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Propagation and breeding of Lilium spp. by means of tissue, organ and cell suspension culture

百合属植物的繁殖和育种

基本信息

批准号：
09460017
负责人：
NIIMI Yoshiji
金额：
$ 7.55万
依托单位：
Niigata University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B).
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 2000
项目状态：
已结题

项目摘要

(1) With a few exceptions, more bulblets were regenerated at 25℃ than 15℃, and the regenerated bulblets grew well under light condition at 25℃.(2) All plantlets regenerated in anther culture of L.× 'Enchantment' were diploid, and ca. 40% of the plantlets were virus-free as shown in the calli-induced bulblets of L.longiflorum 'Georgia'.(3) Liquid medium and renewal of the medium 8-12 weeks after culture promoted growth of L.rubellum bulblets.(4) Treatmnent of 0.25 and 0.5 mM colchicine for 48 or 72 hrs induced double haploid cells and shoots from haploid calli of L.×'Connecticut King'. (5) A hap loid callus line from anther cultures of L.×'Connecticut King' was maintained for a long term in MS medium with picloram.(6) A viability of microspores was maintained only in a mediumn with maltose, in which cell divisions and callus formation occurred.(7) In reciprocal pollination between L.nobilissimum and L.regale, ovules excised at 30 and 40 DPA developed into seedlings, whereas those excised at 40 DAP developed into seedlings in L.nobilissimum x L.regale. In L.× 'Connecticut King' x L.× 'Enchantment', ovules excised at 3 DAP developed into seedlings, when embryo culture was used together.(8) Application of nitrous oxide (NOx) to pollen mother cells at the middle stage of the first division was effective to the production of non-reduced microspores. Treatment of 0.5 mM colchicine on in vitro cultured bulbscales for 3 or 5 days induced tetraploid plantlets.(9) Almost all cell clumps from the cell suspension cultures of L.formosanum maintained regeneration potential for more than 54 months in hormone-free medium and developed many somatic embryos.(10) Hygromycin, G418 and bialaphos were suitable agents for selecting transformed calli and/or organs of L.formosanu.(11) Some calli of L.formosanum, co-cultivated with Agrobacterium tumefaciens, showed several blue spots resulted from transient or stable expression of the gusA gene.

(1)25℃条件下再生的小鳞茎数多于15℃条件下，且25℃光照条件下再生的小鳞茎生长良好。(2)L.×‘Enchantment’花药培养再生植株均为二倍体，约为100%。40%的试管苗是无病毒的，如在百合'Georgia'的Calli诱导的小鳞茎中所示。(3)液体培养基和培养8-12周后更新培养基有利于红花百合鳞茎的生长。(4)用0.25和0.5mM秋水仙素处理48或72小时，从‘康涅狄格王’百合单倍体愈伤组织中诱导出双单倍体细胞和芽。(5)从‘康乃狄克王’百合花药培养中获得的单倍体愈伤组织系，在MS+毒莠定培养基上长期保持。(6)小孢子只有在麦芽糖培养基中才能保持活力，细胞分裂和愈伤组织形成。(7)在百合与帝王百合的相互授粉中，在30和40 DPA切除的胚珠发育成幼苗，而在40 DAP切除的胚珠发育成幼苗。在L.×‘Connecticut King’× L.×‘Enchantment’中，当胚培养与离体培养同时进行时，在第3天切下的胚珠发育成幼苗。(8)在第一次分裂中期对花粉母细胞施加一氧化二氮（NOx），对非还原小孢子的产生是有效的。用0.5mM秋水仙素处理离体培养的鳞茎3或5天，诱导出四倍体植株。(9)几乎所有的台湾乳杆菌悬浮培养细胞团在无菌培养基中保持再生能力超过54个月，并发育出许多体细胞胚。(10)潮霉素、G418和双丙氨磷是筛选台湾乳杆菌转化愈伤组织和/或器官的合适试剂。(11)在台湾乳杆菌与农杆菌共培养的愈伤组织中，gusA基因的瞬时表达或稳定表达使愈伤组织出现蓝色斑点。