小粒咖啡CaFAB2基因家族的鉴定及其在种子脂肪酸合成中的潜在作用OA北大核心
Identification of the CaFAB2 Gene Family in Coffea arabica and Its Potential Role in Seed Fatty Acid Synthesis
小粒咖啡豆的品质是选育优质小粒咖啡(Coffea arabica)新品种的首要目标,其中,咖啡豆香气是评价小粒咖啡豆品质优劣的关键要素,而脂肪酸是形成小粒咖啡豆挥发性香气成分的前体之一.硬脂酰基-ACP去饱和酶(stearoyl-ACP desaturase,FAB)是植物不饱和脂肪酸合成的关键酶,植物体内脂肪酸各组分的比例及不饱和度与FAB的去饱和作用息息相关.本研究通过BLASTP比对,在小粒咖啡基因组中鉴定出4个CaFAB2基因家族成员.进一步分析表明,这4个CaFAB2基因的蛋白质氨基酸数目为381~415 aa;分子量为43 936.6~47 693.4 D;等电点均大于7;不稳定系数为29.55~42.67,脂肪系数均小于100,比较亲水;总平均亲水系数均为负值,为亲水蛋白;CaFAB2家族蛋白成员均定位于叶绿体上.在进行基因家族分析时发现,小粒咖啡的CaFAB2基因家族成员与拟南芥(Arabidopsis thaliana)的AtFAB2基因家族的保守基序完全相同且分布的排列位置高度相似,结构域均为PLN00179.小粒咖啡的CaFAB2基因与香樱桃咖啡(C.eugenoedes)和中粒咖啡(C.canephora)之间出现子代2个基因对应亲本1个基因的现象.此外,对小粒咖啡种子4个发育阶段脂肪酸含量测定发现,其所含脂肪酸主要以亚油酸、棕榈酸、油酸以及硬脂酸为主,并且在其发育阶段中变化趋势一致,含量先减少后逐渐升高.对小粒咖啡种子4个发育阶段的转录组测序及实时荧光定量PCR分析结果表明,CaFAB2.3基因与油酸合成具有正相关性,CaFAB2.1基因和CaFAB2.3基因主要参与前期棕榈油酸的合成.本研究结果为深入研究小粒咖啡种子的脂肪酸变化趋势及脂肪酸生物合成提供理论依据,对小粒咖啡CaFAB2基因的鉴定以及调控机理提供新的见解.
The quality of Coffea arabica beans is the primary objective of breeding new varieties of C.arabica with high quality,among these,the aroma of coffee bean is the key factor to evaluate the quality of C.arabica bean,and fatty acid is one of the precursors to form volatile aroma components of C.arabica bean.Stearoyl-ACP desaturase(FAB)is an essential enzyme involved in unsaturated fatty acid synthesis in plants,and its desaturation function significantly influences the proportion and unsaturation level of fatty acids.Through Blast comparison analysis,4 members of the CaFAB2 gene family within the C.arabica genome were identified in this study.Further investigation illuminated that these 4 CaFAB2 genes consisted of 381~415 amino acids with molecular weights ranging from 43 936.6 to 47 693.4 D.Besides,these 4 genes also had isoelectric points greater than 7 and exhibited hydrophilic characteristics with instability indices from 29.55 to 42.67 but less than 100 fat index values.And the total average coefficients were all negative,so they were hydrophilic proteins.The proteins of 4 CaFAB2 genes were localized in chloroplasts.Through gene family analysis,it was found that the conserved motif of the CaFAB2 gene family was identical to that of the FAB2 gene family of Arabidopsis thaliana,and the distribution position was highly similar,with the domain PLN00179.Additionally,it was observed that 2 offspring genes corresponded to 1 parental gene between C.eugenoedes and Coffea canephora regarding the presence of CaFAB2 genes in the genome structure of C.arabica.Furthermore,the fatty acid contents were mainly composed of linoleic acid,palmitic acid,oleic acid,and stearic acid during 4 developmental stages of C.arabica seeds,and these 4 compounds indicated the same trend characterized by initial decrease followed by gradual increase.The transcriptome sequencing and real-time fluorescence quantitative PCR analyse revealed a positive correlation between CaFAB2.3 and oleic acid synthesis,while the CaFAB2.1 and CaFAB2.3 primarily participated in early palmitic acid synthesis in the 4 developmental stages of C.arabica seeds.This study provides a theoretical foundation for further investigation on variation tendency and biosynthesis of fatty acids in C.arabica,and offered novel insights into the identification and regulatory mechanisms of CaFAB2 genes in C.arabica.
张振伟;董相书;杨婧;杞美军;王步天;施学东;黄文龙;李学俊;葛宇
云南农业大学热带作物学院,普洱 665099云南大学资源植物研究院,昆明 650504
农业科学
小粒咖啡硬脂酰基-ACP去饱和酶2(FAB2)基因表达模式
Coffea arabicaStearoyl-ACP desaturase 2(FAB2)geneExpression pattern
《农业生物技术学报》 2025 (001)
79-91 / 13
云南省基础研究专项面上项目(202301AT070493);云南省"兴滇英才支持计划"青年人才(XDYC-QNRC-2022-0711);云南省咖啡重点实验室(云南农业大学)(202449CE340030);云南省重点研发计划项目(202303AP140010);云南省专家基层科研工作站(2021RYZJGZZ002);云南农业大学教育教学改革研究与实践项目(2024SJJX036);云南农业大学热带作物学院教育教学改革研究项目(202405);云南农业大学大学生创新创业项目(S202310676059)
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