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刺五加甲羟戊酸焦磷酸脱羧酶基因的克隆与表达分析 总被引:1,自引:0,他引:1
利用RACE技术克隆刺五加甲羟戊酸焦磷酸脱羧酶(mevalonate diphosphate decarboxylase,MDD)基因的全长cDNA序列,运用生物信息学方法对该基因进行分析,并通过RT-PCR法检测MDD在刺五加不同生长发育时期和不同器官中的表达情况。结果表明:(1)刺五加MDD基因cDNA序列全长1 769bp(GenBank登录号为JQ905594),开放阅读框全长1 263bp,编码420个氨基酸残基,包含GHMP激酶超家族的特异性识别序列;刺五加MDD蛋白的二级结构中含有161个α螺旋,占38.33%;68个延伸链,占16.19%;19个β折叠,占4.52%;172个无规则卷曲,占40.95%;刺五加MDD蛋白无跨膜区域,定位于膜外。(2)刺五加MDD基因在不同生长发育时期和器官中均有表达,但表达量具有显著差异(P<0.05)。在整个生长期中,MDD的表达呈现高-低-高-低的变化趋势,第一个表达高峰出现在萌芽期至叶片完全展开时,第二个高峰出现在果实体积快速增长期,最高表达量(叶片完全展开期)为最低表达量(叶片衰老期)的4.51倍;不同器官中,幼茎的表达量最高,为最低表达量(叶片)的7.22倍,但叶片、叶柄和根中的表达量差异不显著。研究结果为阐明刺五加皂苷的生物合成及对其进行表达调控奠定了基础。 相似文献
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Linjun Cai Junhua Liu Han Yun Dan Du Xiaolong Zhong Zhenlin Yang Xianchun Sang Changwei Zhang 《Phyton》2021,90(4):1103-1117
Leaf color mutants are ideal materials for studying many plant physiological and metabolic processes such as photosynthesis, photomorphogenesis, hormone physiology and disease resistance. In this study, the genetically stable yellow-green leaf mutant ygl16 was identified from mutated “Xinong 1B”. Compared with the wild type, the pigment concentration and photosynthetic capacity of the ygl16 decreased significantly. The ultrastructural observation showed that the distribution of thylakoid lamellae was irregular in ygl16 chloroplasts, and the grana and matrix lamellae were blurred and loose in varied degrees, and the chloroplast structure was disordered, while the osmiophilic corpuscles increased. The results of the genetic analysis and mapping showed that the phenotype of ygl16 was controlled by a pair of recessive nuclear gene. The gene located in the 56Kb interval between RM25654 and R3 on the long arm of chromosome 10. The sequencing results showed that the 121st base of the first intron of the candidate gene OsPORB/FGL changed from A to T in the interval. qRT-PCR results showed that the expression of chlorophyll synthase-related genes in the mutant decreased. 相似文献
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The lignocellulosic crop Miscanthus spp. has been identified as a good candidate for biomass production. The
responses of Miscanthus sinensis Anderss. to salinity were studied to satisfy the needs for high yields in marginal
areas and to avoid competition with food production. The results indicated that the relative advantages of the
tolerant accession over the sensitive one under saline conditions were associated with restricted Na+ accumulation
in shoots. Seedlings of two accessions (salt-tolerant ‘JM0119’ and salt-sensitive ‘JM0099’) were subjected to 0
(control), 100, 200, and 300 mM NaCl stress to better understand the salt-induced biochemical responses of genes
involved in Na+ accumulation in M. sinensis. The adaptation responses of genes encoding for Na+
/H+ antiporters,
NHX1 and SOS1 to NaCl stress were examined in JM0119 and JM0099.The cDNA sequences of genes examined
were highly conserved among the relatives of M. sinensis based on the sequencing on approximate 600 bp-long
cDNA fragments obtained from degenerate PCR. These salt-induced variations of gene expression investigated by
quantitative real-time PCR provided evidences for insights of the molecular mechanisms of salt tolerance in
M. sinensis. The expression of NHX1 was up-regulated by salt stress in JM0119 shoot and root tissues. However,
it was hardly affected in JM0099 shoot tissue except for a significant increase at the 100 mM salt treatment, and it
was salt-suppressed in the JM0099 root tissue. In the root tissue, the expression of SOS1 was induced by the high
salt treatment in JM0119 but repressed by all salt treatments in JM0099. Thus, the remarkably higher expression
of NHX1 and SOS1 were associated with the resistance to Na+ toxicity by regulation of the Na+ influx, efflux, and
sequestration under different salt conditions. 相似文献
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Gibberellic acid controlled the key developmental processes of the life cycle of landing plants, and regulated the growth and development of plants. In this study, a novel gibberellin receptor gene EuGID1 was obtained from Eucommia ulmoides Oliver. The cDNA of EuGID1 was 1556 bp, and the open reading frame was 1029 bp, which encoded 343 amino acids. EuGID1 had the homology sequence with the hormone-sensitive lipase family. Amino acid sequence alignment confirmed EuGID1 protein had the highest homology with the GID1 protein of Manihot esculenta. EuGID1 was located in the nucleus and cell membrane and had expression in four plant organs. Overexpression of EuGID1 in transgenic Arabidopsis plants promoted plant elongation and increased siliques yield. 相似文献
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Jing Qu Shuang Liu Peng Jiao Zhenzhong Jiang Jianbo Fei Shuyan Guan Yiyong Ma 《Phyton》2022,91(8):1709-1719
To develop cold-tolerant maize germplasms and identify the activation of INDUCER OF CRT/DRE-BINDING FACTOR EXPRESSION (ICE1) expression in response to cold stress, RT-PCR was used to amplify the complete open reading frame sequence of the ICE1 gene and construct the plant expression vector pCAMBIA3301-ICE1-Bar. Immature maize embryos and calli were transformed with the recombinant vector using Agrobacterium tumefaciens-mediated transformations. From the regenerated plantlets, three T1 lines were screened and identified by PCR. A Southern blot analysis showed that a single copy of the ICE1 gene was integrated into the maize (Zea mays L.) genomes of the three T1 generations. Under low temperature-stress conditions (4°C), the relative conductivity levels decreased by 27.51%–31.44%, the proline concentrations increased by 12.50%–17.50%, the malondialdehyde concentrations decreased by 16.78%–18.37%, and the peroxidase activities increased by 19.60%–22.89% in the T1 lines compared with those of the control. A real-time quantitative PCR analysis showed that the ICE1 gene was ectopically expressed in the roots, stems, and leaves of the T1 lines. ICE1 positively regulates the expression of the CBF genes in response to cold stress. Thus, this study showed the successful transformation of maize with the ICE1 gene, resulting in the generation of a new maize germplasm that had increased tolerance to cold stress. 相似文献
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Chromatin structure, determined in part by DNA methylation, is established during differentiation and prevents expression
of genes unnecessary for the function of a given cell type. We reported that DNA methylation and chromatin structure contributes
to lymphoidspecific ITGAL (CD11a) and PRF1 (perforin) expression. We used bisulfite sequencing to compare methylation patterns in the ITGAL promoter and 5′ flanking region of T cells and fibroblasts, and in the PRF1 promoter and upstream enhancer of CD4+ and CD8+ T cells with fibroblasts. The effects of methylation on promoter function
were tested using regional methylation of reporter constructs, and confirmed by DNA methyltransferase inhibition. The relationship
between DNA methylation and chromatin structure was analyzed by DNaseI hypersensitivity. Herein we described the methods and
results in greater detail.
Published: September 16, 2004. 相似文献
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Ziyang Feng Xinqi Cheng Tianwen Wang Yongchao Han Haihong Chen Xinyu Zhang Jie Sun Wei Zhang Feng Liu 《Phyton》2023,92(5):1633-1647
Fatty Acyl-ACP thioesterase (FAT) is a key enzyme controlling oil biosynthesis in plant seeds. FATs can be divided into two subfamilies, FATA and FATB according to their amino acid sequences and substrate specificity. The Upland cotton genome contains 20 GhFAT genes, amongst which 6 genes were of the GhFATA subfamily and 14 of the GhFATB subfamily. The 20 GhFAT genes are unevenly distributed on 14 chromosomes. The GhFATA genes have 5 or 7 exons and the GhFATB genes have 6 or 7 exons. All GhFAT proteins have the conserved Acyl-ACP_TE domain and PLN02370 super family, the typical characteristics of plant thioesterases. Analyses of the expression level of GhFATs and the compositions of fatty acid in 5–60 days-post-anthesis seeds showed that the ratio of saturated fatty acids to unsaturated fatty acids was consistent with the expression profile of GhFATB12, GhFATB3, and GhFATB10; the ratio of monounsaturated fatty acid to polyunsaturated fatty acids was consistent with the expression profile of GhFATA3. The oil contents of mature cottonseeds were positively correlated with the contents of palmitic acid and linolenic acid as well as seed vigor. These results provide essential information for further exploring the role(s) of the specific GhFATs in determining oil biosynthesis and cottonseed compositions. 相似文献
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Phytophthora nicotianae causes substantial economic losses in most countries where tobacco is produced. At present, the control of P. nicotianae mainly depends on chemical methods, with considerable environmental and
health issues. We investigated the effects of ethanol extracts from Scutellaria baicalensis Georgi (SBG) and Magnolia officinalis (MO). On mycelial growth, sporangium formation, and zoospore release of P. nicotianae. Both
extracts inhibited the growth of P. nicotianae, with mycelial growth inhibition rates of 88.92% and 93.92%, respectively, at 40 mg/mL, and EC50 values of 5.39 and 5.74 mg/mL, respectively. The underlying mechanisms were the
inhibition of sporangium formation, the reduction of zoospore number, and the destruction of the mycelium
structure. At an SBG extract concentration of 16.17 mg/mL, the inhibition rates for sporangia and zoospores were
98.66% and 99.39%, respectively. At an MO extract concentration of 2.87 mg/mL, the production of sporangia
and zoospores was completely inhibited. The hyphae treated with the two plant extracts showed different degrees
of deformation and damage. Hyphae treated with SBG extract showed adhesion and local swelling, whereas treatment with MO extract resulted in broken hyphae. Mixture of the extracts resulted in a good synergistic effect. 相似文献
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Xiaofang Zeng Guangzheng Li Nu’an Liu Yan Li Jianrong Li Xiaozhen Huang Degang Zhao 《Phyton》2022,91(11):2419-2437
Leaf-color mutants play an important role in the study of chlorophyll metabolism, chloroplast development, and
photosynthesis system. In this study, the yellow leaf 1 (yl1) rice mutant was identified from the ethyl methane
sulfonate-treated mutant progeny of Lailong, a glutinous japonica rice landrace cultivated in Guizhou Province,
China. Results showed that yl1 exhibited yellow leaves with decreased chlorophyll content throughout the growth
period. Chloroplast development in the yl1 mutant was disrupted, and the grana lamellae was loosely packed and
disordered. RNA sequencing and real-time quantitative polymerase chain reaction (qRT-PCR) analysis revealed
that the chlorophyll synthesis-related genes OsCHLH, OsCHLM, OsCHLG, PORB, and YGL8, as well as the chloroplast development-related genes FtsZ, OsRpoTp, and RbcL, were down-regulated in the yl1 mutant. Genetic analysis revealed that the yellow leaf phenotype of yl1 was controlled by recessive nuclear gene. By employing the
MutMap method, the mutation responsible for the phenotype was mapped to a 6.17 Mb region between
17.34 and 23.51 Mb on chromosome 3. Two non-synonymous single-nucleotide polymorphisms (SNPs) located
in the gene locus LOC_Os03g31210 and LOC_Os03g36760 were detected in this region. The two SNPs were
further confirmed by PCR and Sanger sequencing. The expression patterns of the two candidate genes indicated
that LOC_Os03g36760 showed greater potential for functional verification. Subcellular protein localization
revealed that the encoded product of LOC_Os03g36760 was localized in the nucleus, cytoplasm, and plasma membrane. These results will be useful for further characterization and cloning of the yl1 gene, and for research on the
molecular mechanisms controlling biogenesis and chloroplast biochemical processes. 相似文献