5-azacytidine Reduces Regeneration Frequency of Wheat Mature Embryo

doi:10.11924/j.issn.1000-6850.2020-00006

Abstract

Abstract:

To study the effect of histone acetylation on wheat regeneration, mature embryos of CB037, Fielder and Chinese Spring (CS) were used as explants, different concentrations of 5-azacytidine (5-azaC) were added into the culture medium. The embryonic callus induction rate, callus differentiation rate and green shoot induction rate were analyzed. The results showed that with the addition of 50 μmol/L 5-azaC, the green shoot induction rate of CB037 was 54.72% which reduced significantly compared with the control group of 79.62%. With the addition of 10 μmol/L and 50 μmol/L 5-azaC, the embryonic callus induction rates of Fielder were 39.13% and 35.71% respectively, and the green shoot induction rates were 26.09% and 25.14% respectively, which decreased significantly compared with those of the control. While CS was treated with 50 μmol/L, the embryonic callus induction rate was 23.89% and reached significant difference compared with the control group of 30.95%. Adding 5-azaC could reduce the regeneration frequency of wheat mature embryo; with the increase of additive concentration, the reduction effect becomes more obvious and the effects of 5-azaC on the regeneration frequency are different among genotypes.

Key words: wheat, 5-azacytidine, mature embryo, regeneration frequency, tissue culture

CLC Number:

S512.12

Dong Luhao, Li Xiaohui, Song Yuning, Wang He, Li Xingguo, Bie Xiaomin. 5-azacytidine Reduces Regeneration Frequency of Wheat Mature Embryo[J]. Chinese Agricultural Science Bulletin, 2021, 37(3): 26-30.

Figures/Tables 5

References 25

[1]	Diez C M, Roessler K, Gaut B S. Epigenetics and plant genome evolution[J]. Current Opinion in Plant Biology, 2014,18:1-8. doi: 10.1016/j.pbi.2013.11.017 URL pmid: 24424204
[2]	Zhong X. Comparative epigenomics: A powerful tool to understand the evolution of DNA methylation[J]. New Phytologist, 2016,210(1):76-80. doi: 10.1111/nph.13540 URL
[3]	Friedman S. The inhibition of DNA (cytosine-5) methylases by 5-azacytidine. The effect of azacytosine-containing DNA[J]. Molecular Pharmacology, 1981,19(2):314-320. URL pmid: 6164911
[4]	LoSchiavo F, Pitto L, Giuliano G, et al. DNA methylation of embryogenic carrot cell cultures and its variations as caused by mutation, differentiation, hormones and hypomethylating drugs[J]. Theoretical and Applied Genetics, 1989,77(3):325-331. doi: 10.1007/BF00305823 URL pmid: 24232608
[5]	QuingaL A P, de Freitas Fraga H P, do Nascimento Vieira L, et al. Epigenetics of long-term somatic embryogenesis in Theobroma cacao L.: DNA methylation and recovery of embryogenic potential[J]. Plant Cell, Tissue and Organ Culture, 2017,131(2):295-305. doi: 10.1007/s11240-017-1284-6 URL
[6]	Klimaszewska K, Noceda C, Pelletier G, et al. Biological characterization of young and aged embryogenic cultures of Pinus pinaster (Ait.)[J]. In Vitro Cellular & Developmental Biology-Plant, 2009,45(1):20-33.
[7]	Osorio-Montalvo P, Sáenz-Carbonell L, De-la-Peña C. 5-azacytidine: A promoter of epigenetic changes in the quest to improve plant somatic embryogenesis[J]. International Journal of Molecular Sciences, 2018,19(10):3182. doi: 10.3390/ijms19103182 URL
[8]	刘艳, 沈海龙, 丛建民. 5-氮胞苷对水曲柳合子胚外植体状态及体胚发生的影响[J]. 东北林业大学学报, 2011,39(8):25-27,32.
[9]	Teyssier C, Maury S, Beaufour M, et al. In search of markers for somatic embryo maturation in hybrid larch (Larix × eurolepis): global DNA methylation and proteomic analyses[J]. Physiologia Plantarum, 2014,150(2):271-291. doi: 10.1111/ppl.12081 URL pmid: 23789891
[10]	Santos D, Fevereiro P. Loss of DNA methylation affects somatic embryogenesis in Medicago truncatula[J]. Plant Cell, Tissue and Organ Culture, 2002,70(2):155-161. doi: 10.1023/A:1016369921067 URL
[11]	王子成, 聂丽娟, 何艳霞. 离体条件下5-氮杂胞嘧啶核苷对菊花DNA甲基化和表型性状的影响[J]. 园艺学报, 2009,36(12):1783-1790.
[12]	Solís M T, EI-Tantawy A A, Cano V, et al. 5-azacytidine promotes microspore embryogenesis initiation by decreasing global DNA methylation, but prevents subsequent embryo development in rapeseed and barley[J]. Frontiers in Plant Science, 2015,6:472. doi: 10.3389/fpls.2015.00472 URL pmid: 26161085
[13]	Godfray H C, Beddington J R, Crute I R, et al. Food security: the challenge of feeding 9 billion people[J]. Science, 2010,327(5967):812-818. doi: 10.1126/science.1185383 URL pmid: 20110467
[14]	赵广才, 常旭虹, 王德梅, 等. 小麦生产概况及其发展[J]. 作物杂志, 2018(4):1-7.
[15]	Tester M, Langridge P. Breeding technologies to increase crop production in a changing world[J]. Science, 2010,327(5967):818-822. doi: 10.1126/science.1183700 URL pmid: 20150489
[16]	Wang K, Liu H Y, Du L P, et al. Generation of marker-free transgenic hexaploid wheat via an Agrobacterium-mediated co-transformation strategy in commercial Chinese wheat varieties[J]. Plant Biotechnology Journal, 2017,15(5):614-623. doi: 10.1111/pbi.12660 URL pmid: 27862820
[17]	Bie X M, Wang K, Liu C, et al. Effects of soil drought stress on plant regeneration efficiency and endogenous hormone levels of immature embryos in wheat (Triticum aestivum L.)[J]. Pakistan Journal of Botany, 2017,49(5):1673-1679.
[18]	张伟, 尹米琦, 赵佩, 等. 我国部分主推小麦品种组织培养再生能力评价[J]. 作物学报, 2018,44(2):208-217.
[19]	姚富泉, 董鲁浩, 王悦, 等. 植物生长调节剂优化不同品种冬小麦成熟胚再生体系[J]. 山东农业科学, 2019,51(5):19-23.
[20]	董鲁浩, 姚富泉, 詹悠, 等. 春小麦成熟胚愈伤组织诱导及再生体系的优化[J]. 分子植物育种, 2020,18(4):1244-1249.
[21]	别晓敏, 杜丽璞, 徐惠君, 等. 培养基中CuSO₄和Fe盐浓度对小麦胚培养再生效果的影响[J]. 植物遗传资源学报, 2011,12(6):975-981.
[22]	Grzybkowska D, Morończyk J, Wójcikowska B, et al. Azacitidine (5-AzaC)-treatment and mutations in DNA methylase genes affect embryogenic response and expression of the genes that are involved in somatic embryogenesis in Arabidopsis[J]. Plant Growth Regulation, 2018,85:243-256. doi: 10.1007/s10725-018-0389-1 URL
[23]	Betekhtin A, Milewska-Hendel A, Chajec L, et al. 5-azacitidine induces cell death in a tissue culture of Brachypodium distachyon[J]. International Journal of Molecular Sciences, 2018,19(6):1806. doi: 10.3390/ijms19061806 URL
[24]	Noyer J L, Causse S, Tomekpe K, et al. A new image of plantain diversity assessed by SSR, AFLP and MSAP markers[J]. Genetica, 2005,124(1):61-69. doi: 10.1007/s10709-004-7319-z URL pmid: 16011003
[25]	陈兆贵, 黄学林. 植物离体培养过程中DNA甲基化变异研究进展[J]. 植物遗传资源学报, 2011,12(4):575-580.

培养基	成分
预培养基	MS基本培养基（不含MS维生素）+0.75 g/L MgCl₂+15.0 g/L甘露醇+15.0 g/L山梨醇+5.0 mg/L谷氨酰胺+0.5 g/L水解酪蛋白+12.0 g/L葡萄糖+10 mg/L B₁维生素+1.0 mg/L B₆维生素+1.0 mg/L B₃维生素+2.0 mg/L甘氨酸+39 mg/L乙酰丁香酮+2.0 mg/L dicamba+4.0 mg/L AgNO₃+40.0 mg/L半胱氨酸+100.0 g/L维生素C,pH 5.8,8 g/L琼脂
诱导培养基	MS基本培养基+3.0%蔗糖+2.0 mg/L dicamba+4.0 mg/L AgNO₃,pH 5.8,2.4 g/L植物凝胶
分化培养基	MS基本培养基（不含MS维生素）+10.0 mg/L B₁维生素+1.0 mg/L B₃维生素+1.0 mg/L B₆维生素+2.0 mg/L甘氨酸+5.0 mg/L谷氨酰胺+2.0%蔗糖+0.2 mg/L IAA,pH 5.8,2.4 g/L植物凝胶

培养基	成分
预培养基	MS基本培养基（不含MS维生素）+0.75 g/L MgCl₂+15.0 g/L甘露醇+15.0 g/L山梨醇+5.0 mg/L谷氨酰胺+0.5 g/L水解酪蛋白+12.0 g/L葡萄糖+10 mg/L B₁维生素+1.0 mg/L B₆维生素+1.0 mg/L B₃维生素+2.0 mg/L甘氨酸+39 mg/L乙酰丁香酮+2.0 mg/L dicamba+4.0 mg/L AgNO₃+40.0 mg/L半胱氨酸+100.0 g/L维生素C,pH 5.8,8 g/L琼脂
诱导培养基	MS基本培养基+3.0%蔗糖+2.0 mg/L dicamba+4.0 mg/L AgNO₃,pH 5.8,2.4 g/L植物凝胶
分化培养基	MS基本培养基（不含MS维生素）+10.0 mg/L B₁维生素+1.0 mg/L B₃维生素+1.0 mg/L B₆维生素+2.0 mg/L甘氨酸+5.0 mg/L谷氨酰胺+2.0%蔗糖+0.2 mg/L IAA,pH 5.8,2.4 g/L植物凝胶