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Disruption of carotene biosynthesis leads to abnormal plastids and variegated leaves in Brassica napus.

ABSTRACT: Leaf color is an important characteristic of normal chloroplast development. Variegated plants have green- and white-sectored leaves, which can be used to identify important pathways and molecular mechanisms of chloroplast development. We studied two Brassica napus variegation mutants from same one variegated ancestor, designated ZY-4 and ZY-8, which have different degrees of variegation. When grown in identical conditions, the ratio of white sectors in ZY-4 leaves is higher than in ZY-8. In both mutants, the cells in green sectors contain normal chloroplasts; while, the cells in white sectors contain abnormal plastids. Seedling chloroplasts ultrastructure of both mutants showed that the biogenesis of chloroplasts was blocked in early stages; delayed development and structual damage in ZY-4 were more serious than in ZY-8. Employing bulked segregant analysis(BSA), two bulks (BY142 and BY137) from BC2F1 lines derived from ZY-4 and ZS11, and one bulk (BY56) from BC2F1 lines derived from ZY-8 and ZS11, and screening by Brassica 60K SNP BeadChip Array, showed the candidate regions localized in chromosome A08 (BY142), C04 (BY137), and A08 (BY56), respectively. Transcriptome analysis of five seedling development stages of ZY-4, ZY-8, and ZS11 showed that photosynthesis, energy metabolism-related pathways and translation-related pathways were important for chloroplast biogenesis. The number of down- or up-regulated genes related to immune system process in ZY-4 was more than in ZY-8. The retrograde signaling pathway was mis-regulated in both mutants. DEG analysis indicated that both mutants showed photooxidative damages. By coupling transcriptome and BSA CHIP analyses, some candidate genes were identified. The gene expression pattern of carotene biosynthesis pathway was disrupted in both mutants. However, histochemical analysis of ROS revealed that there was no excessive accumulation of ROS in ZY-4 and ZY-8. Taken together, our data indicate that the disruption of carotene biosynthetic pathways leads to the variegation phenotypes of ZY-4 and ZY-8 and there are some functions that can compensate for the disruption of carotene biosynthesis in ZY-4 and ZY-8 to reduce ROS and prevent seedling mortality.

SUBMITTER: Zhao X

PROVIDER: S-EPMC7297816 | biostudies-literature | 2020 Jul

REPOSITORIES: biostudies-literature

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Disruption of carotene biosynthesis leads to abnormal plastids and variegated leaves in Brassica napus.

Zhao Xiaobin X Hu Kaining K Yan Mengjiao M Yi Bin B Wen Jing J Ma Chaozhi C Shen Jinxiong J Fu Tingdong T Tu Jinxing J

Molecular genetics and genomics : MGG 20200418 4

Leaf color is an important characteristic of normal chloroplast development. Variegated plants have green- and white-sectored leaves, which can be used to identify important pathways and molecular mechanisms of chloroplast development. We studied two Brassica napus variegation mutants from same one variegated ancestor, designated ZY-4 and ZY-8, which have different degrees of variegation. When grown in identical conditions, the ratio of white sectors in ZY-4 leaves is higher than in ZY-8. In bot ...[more]

PMID: 32306107

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Project description:Brassica napus seed is a lipid storage organ containing approximately 40% oil, while its leaves contain many kinds of lipids for many biological roles, but the overall amounts are less than in seeds. Thus, lipid biosynthesis in the developing seeds and the leaves is strictly regulated which results the final difference of lipids. However, there are few reports about the molecular mechanism controlling the difference in lipid biosynthesis between developing seeds and leaves. In this study, we tried to uncover this mechanism by analyzing the transcriptome data for lipid biosynthesis. The transcriptome data were de novo assembled and a total of 47,216 unigenes were obtained, which had an N50 length and median of 1271 and 755 bp, respectively. Among these unigenes, 36,368 (about 77.02%) were annotated and there were 109 up-regulated unigenes and 72 down-regulated unigenes in the developing seeds lipid synthetic pathway after comparing with leaves. In the oleic acid pathway, 23 unigenes were up-regulated and four unigenes were down-regulated. During triacylglycerol (TAG) synthesis, the key unigenes were all up-regulated, such as phosphatidate phosphatase and diacylglycerol O-acyltransferase. During palmitic acid, palmitoleic acid, stearic acid, linoleic acid and linolenic acid synthesis in leaves, the unigenes were nearly all up-regulated, which indicated that the biosynthesis of these particular fatty acids were more important in leaves. In the developing seeds, almost all the unigenes in the ABI3VP1, RKD, CPP, E2F-DP, GRF, JUMONJI, MYB-related, PHD and REM transcript factor families were up-regulated, which helped us to discern the regulation mechanism underlying lipid biosynthesis. The differential up/down-regulation of the genes and TFs involved in lipid biosynthesis in developing seeds and leaves provided direct evidence that allowed us to map the network that regulates lipid biosynthesis, and the identification of new TFs that are up-regulated in developing seeds will help us to further elucidate the lipids biosynthesis pathway in developing seeds and leaves.

Project description:BackgroundMost studies of photosynthesis in chlorenchymas under potassium (K) deficiency focus exclusively on leaves; however, little information is available on the physiological role of K on reproductive structures, which play a critical role in plant carbon gain. Brassica napus L., a natural organ-succession species, was used to compare the morphological, anatomical and photo-physiological differences between leaves and siliques exposed to K-deficiency.ResultsCompared to leaves, siliques displayed considerably lower CO2 assimilation rates (A) under K-deficient (-K) or sufficient conditions (+K), limited by decreased stomatal conductance (g s), apparent quantum yield (α) and carboxylation efficiency (CE), as well as the ratio of the maximum rate of electron transport (J max) and the maximum rate of ribulose 1,5-bisphosphate (RuBP) carboxylation (V cmax). The estimated J max, V cmax and α of siliques were considerably lower than the theoretical value calculated on the basis of a similar ratio between these parameters and chlorophyll concentration (i.e. J max/Chl, V cmax/Chl and α/Chl) to leaves, of which the gaps between estimated- and theoretical-J max was the largest. In addition, the average ratio of J max to V cmax was 16.1% lower than that of leaves, indicating that the weakened electron transport was insufficient to meet the requirements for carbon assimilation. Siliques contained larger but fewer stoma, tightly packed cross-section with larger cells and fewer intercellular air spaces, fewer and smaller chloroplasts and thin grana lamellae, which might be linked to the reduction in light capture and CO2 diffusion. K-deficiency significantly decreased leaf and silique A under the combination of down-regulated stomatal size and g s, chloroplast number, α, V cmax and J max, while the CO2 diffusion distance between chloroplast and cell wall (D chl-cw) was enhanced. Siliques were more sensitive than leaves to K-starvation, exhibiting smaller reductions in tissue K and parameters such as g s, V cmax, J max and D chl-cw.ConclusionSiliques had substantially smaller A than leaves, which was attributed to less efficient functioning of the photosynthetic apparatus, especially the integrated limitations of biochemical processes (J max and V cmax) and α; however, siliques were slightly less sensitive to K deficiency.

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Disruption of carotene biosynthesis leads to abnormal plastids and variegated leaves in Brassica napus.

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Disruption of carotene biosynthesis leads to abnormal plastids and variegated leaves in Brassica napus.

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