朱顶红幼嫩花器官离体诱导试管小鳞茎的研究  

A Study on Bulblets Induced from Immature Floral Organs of Hippeastrum hybridum In vitro

作　　者：于波[1] 黄丽丽[1] 孙映波[1] 朱根发[1] 

机构地区：[1]广东省农业科学院环境园艺研究所,广东省园林花卉种质创新综合利用重点实验室,广州510640

出　　处：《农业科技与信息（现代园林）》2015年第4期286-287,共2页Modern Landscape Architecture

基　　金：广东省重点实验室建设支撑项"石蒜科重要花卉资源收集;鉴评与关键产业技术研究";编号2012A061100007

摘　　要：以荷兰进口大花朱顶红品种"美丽参半"(Half&Half)盆栽植株为材料,对幼嫩的子房和小花梗进行体外培养,建立了高效的植株再生体系。试验于2014年2月份采集刚刚抽薹而花苞尚未打开时的花蕾作为外植体。简化了表面消毒程序:在超净台中将包裹着外苞片的花蕾先用70%乙醇浸泡10min,然后用无菌水清洗3次即可。先去除外苞片,然后在无菌滤纸上用手术刀将小花梗和子房切片,厚度约1 mm,平放于诱导培养基上进行黑暗培养,温度25±1℃。在TDZ(0.25、0.5、1.0或2.0mg·L-1)+2,4-D1.0mg·L-1系列培养基上的比较试验后,结果表明:小花梗不定芽诱导适宜培养基为MS+0.5mg·L-1TDZ+1.0mg·L-12,4-D+30g·L-1蔗糖,p H5.8;子房不定芽诱导适宜培养基为MS+1.0mg·L-1TDZ+1.0mg·L-12,4-D+30g·L-1蔗糖,p H5.8。培养过程中,小花梗和子房的外表面陆续形成突状物,8周后形成不定芽,不定芽诱导率均达100%。为降低生长调节剂对试管鳞茎生根的影响,将带有不定芽的外植体转移至MS+蔗糖30g·L-1+琼脂8.0g·L-1(p H6.0)培养基上培养,温度25±1℃,光照强度54μmol·m-2·s-1,光周期为光照14h/黑暗10h。8周后,每个小花梗外植体平均可形成11.0个小鳞茎,每个子房外植体平均可形成9.3个小鳞茎。利用扫描电镜技术观察小鳞茎淀粉粒分布和形态特征;同时利用组织切片(苏木精和碘-碘化钾复染)对不同层次的小鳞茎淀粉分布及特征进行分析,结果表明:淀粉粒集中分布于鳞茎外层细胞,内层细胞较少;成熟淀粉粒呈圆球形,直径约25μm,表面较光滑。本研究为朱顶红种球生产提供支持,并为其鳞茎膨大机理(鳞茎和叶片的"库"和"源"功能)研究提供离体模型。This study established plant regeneration system of Hippeastrum hybridum through bulblets induced from tender ovaries and peduncles In vitro. The results suggested that the most optimal conditions of cultivation included： sterilizing explants by soaking in 70% ethyl alcohol for 10min; adventitious buds from tender peduncles were induced in Murashige and Skoog（MS） medium supplemented with 0.5mg·L-1N-phenyl-N＇-1,2,3-thiadiazol-5-ylurea（TDZ）, 1.0mg·L-12,4-Dichlorophenoxyacetic acid（2,4-D）, and 30g·L-1 sucrose（ p H 6.0）; adventitious buds from ovaries were induced in MS medium supplemented with 1.0mg·L-1 TDZ and 1.0mg·L-1 2,4-D, and 30g·L-1 sucrose（ p H 6.0）. Adventitious buds could root and further expand into bulblets in culture medium without growth regulators. Under these circumstances, 11.0 bulblets were formed from each tender peduncle, and 9.3 for each ovary in average. Through scanning electron microscope and histological section technique, moreover, the distribution and morphological characteristics of starch grains in bulblets were analyzed. The results showed that starch grains centrally distributed in outer bulblet cells but fewer in inner cells; mature starch grains shaped in spherosomes with a diameter in 25μm and smoother surfaces. These results provided good technique supports for effecient production of bulblets with tissue culture, as well as in studying the bulblets development and expanding mechanism in Hippeastrum hybridum.

关 键 词：美朱顶红 花器官 试管小鳞茎 再生 

分 类 号：S682.36[农业科学—观赏园艺]