百合未授粉子房离体培养胚胎形成及植株再生
? 共同第一作者
收稿日期: 2014-05-13
录用日期: 2014-08-25
网络出版日期: 2015-04-08
基金资助
国家自然科学基金(No.31301804)、国家科技支撑计划(No.2012BAD01B0701)、北京市花卉重点项目(No.YLHH201000101, No.YLHH201400101)和农业部园艺作物生物学与种质创制重点实验室项目
Embryogenesis and Plant Regeneration from Unpollinated Ovary Culture of Lily
? These authors contributed equally to this paper
Received date: 2014-05-13
Accepted date: 2014-08-25
Online published: 2015-04-08
未受精子房离体培养是诱导雌核产生单倍体的技术之一。以1个野生种和3个杂种系共7个百合(Lilium)基因型为实验材料, 探讨了基础培养基、花蕾取样时期和外源激素等因素对百合未授粉子房离体培养胚胎形成的影响。结果表明, CBM、MS和BDS三种基础培养基均可诱导百合未授粉子房胚胎形成, 但以BDS培养基诱导效果最佳; 开花前1天的花蕾较适于百合未授粉子房离体培养; 2 mg·L-1 2,4-D + 2 mg·L-1 6-BA和2 mg·L-1 2,4-D + 2 mg·L-1 KT两种外源激素配方均适用于百合未授粉子房离体培养诱导胚胎形成。在培养过程中, 大多数胚性胚珠中只含有1个胚胎, 位于珠孔端、合子端或极核处, 少数胚性胚珠中含有双胚胎。通过百合未授粉子房离体培养, 从5个基因型材料中共获得146株再生植株。采用根尖染色体计数法对其中的62株进行了倍性测定, 其中43株与母体植株染色体倍性不同。
袁素霞, 李佳, 明军, 刘春, 徐雷锋, 袁迎迎 . 百合未授粉子房离体培养胚胎形成及植株再生[J]. 植物学报, 2015 , 50(3) : 378 -387 . DOI: 10.3724/SP.J.1259.2015.00000
Unfertilized ovary or ovule culture is an effective technology for producing homozygous materials (doubled haploids) via gynogenesis. We studied the effects of basal medium and bud developmental stage on the genogenesis in seven lily genotypes, including one wild species and six cultivars. Gynogenesis occurred in all genotypes, but BDS was the optimal basal medium as compared with CBM and MS. When ovary sections from buds collected 1 d before anthesis were cultured in BDS medium, more embryogenic ovules or responding ovules were obtained. To estimate the effect of phytohormones on lily gynogenic embryogenesis, two genotypes were tested. An amount of 2 mg·L-1 2,4-dichloro- phenoxyacetic acid (2,4-D) and 2 mg·L-1 6-benzylaminopurine (6-BA) or kinetin (KT) was suitable for embryo induction from unfertilized ovaries. During culture, embryo development was observed by the cleaning-squash technique; most ovules contained one embryo, which lay on the micropylar end, zygote end or polar nucleus position. In addition, a few ovules with two embryos were observed. In total, 146 regenerants were obtained from unpollinated ovary culture of five genotypes; for 62, the ploidy level was determined by chromosome counting. We found that 43 plantlets had a chromosome number different from maternal plants.
1 | 谷祝平, 郑国锠 (1983). 百合未授粉子房的培养及其胚胎学观察. 植物学报 25, 24-30. |
2 | 韩秀丽, 田晓明, 贾桂霞 (2010). 新铁炮百合单倍体植株的诱导. 园艺学报 37, 263-268. |
3 | 李佳, 袁素霞, 刘春, 田梅, 于娜丽 (2012). 影响百合游离小孢子活力与发育因素分析. 北方园艺 (3), 59-63. |
4 | 李克虎, 周桂雪, 任贵玲, 张线线, 郭方其, 周树军 (2011). 百合品种染色体倍性观察. 园艺学报 38, 970-976. |
5 | 赵珺 (2007). 百合未授粉子房离体培养及胚胎学研究. 硕士论文. 沈阳: 沈阳农业大学. pp. 1-2. |
6 | 褚云霞, 黄燕文, 陈龙清, 张永春 (2001). 百合的花药培养研究. 园艺学报 28, 472-474. |
7 | Bhat JG, Murthy HN (2007). Factors affecting in-vitro gynogenic haploid production in niger (Guizotia abyssinica (L. f.) Cass.).Plant Growth Regul 52, 241-248. |
8 | Bohanec B (2009). Doubled haploids via gynogenesis. In: Touraev A, Forster BP, Moha S, eds. Advances in Haploid Production in Higher Plants. United Kingdom: Springer. pp. 35-46. |
9 | Bohanec B, Jakše M, Havey MJ (2003). Genetic analyses of gynogenetic haploid induction in onion.J Amer Soc Hort Sci 128, 571-574. |
10 | Ferrant V, Bouharmont J (1994). Origin of gynogenetic embryos of Beta vulgaris L.Sex Plant Rep 7, 12-16. |
11 | Ferrie AMR, Caswell KL (2011). Isolated microspore culture techniques and recent progress for haploid and doubled haploid plant production.Plant Cell Tissue Org Cult 104, 301-309. |
12 | Forster BP, Heberle-Bors E, Kasha KJ, Touraev A (2007). The resurgence of haploids in higher plants.Trends Plant Sci 12, 368-375. |
13 | Gelebart P, San LH, Martin MM (1987). Obtention de plantes haploïdes par culture in vitro d'ovaires et d'ovules non fécondés de tournesol (Helianthus annuus L.).Agronomie 7, 81-86. |
14 | Gémes-Juhász A, Balogh P, Ferenczy A, Kristóf Z (2002). Effect of optimal stage of female gametophyte and heat treatment on in vitro gynogenesis induction in cucumber (Cucumis sativus L.).Plant Cell Rep 21, 105-111. |
15 | Han DS, Niimi Y, Nakano M (1997). Regeneration of haploid plants from anther cultures of the Asiatic hybrid lily ‘Connecticut King’.Plant Cell Tissue Org Cult 47, 153-158. |
16 | Han DS, Niimi Y, Nakano M (2000). Formation of calli from isolated microspore cultures of Asiatic hybrid lily ‘Connecticut King’.J Japan Soc Hort Sci 69, 52-56. |
17 | Huang QF, Yang HY, Zhou C (1982). Embryological observations on ovary culture of unpollinated young flowers in Hordeum vulgare L.Acta Bot Sin 24, 295-300. |
18 | Murashige T, Skoog F (1962). A revised medium for rapid growth and bio assays with tobacco tissue cultures.Plant Physiol 15, 473-497. |
19 | Muren RC (1989). Haploid plant induction from unpollinated ovaries in onion.HortScience 24, 833-834. |
20 | Musial K, Bohanec B, Jakše M, Przywara L (2005). The development of onion (Allium cepa L.) embryo sacs in vitro and gynogenesis induction in relation to flower size.In vitro Cell Dev Biol Plant 41, 446-452. |
21 | Musial K, Bohanec B, Przywara L (2001). Embryological study on gynogenesis in onion (Allium cepa L.).Sex Plant Rep 13, 335-341. |
22 | Prakash J, Giles KL (1986). Production of doubled haploids in oriental lilies. In: Horn W, Jensen CJ, Odenbach W, Schieder O, eds. Genetic Manipulation in Plant Breeding. Berlin: Walter de Gruyter. pp. 335-337. |
23 | Ramsay JL, Galitz DS, Lee CW (2003). Basal medium and sucrose concentration influence regeneration of Easter Lily in ovary culture.HortScience 38, 404-406. |
24 | San Noeum LH (1979). In vitro induction of gynogenesis in higher plants. In: Zeven AC, van Harten AM, eds. Broadening Genetic Base of Crops. The Netherlands: Kluwer Academic Publishers. pp. 327-329. |
25 | Sharp WR, Raskin RS, Sommer HE (1971). Haploidy in Lilium.Phytornorphology 21, 334-335. |
26 | Tanaka I, Ito M (1980). Induction of typical cell division in isolated microspores of Lilium longiflorum and Tulipa gesneriana.Plant Sci Lett 17, 279-285. |
27 | Tanaka I, Ito M (1981). Studies on microspore development in liliaceous plants III. Pollen tube development in lily pollens cultured from the uninucleate microspore stage.Plant Cell Physiol 22, 149-153. |
28 | Van de Bulk RW, de Vries-van Hulten HPJ, Dons JJM (1992). Formation of multinucleate lily microspores in the culture.Acta Hort 325, 649-654. |
29 | Wu BJ, Chen KC (1982). Cytological and embryological studies on haploid plant production from cultured unpollinated ovaries of Nicotiana tabacum L.Acta Bot Sin 24, 125-129. |
30 | Yang H, Zhou C, Cai D, Yan H, Wu Y, Chen X (1986). In vitro culture of unfertilized ovules in Helianthus annuus L. In: Hu H, Yang H, eds. Haploids of Higher Plants In Vitro. Berlin: Springer. pp. 182-191. |
31 | Zhou C, Yang H, Tian H, Liu Z, Yan H (1986). In vitro culture of unpollinated ovaries in Oryza sativa. In: Hu H, Yang H, eds. Haploids of Higher Plants In Vitro. Berlin: Springer. pp. 167-181. |
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