Knowledge on genes related to plant responses to adverse growth conditions and development is essential for germplasm improvement. In this study, a chrysanthemum R2R3-MYB transcription factor gene, designated CmMYB2 (GenBank accession No. JF795918), was cloned and functionally characterized. Expression of CmMYB2 in chrysanthemum leaves was up-regulated in response to drought, salinity and cold stress, as well as by treatment with exogenous abscisic acid (ABA). When the gene was constitutively expressed in Arabidopsis thaliana, it increased plant sensitivity to ABA and reduced stomatal aperture. Plant survival under drought was improved than in the wild type, as was the plants' salinity tolerance. The level of expression of a number of genes associated with the stress response, including RD22, RD29A, RAB18, COR47, ABA1 and ABA2, was raised in the CmMYB2 transgenic Arabidopsis plants. CmMYB2 transgenic Arabidopsis plants were also delayed in flowering. The expression of CONSTANS (CO), FLOWERING LOCUS T (FT), SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1 (SOC1), LEAFY (LFY) and APETALA1 (AP1) genes involved in flowering was down-regulated in the CmMYB2 transgenics. Together, these results suggest that CmMYB2 may be a promising gene for the drought and salt tolerance improvement and flowering-time modulation.
Heat shock proteins are associated with protection against various abiotic stresses. Here, the isolation of a chrysanthemum cDNA belonging to the HSP70 family is reported. The cDNA, designated CgHSP70, encodes a 647-residue polypeptide, of estimated molecular mass 70.90 kDa and pI 5.12. A sub-cellular localization assay indicated that the cDNA product is deposited in the cytoplasm and nucleus. The performance of Arabidopsis thaliana plants constitutively expressing CgHSP70 demonstrated that the gene enhances tolerance to heat, drought and salinity. When CgHSP70 was stably over-expressed in chrysanthemum, the plants showed an increased peroxidase (POD) activity, higher proline content and inhibited malondialdehyde (MDA) content. After heat stress, drought or salinity the transgenic plants were better able to recover, demonstrating CgHSP70 positive effect.
Zinc finger proteins (ZFPs) play an important role in plant growth and development. Here, we describe the isolation of CgZFP1, a Cys2/His2 (C(2)H(2)) ZFP gene, using RACE PCR from chrysanthemum, and the investigation of its function with ectopic expression in Arabidopsis. CgZFP1 contains two conserved C(2)H(2) regions, a nuclear localization domain (B box), a Leu-rich domain (L box) and a conserved DLN sequence (DLN box) close to its C-terminus. Its expression in the chrysanthemum leaf was strongly induced by salinity or drought, but not by ABA. Subcellular localization assay indicated that CgZFP1 protein is localized in nucleus in vivo. Yeast-one hybrid assay showed that CgZFP1 possesses transcriptional activation ability, heterologous expression of CgZFP1 conferred tolerance of transgenic Arabidopsis plants to both salinity and drought stresses. Under salinity stress, genes involved in osmotic adjustment, ROS scavenging, and ion homeostasis: Atlea3, AtP5CS2, AtProT1, and AtMnSOD, AtPOD, AtAPX1, and AtSOS1, AtSOS2, AtSOS3, AtNHX1 were enhanced in CgZFP1 transgenic Arabidopsis plants. Moreover, genes involved in the osmotic adjustment and oxidative stress responses: Atlea3, AtP5CS2, AtProT1, the aquaporin AtPIP2A, and AtMnSOD, AtPOD, AtAPX1 were induced in CgZFP1 transgenic Arabidopsis under drought stress. These results indicate CgZFP1 is an important regulator involved in the salt and drought stress response in plants.
Omics research has indicated that heat shock protein 70 (HSP70) is a potential biomarker of meat quality. However, the specific changes and the potential role of HSP70 in postmortem meat quality development need to be further defined. In this study, Arbor Acres broiler chickens (n = 126) were randomly categorized into three treatment groups of unstressed control (C), 0.5-h transport (T) and subsequent water shower spray following transport (T/W). Each treatment consisted of six replicates with seven birds each. The birds were transported according to a designed protocol. The pectoralis major (PM) muscles of the transport-stressed broilers were categorized as normal and pale, soft and exudative (PSE)-like muscle samples according to L* and pH 24 h values to test the expression and location of HSP70. Results revealed that the activities of plasma creatine kinase and lactate dehydrogenase increased significantly ( P < 0.05) in normal and PSE-like muscle samples after transportation. The mRNA expression of HSP70 in normal muscle samples increased significantly ( P < 0.05) compared with that in the controls after stress. The protein expression of HSP70 increased significantly in normal muscle samples and decreased significantly ( P < 0.05) in PSE-like muscles. Immuno-fluorescence showed that HSP70 was present in the cytoplasm and on surface membranes of PM muscle cells in the normal samples following stress. Meanwhile, HSP70 was present on the surface membranes and extracellular matrix but was barely visible in the cytoplasm of the PSE-like samples. Principal component analysis showed high correlations between HSP70 and meat quality and stress indicators. In conclusion, this research suggests that the variation in HSP70 expression may provide a novel insight into the pathways underlying meat quality development.
Streptococcus thermophilus is one of the economically most representatives of lactic acid bacteria, which is widely used as a starter to produce fermented milk products. In this study, 22 S. thermophilus strains were isolated from 26 fermented milk samples. Most isolates showed the ability to ferment a broad range of carbohydrates. Interestingly, eight strains are galactose positive, which is a desirable property in various industrial dairy fermentations. Four different nucleotide sequences were found in the galR-galK intergenic regions. The 16S-23S intergenic spacer region sequences of most isolates were determined as ITS-St-II type, which are related with protease positive and fast acidification. CS18 presented excellent technological performances, and showed potential as a promising starter candidate. To gain a comprehensive view of stress response mechanisms of strains, the distribution of all the two-component systems (TCSs) in strains were investigated. TCS analysis indicated that the nucleotide sequence of TCSs have obvious differences in different strains. And the strains with the special nucleotide sequences of TCS have distinctive traits. Therefore, it was speculated that there is a certain connection between the traits' difference and the TCS difference of strains.
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