos and plant Regeneration Via Somatic Embryogenesis in Loquat[J].China Horticulture,1983,29(4):343-345.何伟其.枇杷幼胚及茎顶培养诱导体细胞胚胎发生的研究[J].中国园艺(台湾),1983,29(4):343-345.
- [7]YANG Feng-ling,LIN Shun-quan.Establishment of the first generation of in Vitro culture from loquat stem apex[J].South China Fruits,2005,34(6):36-38.杨凤玲,林顺权.枇杷茎尖组织培养初代培养物的建立[J].中国南方果树,2005,34(6):36-38.
- [8]YANG Qin,FU Yan,WANG Yong-qing,TAO Lian,DENG Qunxian,FAN Jian-xin,DENG Ren-ju.Establishment and optimization of AS-PCR reaction system for Eriobotrya[J].Journal of Fruit Science,2013,30(1):62-68.杨芩,付燕,王永清,陶炼,邓群仙,范建新,邓仁菊.枇杷ASPCR反应体系的建立和优化[J].果树学报,2013,30(1):62-68.
- [9]ZHANG Ling,LIN Shun-quan,HU You-li,ZHENG Ting-ting,YANG Xiang-hui.Cloning of DXR gene and its expression analysis during fruit ripening in loquat[J].Journal of Fruit Science,2013,30(4):563-566.张玲,林顺权,胡又厘,郑婷婷,杨向晖.枇杷DXR基因的克隆及其在果实成熟过程中的表达分析[J].果树学报,2013,30(4):563-566.
- [10]ZHANG Ling,LIN Shun-quan,YANG Xiang-hui.Extraction of total RNA from loquat fruit and cloning of phytoene synthase gene fragment[J].Journal of Fruit Science,2012,29(4):577-582.张玲,林顺权,杨向晖.枇杷果实总RNA的提取及八氢番茄红素合成酶基因(PSY)片段的克隆[J].果树学报,2012,29(4):577-582.
- [11]LIN Shun-quan.Biotechnology in horticultural plants[M].Beijing:Chinese Agricultural Press,2008:55-56.林顺权主编.园艺植物生物技术[M].北京:中国农业出版社,2008:55-56.
- [12]SHAO Ju-fang,ZHU Hong-wei,YU Wen-feng,ZHANG Wei.Study on growth promotion and browning prevention of ginkgo callus with antioxidant and sorbent[J].Jour of Fujian Forestry Sci and Tech,2009,36(3):117-122.邵菊芳,朱红威,于文峰,张伟.抗氧化剂和吸附剂促进银杏愈伤组织生长及防止褐化作用研究[J].福建林业科技,2009,36(3):117-122.
- [13]ZHANG Zhi-ke,CHEN Bin,YANG Feng-ling,LIN Shun-quan.Preliminary studies on germplasm preservation in vitro of Eriobotrya[J].Fujian Fruits,2008(3):28-31.张志珂,陈彬,杨凤玲,林顺权.枇杷野生种的种质资源离体保存初探[J].福建果树,2008(3):28-31.
- [14]ZHOU Jun-hui,ZHOU Jia-rong,CENG Hao-sen,WANG Guo-bin,ZHU Zhan-ping.Advance of studies on browning and antibrowning techniques in the tissue culture of horticultural plants[J].Acta Horticulturae Sinica,2000,27(1):481-486.周俊辉,周家容,曾浩森,王国彬,祝展平.园艺植物组织培养中的褐化现象及抗褐化研究进展[J].园艺学报,2000,27(1):481-486.
- [15]ZOU Ying-ning,WU Qiang-sheng.Research progress in tissue culture of fruit browning and its suppression[J].Journal of Yangtze University:Natural Science Edition,Agricultural Science Volume,2007(3):90-94.邹英宁,吴强盛.果树组织培养中褐变现象及其抑制研究进展[J].长江大学学报:自科版农学卷,2007(3):90-94.