Cloning and Expression Analysis of Porphobilinogen Deaminase Genes in Upland Cotton (Gossypium hirsutum)
CHEN Yi-Zhen, LI Hao, FU Ming-Chuan, WANG Li-Guo, LIU Ren-Zhong, LIU Zhan-Ji*
Key Laboratory of Cotton Breeding and Cultivation in Huang-Huai-Hai Plain, Ministry of Agriculture/Shandong Cotton Research Center, Jinan 250100, China
Abstract：Leaf color mutants are ideal materials for studying a series of physiological metabolic processes such as chlorophyll metabolism, chloroplast development, photosynthesis, and hormone physiology in higher plants, as well as important materials in genetics and breeding researches. Porphobilinogen deaminase (PBGD) is the key enzyme in the process of plant chlorophyll biosynthesis. In this study, 4 PBGD genes were isolated using mottled leaf mutant of upland cotton (Gossypium hirsutum) 'Lumian 418', and named as GhPBGD1A (GenBank No. MN849924), GhPBGD1D (GenBank No. MN849925), GhPBGD2A (GenBank No. MN849926) and GhPBGD2D (GenBank No. MN849927), respectively. The sequence analysis showed that each of the 4 GhPBGD genes had no difference in sequences between the normal and mottled leaves. All GhPBGD genes had a similar gene structure with 5 exons and 4 introns, and were predicted to be located in chloroplasts. Phylogenetic analysis indicated that GhPBGDs had close relationship with the PBGD proteins from Arabidopsis thaliana and Brassica napus. In addition, the cis-elements analysis revealed that many elements involved in light, anaerobic and hormone responsiveness were identified in the promoter regions of GhPBGD genes. Furthermore, qRT-PCR analysis showed that GhPBGD genes expressed in leaves, flowers, bolls and sepals, and had the highest expression in the leaves. Compared to normal leaves, the expression of GhPBGD genes were significantly down-regulated in the mottled leaves. The above results suggested that GhPBGD genes might be involved in the regulation of leaf color mutation. This study could provide reference data for further exploration of the molecular mechanism of leaf color deletion mutants.
陈义珍, 李浩, 傅明川, 王立国, 刘任重, 柳展基. 陆地棉胆色素原脱氨酶基因的克隆及表达分析[J]. 农业生物技术学报, 2020, 28(7): 1221-1229.
CHEN Yi-Zhen, LI Hao, FU Ming-Chuan, WANG Li-Guo, LIU Ren-Zhong, LIU Zhan-Ji. Cloning and Expression Analysis of Porphobilinogen Deaminase Genes in Upland Cotton (Gossypium hirsutum). 农业生物技术学报, 2020, 28(7): 1221-1229.
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