菠萝AcKNOX1影响叶缘发育的功能研究  

Functional Studies of Pineapple AcKNOX1 on Leaf Margin Development

作　　者：荆永琳 陈浪欣 李俊国 王加宾[1,2] 王小冰 杨庆全 孟春阳 徐立[1,2] JING Yonglin;CHEN Langxin;LI Junguo;WANG Jiabin;WANG Xiaobing;YANG Qingquan;MENG Chunyang;XU Li(Key Laboratory of Crop Gene Resources and Germplasm Enhancement in Southern,Ministry of Agriculture and Rual Affairs,Key Laboratory of Tropical Crops Germplasm Resources Genetic Improvement and Innovation of Hainan Province,Institute of Tropical Crop Genetic Resources,Chinese Academy of Tropical Agricultural Sciences,Haikou 571101,China;National Gene Bank of Tropical Crops,Danzhou,Hainan 571700,China)

机构地区：[1]中国热带农业科学院热带作物品种资源研究所,农业农村部华南作物基因资源与种质创制重点实验室,海南省热带作物资源遗传改良与创新重点实验室,海口571101 [2]国家热带作物中期库,海南儋州571700

出　　处：《园艺学报》2025年第2期292-308,共17页Acta Horticulturae Sinica

基　　金：海南省自然科学基金项目(321QN322);国家自然科学基金项目(31372106)。

摘　　要：以菠萝‘台农16’为材料,克隆得到基因AcKNOX1。AcKNOX1是同源异型盒基因,属于3个氨基酸环延伸蛋白超家族。AcKNOX1的开放阅读框为990 bp,编码329个氨基酸,编码蛋白定位于细胞核中。AcKNOX1启动子区域除核心启动子元件TATA-box和CAAT-box外,还包含许多与光响应、激素响应和非生物胁迫响应相关的元件。AcKNOX1在不同组织中均有表达,但具有明显的组织表达特异性,在茎的表达量最高,其次是叶片,且有刺叶片显著高于无刺叶片中。AcKNOX1响应GA3、ABA和6-BA信号诱导其表达上调。异源表达拟南芥,叶缘裂刻加深。利用酵母单双杂进行互作蛋白筛选,发现AcKNOX1可以与AcCUC1和AcCUC2启动子区域结合激活其表达;与AcBLH1、AcBLH4可形成异源二聚体,在细胞核中互作,且荧光素酶互补和双分子荧光互补实验验证了其互作的准确性。研究结果表明AcKNOX1在菠萝叶缘发育中起重要作用。In this study,AcKNOX1 was cloned from pineapple‘Tainong 16’,which is a homeobox gene,belongs to the three amino acid loop extension superfamily.The gene’s open reading frame was 990 bp with 329 amino acids.The protein was localized in the nucleus.In addition to the core promoter elements TATA-box and CAAT-box,the AcKNOX1 promoter region contained many elements related to light,hormonal and abiotic stress responses.AcKNOX1 was expressed in different tissues with the highest expression in the stem,followed by leaves,which suggested a certain tissue specificity.AcKNOX1 showed higher expression in spiny leaf materials than in spineless leaf materials.The expression of AcKNOX1 was up-regulated by short-term GA3,ABA,and 6-BA stress.Ectopic overexpression of AcKNOX1 in Arabidopsis thaliana suggested that transformants showed larger and deeper serrations.Analysis of interaction proteins obtained by Yeast one and two hybrid system showed that AcKNOX1 could interact and activate AcCUC1 and AcCUC2 promoter regions.And AcKNOX1 interacted with AcBLH1 and AcBLH4 to form a heterodimer.And the inteaction was verified by Luciferase complementation imaging assay(LCI)and Bimolecular fluorescence complementation(BIFC).The results revealed that AcKNOX1 gene played a vital role in the regulation of pineapple leaf margin development.

关 键 词：菠萝 叶缘发育 AcKNOX1 蛋白互作 

分 类 号：S668.3[农业科学—果树学]