Macrophage migration inhibitory factor in mud crab Scylla paramamosain: Molecular cloning, expression profiles in various tissues and under Vibrio challenge

As one of the first found cytokines, macrophage migration inhibitory factor (MIF) plays an important role in several physiological processes in crabs. In this study, a full-length MIF cDNA (GenBank accession number: JX131610) from mud crab Scylla paramamosain (Sp) was cloned based on a sequence of S. paramamosain cDNA library. The full length of SpMIF was 734 bp consisting of a 363 bp open reading frame encoding the SpMIF, a 120 amino acid peptide chain. The molecular weight of SpMIF was 13.46 kDa with the pI of 6.82. The alignment analysis showed that SpMIF appeared to be closely related to the counterpart from Eriocheir sinensis (68%). Quantitative real-time PCR analysis revealed that SpMIF was highly expressed in hepatopancreas and hemocytes. In addition, the expression level of SpMIF was increased significantly after a 6-h challenge by Vibrio parahaemolyticus (4.00 × 10⁶ CFU/mL), peaked at 8 h, and then declined to the common level in 48 h. This data indicated that SpMIF was cloned successfully, and suggested that it participated in the immune system of mud crabs.     

Effects of cytokinin on photosynthetic gas exchange, chlorophyll fluorescence parameters, antioxidative system and carbohydrate accumulation in cucumber (Cucumis sativus L.) under low light

The effects of 6-benzylaminopurine (6-BA) on plant growth, net photosynthetic rate, relative chlorophyll content, soluble protein, carbohydrates contents and antioxidant systems of cucumber (Cucumis sativus L.) under low-light environment were investigated using two different cucumber cultivars. The results showed that the weak light resulted in the remarkable decrease in plant net photosynthetic rate, relative chlorophyll content, soluble protein and carbohydrates contents, but promoted the superoxide dismutase and guaiacol peroxidase activities. However, application of 6-BA alleviated the reduction of the correlative parameters and mediated the changes of antioxidant systems. The potential mechanisms may involve the following aspects: 6-BA clearly enhanced the plants’ tolerance to low light by increasing chlorophyll content, reducing the production of superoxide radical (O ₂ ^·? ), and enhancing the quenching of hydrogen peroxide (H₂O₂), consequently alleviating the injury of photosynthetic system, and further increasing the efficiency of CO₂ assimilation, producing more carbohydrates which can meet the growth need of cucumber. Meanwhile, the present study indicated that cucumber of Europe mini type (Chunqiuwang) was more tolerant to low light than HuaNan type (Huza No.3).     

Screening proteins interacting with VpPR10.1 of Chinese wild grapevine using the yeast two-hybrid system

Among the 17 plant pathogenesis-related (PR) protein families, only PR10 family is intracellular and cytosolic. PR proteins are expressed in response to pathogen challenge and abiotic stresses in higher plants. However, the molecular mechanisms of their actions remain poorly understood. In a previous work, we isolated a PR10 gene from Erysiphe necator-resistant Chinese wild Vitis sp. (Baihe-35-1) and it was designated as VpPR10.1. In this study, yeast two-hybrid method was used to screen proteins interacting with VpPR10.1 proteins. Twenty-one ESTs were isolated and sequenced. All sequences were compared using BLASTx to identify presumptive orthologs. Several proteins associated with VpPR10.1 protein were screened, including CNR8, UFGT6, HSP, DEAD-box, Trx h2, Grx C9 and GLOX. These proteins are closely related to defensive action of plants against pathogens and abiotic stresses. Our results reveal that VpPR10.1 gene may be involved in hormone signaling, programmed cell death and defense responses of grapevine.     

Purification and characterization of two extracellular polyhydroxyalkanoate depolymerases from Pseudomonas mendocina

Two polyhydroxyalkanoate depolymerases, PHAase I and PHAase II, were purified to homogeneity from the culture supernatant of an effective PHA-degrading bacterium, Pseudomonas mendocina DS04-T. The molecular masses of PHAase I and PHAase II were determined by SDS-PAGE as 59.4 and 33.8 kDa, respectively. Their optimum pH values were 8.5 and 8, respectively. Enzymatic activity was optimal at 50 °C. Both purified enzymes could degrade PHB, PHBV, and P(3HB-co-4HB). Addition of Na⁺ and K⁺ slightly increased the rate of PHAase II. EDTA significantly inhibited PHAase II but not PHAase I. Mercaptoethanol and H₂O₂ also inhibited the activities of both enzymes.     

Growth characteristics of Adiantum reniforme var. sinensis and A. capillus‐veneris in response to light and soil moisture

Adiantum reniforme var. sinensis (Adiantaceae) is an endangered fern endemic to the Three Gorges region in southwest China. To elucidate possible ecophysiological mechanisms restricting its distribution, effects of the availability of light (28%, 14% and 7% of open field) and soil moisture (60% and 40% of field capacity) on dry matter production and allocation, leaf morphology and water use efficiency (WUE) were examined in A. reniforme var. sinensis and its widespread congener A. capillus‐veneris. Both species had lower root/shoot ratio (R/S) and higher specific leaf area (SLA) when grown at low light. However, A. reniforme var. sinensis showed less plasticity for total leaf area (LA) and leaf area ratio (LAR) than A. capillus‐veneris, and its root mass, total mass and WUE decreased as light availability decreased. Under water stress, all traits of both species except WUE were significantly affected. However, drought stress decreased total mass of A. capillus‐veneris but did not have a significant effect on A. reniforme var. sinensis. Compared with A. capillus‐veneris, A. reniforme var. sinensis had significantly higher R/S but lower values for other analyzed traits. These results suggest that A. reniforme var. sinensis is relatively superior in drought tolerance but inferior at low light, allowing it to persist in habitats with low soil moisture and high light availability but with few coexisting species present.     

CmPEX6, a Gene Involved in Peroxisome Biogenesis,Is Essential for Parasitism and Conidiation by the Sclerotial Parasite Coniothyrium minitans

Coniothyrium minitans is a sclerotial parasite of the plant-pathogenic fungus Sclerotinia sclerotiorum, and conidial production and parasitism are two important aspects for commercialization of this biological control agent. To understand the mechanism of conidiation and parasitism at the molecular level, we constructed a transfer DNA (tDNA) insertional library with the wild-type strain ZS-1. A conidiation-deficient mutant, ZS-1TN22803, was uncovered through screening of this library. This mutant could produce pycnidia on potato dextrose agar (PDA), but most were immature and did not bear conidia. Moreover, this mutant lost the ability to parasitize or rot the sclerotia of S. sclerotiorum. Analysis of the tDNA flanking sequences revealed that a peroxisome biogenesis factor 6 (PEX6) homolog of Saccharomyces cerevisiae, named CmPEX6, was disrupted by the tDNA insertion in this mutant. Targeted gene replacement and gene complementation tests confirmed that a null mutation of CmPEX6 was responsible for the phenotype of ZS-1TN22803. Further analysis showed that both ZS-1TN22803 and the targeted replacement mutants could not grow on PDA medium containing oleic acid, and they produced much less nitric oxide (NO) and hydrogen peroxide (H₂O₂) than wild-type strain ZS-1. The conidiation of ZS-1TN22803 was partially restored by adding acetyl-CoA or glyoxylic acid to the growth media. Our results suggest that fatty acid β-oxidation, reactive oxygen and nitrogen species, and possibly other unknown pathways in peroxisomes are involved in conidiation and parasitism by C. minitans.     

Abundance and Diversity of Ammonia-Oxidizing Bacteria and Archaea in Cold Springs on the Qinghai-Tibet Plateau

The cold springs underlain by gas hydrates on the Qinghai-Tibet Plateau (QTP) are similar to deep-sea cold seeps with respect to methane biogeochemistry. Previous studies have shown that ammonia oxidizing bacteria (AOB) and archaea (AOA) are actively present and play important roles in the carbon/nitrogen cycles in cold seeps. Studying AOA and AOB communities in the QTP cold springs will be of great importance to our understanding of carbon and nitrogen cycling dynamics related to the underlying gas hydrates on the QTP. Thus, the abundance and diversity of AOB and AOA in sediments of four cold springs underlain by gas hydrates on the QTP were determined by using quantitative polymerase chain reaction and amoA gene (encoding ammonia monooxygenase involved in ammonia oxidation) phylogenetic analysis. The results showed that the AOB and AOA amoA gene abundances were at 10³–10⁴ copies per gram of the sediments in the investigated cold springs. The AOB population consisted of Nitrosospira and Nitrosomonas in contrast with the mere presence of Nitrosospira in marine cold seeps. The AOB diversity was higher in cold springs than in cold seeps. The AOA population was mainly composed of Nitrososphaera, in contrast with the dominance of Nitrosopumilus in cold seeps. The terrestrial origin and high level of dissolved oxygen of the cold springs may be the main factors accounting for the observed differences in AOB and AOA populations between the QTP cold springs and marine cold seeps.