胡萝卜转录因子DcBZR1基因的克隆及组织表达特异性分析

王欣蕊,王雅慧,张榕蓉,徐志胜,熊爱生*
南京农业大学园艺学院, 作物遗传与种质创新国家重点实验室, 农业农村部华东地区园艺作物生物学与种质创 制重点实验室, 南京210095

通信作者:熊爱生;E-mail: xiongaisheng@njau.edu.cn

摘 要:

本研究以胡萝卜(Daucus carota ssp. sativus) ‘黑田五寸为试验材料克隆得到DcBZR1基因。该基因开放阅读框长939 bp, 编码312个氨基酸。氨基酸序列比对和保守结构域分析结果表明DcBZR1含有bHLH结构域, 属于BZR1/BES1家族。利用实时荧光定量PCR技术检测黑田五寸’ (橙色)紫宙’ (紫色)白玉’(白色)胡萝卜不同组织中和不同发育阶段下DcBZR1基因的相对表达水平, 结果表明: 不同组织中, DcBZR1基因均有表达且具有组织表达特异性, 在根中的表达量最高; 不同发育阶段下, 各品种胡萝卜DcBZR1基因的相对表达水平变化规律基本一致, 在播种后75 d时达到峰值; 不同根色胡萝卜材料中, DcBZR1基因在橙色胡萝卜黑田五寸中的表达水平较高。研究结果表明DcBZR1可能与类胡萝卜素合成相关并在胡萝卜肉质根发育过程中发挥重要作用。

关键词:胡萝卜; BZR1; 基因克隆; 表达分析; 类胡萝卜素

收稿:2020-08-12   修定:2020-12-16

资助:国家自然科学基金(31872098)和江苏高校优势学科建设工程项目(PAPD)

Cloning and tissue-specifc expression profle analysis of DcBZR1 gene, a transcription factor from Daucus carota ssp. sativus

WANG Xinrui, WANG Yahui, ZHANG Rongrong, XU Zhisheng, XIONG Aisheng*
College of Horticulture, Nanjing Agricultural University, State Key Laboratory of Crop Genetics and Germplasm Enhancement, Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China, Ministry of Agriculture and Rural Affairs, Nanjing 210095, China

Corresponding author: XIONG Aisheng; E-mail: xiongaisheng@njau.edu.cn

Abstract:

In this study, DcBZR1 gene was cloned from carrot (Daucus carota subsp. sativus) variety ‘Kurodagosun’. The fragment of DcBZR1 includes a open reading frame of 939 bp, which encodes a protein product of 312 amino acids. Multiple sequence alignment and conserved domain analysis showed that DcBZR1 protein contains a basic bHLH domain, which belongs to BZR1/BES1 family. The relative expression levels of DcBZR1 gene in different tissues and different developmental stages of ‘Kurodagosun’ (orange), ‘Zizhou’(purple) and ‘Baiyu’ (white) carrots were detected and analyzed by real-time quantitative PCR. The results show that DcBZR1 was tissue-specifc and highly expressed in carrot root. The gene expression in different varieties was basically the same at different developmental stages, which reached the peak at 75 days after sowing. Comparison among varieties with different root colors indicates that the expression level of DcBZR1 gene in orange cultivar ‘Kurodagosun’ was the highest. The results demonstrate that DcBZR1 may be related to the biosynthesis of carotenoids and may play an important role in the development of carrot fleshy roots.

Key words: Daucus carota ssp. sativus; BZR1; gene cloning; expression analysis; carotenoid

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