|
|
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.
|
Received: 25 December 2019
|
|
Corresponding Authors:
* scrcliuzhanji@sina.com
|
|
|
|
[1] 曹璐, 于旭东, 蔡泽坪, 等. 2019. 植物叶色白化的研究进展[J]. 分子植物育种, 17(16): 5390-5397. (Cao L, Yu X D, Cai Z P, et al.2019. Research progress of plant leaf albino[J]. Molecular Plant Breeding, 17(16): 5390-5397.) [2] 狄佳春, 赵亮, 陈旭升. 2016. 棉花叶片突变体的研究进展[J]. 中国农学通报, 32(36): 78-86. (Di J C, Zhao L, Chen X S.2016. Research progress of cotton leaf mutant[J].Chinese Agricultural Science Bulletin, 32(36): 78-86.) [3] 李佳佳, 于旭东, 蔡泽坪, 等. 2019. 高等植物叶绿素生物合成研究进展[J]. 分子植物育种, 17(18): 6013-6019. (Li J J, Yu X D, Cai Z P, et al.2019. An overview of chlorophyll biosynthesis in higher plants[J]. Molecular Plant Breeding, 17(18): 6013-6019.) [4] 阂留芳, 何金龙, 肖松华, 等. 1996. 陆地棉芽黄品系在棉花杂种优势上的利用研究[J]. 棉花学报, 8(3): 113-119. (Min L F, He J L, Xiao S H, et al.1996. Studies on utilization of heterosis of virescent strains in upland cotton[J]. Cotton Science, 8(3): 113-119.) [5] 邵勤. 2013. 一个新的甜瓜叶色黄化突变体研究[D]. 博士学位论文, 东北农业大学, 导师:于泽源, pp. 107. (Shao Q.2013. Characterization and proteomics of a novel xantha mutant in muskmelon[D]. Thesis for Ph.D., Northeast Agriculture University, Supervisor: Yu Z Y, pp. 107.) [6] 苏小静, 汪沛洪, 陈毓荃. 1990. 小麦突变体返白系返白机理的研究—Ⅱ、返白阶段叶绿素代谢变化研究[J]. 西北农林科技大学学报(自然科学版), 11(3): 80-84. (Su X J, Wang P H, Chen Y Q.1990. Studies on the albrescent mechanism in mutant "stage albinism line of winter wheat"Ⅱ. Changes in the chlorophyll metabolism during the albescent stage[J]. Journal of Northwest A&F University (Natural Science Edition), 11(3): 80-84.) [7] Bae C H, Abe T, Matsuyama T, et al.2001. Regulation of chloroplast gene expression is affected in ali, a novel tobacco albino mutant[J]. Annals of Botany, 88(4): 543-553. [8] Borkird C, Sink K C.1983. Medium components for shoot cultures of chlorophyll-deficient mutants of Petunia inflata[J]. Plant Cell Reports, 2(1): 1-4. [9] Chen N, Wang P, Li C.2018. A single nucleotide mutation of the IspE gene participating in the MEP pathway for isoprenoid biosynthesis causes a green-revertible yellow leaf phenotype in rice[J]. Plant Cell Physiology, 59(9): 1905-1917. [10] Fambrini M, Pugliesi C, Vernieri P, et al.1993. Characterization of a sunflower (Helianthus annuus L.) mutant, deficient in carotenoid synthesis and abscisic-acid content, induced by in vitro tissue culture[J]. Theoretical and Applied Genetics, 87(1-2): 65-69. [11] Huang C, Yu Q B, Li Z R, et al.2017. Porphobilinogen deaminase HEMC interacts with the PPR protein AtECB2 for chloroplast RNA editing[J].The Plant Journal, 92(4): 546-556. [12] Han S H, Sakuraba Y, Koh H J, et al.2012. Leaf variegation in the rice zebra2 mutant is caused by photoperiodic accumulation of tetra-cis-lycopene and singlet oxygen[J]. Molecules and Cells, 33(1): 87-97. [13] Hein P, Stöckel J, Bennewitz S, et al.2009. A protein related to prokaryotic UMP kinases is involved in psaA/B transcript accumulation in Arabidopsis[J]. Plant Molecular Biology, 69: 517-528. [14] Huang M, Slewinski T L, Baker, R F, et al.2009. Camouflage patterning in maize leaves results from a defect in porphobilinogen deaminase[J]. Molecular Plant, 2(4): 773-789. [15] Lewis C F.1958. Genetic studies of a mosaic leaf mutant: Causing somatic instability in cotton[J]. Journal of Heredity, 49(6): 267-271. [16] Lewis C F.1960. The inheritance of mottled leaf in cotton[J]. Journal of Heredity, 51(5): 209-212. [17] Li X, Kanakala S, He Y, et al.2017. Physiological characterization and comparative transcriptome analysis of white and green leaves of Ananascomosus var. bracteatus[J]. PLOS ONE, 12(1): e0169838. [18] Livak K J, Schmittgen T D.2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method[J]. Methods, 25(4): 402-408. [19] Nott A, Jung H S, Koussevitzky S, et al.2006. Plastid-to-nucleus retrograde signaling[J]. Annual Review of Plant Biology, 57(1): 739-759. [20] Parks B M, Quail P H.1991. Phytochrome-deficient hy1 and hy2 long hypocotyl mutants of Arabidopsis are defective in phytochrome chromophore biosynthesis[J]. The Plant Cell, 3(11): 1177-1186. [21] Schmitz-Linneweber C, Williams-Carrier R E., Williams-Voelker P M, et al.2006. A pentatricopeptide repeat protein facilitates the trans-splicing of the maize chloroplast rps12 pre-mRNA[J]. The Plant Cell, 18(10): 2650-2663. [22] Sundberg E, Slagter J G, Fridborg I, et al.1997. ALBINO3, an Arabidopsis nuclear gene essential for chloroplast differentiation, encodes a chloroplast protein that shows homology to proteins present in bacterial membranes and yeast mitochondria[J]. The Plant Cell, 9(5):717-730. [23] Terry M J, Kendrick E R.1996. The aurea and yellow-green-2 mutants of tomato are deficient in phytochrome chromophore synthesis[J]. The Journal of Biological Chemistry, 271(35): 21681-21686. [24] Xue Y B, Ma J, He Y, et al.2019. Comparative transcriptomic and proteomic analyses of the green and white parts of chimeric leaves in Ananas comosus var. bracteatus[J]. PeerJ, 7: e7261. [25] Vicente-Dólera N, Troadec C, Moya M, et al.2014. First TILLING platform in Cucurbita pepo: A new mutant resource for gene function and crop improvement[J]. PLOS ONE, 9(11): e112743. [26] Yaronskaya E, Ziemann V, Walter G, et al.2003. Metabolic control of the tetrapyrrole biosynthetic pathway for porphyrin distribution in the barley mutant albostrians[J]. The Plant Journal, 35: 512-522. [27] Zhang F, Tang W J, Hedtke B.2014. Tetrapyrrole biosynthetic enzyme protoporphyrinogen Ⅸ oxidase 1 is required for plastid RNA editing[J]. Proceedings of the National Academy of Sciences of the USA, 111(5): 2023-2038. [28] Zhu X, Guo S, Wang Z, et al.2016. Map-based cloning and functional analysis of YGL8, which controls leaf colour in rice (Oryza sativa)[J]. BMC Plant Biology, 16(1):134. [29] Zhu X B, Liang S H, Yin J J, et al.2015. The DnaJ OsDjA7/8 is essential for chloroplast development in rice (Oryza sativa)[J]. Gene, 574(1): 11-19. |
|
|
|