Cloning of GbAGL15 and GbAIL5 from Gossypium barbadense and Its Expression during Synchronization of XH16 Somatic Embryogenesis Induced by Low Temperature
Abstract Synchronization of somatic embryogenesis helps accelerate the process of crop molecular breeding. The specific expression of key genes such as Agamous-like15 (AGL15) and Aintegumenta-like5 (AIL5) plays a role similar to switch during somatic embryogenesis. To further explore the molecular mechanism of hypothermia induced somatic embryo synchronization, in this study, GbAGL15 (GenBank No. MK580460) and GbAIL5 (GenBank No. MK591943) were cloned by homologous sequences methods from the embryonic callus of Gossypium barbadense XH16, and the expression pattern of GbAGL15 and GbAIL5 genes in low temperature treated embryogenic cells were checked by qRT-PCR. The results showed that the coding region of GbAGL15 gene was 753 bp and encoded 250 amino acids, which contained the MADS-MEF2-like domain and the K-box conserved region. Phylogenetic tree analysis showed that there was a close relationship among GbAGL15 together with GrAGL15 (G. raimondii) and GaAGL15 (G. arboretum). The coding region of GbAIL5 gene was 1 626 bp and encoded 541 amino acid in the coding segment, and it contained two AP2 domains. Phylogenetic tree analysis showed that GbAIL5 was located in the same evolutionary branch with GhAIL5 (G. hirsutum). qRT-PCR results showed that the expression levels of GbAGL15 and GbAIL5 in embryogenic callus were significantly lower than those of the control group after low temperature treatment. It was speculated that the GbAGL15 and GbAIL5 genes might be involved in somatic embryogenesis and played regulatory roles. Under low temperature conditions, the two gene expression might be inhibited, resulting in the development and differentiation of hypocotyl callus tissue at low temperature, which might promotes the synchronization of somatic embryos. And after the stress conditions were removed, the embryo development could be swithced on in synchronization pattern. The study partially elucidates the mechanism of hypothermia induced somatic embryo synchronization, promoting the foundation of high efficient system of somatic embryogenesis-dependent genetic transformation in G. barbadense.
ZHOU Jing,GUO Jia-Yan,YANG Rui-Si, et al. Cloning of GbAGL15 and GbAIL5 from Gossypium barbadense and Its Expression during Synchronization of XH16 Somatic Embryogenesis Induced by Low Temperature[J]. 农业生物技术学报, 2019, 27(7): 1190-1198.
[1] 陈凡国, 张华, 支大英, 等. 2004. 普通小麦的体细胞胚发生、发育相关蛋白质的毛细管电泳法鉴定[J]. 山东大学学报(理学版), 39(3): 99-103. (Chen F G, Zhang H, Zhi D Y, et al.2004. Identification of specific proteins related to somatic embryogenesis and embryo development of wheat by capillary electrophoresis[J]. Journal of Shandong University, 39(3): 99-103.) [2] 郭家雁, 张霞, 丁喜莲, 等. 2017. 不同处理诱导XH16号体细胞胚胎同步化发生[J]. 棉花学报, 29(4): 385-392. (Guo J Y, Zhang X, Ding X L, et al.2017. Synchronized somatic embryogenesis of "Xinhai 16" induced by different treatments[J]. Cotton Science, 29(4): 385-392.) [3] 姜福星, 魏丕伟, 寇亚平, 等. 2017. 探析植物体胚发生的关键基因[J]. 分子植物育种, 4: 1304-1311. (Jiang F X, Wei P W, Kou Y P, et al.2017. Evaluation and analysis of key genes in plant somatic embryogenesis[J]. Molecular Plant Breeding, 4: 1304-1311.) [4] 毛春娜, 张爱民, 薛建平, 等. 2011. 低温处理对半夏悬浮培养细胞同步化的影响[J]. 中国中药杂志, 36(8): 959-962. (Mao C N, Zhang A M, Xue J P, et al.2011. Effect of low temperature on cell synchronization division in suspension cultore cells of Pinellia ternata[J]. China Journal of Chinese Materia Medica, 36(8): 959-962.) [5] 梅兴国, 周爱文. 2001. 红豆杉细胞同步化的研究[J]. 生物技术, 11(3): 9-10. (Mei X G, Zhou A W.2001. Studies on synchronization of taxus cells[J]. Biotechnology, 11(3): 9-10.) [6] 王兴春, 李宏, 王敏, 等. 2010. 植物体细胞胚胎发生的调控网络[J]. 生物工程学报, 26(2): 141-146. (Wang X C, Li H, Wang M, et al.2010. Regulatory networks of somatic embryogenesis in plant[J]. Chinese Journal of Biotechnology, 26(2): 141-146.) [7] 王哲之, 张大力. 1990. 陆地棉愈伤组织产生及胚胎发生的细胞学研究[J]. 西北植物学报, 10(2): 77-83. (Wang Z Z, Zhang D L.1990. Cat cytological observation on callus production and embryogenesis of Gossypium hirsutum L[J]. Acta Botanica Boreali-Occidentalia Sinica, 10(2): 77-83.) [8] 张宇, 沈海龙. 2007. 植物体细胞胚同步化发生的控制[J]. 植物生理学通讯, (3): 583-587. (Zhang Y, Shen H L. 2007. Control of synchronization for plant somatic embryogenesis[J]. Plant Physiology Communications, (3): 583-587) [9] Ashish A, Ganesan P, Gary B, et al.2009. Identification and analysis of transcription factor family-specific features derived from DNA and protein information[J].Pattern Recognition Letters, 31(14): 2097-2102. [10] Fernandez D E, Heck G R, Perry S E, et al.2000. The embryo MADS domain factor AGL15 acts postembryonically: Inhibition of perianth senescence and abscission via constitutive expression[J]. The Plant Cell, 12(2): 183-197. [11] Harding E W, Tang W, Nichols K W, et al.2003. Expression and maintenance of embryogenic potential is enhanced through constitutive expression of Agamous-like 15[J]. Plant Physiology,133(2): 653-663. [12] Jofuku K D, Omidyar P K, Gee Z, et al.2005. Control of seed mass and seed yield by the floral homeotic gene APETALA2[J]. Proceedings of the National Academy of Sciences of the USA, 102(8): 3117-3122. [13] Ouakfaoui S E, Schnell J, Abdeen A, et al.2010. Control of somatic embryogenesis and embryo development by AP2 transcription factors[J]. Plant Molecular Biology, 74(4-5): 313-326. [14] Quiroz-Figueroa F R, Rojas-Herrear R, Galaz-Avalos R M.2006. Embryo production through somatic embryogenesis can be used to study cell differentiation in plants[J]. Plant Cell, Tissue and Organ Culture, 86(3): 285-301. [15] Nole-Wilson S, Tranby T L, Krizek B A.2005. Aintegumenta-like (AIL) genes are expressed in young tissues and may specify meristematic or division-competent states[J]. Plant Molecular Biology, 57(5): 613-28. [16] Tsuwamoto R, Yokoi S, Takahata Y.2010. Arabidopsis EMBRYOMAKER encoding an AP2 domain transcription factor plays a key role in developmental change from vegetative to embryonic phase[J]. Plant Molecular Biology, 73: 481-492. [17] Wang H, Caruso L V, Downie A B, et al.2004. The embryo MADS domain protein Agamous-Like 15 directly regulates expression of a gene encoding an enzyme involved in gibberellin metabolism[J]. The Plant Cell, 16(5): 1206-1219. [18] Yang Z, Li C, Wang Y, et al.2014. GhAGL15s, preferentially expressed during somatic embryogenesis, promote embryogenic callus formation in Cotton (Gossypium hirsutum L.)[J]. Molecular Genetics and Genomics, 289(5): 873-883. [19] Zheng Q L, Zheng Y M, Perry S E, et al.2013. Decreased GmAGL15 expression and reduced ethylene synthesis may contribute to reduced somatic embryogenesis in a poorly embryogenic cultivar of glycine max[J]. Plant Signaling and Behavior, 8(9): e25422. [20] Zheng Q, Perry S E.2014. Alterations in the transcriptome of soybean in response to enhanced somatic embryogenesis promoted by orthologs of Agamous-like15 and Agamous-like18[J]. Plant Physiology, 164(3): 1365-1377. [21] Zheng Q, Zheng Y, Ji H, et al.2016. Gene regulation by the AGL15 transcription factor reveals hormone interactions in somatic embryogenesis[J]. Plant Physiology, 172(4): 2374-2387.
