Chinese Bulletin of Botany ›› 2015, Vol. 50 ›› Issue (3): 378-387.DOI: 10.3724/SP.J.1259.2015.00000
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Suxia Yuan, Jia Li, Jun Ming*, Chun Liu, Leifeng Xu, Yingying Yuan
Received:
2014-05-13
Accepted:
2014-08-25
Online:
2015-05-01
Published:
2015-04-08
Contact:
Ming Jun
About author:
? These authors contributed equally to this paper
Suxia Yuan, Jia Li, Jun Ming, Chun Liu, Leifeng Xu, Yingying Yuan. Embryogenesis and Plant Regeneration from Unpollinated Ovary Culture of Lily[J]. Chinese Bulletin of Botany, 2015, 50(3): 378-387.
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URL: https://www.chinbullbotany.com/EN/10.3724/SP.J.1259.2015.00000
Genotype | Medium | Number of the expand ovules per bud (means±SD) | Number of ovules with embryo per bud (means±SD) | Rate of embryogenic ovules (%) |
---|---|---|---|---|
Siberia | BDS | 132.7 ± 7.6 a | 108.7 ± 5.9 a | 81.9 |
CBM | 119.0 ± 16.4 ab | 94.0 ± 12.8 ab | 79.0 | |
MS | 101.3 ± 3.5 b | 82.0 ± 3.0 b | 80.9 | |
Sorbonne | BDS | 136.3 ± 24.6 a | 116.0 ± 20.8 a | 85.1 |
CBM | 88.7 ± 4.7 b | 71.7 ± 3.8 b | 80.8 | |
MS | 82.3 ± 5.7 b | 70.0 ± 4.6 b | 85.1 | |
Conca'dor | BDS | 150.0 ± 11.1 a | 127.3 ± 9.6 a | 84.9 |
CBM | 136.3 ± 13.7 ab | 105.0 ± 10.4 ab | 77.0 | |
MS | 113.0 ± 18.5 b | 91.7 ± 14.7 b | 81.2 | |
Robina | BDS | 235.3 ± 6.0 a | 223.7 ± 5.5 a | 95.1 |
CBM | 147.7 ± 10.4 b | 118.3 ± 8.3 c | 80.1 | |
MS | 182.7 ± 19.4 c | 157.0 ± 16.5 Bb | 85.9 | |
Yellow Ween | BDS | 184.3 ± 6.0 a | 169.3 ± 6.0 a | 91.9 |
CBM | 171.7 ± 5.7 b | 152.7 ± 4.7 ab | 88.9 | |
MS | 161.0 ± 12.1 c | 137.0 ± 10.1 b | 85.1 | |
Ceb Dazzle | BDS | 242.3 ± 8.3 a | 201.0 ± 7.2 a | 83.0 |
CBM | 131.7 ± 8.6 b | 92.3 ± 6.1 c | 70.1 | |
MS | 172.0 ± 12.1 c | 137.7 ± 9.6 b | 80.0 | |
Lilium regale | BDS | 175.0 ± 15.5 a | 152.0 ± 13.5 a | 86.9 |
CBM | 108.3 ± 5.5 b | 82.3 ± 4.5 c | 76.0 | |
MS | 131.0 ± 9.2 c | 119.0 ± 8.2 b | 90.8 |
Table 1 Induction of embryogenic ovules via lily gynogenesis in different mediums
Genotype | Medium | Number of the expand ovules per bud (means±SD) | Number of ovules with embryo per bud (means±SD) | Rate of embryogenic ovules (%) |
---|---|---|---|---|
Siberia | BDS | 132.7 ± 7.6 a | 108.7 ± 5.9 a | 81.9 |
CBM | 119.0 ± 16.4 ab | 94.0 ± 12.8 ab | 79.0 | |
MS | 101.3 ± 3.5 b | 82.0 ± 3.0 b | 80.9 | |
Sorbonne | BDS | 136.3 ± 24.6 a | 116.0 ± 20.8 a | 85.1 |
CBM | 88.7 ± 4.7 b | 71.7 ± 3.8 b | 80.8 | |
MS | 82.3 ± 5.7 b | 70.0 ± 4.6 b | 85.1 | |
Conca'dor | BDS | 150.0 ± 11.1 a | 127.3 ± 9.6 a | 84.9 |
CBM | 136.3 ± 13.7 ab | 105.0 ± 10.4 ab | 77.0 | |
MS | 113.0 ± 18.5 b | 91.7 ± 14.7 b | 81.2 | |
Robina | BDS | 235.3 ± 6.0 a | 223.7 ± 5.5 a | 95.1 |
CBM | 147.7 ± 10.4 b | 118.3 ± 8.3 c | 80.1 | |
MS | 182.7 ± 19.4 c | 157.0 ± 16.5 Bb | 85.9 | |
Yellow Ween | BDS | 184.3 ± 6.0 a | 169.3 ± 6.0 a | 91.9 |
CBM | 171.7 ± 5.7 b | 152.7 ± 4.7 ab | 88.9 | |
MS | 161.0 ± 12.1 c | 137.0 ± 10.1 b | 85.1 | |
Ceb Dazzle | BDS | 242.3 ± 8.3 a | 201.0 ± 7.2 a | 83.0 |
CBM | 131.7 ± 8.6 b | 92.3 ± 6.1 c | 70.1 | |
MS | 172.0 ± 12.1 c | 137.7 ± 9.6 b | 80.0 | |
Lilium regale | BDS | 175.0 ± 15.5 a | 152.0 ± 13.5 a | 86.9 |
CBM | 108.3 ± 5.5 b | 82.3 ± 4.5 c | 76.0 | |
MS | 131.0 ± 9.2 c | 119.0 ± 8.2 b | 90.8 |
Genotype | Bud developmental stage | Number of the expand ovules per bud (means±SD) | Number of ovules with embryo per bud (means±SD) | Rate of embryo- genic ovules (%) |
---|---|---|---|---|
Siberia | 6 cm | 97.3 ± 5.1 c | 59.7 ± 3.1 c | 61.4 |
8 cm | 116.0 ± 8.9 b | 92.7 ± 7.4 b | 79.9 | |
1 d before anthesis (10 cm) | 132.3 ± 7.4 a | 108.3 ± 6.4 a | 81.9 | |
3 d after anthesis | 122.0 ± 6.6 ab | 104.0 ± 5.6 ab | 85.2 | |
Sorbonne | 6 cm | 87.7 ± 4.0 b | 58.7 ± 3.1 b | 66.9 |
8 cm | 110.3 ± 26.0 ab | 87.3 ± 20.4 ab | 79.1 | |
1 d before anthesis (10 cm) | 136.2 ± 24.6 a | 116.0 ± 20.8 a | 85.2 | |
3 d after anthesis | 122.3 ± 14.4 ab | 101.3 ± 11.7 a | 82.8 | |
Conca'dor | 7 cm | 152.7 ± 8.1 b | 119.0 ± 6.1 a | 77.9 |
9 cm | 124.7 ± 13.4 ab | 90.0 ± 9.6 b | 72.2 | |
1 d before anthesis (11 cm) | 148.0 ± 11.5 a | 126.0 ± 9.5 a | 85.1 | |
3 d after anthesis | 132.3 ± 11.7 a | 110.0 ± 9.6 a | 83.1 | |
Robina | 7 cm | 164.3 ± 7.1 b | 125.0 ± 5.6 d | 76.1 |
9 cm | 225.0 ± 8.7 a | 189.0 ± 7.0 b | 84.0 | |
1 d before anthesis (11 cm) | 234.3 ± 10.1 a | 223.0 ± 9.5 a | 95.2 | |
3 d after anthesis | 179.0 ± 19.2 b | 161.0 ± 17.4 c | 89.9 | |
Yellow Ween | 6 cm | 115.0 ± 4.6 c | 91.0 ± 3.6 c | 79.1 |
8 cm | 152.0 ± 5.0 b | 126.0 ± 4.0 b | 82.9 | |
1 d before anthesis (10 cm) | 213.7 ± 12.1 a | 196.7 ± 11.1 a | 92.0 | |
3 d after anthesis | 97.7 ± 4.7 d | 87.0 ± 4.4 d | 89.0 | |
Ceb Dazzle | 5 cm | 104.0 ± 11.0 c | 77.0 ± 8.0 c | 74.0 |
7 cm | 125.3 ± 7.5 bc | 99.0 ± 6.0 bc | 79.0 | |
1 d before anthesis (9 cm) | 242.0 ± 19.5 a | 201.0 ± 16.5 a | 83.1 | |
3 d after anthesis | 148.7 ± 20.2 b | 122.0 ± 16.7 b | 82.0 | |
Lilium regale | 6 cm | 164.0 ± 9.0 b | 128.0 ± 7.0 b | 78.0 |
8 cm | 166.7 ± 8.7 b | 135.0 ± 7.2 b | 81.0 | |
1 d before anthesis (10 cm) | 169.7 ± 18.8 b | 148.0 ± 16.5 ab | 87.2 | |
3 d after anthesis | 236.3 ± 28.9 a | 168.0 ± 20.8 a | 71.1 |
Table 2 Embryo production of different lily flower buds in BDS mediums
Genotype | Bud developmental stage | Number of the expand ovules per bud (means±SD) | Number of ovules with embryo per bud (means±SD) | Rate of embryo- genic ovules (%) |
---|---|---|---|---|
Siberia | 6 cm | 97.3 ± 5.1 c | 59.7 ± 3.1 c | 61.4 |
8 cm | 116.0 ± 8.9 b | 92.7 ± 7.4 b | 79.9 | |
1 d before anthesis (10 cm) | 132.3 ± 7.4 a | 108.3 ± 6.4 a | 81.9 | |
3 d after anthesis | 122.0 ± 6.6 ab | 104.0 ± 5.6 ab | 85.2 | |
Sorbonne | 6 cm | 87.7 ± 4.0 b | 58.7 ± 3.1 b | 66.9 |
8 cm | 110.3 ± 26.0 ab | 87.3 ± 20.4 ab | 79.1 | |
1 d before anthesis (10 cm) | 136.2 ± 24.6 a | 116.0 ± 20.8 a | 85.2 | |
3 d after anthesis | 122.3 ± 14.4 ab | 101.3 ± 11.7 a | 82.8 | |
Conca'dor | 7 cm | 152.7 ± 8.1 b | 119.0 ± 6.1 a | 77.9 |
9 cm | 124.7 ± 13.4 ab | 90.0 ± 9.6 b | 72.2 | |
1 d before anthesis (11 cm) | 148.0 ± 11.5 a | 126.0 ± 9.5 a | 85.1 | |
3 d after anthesis | 132.3 ± 11.7 a | 110.0 ± 9.6 a | 83.1 | |
Robina | 7 cm | 164.3 ± 7.1 b | 125.0 ± 5.6 d | 76.1 |
9 cm | 225.0 ± 8.7 a | 189.0 ± 7.0 b | 84.0 | |
1 d before anthesis (11 cm) | 234.3 ± 10.1 a | 223.0 ± 9.5 a | 95.2 | |
3 d after anthesis | 179.0 ± 19.2 b | 161.0 ± 17.4 c | 89.9 | |
Yellow Ween | 6 cm | 115.0 ± 4.6 c | 91.0 ± 3.6 c | 79.1 |
8 cm | 152.0 ± 5.0 b | 126.0 ± 4.0 b | 82.9 | |
1 d before anthesis (10 cm) | 213.7 ± 12.1 a | 196.7 ± 11.1 a | 92.0 | |
3 d after anthesis | 97.7 ± 4.7 d | 87.0 ± 4.4 d | 89.0 | |
Ceb Dazzle | 5 cm | 104.0 ± 11.0 c | 77.0 ± 8.0 c | 74.0 |
7 cm | 125.3 ± 7.5 bc | 99.0 ± 6.0 bc | 79.0 | |
1 d before anthesis (9 cm) | 242.0 ± 19.5 a | 201.0 ± 16.5 a | 83.1 | |
3 d after anthesis | 148.7 ± 20.2 b | 122.0 ± 16.7 b | 82.0 | |
Lilium regale | 6 cm | 164.0 ± 9.0 b | 128.0 ± 7.0 b | 78.0 |
8 cm | 166.7 ± 8.7 b | 135.0 ± 7.2 b | 81.0 | |
1 d before anthesis (10 cm) | 169.7 ± 18.8 b | 148.0 ± 16.5 ab | 87.2 | |
3 d after anthesis | 236.3 ± 28.9 a | 168.0 ± 20.8 a | 71.1 |
Genotype | Treatment | Number of the expand ovules per bud (means±SD) | Number of ovules with embryo per bud (means±SD) | Rate of embryogenic ovules (%) |
---|---|---|---|---|
Siberia | 2 mg·L-1 2,4-D + 2 mg·L-1 6-BA | 164.7 ± 13.2 ab | 133.3 ± 10.7 ab | 80.9 |
4 mg·L-1 2,4-D + 4 mg·L-1 6-BA | 143.7 ± 8.02 b | 112.3 ± 6.5 b | 78.1 | |
2 mg·L-1 2,4-D + 2 mg·L-1 KT | 175.3 ± 12.7 a | 143.7 ± 10.6 a | 82.0 | |
4 mg·L-1 2,4-D + 4 mg·L-1 KT | 158.0 ± 10.5 ab | 131.0 ± 8.5 ab | 82.9 | |
Sorbonne | 2 mg·L-1 2,4-D + 2 mg·L-1 6-BA | 135.3 ± 9.5 a | 104.3 ± 6.8 ab | 77.1 |
4 mg·L-1 2,4-D + 4 mg·L-1 6-BA | 129.0 ± 13.0 a | 98.0 ± 10.0 b | 76.0 | |
2 mg·L-1 2,4-D + 2 mg·L-1 KT | 141.0 ± 12.0 a | 114.3 ± 9.5 a | 81.1 | |
4 mg·L-1 2,4-D + 4 mg·L-1 KT | 131.3 ± 5.5 a | 105.3 ± 4.5 ab | 80.2 |
Table 3 Effect of hormone concentration and proportion on the embryo induction in lily
Genotype | Treatment | Number of the expand ovules per bud (means±SD) | Number of ovules with embryo per bud (means±SD) | Rate of embryogenic ovules (%) |
---|---|---|---|---|
Siberia | 2 mg·L-1 2,4-D + 2 mg·L-1 6-BA | 164.7 ± 13.2 ab | 133.3 ± 10.7 ab | 80.9 |
4 mg·L-1 2,4-D + 4 mg·L-1 6-BA | 143.7 ± 8.02 b | 112.3 ± 6.5 b | 78.1 | |
2 mg·L-1 2,4-D + 2 mg·L-1 KT | 175.3 ± 12.7 a | 143.7 ± 10.6 a | 82.0 | |
4 mg·L-1 2,4-D + 4 mg·L-1 KT | 158.0 ± 10.5 ab | 131.0 ± 8.5 ab | 82.9 | |
Sorbonne | 2 mg·L-1 2,4-D + 2 mg·L-1 6-BA | 135.3 ± 9.5 a | 104.3 ± 6.8 ab | 77.1 |
4 mg·L-1 2,4-D + 4 mg·L-1 6-BA | 129.0 ± 13.0 a | 98.0 ± 10.0 b | 76.0 | |
2 mg·L-1 2,4-D + 2 mg·L-1 KT | 141.0 ± 12.0 a | 114.3 ± 9.5 a | 81.1 | |
4 mg·L-1 2,4-D + 4 mg·L-1 KT | 131.3 ± 5.5 a | 105.3 ± 4.5 ab | 80.2 |
Figure 1 Gynogenic embryogenesis in lily (A) Globular embryo on micropylar end; (B) Globular embryo on polar nucleus position; (C) Globular embryo on zygote end; (D) Cotyledonary embryo; (E) Clavate embryo; (F) Ovule with two embryos (on micropylar end and polar nucleus position, respectively); (G) Ovule with two embryos (on micropylar end and zygote end, respectively); (H) Embryogenic ovule (a) and ovule without embryo (b)
Genotype | Number of population plants | Haploid | Diploid | Triploid | Tetraploid | Aneuploid |
---|---|---|---|---|---|---|
Sorbonne (2n=2x=24) | 23 | 6 | 10 | 0 | 1 | 6 |
Conca'dor (2n=3x=36) | 39 | 4 | 14 | 9 | 0 | 12 |
Table 4 Chromosome ploidy level of population derived from microspores in Sorbonne and Conca'dor
Genotype | Number of population plants | Haploid | Diploid | Triploid | Tetraploid | Aneuploid |
---|---|---|---|---|---|---|
Sorbonne (2n=2x=24) | 23 | 6 | 10 | 0 | 1 | 6 |
Conca'dor (2n=3x=36) | 39 | 4 | 14 | 9 | 0 | 12 |
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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