Molecular cloning, characterization, and expression of an alfalfa (Medicago sativa L.) heme oxygenase-1 gene, MsHO1, which is pro-oxidants-regulated

Fu, Guang-Qing; Xu, Sheng; Xie, Yanjie; Han, Bin; Nie, Li; Shen, Wenbiao; Ren, Wei

doi:10.1016/j.plaphy.2011.01.018

Cited by 35 publications

(30 citation statements)

References 32 publications

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“…After electrophoresis, the gel was stained with Coomassie brilliant blue and photographed. Western blotting analysis was carried out as Fu et al (2011) described with small modifications. Briefly, separated proteins were transferred to polyvinylidene difluoride (PVDF) membranes using a semi-dry transfer system.…”

Section: Prokaryotic Expression and Western Blotting Of Cshis Proteinmentioning

confidence: 99%

Overexpression of Camellia sinensis H1 histone gene confers abiotic stress tolerance in transgenic tobacco

Wang

et al. 2014

Plant Cell Rep

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Overexpression of CsHis in tobacco promoted chromatin condensation, but did not affect the phenotype. It also conferred tolerance to low-temperature, high-salinity, ABA, drought and oxidative stress in transgenic tobacco. H1 histone, as a major structural protein of higher-order chromatin, is associated with stress responses in plants. Here, we describe the functions of the Camellia sinensis H1 Histone gene (CsHis) to illustrate its roles in plant responses to stresses. Subcellular localization and prokaryotic expression assays showed that the CsHis protein is localized in the nucleus, and its molecular size is approximately 22.5 kD. The expression levels of CsHis in C. sinensis leaves under various conditions were investigated by qRT-PCR, and the results indicated that CsHis was strongly induced by various abiotic stresses such as low-temperature, high-salinity, ABA, drought and oxidative stress. Overexpression of CsHis in tobacco (Nicotiana tabacum) promoted chromatin condensation, while there were almost no changes in the growth and development of transgenic tobacco plants. Phylogenetic analysis showed that CsHis belongs to the H1C and H1D variants of H1 histones, which are stress-induced variants and not the key variants required for growth and development. Stress tolerance analysis indicated that the transgenic tobacco plants exhibited higher tolerance than the WT plants upon exposure to various abiotic stresses; the transgenic plants displayed reduced wilting and senescence and exhibited greater net photosynthetic rate (Pn), stomatal conductance (Gs) and maximal photochemical efficiency (Fv/Fm) values. All the above results suggest that CsHis is a stress-induced gene and that its overexpression improves the tolerance to various abiotic stresses in the transgenic tobacco plants, possibly through the maintenance of photosynthetic efficiency.

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Section: Prokaryotic Expression and Western Blotting Of Cshis Proteinmentioning

confidence: 99%

Overexpression of Camellia sinensis H1 histone gene confers abiotic stress tolerance in transgenic tobacco

Wang

et al. 2014

Plant Cell Rep

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“…The nucleotide sequence was 53.59% identical to the AtHO2 gene within the predicted coding region, and the putative translated product was 50.9% similar to the AtHO2 protein. Similar to that of MsHO1 [20], MsHO2 protein also has an N-terminal transit peptide sequence for import into chloroplast or other cell organelle. A proposed N-terminal transit peptide of 28 amino acid residues was predicted by the ChloroP algorithm ( [24]; http://www.cbs.dtu.dk/services/ ChloroP/).…”

Section: Resultsmentioning

confidence: 99%

“…HO activity was analyzed using the method described in our previous reports [20][21][22]. In the HO activity test, the concentration of biliverdin IX (BV) was estimated using a molar absorption coefficient at 650 nm of 6.25 mM −1 cm −1 in 0.1 M HEPES-NaOH buffer (pH 7.2).…”

Section: Ho Activity Determinationmentioning

confidence: 99%

“…Further results showed that AtHO2 exhibited strong binding of proto IX (protoporphyrin IX), a precursor for both heme and chlorophyll biosynthesis, thus suggesting that HO2 may play a role in the regulation of tetrapyrrole metabolism [19]. Previously, we cloned and characterized an alfalfa (Medicago sativa L.) HO-1 gene, MsHO1 [20]. Further experiments showed that MsHO-1 is pro-oxidantregulated.…”

Section: Introductionmentioning

confidence: 99%

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Cloning and Characterization of a Heme Oxygenase-2 Gene from Alfalfa (Medicago sativa L.)

Jin

Lin

et al. 2011

Appl Biochem Biotechnol

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Heme oxygenase (HO, EC 1.14.99.3) catalyzes the oxidation of heme and performs vital roles in plant development and stress responses. Two HO isozymes exist in plants. Between these, HO-1 is an oxidative stress-response protein, and HO-2 usually exhibited constitutive expression. Although alfalfa HO-1 gene (MsHO1) has been investigated previously, HO2 is still poorly understood. In this study, we report the cloning and characterization of HO2 gene, MsHO2, from alfalfa (Medica sativa L.). The full-length cDNA of MsHO2 contains an ORF of 870 bp and encodes for 290 amino acid residues with a predicted molecular mass of 33.3 kDa. Similar to MsHO1, MsHO2 also appears to have an N-terminal transit peptide sequence for chloroplast import. Many conserved residues in plant HO were also conserved in MsHO2. However, unlike HO-1, the conserved histidine (His) required for heme-iron binding and HO activity was replaced by tyrosine (Tyr) in MsHO2. Further biochemical activity analysis of purified mature MsHO2 showed no HO activity, suggesting that MsHO2 may not be a true HO in nature. Semi-quantitative RT-PCR confirmed its maximum expression in the germinating seeds. Importantly, the expression levels of MsHO2 were up-regulated under sodium nitroprusside (SNP) and H(2)O(2) (especially) treatment, respectively.

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“…Heme oxygenase is the rate-limiting enzyme in the breakdown of heme changing into carbon monoxide (CO), iron etc (Shekhawat & Verma, 2010) and it plays a vital role in stress responses. The expression and protein levels of MsHO1 are higher in alfalfa stems and leaves than in germinating seeds and roots and are induced significantly by some pro-oxidant compounds including hemin and nitric oxide donor sodium nitroprusside (Fu et al, 2011).…”

Section: Signal Transductionmentioning

confidence: 99%

Molecular and Genetic Analysis of Abiotic Stress Resistance of Forage Crops

Wang¹,

Gao²,

Li³

et al. 2011

Plants and Environment

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Molecular cloning, characterization, and expression of an alfalfa (Medicago sativa L.) heme oxygenase-1 gene, MsHO1, which is pro-oxidants-regulated

Cited by 35 publications

References 32 publications

Overexpression of Camellia sinensis H1 histone gene confers abiotic stress tolerance in transgenic tobacco

Overexpression of Camellia sinensis H1 histone gene confers abiotic stress tolerance in transgenic tobacco

Cloning and Characterization of a Heme Oxygenase-2 Gene from Alfalfa (Medicago sativa L.)

Molecular and Genetic Analysis of Abiotic Stress Resistance of Forage Crops

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