Abstract:
Doubled haploids (DHs) production through androgenesis is a biotechnological method for genetic improvement of crops. Biotechnological DH line production offers advantages to plant breeders, including the possibility to obtain homozygous lines within a year in contrast to common inbreeding methods, which may take 6–12 years. The greatest success in androgenesis has been achieved in some varieties of rapeseed (Brassica napus L.). However, the efficiency of androgenesis in other Brassica species is still poor. Induction of microspore embryogenesis is usually induced by many factors such as conditions of donor plant growth, genotype, microspore developmental stage, culture medium composition, and culture conditions. The reprogramming of microspores from the gametophytic to the sporophytic habit of development depends on various stress factors. Certain pretreatments of microspores, such as high temperature and colchicine, can favor androgenesis in Brassica species. Plant regeneration from microspores can be improved by proper application of different growth regulators (ethylene, abscisic acid, and indole acetic acid). Optimal combinations of these factors are mandatory for efficient androgenesis. In this review, we summarize the experience of our colleagues in DH-technology in the Brassica genus. Attention is focused on some factors influencing the development of doubled haploid plants and their impact on enhancing the efficiency of androgenesis in Brassica species.
About The Authors:
N. A. Shmykova. All-Russia Research Institute of Vegetable Breeding and Seed Production, VNIISSOK village, Odintsovo district, Moscow region, Russia, Russian Federation
References:
1. Bunin M.S., Shmykova N.A., Stepanov V.A. Metody reproduktivnoĭ biologii v selektsii ovoshchnykh kul’tur roda Brassica L. M.: VO Minsel’khoza Rossii, 2003:3-21.
2. Zubareva I.A., Goloveshkina E.N., Vinogradova S.V., Gribova T.N., Monakhos S.G., Ignatov A.N. Sozdanie digaploidnykh liniĭ Brassica napus L. — donorov ustoĭchivosti k virusu mozaiki turnepsa. S.-kh. biologiya. 2013;5:122-124.
3. Kalashnikova E.A., Chung M.D., Solov’ev A.A., Shevelukha V.S. Tekhnologiya polucheniya digaploidnykh rasteniĭ roda Brassica in vitro. Dokl. RASKhN. 2011;1:5-8.
4. Shmykova N.A. Razrabotka sistemy biotekhnologicheskikh metodov, napravlennykh na uskorenie selektsionnogo protsessa ovoshchnykh kul’tur: Avtoref. dis. … d-ra s.-kh. nauk. M., 2006.
5. Agarwal P.K., Agarwal P., Custers J.B.M., Liu C., Bhojwan S.S. PCIB an antiauxin enhances microspore embryogenesis in microspore culture of Brassica juncea. Plant Cell Tiss. Organ Cult. 2006;86:201-210. DOI10.1007/s11240-006-9108-0.
6. Ahmadi B., Alizadeh K., da Silva J.A.T. Enhanced regeneration of haploid plantlets from microspores of Brassica napus L. using bleomycin, PCIB, and phytohormones. Plant Cell Tiss. Organ Cult. 2012;109:525-533. DOI 10.1007/s11240-012-0119-8.
7. Baillie A.M.R., Epp D.J., Hutcheson D., Keller W.A. In vitro culture of isolated microspores and regeneration of plants in Brassica campestris. Plant Cell Rep. 1992;11:234-237.
8. Barbulescu D.M., Burton W.A., Salisbury P.A. Pluronic F-68: an answer for shoot regeneration recalcitrance in microspore-derived Brassica napus embryos. In Vitro Cell Dev. Biol. Plant. 2011;47:282-288. DOI10.1007/s11627-011-9353-8
9. Belmonte M., Elhiti M., Waldner B., Stasolla C. Depletion of cellular brassinolide decreases embryo production and disrupts the architecture of the apical meristems in Brassica napus microspore-derived embryos. J. Exp. Bot. 2010;61:2779-2794. DOI: 10.1093/jxb/erq110
10. Bhowmik P., Dirpaul J., Polowick P., Ferrie A.M.R. A high throughput Brassica napus microspore culture system: influence of percoll gradient separation and bud selection on embryogenesis. Plant Cell Tissue Org. Cult. 2011;106:359-362. DOI: 10.1007/s11240010-9913-3
11. Binarova P., Straatman K., Hause B., Hause G., van Lammeren A.A.M. Nuclear DNA synthesis during the induction of embryogenesis in cultured microspores and pollen of Brassica napus L. Theor. Appl. Genet. 1993;87:9-16.
12. Cao M.Q., Li Y., Liu F., Dore C. Embryogenesis and plant regeneration of pakchoi (Brassica rapa L. ssp. chinensis) via in vitro isolated microspore culture. Plant Cell Rep. 1994;13:447-450.
13. Cegielska-Taras T., Tykarska T., Szala T., Kuras M., Krzymanski J. Direct plant development from microspore-derived embryos of winter oilseed rape Brassica napus L. ssp. oleifera (DC.) Metzger. Euphytica. 2002;124:341-347.
14. Chanana N.P., DhawanV.V., Bhojwani S.S. Morphogenesis in isolated microspore cultures of Brassica juncea. Plant Cell Tiss. Organ Cult. 2005;83:169-177. DOI: 10.1007/s11240-005-4855-x
15. Charne D.G., Beversdorf W.D. Improving microspore culture as a rapeseed breeding tool: the use of auxins and cytokinins in an induction medium. Can. J. Bot. 1988;66:1671-1675.
16. Chen J.T., Chang W.C. 1-Aminocyclopropane-1-carboxylic acid enhanced direct somatic embryogenesis from Oncidium leaf cultures. Biol. Plant. 2003;46:455-458.
17. Chun C., Park H., Na H. Microspore-derived embryo formation in radish (Raphanus sativus L.) according to national and environ mental conditions. Hort. Environ. Biotechnol. 2011;52:530-535. DOI: 10.1007/s13580-011-0080-1
18. Cordewener J.H.G., Hause G., Gorgen E., Busink R., Hause B., Hans J.M., Dons H.J.M., van Lammeren A.A.M., van Lookeren Campagne M.M., Pechan P. Changes in synthesis and localization of members of the 70-kDa class of heat-shock proteins accompany the induction of embryogenesis in Brassica napus L. microspores. Planta. 1995;196:747-755.
19. Custers J.B.M., Cordewener J.H.G, Nollen Y., Dons H.J.M., van Lookeren Campagne M.M. Temperature controls both gametophytic and sporophytic development in microspore cultures of
20. Brassica napus. Plant Cell Rep. 1994;13:267-271.
21. Dias J.C.S. Effect of activated charcoal on Brassica oleracea microspore culture embryogenesis. Euphytica. 1999;108:65-69.
22. Dias J.C.S. Effect of incubation temperature regimes and culture medium on broccoli microspore culture embryogenesis. Euphytica. 2001;119:389-394.
23. Dias J.C.S., Martins M.G. Effect of silver nitrate on anther culture embryo production of different Brassica oleracea morphotypes. Sci. Hort. 1999;82:299-307.
24. Duijs J.C., Voorrips R.E., Visser D.L., Custers J.B.M. Microspore culture is successful in most crop types of Brassica oleracea L. Euphytica. 1992;60:45-55.
25. Dunwell J.M. Haploids in flowering plants: origins and exploitation. Plant Biotechnol. J. 2010;8:377-424. DOI: 10.1111/j.1467-7652.2009.00498.x
26. Dunwell J.M., Cornish M., De Courcel A.G.L. Influence of genotype, plant growth temperature and anther incubation temperature on microspore embryo production in Brassica napus ssp. oleifera. J. Exp. Bot. 1985;36:679-689.
27. Feher A., Pasternak T.P., Dudits D. Transition of somatic plant cells to an embryogenic state. Plant Cell Tissue Organ Cult. 2003;74: 201-228.
28. Ferrie A.M.R. Microspore culture of Brassica species. Doubled haploid production in crop plants (Ed. M. Maluszynski et al.). Kluwer, Dordrecht. 2003:205-215.
29. Ferrie A.M.R., Dirpaul J., Krishna P., Krochko J., Keller W.A. Effects of brassinosteroids on microspore embryogenesis in Brassica species. In Vitro Cell Dev. Biol. Plant. 2005;41:742-745. DOI:10.1079/IVP2005690
30. Ferrie A.M.R., Taylor D.C., MacKenzie S.L., Keller W.A. Microspore embryogenesis of high sn-2 erucic acid Brassica oleracea germplasm. Plant Cell Tissue Organ Cult. 1999;57:79-84.
31. Fridborg G., Pedersen M., Landstrom A., Eriksson T. The effect of charcoal on tissue cultures: adsorption of metabolites inhibiting morphogenesis. Physiol Plant. 1978;43:104-106.
32. Gland F., Lichter R., Schweiger H.G. Genetic and exogenous factors affecting embryogenesis in microspore cultures of Brassica napus L. J. Plant Physiol. 1988;132:613-617.
33. Goldberg R.B., Beals T.P., Sanders P.M. Anther development: basic principles and practical applications. Plant Cell. 1993;5: 1217-1229.
34. Gu H.H., Hagberg P., Zhou W.J. Cold pretreatment enhances microspore embryogenesis in oilseed rape (Brassica napus L.). Plant Growth Regul. 2004;42:137-143.
35. Guo Y.D., Pulli S. High-frequency embryogenesis in Brassica campestris microspore culture. Plant Cell Tiss Org Cult. 1996;46: 219-225.
36. Halkjaer M., Ringgaard S. Microspore culture in B. oleracea L. COST-824: Gametic Embryogenesis Workshop, Book of Abstracts. Sjusjoen, Norway, 1997.
37. Hause G., Hause P., Pehan P., van Lammeren A.A.M. Cytoskeleton changes and induction of embryogenesis in microspore and pollen cultures of Brassica napus L. Cell Biol. Int. 1993;17:153-166.
38. Hays D.B. Mandel R.M., Pharis R.P. Hormones in zugotic and microspore embryos of Brassica napus. Plant Growth Regul. 2001;35:47-58.
39. Hays D.B., Reid D.M., Yeung E.C., Pharis R.P. Role of ethylene in cotyledon development of microspore-derived embryos of Brassica napus. J. Exp. Bot. 2000;51:1851-1859. DOI: 10.1093/jexbot/51.352.1851
40. Huang B., Bird S., Kemble R., Miki B., Keller W. Plant regeneration from microspore-derived embryos of Brassica napus: effect of embryo age, culture temperature, osmotic pressure, and abscisic acid. In Vitro Cell Dev. Biol. 1991;27:28-31.
41. Huang B., Bird S., Kemble R., Simmonds D., Keller W., Miki B. Effects of culture density, conditioned medium feeder cultures on microspore embryogenesis in Brassica napus L. cv. Topas. Plant Cell Rep. 1990;8:594-597.
42. Ilic-Grubor K., Attree S.M., Fowke L.C. Comparative morphological study of zygotic and microspore-derived embryos of Brassica napus L. as revealed by scanning electron microscopy. Ann. Bot. 1998;82:157-165.
43. Johansson L. Effects of activated charcoal in anther cultures. Physiol. Plant. 1983;59:397-403.
44. Keller W.A., Armstrong K.C. Stimulation of embryogenesis and haploid production in Brassica campestris anther cultures by elevated temperature treatments. Theor. Appl. Genet. 1979;55:65-67.
45. Keller W.A., Armstrong K.C. Production of anther-derived dihaploid plants in autotetraploid marrowstem kale (Brassica oleracea var. acephala). Can. J. Genet. Cytol. 1981;23:259-265.
46. Keller W.A., Armstrong K.C. Production of haploids via anther culture in Brassica oleracea var. italica of Broccoli. Plant Breed. 1983;32:151-159.
47. Kott L., Beversdorf W.D. Enhanced plant regeneration from microspore-derived embryos of Brassica napus by chilling, partial desiccation and age selection. Plant Cell Tiss. Org. Cult. 1990;23:187-192.
48. Kott L.S., Polsoni L., Beversdorf W.D. Cytological aspects of isolated microspore culture in Brassica napus. Can. J. Bot. 1988;66: 1658-1664.
49. Langowska T. Effect of activated carbon and brown coal suspensions for growth and development of Scenedes musobliquus (Sc.449). Pol. Arch. Hydrobiol. 1980;27:125-136.
50. Lee S.S., Kim A.J. Effect of cultural vessel, plant growth regulator, illuminating and shaking on embryo induction and growth in microspore culture of heading Chinese cabbage. J. Kor. Soc. Hort. Sci. 2000;41:16-20.
51. Lichter R. Induction of haploid plants from isolated pollen of Brassica napus. Z. Pflanzenphysiol. 1982;105:427-434.
52. Lichter R. Efficient yield of embryoids by culture of isolated microspores of different Brassicacea species. Plant Breeding. 1989;103:119-123.
53. Lionneton E., Beuret W., Delaitre C., Rancillac M. Improved microspore culture and doubled-haploid plant regeneration in the brown condiment mustard (Brassica juncea). Plant Cell Rep. 2001;20:126-130.
54. Lo K.H., Pauls K.P. Plant growth environment effects on rapeseed microscope development and culture. Plant Physiol. 1992;99: 468-472.
55. Margale E., Chevre A.M. Factors effecting embryo production from microspore culture of Brassica nigra (Koch). Cruciferae Newslett. 1991;14(15):100-101.
56. Mathias R. An improved in vitro procedure from embryoids derived from isolated microspores of rape (Brassica napus L.). Plant Breeding. 1988;100:320-322.
57. Mollers C., Iqbal M.C.M., Robbelen G. Efficient production of doubled haploid Brassica napus plants by colchicines treatment of microspores. Euphytica. 1994;75:95-104.
58. Na H., Kwak J.H., Chun C. The effect of plant growth regulators, activated charcoal and AgNO3 on microspore derived embryo formation in broccoli (Brassica oleracea L. var. italica). Hort. Environ. Biotechnol. 2011;52(5):524-529. DOI: 10.1007/s13580011-0034-7
59. Ockendon D.J. Anther culture in Brussels sprouts (Brassica oleracea var. gemmifera). I. Embryo yields and plant regeneration. Ann. Appl. Biol. 1984;105:285-291.
60. Ockendon D.J. Anther culture in Brussels sprouts (Brassica oleracea var. gemmifera). II. Effect of genotype on embryo yields. Ann. Appl. Biol. 1985;107:101-104.
61. Ockendon D.J. The ploidy of plants obtained from anther culture of cauliflowers (Brassica oleracea var. botrytis). Ann. Appl. Biol. 1988;113:319-325.
62. Orton T.J., Browers M.A. Segregation of genetic markers among plants regenerated from cultured anthers of broccoli (Brassica oleracea var. italica). Theor. Appl. Genet. 1985;69:637-643.
63. Osolnik B., Bohanes B., Jelaska S. Stimulation of androgenesis in white cabbage (Brassica oleracea var. capitata) anthers by low temperature and anther dissection. Plant Cell Tiss. Org. Cult. 1993;32:241-246.
64. Palmer C.E., Keller W.A., Arnison P.G. Experimental haploidy in Brassica species. In vitro haploid production in higher plants (Ed. S.M. Jain et al.). V. 3: Important selected plants. Kluwer. Dordrecht. 1996:143-171.
65. Pechan P.M., Bartels D., Brown D.C.W., Schell J. Messenger-RNA and protein changes associated with induction of Brassica microspore embryogenesis. Planta. 1991;184:161-165.
66. Pechan P.M., Keller W.A. Identification of potentially embryogenic microspores in Brassica napus. Physiol. Plant. 1988;74:377-384.
67. Pechan P.M., Smykal P. Androgenesis: affecting the fate of the male gametophyte. Physiol. Plant. 2001;111:1-8. DOI: 10.1034/j.13993054.2001.1110101.x
68. Prem D., Gupta K., Agnihotri A. Effect of various exogenous and endogenous factors on microspore embryogenesis in Indian mustard (Brassica juncea [L.] Czern & Coss). In vitro Cell Dev. Biol. Plant. 2005;41:266-273. DOI: 10.1079/IVP2005636
69. Prem D., Gupta K., Sarkar G., Agnihotri A. Activated charcoal induced high frequency microspore embryogenesis and efficient doubled haploid production in Brassica juncea. Plant Cell Tiss. Org. Cult. 2008;93:269-282. DOI: 10.1007/s11240-008-9373-1
70. Rudolf K., Bohanec B., Hansen M. Microspore culture of white cabbage, Brassica oleracea var. capitata L.: genetic improvement of non-responsive cultivars and effect of genome doubling agents. Plant Breeding. 1999;118:237-241.
71. Sato S., Katoh N., Iwai S., Hagimori M. Effect of low temperature pretreatment of buds or inflorescence on isolated microspore culture in Brassica rapa (syn. B. campestris). Breed. Sci. 2002;52: 23-26. DOI:10.1270/jsbbs.52.23
72. Simmonds D.H., Keller W.A. Significance of pre-prophase bands of microtubules in the induction of microspore embryogenesis of Brassica napus. Planta. 1999;208:383-391.
73. Smith D.L., Krikorian A.D. Somatic pro-embryo production from excised wounded zygotic carrot embryos on hormone free medium; evaluation of the effects of pH, ethylene and activated charcoal. Plant Cell Rep. 1990;9:34-37.
74. Sopory S., Munshi M. Anther culture. In vitro haploid production in higher plants (Ed. J.M. Mohan et al.). Dordrecht: Kluwer, 1996;1:145-176.
75. Takahata Y., Keller W.A. High frequency embryogenesis and plant regeneration in isolated microspore cultures of Brassica oleracea L. Plant Sci. 1991;74:235-242.
76. Takahata Y., Komatsu H., Kaizuma N. Microspore culture of radish (Raphanus sativus L.): influence of genotype and culture conditions on embryogenesis. Plant Cell Rep. 1996;16:163-166.
77. Takahashi Y., Yokoi S., Takahata Y. Improvement of microspore culture method for multiple samples in Brassica. Breed. Sci. 2012;61:96-98. DOI: 10.1270/jsbbs.61.96.
78. Telmer C.A., Simmonds D., Newcomb W. Determination of developmental stage to obtain high frequencies of embryogenic microspores in Brassica napus. Physiol. Plantarum. 1992;84:417-424.
79. Wang M., Farnham M.W., Nannes J.S.P. Ploidy of broccoli regenerated from microspore culture versus anther culture. Plant Breed. 1999;118:249-252.
80. Wang T., Li H., Zhang J., Ouyang B., Lu Y.G., Ye Z.B. Initiation and development of microspore embryogenesis in recalcitrant purple flowering stalk (Brassica campestris ssp. chinensis var. purpurea Hort.) genotypes. Sci. Hortic. 2009;121:419-424. DOI: 10.1016/j.scienta.2009.03.012
81. Weatherhead M.A., Burdon J., Henshaw G.G. Effects of activated charcoal as an additive to plant tissue culture media. Z. Pflanzenphysiol. 1979;94:399-406.
82. Winarto B., Teixeira da Silva J.A. Microspore culture protocol for Indonesian Brassica oleracea. Plant Cell Tiss. Org. Cult. 2011;107:305-315. DOI: 10.1007/s11240-011-9981-z
83. Yadollani A., Abdollani M.R., Moieni A. Effects of carbon source, polyethylene glycol and abscisic acid on secondary embryo induction and maturation in rapeseed (Brassica napus L.) microspore-derived embryos. Acta Physiol. Plant. 2011;33:1905-1912. DOI: 10.1007/s11738-011-0738-4
84. Yeung E.C., Ramesar-Fortner N.S. Physiological influences in development and function of shoot apical meristem of microspore-derived embryos of Brassica napus «Topas». Can. J. Bot. 2006;84:371-384.
85. Yuan S.X., Su Y.B., Liu Y.M., Fang Z.Y., Yang L.M., Zhuang M., Zhang Y.Y., Sun P.T. Effects of pH, MES, arabinogalactan-proteins on microspore cultures in white cabbage. Plant Cell Tiss. Org. Cult. 2012;110:69-76. DOI: 10.1007/s11240-012-0131-z
86. Zaki M., Dickinson H. Modification of cell development in vitro — the effects of colchicines on anther and isolated microspore culture in Brassica napus. Plant Cell Tiss. Org. Cult. 1995;40:255-270.
87. Zaki M., Dickinson H.G. Structural changes during the first division of embryos resulting from anther and microspore culture in Brassica napus. Protoplasma. 1990;156:149-162.
88. Zhang G., Zhang D., Tang G., He Y., Zhou W. Plant development from microspore-derived embryos in oilseed rape as affected by chilling, desiccation and cotyledon excision. Biol. Plant. 2006;50:180-186.
89. Zhang W., Qiang F., Xigang D., Manzhu B. The culture of isolated microspores of ornamental kale (Brassica oleracea var. acephala) and the importance of genotype to embryo regeneration. Sci. Hortic. 2008;117:69-72. DOI: 10.1016/j.scienta.2008.03.023
90. Zhou W.J., Hagberg P., Tang G.X. Increasing embryogenesis and doubling efficiency by immediate colchicine treatment of isolated microspores in spring Brassica napus. Euphytica. 2002;128:27-34.
