Elementary study of differential expression of total soluble proteins in different life phases of Porphyra yezoensis

Total soluble proteins of different life stages, filamentous sporophytes cultivated in high temperatures, and blade gametophytes harvested in different seasons, were identified by SDS-PAGE. The types and amounts of expressed proteins also varied amongst the samples. The fewest soluble proteins were present in filamentous sporophytes. There were more types and amounts of soluble protein in conchospores than in filamentous sporophytes, but fewer than in bulgy sporophytes. More types of protein were detected in filamentous sporophytes cultivated in high temperatures than in those growing in normal situations. The most types and amounts of protein were found in blade gametophytes in all samples. Blade gametophytes harvested last year and stored at -20 ℃ showed only minor differences in expression of proteins when compared with those harvested in different seasons.

The Relationship Between Developmental Accumulation of Leaf Soluble Proteins and Vernalization Response of Wheat(Triticum aestivum L.em. Thell)

The relationship between vernalization requirement and quantitative and qualitative changes in total leaf soluble proteins were determined in one spring （cv. Kohdasht） and two winter （cvs. Sardari and Norstar） cultivars of wheat （Triticum aestivum L.） exposed to 4℃. Plants were sampled on days 2, 14, 21 and 35 of exposure to 4℃. The final leaf number （FLN） was determined throughout the vernalization periods （0, 7, 14, 24, and 35 d） at 4℃. The final leaf number decreased until days 24 and 35 in Sardari and Norstar eultivars, respectively, indicating the vernalization saturation at these times. No clear changes were detected in the final leaf number of Kohdash cultivar, verifying no vernalization requirement for this spring wheat cultivar. Comparing with control, clear cold-induced 2-fold increases in proteins quantity occurred after 48 h following the 4℃-treatment in the leaves of the both winter wheat cultivars but, such response was not detected in the spring cultivar. However, the electrophoretic protein patterns showed between-cultivar and between-temperature treatment differences. With increasing exposure time to 4℃, the winter cultivars tended to produce more HMW polypeptides than the spring cultivar. Similar proteins were induced in both Sardari and Norstar winter wheat cultivars, however, the long vernalization requirement in Norstar resulted in high level and longer duration of expression of cold-induced proteins compared to Sardari with a short vernalization requirement. These observations indicate that vernalization response regulates the expression of low temperature （LT） tolerance proteins and determines the duration of expression of LT- induced proteins.

Effect of Light Emitting Diodes (LEDs) on Growth, Mineral Composition, and Nutritional Value of Wheat & Lentil Sprouts

Sprouts are ready-to-eat and are recognized worldwide as functional components of the human diet.Recent advances in innovative agricultural techniques could enable an increase in the production of healthy food.The use of light-emitting diode(LED)in indoor agricultural production could alter the biological feedback loop,increasing the functional benefits of plant foods such as wheat and lentil sprouts and promoting the bioavailability of nutrients.The effects of white(W),red(R),and blue(B)light were investigated on the growth parameters and nutritional value of wheat and lentil sprouts.In the laboratory,seeds were sown under three different LED treat-ments:white,red,and blue light,while normal incandescent light served as a control.Percentage seed germina-tion improved by 18.34%and 12.67%for wheat and 18.34%and 12.67%for lentil sprouts under LED treatments R and B,respectively.An increase in total soluble protein and sugar by 33.4%and 9.23%in wheat and by 31.5%and 5.87%in lentils was observed under the R LED treatment.Vitamin C concentrations in wheat and lentils were significantly increased by R LED compared to all other treatments.Other parameters,including potassium and sodium concentrations,were significantly increased under red and blue light compared to the control;white light,on the other hand,significantly decreased all these parameters.According to the experimental data,red and blue LED light could be beneficial in the production of functional wheat and lentil sprouts with high nutrient concentrations.

Effects of Cold Acclimation and CaCl_2 on Total Soluble Protein, CaM and Freezing Resistance of Populus tomentosa Seedlings

Populus tomentosa seedlings used for cold acclimation at -3℃ were pretreated with or without 10?mmol·L -1 CaCl 2, 3?mmol·L -1 of Ca 2+ chelator EGTA, 0 1?mmol·L -1 of Ca 2+ channel inhibitor LaCl 3,and 0 05?mmol·L -1 of CaM antagonist CPZ. The changes in the contents of total soluble protein and CaM, and freezing resistance in all pretreated seedlings in various periods ( viz: following cold acclimation, chilling stress and recovery) were investigated. Results showed that cold acclimation increased the contents of total soluble protein and CaM, and freezing resistance of seedlings, which could be strongly reduced by the pretreatments of EGTA CPZ and LaCl 3 Cold acclimation combined with CaCl 2 pretreatment enhanced the effect of cold acclimation on freezing resistance, and obviously increased the contents of total soluble protein and CaM, reduced the declining degree of the contents of total soluble protein and CaM caused by chilling stress as compared with cold acclimation, augmented the increase in the level of total soluble protein and CaM during the recovery periods. Further analysis found that an increase in total soluble protein content during cold acclimation with or without CaCl 2 pretreatment mainly resulted from the increase in content of heat stable protein in total soluble protein. It is suggested that Ca 2+ calmodulin may be involved in the synthesis of total soluble protein, and the induction of freezing resistance of seedlings.

Cold Acclimation-Induced Changes in Total Soluble Protein, RNA, DNA, RNase and Freezing Resistance in Populus tomentosa Cuttings

The changes in the contents of total soluble protein and RNA, the activity of RNase in leaves and branches of Populus tomentosa cuttings at various periods (viz: cold acclimation, deacclimation, chilling stress and the recovery after chilling stress), and the survival rate and the freezing resistance of cuttings during cold acclimation at -3℃ were investigated. Results showed that cold acclimation not only increased the contents of total soluble protein and RNA, the survival rates and the freezing resistance of cuttings, decreased the activity of RNase, but also reduced the declining degree of total soluble protein and RNA contents, and the increasing level of RNase caused by chilling stress as compared with the controls. In addition, cold acclimation augmented the increase in the level of total soluble protein and RNA, and facilitated the decrease of RNase during the recovery periods. Further analysis found that the DNA content of all treatments kept relative stability at various periods. The changes in total soluble protein, RNA and RNase were closely related to the freezing resistance of cuttings. It appears that the increase of RNA content caused by cold acclimation induced decrease of RNase activity may be involved in the accumulation of total soluble protein and the induction of freezing resistance of cuttings.

A Comparative Study of Soluble Protein Extractions of Populus deltoides × （ Trichocarpa × Deltoides） for 2-DE

Background： The disclosure of the poplar genome strengthens its position as well-established model organism. Populus has been subject of several proteome studies, but up to date no comparative study was performed on the extraction method of soluble proteins for this species. The extraction is the most critical step in two-dimensional gel electrophoresis and each extraction method has its advantages, disadvantages and limitations. Therefore protein extraction methods should be optimized for each tissue before starting an experimental setup. In prospect of future DIGE （Differential Gel electrophoresis） experiments for the investigation of the effects of cadmium and inoculation with plant growth promoting bacteria at the proteome level, the aim of this study was to optimize an extraction method for soluble proteins of poplar leaves and roots. Results： The acetone-phenol extraction method was found to be the most suited, rendering a high spot number and low background interference. During further optimization, several critical steps in the extraction method were revealed. Conclusion： Aiming to optimize the extraction of soluble leaf and root proteins of Populus deltoides × （trichocarpa× deltoides） compatible with DIGE analysis, a protocol rendering high reproducibility, low background interference and a high spot number was established, however no novel insights were acquired.

Soluble protein and acid phosphatase exuded by ectomycorrhizal fungi and seedlings in response to excessive Cu and Cd

Fungi and their symbionts can alleviate heavy metal stress by exuding soluble proteins and enzymes. This study examined the role of soluble protein and acid phosphatase （APase） exuded by Xerocomus chrysenteron, an ectomycorrhizal fungus, and the seedlings of its symbiont, Chinese pine （Pinus tabulaeformis）, under conditions of excessive Cu and Cd. The growth type showed that this poorly studied ectomycorrhizal fungus was capable of tolerating high concentrations of Cu, and may be useful in phytoremediation. X. chrysenteron grew well at 80 mg/L Cu, and the EC50 for Cd was 17.82 mg/L. X. chrysenteron also showed enhanced exudation of soluble protein in both isolated and inoculated cultivations under the influence of Cu and Cd. Soluble protein exudation, however, differed under Cu and Cd stress in isolates. In mediums containing Cu, soluble protein exudation increased with concentration, but in mediums containing Cd the content of soluble protein increased to a comparable level at all concentrations. This study demonstrated that soluble protein was related to heavy metal tolerance, although the different ions played different roles. While APase activity in exudates of fungi and seedlings decreased under Cu and Cd stress in comparison to the control, the APase activity in seedlings was maintained by inoculation. Thus, X. chrysenteron facilitated the ability of plant to maintain a normal nutrient uptake, and therefore to protect it from heavy metal toxicity.

Relationship Between Soluble Protein,Chlorophyll and ATP in Drought-Resistant Sweet Potato Under Water Stress

Some indices concerning the metabolism of substance and energy in sweet potato leaves under water stress were studied. The results showed an obvious increase in soluble protein content. Compared with control, Chl a, Chl b, total Chl contents and the ratio of Chl a to Chl b all decreased to some extent. ATP content increased in some varieties and decreased in others, but the stronger the drought resistance of the variety , the higher the ATP content. The correlation coefficient(r)of the soluble protein content, ratio of Chl a to Chl b and ATP content as a percentage of the drought-resistant sweet potato control variety are 0. 8968, - 0. 8509 and 0. 8200, respectively, P<0. 01. So these indices can be used to evaluate the drought resistance of different sweet potato varieties.

An ELISA Based on a Truncated Soluble ORF2 Protein for the Detection of PCV2 Antibodies in Domestic Pigs

Postweaning multisystemic wasting syndrome (PMWS) is an important swine disease that is closely associated with porcine circovirus type 2 (PCV2). The capsid protein (Cap protein) is a major structural protein that has at least three immunoreactive regions, and it can be a suitable candidate antigen for detecting the specific antibodies of a PCV2 infection. In the present study, an indirect enzyme-linked immunosorbent assay (TcELISA) based on a truncated soluble Cap protein produced in Escherichia coli (E.coli) was established and validated for the diagnostic PCV2 antibodies in swine. The TcELISA was validated by comparison with an indirect immunofluorescence assay (IIFA). The diagnostic sensitivity (DSN), specificity (DSP), and accuracy of the TcELISA were 88.6%, 90.7% and 89.4%, respectively. The agreement rate was 89.38% between results obtained with TcELISA and IIFA on 113 field sera. A cross-reactivity assay showed that the method was PCV2-specific by comparison with other sera of viral disease. Therefore ,the TcELISA will be helpful for the development of a reliable serology diagnostic test for large scale detection of PCV2 antibodies and for the evaluation of vaccine against PCV2 in swine.

Dynamic Effects of Cerium on Syntheses of Soluble Protein and Taxol in Suspension Culture of Taxus Chinensis Var. Mairei Cells

The dynamic effects of Ce4+ on the syntheses of soluble protein and taxol in suspension cultures of Taxus chinensis var. mairei cells were studied. The phenomena of 'partition' and 'bifurcation' were observed in studying the dynamic effect of Ce4+ on soluble protein synthesis and cell activity. That is, Ce4+ of low concentration improves the soluble protein synthetic strength and cell activity, while Ce4+ of high concentration is harmful to protein synthesis and cell activity. In addition, Ce4+ of appropriate concentration enhances taxol synthesis.

Analysis of SSLP and Soluble Protein Contents in Leaves of Mutants Induced by High Pressure in Rice (Oryza sativa)

Rice variety Yuexiangzhan and its mutants induced by high pressure were studied using microsatcllite markers and soluble protein content analyses. Eleven of the 88 microsatellite primer pairs showed evident polymorphisms repeatedly, and the polymorphic frequencies were 3.4-11.3% between the mutants and Yuexiangzhan. The polymorphic markers were randomly located on chromosomes. The more similar the plant types of the mutants like their original variety, the less polymorphic loci were detected. In addition, there was variation in the soluble protein contents among the leaves of mutants, and the contents were significantly lower than those of the original variety.

Inhibitory Function of Tregs via Soluble FGL2 in Chronic Hepatitis B

CD4 + CD25 + CD127 dim/regulatory T cells(Tregs) have been implicated in suppressing T cell immune responses to hepatitis B virus(HBV),but the inhibition mechanism has not being clear yet.This study investigated the effects of soluble FGL2(sFGL2) secreted by Tregs on immune suppression in chronic HBV-infected patients.We verified that sFGL2 protein and mRNA were highly expressed in Tregs.The separated Tregs by using magnetic beads from peripheral blood mononuclear cells(PBMCs) in 20 patients with chronic hepatitis B were co-cultured with PBMCs at a ratio of 1:3 with anti-CD3 stimulating antibody or FGL2 blocking antibody.The proliferation index of CD8 + T cells after blocking FGL2 was higher than that in blank group(3.58±0.18 vs.3.28±0.17,P=0.034) in 18 of 20 samples,and lower than that in CD3 stimulation group(3.82±0.19,P=0.026) in 16 of 20 samples.The IFN-γ secreted in the mixed culture in the absence of Tregs was higher than that in the culture in the presence of Tregs,but it could be abolished by FGL2 blocking antibody.These results suggest that sFGL2 protein secreted by Tregs suppresses the proliferation and function of CD8 + T cells in chronic hepatitis B.

Comparison of Some Characteristics Between Phragmites communis and Its Salt Tolerant Variant

R5002_12, a salt tolerant line of Phragmites communis Trin., which was obtained from ethyl methane sulfonate (EMS) treated callus selected under saline stress, was compared with its wild line in respect to their molecular biological, physiological and biochemical characterizations. Five arbitrary primers were screened which showed differences in DNA amplified polymorphism between the variant and its wild line. Some new proteins appeared in the salt tolerant plant under salt stress. Electrophoresis of peroxidase and esterase also showed some differences in isozyme expression between them. The chlorophyll content of the variant was higher than that of the original variety, whether the plants were under salt stress or not.

Effects of Polyamine on Leaf Physiological and Biochemical Indexes of Maize Seedlings under Osmotic Stress

[Objective]The research aimed to probe into the mechanism of polyamine enhancing the tolerance of plants to drought stress and provide theoretical basis for application of polyamine in process of maize drought resistance. [Method]With PEG-6000 simulating natural drought,the change in content of soluble protein and relative water content were investigated in seedling leaves of two maize cultivars,Nongda 108 and Yedan 13 under osmotic stress with exogenous Spd treatment. [Result]On the 7th day,leaf relative water content and the content of soluble protein decreased more significantly in leaves of Yedan 13 （drought-sensitive） than in Nongda 108 （drought-tolerant）. Exogenous Spd treatment not only obviously inhibited the decrease of leaf relative water content,but also increased the content of soluble protein. [Conclusion]Exogenous Spd treatment could enhance the tolerance of maize seedlings to osmotic stress,via improving the content of soluble protein in seedling leaves.

Effects of Two Kinds of Heavy Metals Including Cr^(6+) and Pb^(2+) on Garlic Germination

[Objective] This study aimed to revealing the effects of two kinds of heavy metals including Cr6＋ and Pb2＋ on garlic germination.[Method] Root length,enzyme（SOD and CAT）activity and soluble protein content during garlic germination were determined under the stress of two kinds of heavy metals including Cr6＋ and Pb2＋.[Result] During garlic germination,there was an obvious inhibition on buds and seedlings when the concentration of heavy metals including Cr6＋ and Pb2＋ was more than 20 mg/L,while SOD and CAT had certain buffering ability to the heavy metal toxicity with low concentration.With the stress of Cr6＋ in low concentration,SOD activity increased with the increase of metal ion concentration,and then reached maximum when the treatment concentration was 50 mg/L,while CAT activity gradually increased when Cr6＋ concentration was lower than 20 mg/L.With the high concentration of Cr6＋（more than 50 mg/L）or the Pb2＋ stress,SOD and CAT activity continued to decrease with the increase of concentration of heavy metal ions.With various concentrations of heavy metal（Cr6＋ and Pb2＋）solution,soluble protein content showed the decreasing trend with time prolong,and the decreasing trend was obvious with the increase of concentration.Moreover,the tolerance of garlic on Cr6＋ toxicity was much higher than that on Pb2＋.[Conclusion] Two kinds of heavy metals including Cr6＋ and Pb2＋ have impacts on four factors mentioned above during garlic germination,and this study provides a theoretical basis for further discussion of the toxic mechanism of heavy metals on plants.

Effects of IBA on the rooting of branch cuttings of Chinese jujube (Zizyphus jujuba Mill.) and changes to nutrients and endogenous hormones

‘Zhongqiusucui’ jujube secondary shoots were treated with 3-indolebutyric acid(IBA) at three concentrations, 500, 1000 and 1500 mg/L. Results show that IBA could significantly enhance rooting and root characteristics of cuttings and were best with IBA at 1500 mg/L. In the rooting process, the formation of adventitious roots was related to the consumption and accumulation of nutrients(soluble sugars and proteins) and the changes in endogenous hormones in phloem, leaf tips and leaf bases. The rooting of cuttings had a positive correlation with the consumption of soluble sugars during the period of callus formation and with the accumulation of soluble sugars during adventitious root formation and growth. Rooting was positively related to the breakdown of soluble proteins in the phloem when the callus formed, and had a positive correlation with its accumulation during adventitious root formation and growth. Leaf tips and leaf bases showed a reverse trend in changes of soluble protein. However, together with the phloem, leaf tips and leaf bases regulated and controlled the formation and development of adventitious roots. The main activities of soluble proteins exist in the leaf tips as this was the main source of soluble proteins. The relation between rooting and IAA(indole-3-acetic acid) content in phloem was positive and thus a high concentration of IAA could benefit the induction and formation of adventitious roots. However, rooting was negatively related with ABA(abscisic acid) and GA(gibberellic acid) and a high concentration of both could inhibit the induction and formation of adventitious roots. Rooting had a positive correlation with phloem IAA/ABA ratios, and higher ratios could improve rooting. Low concentrations of ZR(zeatin riboside) triggered the induction of adventitious roots, while higher concentrations promoted root growth.Endogenous hormones in leaf tips and bases had an impact on rooting. The activities of endogenous hormones mainly existed in leaf tips because they play a major role in the production and consumption of IAA and its ABA content increased during rooting. The ZR in leaf tips influenced the rooting of cuttings, especially in the callus formation and rooting stage. Leaf tips were the main source of GA.

Comparison of Biochemical Substances in the Fiber Development of Different Cotton Varieties

Zhongmian 42 and Xinluzao 36 were used as raw materials to determine the contents of soluble sugar and protein, as well as dynamic changes of enzyme activities after flowering during cotton fiber growth. The results showed that contents of soluble protein in the two species sharply declined 7 to 21 days after flowering, as the soluble sugar in Zhongmian 42 leveling off after 21 days of flowering while the soluble sugar in Xinluzao 36 dropped notably after 21 days of flowering before remaining stable after seven days later. The soluble sugar decreased 7 to 14 days after flowering before sharply rising to the maximum seven days later, and then began to decline quickly. The soluble sugar was the minimum after 35 days of flowering and then remaining stable. Peroxidase activity generally increased. Indole-3- acetic acid oxidase activities were low at 7 days after flowering. IAAO activity reached to the peaks on the 14th and 28th day after flowering. IAAO activity of two varieties decreased with the same trend 35 days after flowering.

Identification of Expression of CpTI Gene in Transgenic Poplars at Protein Level

The contents of total soluble protein and cowpea trypsin inhibitor (CpTI) in the browse and metaphylla of transgenic hybrid triploid poplars [ (Populus tomentosa×P.bolleana)×P.tomentosa ] transformed with CpTI gene were determined in order to study the products of CpTI gene expression at protein level. The results indicated that the amount of total soluble protein was greater in transgenic poplars than in non transgenic poplars, but was more in the metaphylla than in browse. The expression of CpTI gene resulted in an obvious increase in CpTI content, whereas CpTI was not detected in non transgenic poplars. It was found that there were high amount of total soluble protein and CpTI in 3 clones of TG07, TG04 and TG71 compared with other transgenic clones. In addition, the analysis of protein by SDS PAGE showed that a specific protein band of about 11.3?kD corresponding to the 80 amino acids encoded by the CpTI gene was observed in transgenic poplars on the gel of protein, which was not detected in non transgenic poplars.

Effect of Low Temperature Stress on Protein Content of Toona Sinensis Seedlings and Expression of Specific Protein

[Objective] The paper was to study the changes of protein content and component in leaves of Toona Sinensis under low-temperature stress.[Method] Potted experiment was adopted,and SDS-PAGE electrophoresis was also used to analyze the dynamic changes of protein in T.Sinensis leaves.[Result] Low temperature stress could change the content and component in leaves of T.Sinensis,but different provenances had different performance under different temperatures,so did the same provenance under different stress periods.SDS-PAGE electrophoresis results indicated that the change law of provenances of Xixia of Henan and Nanjing of Jiangsu with stronger cold resistance was similar,showing the change trend of "increase-decrease-increase".Protein was greatly expressed after stress for 1 d,the color of band became darker;the content gradually decreased in the following second and third day,and the color of bands was lighter;the content began to increase at the forth day.Two provenances induced the specific proteins with molecular weights of 27.6 and 22.5 kD,respectively.The soluble protein content of provenance of Xiapu of Fujian with relatively weak cold resistance was gradually increased,but no new protein bands were induced.The changes of protein band color of various provenances in SDS-PAGE electrophoresis was basically consistent with the changes of protein content,the provenance with stronger cold resistance could induce the production of specific proteins.[Conclusion] The study provided theoretical basis for under the molecular mechanism of plant cold resistance,which had great significance in theory and practice.

Effect of high salinity on cell growth and protein production of Antarctic ice microalgae Chlamydomonas sp. ICE-L

Antarctic ice microalgae Chlamydomonas sp.ICE-L can survive and thrive in Antarctic sea ice.In this study,Chlamydomonas sp.ICE-L could survive at the salinity of 132‰ NaCl.SDS-PAGE showed that the density of 2 bands（26 and 36 kD） decreased obviously at the salinity of 99‰ NaCl compared to at the salinity of 33‰ NaCl.The soluble proteins in Chlamydomonas sp.ICE-L grown under salinity of 33‰ and 99% NaCl were compared by 2-D gel electrophoresis.After shocking with high salinity,8 protein spots were found to disappear,and the density of 28 protein spots decreased.In addition,19 protein spots were enhanced or induced,including one new peptide（51 kD）.The changes of proteins might be correlated with the resistance for Chlamydomonas sp.ICE-L to high salinity.