摘要
组蛋白去甲基化修饰是染色质修饰中组蛋白修饰的一种重要方式.赖氨酸特异的去甲基化酶(lysine-specific demethylase1,LSD1)和Jumonji C(JmjC)结构域包含蛋白作为两种组蛋白去甲基化酶,被报道与植物营养生长、生殖发育和疾病抗性有关.然而,组蛋白去甲基化修饰酶LSD1/JmjC在龙眼中的功能仍不清楚.使用龙眼‘红核子’品种早期体细胞胚胎发生的胚性愈伤组织(embryogenic callus,EC)、不完全胚性紧实结构(incomplete embryogenic compact structures,ICpEC)和球形胚(globular embryos,GE)为材料,通过生物信息学和实时荧光定量(real-time quantitative PCR,qRT-PCR)方法分析DILSD1/DIJmjC基因在龙眼早期体细胞胚胎发生中的作用.结果显示,龙眼基因组存在4个DILSD1基因和18个DIJmjC基因,进化树分析显示这些基因被分为LSD1、KDM3/JMJD1、KDM4/JMJD2、KDM5/JARID和JmjC domain only组.全基因组或片段复制事件被确定在DILSD1/DIJmjC基因家族扩大的进程中发挥重要的作用,而同线性分析和系统发育比较为DILSD1/DIJmjC基因的进化特征提供更深的认识.蛋白-蛋白互作预测显示,DIJMJ11、DIJMJ12和DIJMJ13蛋白具有较强的相关性,启动子分析也发现,D/LDL1、DIJMJ11、DIJMJ12、DIJMJ14、DIJMJ16-1、DIJMJ17和DIJMJ32等基因存在较多的胁迫和激素响应元件,暗示它们在胁迫和激素调控网络中的重要作用.转录组数据和qRT-PCR分析的表达谱显示:从龙眼EC分化到GE,DIJMJ14和DIJMJ17表达显著逐渐上调,但DIJMJ26和D/LDL1显著逐渐下调:DIJMJ11、DIJMJ12、DIJMJ14、DIJMJ17和D/LDL1均响应水杨酸(salicylic acid,SA)和茉莉酸甲酯(methyl jasmonate,MeJA)调控,而DIJMJ12和D/LDL1也被脱落酸(abscisic acid,ABA)和赤霉素(gibberellin A3,GA3)抑制表达,且DIJMJ11、DIJMJ14、DIJMJ17和D/LDL1均在SA和MeJA处理4 h表达量显著上调;DIJMJ11、DIJMJ12、DIJMJ17和D/LDL1均响应低温调控,它们中的DIJMJ11和D/LDL1响应高温调控,DIJMJ12和DIJMJ17被高温抑制表达.本研究表明相比DIJmjC在进化上存在一定保守性,DILSD1存在进化上的高度保守,且DILSD1/DIJmjC可能通过响应激素和非生物胁迫调控来参与龙眼体细胞胚胎发育和发生过程.
Histone demethylation modification is an important method of modifying chromatin.The lysine-specific demethylase1(LSD1)and Jumonji C(JmjC)-containing proteins are two histone demethylases that have been reported as being involved in plant growth,reproductive development,and disease resistance.However,the function of LSD1/JmjC in longan remains unclear.We used the embryogenic callus(EC),incomplete compact proembryogenic cultures(ICpEC),and globe embryos(GE)in early somatic embryogenesis(SE)of the longan cultivar‘Honghezi’,and utilized bioinformatic methods and real-time quantitative PCR to analyze the roles of DlLSD1/DlJmjC genes during early SE in longan.The results showed that there are 4 DlLSD1 and 18 DlJmjC genes on the longan genome;phylogenetic tree analysis showed that these genes were divided into LSD1,KDM3/JMJD1,KDM4/JMJD2,KDM5/JARID and JmjC domain only groups based on their structural and phylogenetic features.Whole genomic duplication(WGD)and segmental duplication events played an important role in the expansion of the longan DlLSD1/DlJmjC gene family,while synteny and phylogenetic analyses provided insights into the evolutionary characteristics of the DlLSD1/DlJmjC genes in longan.Predictions of protein-protein interactions have shown that DlJMJ11,DlJMJ12,and DlJMJ13 have a strong correlation,and promoter analysis found that the genes DlLDL1,DlJMJ11,DlJMJ12,DlJMJ14,DlJMJ16-1,DlJMJ17,and DlJMJ32 have a high number of stress and hormone response elements,suggesting their important roles in stress and hormone regulatory networks.Expression profiles derived from transcriptome data and qRT-PCR analysis showed that the expression of DlJMJ14 and DlJMJ17 was significantly and gradually upregulated during the differentiation of ECs into GEs,but DlJMJ26 and DlLDL1 were significantly and gradually downregulated.DlJMJ11,DlJMJ12,DlJMJ14,DlJMJ17,and DlLDL1 all responded to the regulation of salicylic acid(SA)and methyl jasmonate(MeJA),while the expression of DlJMJ12 and DlLDL1 was inhibited by abscisic acid(ABA)and gibberellin A3(GA3);DlJMJ12 was also inhibited by SA and MeJA.Moreover,the expression levels of DlJMJ11,DlJMJ14,DlJMJ17,and DlLDL1 were significantly upregulated at 4 h of SA and MeJA treatment while DlJMJ11,DlJMJ12,DlJMJ17,and DlLDL1 all responded to the regulation of low temperatures.Of the latter,DlJMJ11 and DlLDL1 both also responded to the regulation of high temperatures while the expression of DlJMJ12 and DlJMJ17 was inhibited under the same conditions.This study indicated that DlLSD1 is highly conserved,compared to DlJmjC,during the process of evolution and that DlLSD1/DlJmjC may be involved in the embryonic development and somatic embryogenesis in longan as they responded to the regulation of hormones and abiotic stressors.
作者
申序
陈晓慧
陈荣珠
徐小萍
李晓斐
倪珊珊
张婧
蒋梦琦
林玉玲
赖钟雄
SHEN Xu;CHEN Xiaohui;CHEN Rongzhu;XU Xiaoping;LI Xiaofei;NI Shanshan;ZHANG Jing;JIANG Mengqi;LIN Yuling;LAI Zhongxiong(Institute of Horticultural Biotechnology,Fujian Agriculture and Forestry University,Fuzhou 350002,China)
出处
《应用与环境生物学报》
CAS
CSCD
北大核心
2021年第5期1341-1353,共13页
Chinese Journal of Applied and Environmental Biology
基金
国家自然科学基金项目(31672127,31572088)
福建省自然科学基金项目(2015 J06004)
福建省高原学科建设经费项目(102/71201801101)
福建农林大学科技创新基金项目(CXZX2017314,CXZX2017189,CXZX2016118,CXZX2020027A,KF2015108)资助。
