金鱼草KNOX基因家族鉴定及调控其雄蕊瓣化的候选基因挖掘  

Genome-wide identification of KNOX gene family and exploration of candidate genes regulating stamen petalization in Antirrhinum majus

作　　者：顾伟卓 汪仲毅 杨洁[1,2] 赵永晶 赵晗茜 谯正林 杨楠 胡慧贞[1,2] CU Wei-huo;AN Zhong-yi;YAN Jie;ZHAO Yong-jing;ZHAO Han-qian;QIAO Zheng-lin;YANG Nan;HU Hui-zhen(College of Landscape Architecture and Horticulture Sciences,Southwest Forestry University,Kunming 650224,China;Yunnan Province Engineering Research Center for Functional Flower Resources and Industrialization,Kunming 650224,China)

机构地区：[1]西南林业大学园林园艺学院,昆明650224 [2]云南省功能性花卉资源及产业化技术工程研究中心,昆明650224

出　　处：《西南农业学报》2024年第6期1329-1339,共11页Southwest China Journal of Agricultural Sciences

基　　金：国家自然科学基金地区科学基金项目(32360070);云南省高层次人才引进青年人才专项竞争性培养支持经费(YNQR-QNRC-2018-122);云南省农业基础联合面上项目(202101BD070001-124、202301BD070001-072);西南林业大学科研启动项目(01102-112109)。

摘　　要：【目的】KNOX(KNOTTED1-like homeobox)转录因子家族在植物生长发育及花器官发育中发挥重要作用。文章旨在鉴定并分析金鱼草(Antirrhinum majus)KNOX转录因子,挖掘出调控金鱼草雄蕊瓣化的关键候选基因。【方法】采用生物信息学方法,在金鱼草全基因组水平鉴定AmKNOX基因,并对其基因结构、蛋白理化性质、亚细胞定位、系统进化关系、染色体定位、启动子顺式作用元件及转录因子结合位点等进行分析,并利用金鱼草雄蕊瓣化和非瓣化材料进行RNA-seq分析和qRT-PCR验证,挖掘出候选基因。【结果】从金鱼草中共鉴定出11个AmKNOX基因,都具有KNOXⅠ、KNOXⅡ、ELK和HOX 4个相对保守的区域,分为ClassⅠ(AmKNOX1、AmKNOX2、AmKNOX3、AmKNOX9和AmKNOX11)和ClassⅡ(AmKNOX5、AmKNOX4、AmKNOX7、AmKNOX10、AmKNOX6和AmKNOX8)2类。AmKNOXs蛋白包含282~406个氨基酸,均定位于细胞核中。启动子顺式作用元件分析结果显示,AmKNOXs家族成员可参与植物生长、激素和非生物胁迫响应,且AmKNOXs存在大量与植物生长发育、器官分化和逆境生理调节相关的转录因子结合位点。RNA-seq和qRT-PCR分析表明AmKNOX家族基因在金鱼草正常雄蕊和瓣化雄蕊中显著差异表达。【结论】本研究共鉴定出11个AmKNOX成员,最终挖掘出5个正向(AmKNOX5、AmKNOX10、AmKNOX6、AmKNOX4和AmKNOX9)以及1个负向(AmKNOX11)调控雄蕊瓣化的AmKNOX候选基因。其中AmKNOX5基因在瓣化雄蕊中的表达量比正常雄蕊中高2703%,推测其功能非常重要,该研究为后期深入解析金鱼草花型发育分子机制和遗传改良奠定了基础。【Objective】The KNOX(KNOTTED1-like homeobox)transcription factor family plays an important role in plant growth and flower organ development.The KNOX transcription factors of Antirrhinum majus were identified and analyzed,and the key candidate genes regulating stamen petalization in A.majus were explored.【Method】The bioinformatics methods were used to identify the AmKNOX genes at the whole genome level,including gene structure,protein physicochemical properties,subcellular localization,phylogenetic relationship,chromosome localization,promoter cis-acting elements and transcription factor binding sites analysis.RNA-seq analysis and qRT-PCR verification were performed to explore AmKNOX candidate genes using petalized and non-petalized stamens of A.majus.【Result】A total of 11 AmKNOX genes were identified from A.majus,all of which had four conserved regions of KNOX I,KNOX II,ELK and HOX and were classified into Class I(AmKNOX1,AmKNOX2,AmKNOX3,AmKNOX9 and AmKNOX11)and Class II(AmKNOX5,AmKNOX4,AmKNOX7,AmKNOX10,AmKNOX6 and AmKNOX8).The AmKNOX protein contained 282-406 amino acids and were localized in the nucleus.Analysis of promoter cis-acting elements showed that AmKNOX family members were involved in plant growth,hormonal and abiotic stress responses.Moreover,AmKNOXs had a large number of transcription factor binding sites related to plant growth and development,organ differentiation,and physiological regulation of stress.RNA-seq and qRT-PCR analysis showed that AmKNOX family genes were significantly differentially expressed in petalized and non-petalized stamens of A.majus.【Conclusion】The study identified 11 AmKNOX members,of which five positive(AmKNOX5,AmKNOX10,AmKNOX6,AmKNOX4 and AmKNOX9)and one negative(AmKNOX11)candidate genes involved in stamen petalization were ultimately identified.Among them,the expression level of AmKNOX5 gene in petalized stamens was 2703%higher than that in normal stamens,and the inference function should be particularly important.The study laid a foundation for in-depth an

关 键 词：金鱼草 KNOX基因家族 生物信息学 雄蕊瓣化 

分 类 号：S681.9[农业科学—观赏园艺]