Analysis of genes specifically expressed under salt stress in salt-tolerant mutant of rice by using DDRT-PCR technique

Differential display reverse transcription-PCR (DDRT-PCR) technique was used to identify those genes that are expressed differentially between wild type rice variety 77-170 (Oryza Sativa vas Japonica) and its salt-tolerant mutant (M-20) under salt stress. Totally 13 salt-inducible cDNA fragments of 200-600 bp were identified and doned, and were designated as SIGR1 - SIGR13 (salt-induced gene in rice). Northern blot analysis showed that expression of SIGR6 and SIGR8 was salt-inducible in both wild type and mutant, and expression of SIGR12 in M-20 was much higher than that in 77-170 under salt stress. It was also shown that expression of SIGR3, SIGR4, SIGR7, SIGRIO and SIGR13 was salt-inducible, and the genes were highly homologous with Rab1d which was an ABA-inducible gene of rice. The great potential application of DDRT-PCR technique in plant molecular biology research may promote the investigation of expression of salt-induced protein in rice.

Enhanced H^+ transport activity of tonoplast vesicles isolated from roots of salt-tolerant mutant of wheat under NaCl stress

Both enhanced were H^+ transport activities of tonoplast vesicles isolated from roots of the salt-tolerant mutant and wild type of wheat with treatment of NaCI, but the activity of the mutant was significantly higher than that of wild type. H^+ transport activity was indicated as the stable value of fluorescence quenching per mg membrane proteins. The H^+ transport activities dependent on ATP of the mutant and wild type were 1099 and 558 respectively and their activities dependent on PPi were 358 and 228 separately.

Reduced Na^+ and K^+ permeability of K^+ channel in plasma membrane isolated from roots of salt-tolerant mutant of wheat

The Na^+ and K^+ permeability of K^+ channel in plasma membrane, isolated from roots of the salt-tolerant mutant of ,wheat, was lower than that of wild type in 100 mmol/L KCI and NaCI solution. The opening frequency of K^+ channel of the mutant reduced more significantly than that of wild type in two kinds of solution mentioned above. It is assumed that the reduction of opening frequency mainly contributes to the Na^+ and K^+ permeability of K^+ channel of the mutant. The electric conductance of single-channel of the mutant was similar to that of wild type and the main difference between them was exhibited as the opening frequency. Their K^+/Ka^+ selectivity of K^+ channel had no significant difference. The K^+/Na^+ selectivity of the mutant and wild type was 3.35 and 3. 18 respectively.

Decolorization of azo dyes with high salt concentration by salt-tolerant mixed cultures under anaerobic conditions

Because the lack of detailed study of biological decolorization in high salt dye wastewater, it is still difficult to evaluate the biological treatment on high-salinity dye wastewater. The experiments were carried out to study the salt-tolerant bacteria, which is useful in the treatment of high-salinity colored wastewater. Simulated wastewater containing 5-150 g/L salt （NaCI） and 50-2000 mg/L Reactive Brilliant Red K-2BP was treated with three salt-tolerant mixed cultures （CAS, TAS, DSAS）, which were under a gradually acclimated procedure. With the increase of concentrations of salt and dye, the decolorization became low. The abilities of decolorization of dyes wastewater by three mixed cultures （CAS, TAS, DSAS） were studied, CAS and DSAS mixed cultures showed more active for the treatment of high-salinity colored wastewater than TAS mixed cultures. The results suggested that there might be a simple process for the high salt wastewater treatment, which could be incorporated into conventional activated sludge plants.

Generation and analysis of expressed sequence tags from the salt-tolerant eelgrass species, Zostera marina

Zostera marina, a monocotyledonous angiosperm, is one of the most important seagrass species. To inves- tigate the salt-tolerance mechanism and discover salt-tolerant genes in Z. marina, a cDNA library was con- structed. Single-pass sequencing of the 5＇ ends of 4 081 clones yielded 4 002 high quality expressed sequence tags （ESTs）, which were assembled into 241 contigs and 1 673 singletons, representing 1 914 unigenes. The average length of the ESTs was 582 bp, with sizes ranging from 100-1 500 bp. Basic Local Alignment Search Tool （BLASTX） analysis revealed that 1 664 unigenes had significant homology to known genes in the Na- tional Center for Biotechnology Information （NCBI） non-redundant （nr） database （E-value≤5-10）. Among them, the two most abundant genes encoded metallothionein （157 ESTs） and chlorophyll a/b-binding pro- tein （38 ESTs）, accounting for 7.1% and 1.7% of the total ESTs, respectively. Using Kyoto Encyclopedia of Genes and Genomes （KEGG）, 1 462 unigenes were assigned to 1 161 pathways （E-value≤5-10）. A total of 938 unigenes were assigned Gene Ontology （GO） terms based on the GO hierarchy analysis, and InterProScan searches recognized 1 003 InterPro families. Three genes for metallothionein in Z. marina that belonged to Class II was identified. Results of this study will improve understanding of the molecular mechanisms of saline tolerance in Z. marina.

A novel salt-tolerant Micrococcus sp. DUT_AHX capable of degrading nitrobenzene

A novel strain of Micrococcus sp.DUT_AHX,which was isolated from the sludge of a nitrobenzene(NB)-manufacturing plant and could utilize NB as the sole carbon source,was identified on the basis of physiological and biochemical tests and 16S ribosomal DNA(rDNA)sequence analysis.It can grow at the temperature up to 40℃or in the presence of NaCl concentration up to 12 g/L in Luria-Bertani(LB)medium.The optimal degradation conditions are as follows:temperature 37℃,pH 7.0,and shaking speed 150 r/min.The strain involves a partial reductive pathway due to the release of ammonia and can also utilize 2-aminophenol as the sole carbon source.Furthermore,the enzyme activity tests show that crude extracts of NB-grown strain DUT_AHX mainly contain 2-aminophenol 1,6-dioxygenase activity.The exploitation of salt-tolerant bacteria will be a remarkable improvement in NB bioremediation and wastewater treatment at high salinity and high temperature.

Salt-tolerant mechanism of marine Aspergillus niger cellulase cocktail and improvement of its activity

The cellulase cocktail produced by marine Aspergillus niger exhibits a property of salt-tolerance,which is of great potential in cellulose degradation in high salt environment.In order to explain the mechanism on the salttolerance of the cellulase cocktail produced by marine A.niger,six cellulase components(AnCel6,AnCel7A,AnCel7B,AnEGL,AnBGL1 and AnBGL2)were obtained by directed expression.Studies on their enzymatic properties revealed that oneβ-glucosidase(AnBGL2)and one endoglucanase(AnEGL)exhibited an outstanding salttolerant property,and one cellobiohydrolase(AnCel7B)exhibited a certain salt-tolerant property.Subsequent study revealed that the salt-tolerant An EGL and AnCel7B endowed the cellulase cocktail with stronger salttolerant property,while the salt-tolerant An BGL2 had no positive effect.Moreover,after overexpression of AnCel6,AnCel7A,AnCel7B and AnEGL,the activity of cellulase cocktail increased by 80%,70%,63%and 68%,respectively.However,the activity of cellulase cocktail was not improved after overexpression of AnBGL1 and AnBGL2.After mixed-strain fermentation with cellobiohydrolase recombinants(cel6 a,cel7a and cel7b recombinants)and endoglucanase recombinant(egl recombinant),the the activity of cellulase cocktail increased by 114%,102%and91%,respectively.

Rheological properties of salt-tolerant HPAM solutions with ultrahigh molecular weight

The rheological properties of salt-tolerant partially hydrolyzed polyacrylamide(HPAM)solutions with molecular of 2.5×107 g/mol at different concentrations were measured in steady-state shear flow mode by Haake Rheostress 150 rheometer.Three constitutive equations(Oldroyd four constant model,Guesekus model and FENE-P model) were used for describing the apparent viscosity and first normal stress difference.The apparent viscosity of salt-tolerant HPAM solutions appears a first Newtonian zone when the shear rate is approximately lower than 0.2 s-1.At high shear rate,the HPAM solutions show shear-thinning and elasticity.The results show that the FENE-P model has the best agreement between theoretical and experimental data within the available shear rate range.The material parameters are useful for numerical analysis of polymer solution flow fields.

High-yield Cultivation Technique of Salt-tolerant Spartina and Rice in Coastal Mudflat

Directing at the characteristics of coastal mudfiat saline and alkaline land, the yield of salt-tolerant Spartina and rice could reach 5 925-8 280 kg/hm^2 by the techniques of land and water resource utilization, farming improvement, construction of matched water system, seed selection and treatment, water direct seeding, seedling raising on seedbed, field transplanting, weeding, nutrient and water management, and disease and pest control, providing a scientific basis for the cultivation of Spartina and Rice in coast mudflat.

Introduction and Selection of Salt-tolerant Wheat in Yellow River Delta

[Objective] This study was to screen a salt-tolerant wheat variety in Dongying, a city in the center of the Yellow River Delta with a large area of coastal saline soil. [Method] Total 9 salt-tolerant, stress-resistant and high-yielding wheat varieties （lines） were introduced, and they were cultivated in the saline soil with total salt content of 3-4 g/kg with Dekang 961 as the control. [Result] The yields of Jinan 18, Yanjian 14 and Shanrong 3 were all significantly higher than that of Dekang 961 （P〈0.05）. These three varieties （lines） all ripen before June 13 with moderate growth period that does not affect the seeding of next-season crop. [Conclusion] Jinan 18, Yanjian 14 and Shanrong 3 are suitable for planting in light and median saline soil in the Yellow River Delta.

Response of Salt-Tolerant Rice Varieties to Biocompost Application in Sodic Soil of Eastern Uttar Pradesh

Sodic soils have immense productivity potential, if managed through proper technology interventions. Biocompost is prepared by composting pressmud (a sugar industry byproduct) received from cane juice filtration and spent wash received from distilleries through microbial aerobic decomposition and can be used to reclaim sodic soils. Field experiments were conducted during the wet season of 2011 and 2012 to study the effect of incorporation of biocompost in sodic soil with four treatments: T1—Control, T2—Biocompost at 2 t ha-1, T3—Biocompost at 4 t ha-1 and T4—Biocompost at 6 t ha-1. The two promising salt tolerant rice varieties preferred by farmers, Narendra usar 3 and NDR 359 were used as test crops, which can produce yields ranging between 2-4 t ha-1 in soil having a pH range of 9.2 to 10.5. Among the different doses of biocompost tested, application of biocompost at 6 t ha-1 registered highest yields, enabled by a higher biomass, ear bearing tiller (EBT), and grain fertility in both varieties. Narendra usar 3 was more responsive to treatments even at lower doses of biocompost than NDR 359, but NDR 359 yielded slightly higher than Narendra usar 3 in all treatments. Soil health was also improved evidently on better fertility and low soil pH and EC at harvest. Thus, biocompost can be considered as a commercially viable, environmentally acceptable and practically enforceable option for improving the crop productivity and soil fertility status.

Analysis of nutrient content and assessment of economic value for major salt-tolerant plants in coastal areas:a case study of Dongying City, Shandong Province

Salinization of soil is a worldwide problem concerning resources and ecology,especially serious in coastal areas.Testing of 26 sorts of data or parameters are carried out on leaves of 22 plant species(in 24 plant variety) of existing main salt-tolerant plant of the Yellow River Delta region.Data or parameters include the following ele-ments:contents of K+,Na+,Ca2+,Mg2+ and Cl-,contents of protein,fat,total energy,ash and contents of 17 amino acids.The results show that these tested plants have economic values.For example,according to their uses,they can be divided into edible plants,forage plants,medicine or health plants,and some of them can be used for multipurposes.These plants have played important roles in the sustainable utilization of plant resources in coastal areas.This paper has taken evaluations on the economic uses of salt-tolerant plants and given suggestions for saline soil improvement and resource utilization in coastal areas.Based on the results of investigation and experiments,we suppose that the salt-tolerant plants in coastal areas can be grouped into 9 main groups according to their economic value:pioneer plants for saline land improvement,medicine,edible and forage plants,industry material,forestation,breeding material,energy plants and eco-tourism resources.

Chemical Composition and Ruminal Digestibility of Ceratoides, a Salt-tolerant Perennial Plant,at Different Years

It has been proposed that salt-tolerant plant could be used as a feed resource for ruminants whereby salt would be removed from salinized land (Asian -Aust. J. Anim. Sci. (2002) 15:998 -1001). Ceratoides arborescens (Losinsk.) Tsien et C. G. Ma is known as a drought-and salt-tolerant plant,a kind of shrubs, growing in semi-arid land of Inner Mongolia. Because the covering effect of the perennial plant as a mulch over the soil might be expected, the optimum covering effect would be obtained after the growth period.The perennial plant produces seeds around summer and end its growth thereafter. Nutrient value of the perennial salt-tolerant plant, however,had not been reported in flowering period at different year. It is necessary to know the ruminal degradability of the plants of each growing year in order to determine the regimen to diet for ruminants. The present experiment,therefore,was undertaken to analyze the digestibility and chemical composition of Ceratoides arborescens as feed for ruminants.

A novel transcription factor FnMYB4 regulates pigments metabolism of yellow leaf mutants in Fragaria nilgerrensis

The strawberry species Fragaria nilgerrensis Schlechtendal ex J.Gay,renowned for its distinctive white,fragrant peach-like fruits and strong disease resistance,is an exceptional research material.In a previous study,an ethyl methane sulfonate(EMS)mutant library was established for this species,resulting in various yellow leaf mutants.Leaf yellowing materials are not only the ideal materials for basic studies on photosynthesis mechanism,chloroplast development,and molecular regulation of various pigments,but also have important utilization value in ornamental plants breeding.The present study focused on four distinct yellow leaf mutants:mottled yellow leaf(MO),yellow green leaf(YG),light green leaf(LG),and buddha light leaf(BU).The results revealed that the flavonoid content and carotenoid-to-chlorophyll ratio exhibited a significant increase among these mutants,while experiencing a significant decrease in chlorophyll and carotenoid contents compared to the wild type(WT).To clarify the regulatory mechanisms and network relationships underlying these mutants,the RNA-seq and weighted gene coexpression network(WGCNA)analyses were employed.The results showed flavonoid metabolism pathway was enriched both in MO and YG mutants,while the chlorophyll biosynthesis pathway and carotenoid degradation pathway were only enriched in MO and YG mutants,respectively.Subsequently,key structural genes and transcription factors were identified on metabolic pathways of three pigments through correlation analyses and quantitative experiments.Furthermore,a R2R3-MYB transcription factor,FnMYB4,was confirmed to be positively correlated with flavonoid synthesis through transient overexpression,virus-induced gene silencing(VIGS),and RNA interference(RNAi),accompanying by reoccurrence and attenuation of mutant phenotype.Finally,dual-luciferase(LUC)and yeast one-hybrid assays confirmed the binding of FnMYB4 to the FnFLS and FnF3H promoters,indicating that FnMYB4 positively regulates flavonoid synthesis.In addition,correlation analyses suggested that FnMYB4 also might be involved in chlorophyll and carotenoid metabolisms.These findings demonstrated the pivotal regulatory role of FnMYB4 in strawberry leaf coloration.

Detection of ARV-Resistant Mutants in HIV-1-Infected Individuals in a Context of Systematic Switching to an Association Based on Dolutegravir in Abidjan, Côte d’Ivoire

The emergence of antiretroviral resistance mutations represents a major threat to the achievement of national and global goals for the elimination of HIV-1 infection. The global strategy in 2019 in Cte d'Ivoire is a new national policy for the management of people living with HIV with the administration of dolutegravir (DTG)-based fixed-dose combination. The aim of our study was to evaluate HIV-1 resistance to antiretrovirals (ARVs) in infected adult subjects in Cte d’Ivoire in the context of a systematic switch to a DTG-based combination. Between February 2022 and October 2023, a cross-sectional survey with random sampling was conducted in 06 services caring for people living with HIV. A total of 139 participants were included in the study. Adults with a viral load ≥ 1000 copies/mL were tested for HIV-1 ARV resistance mutations. Molecular analyses were performed using protocol of ANRS-MIE (National Agency for Research on AIDS and emerging infectious diseases). The interpretation is performed by HIVGRAD (https://www.hiv-grade.de/cms/grade/). The frequencies of HIV-1 resistance to non-nucleotide reverse transcriptase inhibitors (NNRTIs), nucleotide reverse transcriptase inhibitors (NRTIs), integrase inhibitors (IINTs) and protease inhibitors (PIs) were 82%, 73%, 19% and 11% respectively. The main mutations observed in the different classes were K103N (45%), M184V (64%), E157Q (19%) and L10V/M46I/A71V/I54V (6%) respectively. This study reveals the emergence of resistance to DTG-based fixed-dose combinations, favored by high rates of resistance to NRTIs and NNRTIs. This finding underlines the need for enhanced viral load monitoring and HIV-1 genotyping tests to guide the choice of NRTIs for combination therapy. In addition, monitoring for mutations to second-generation NRTIs is essential, given the scale-up of DTG-based regimens currently underway in Cte d’Ivoire.

An efficient method for constructing a random insertional mutant library for forward genetics in Nannochloropsis oceanica

Insertional mutation,phenotypic evaluation,and mutated gene cloning are widely used to clone genes from scratch.Exogenous genes can be integrated into the genome during non-homologous end joining(NHEJ)of the double-strand breaks of DNA,causing insertional mutation.The random insertional mutant library constructed using this method has become a method of forward genetics for gene cloning.However,the establishment of a random insertional mutant library requires a high transformation efficiency of exogenous genes.Many microalgal species show a low transformation efficiency,making constructing random insertional mutant libraries difficult.In this study,we established a highly efficient transformation method for constructing a random insertional mutant library of Nannochloropsis oceanica,and tentatively tried to isolate its genes to prove the feasibility of the method.A gene that may control the growth rate and cell size was identified.This method will facilitate the genetic studies of N.oceanica,which should also be a reference for other microalgal species.

Roles of Mutant TP53 Gene in Cancer Development and Progression

TP53 is a tumor suppressor gene that is mutated in most cancer types and has been extensively studied in cancer research.p53 plays a critical role in regulating the expression of target genes and is involved in key processes such as apoptosis,cell cycle regulation,and genomic stability,earning it the title“guardian of the genome.”Numerous studies have demonstrated p53’s influence on and regulation of autophagy,ferroptosis,the tumor microenvironment,and cell metabolism,all of which contribute to tumor suppression.Alterations in p53,specifically mutant p53(mutp53),not only impair its tumor-suppressing functions but also enhance oncogenic characteristics.Recent data indicate that mutp53 is strongly associated with poor prognosis and advanced cancers,making it an ideal target for the development of novel cancer therapies.This review summarizes the post-translational modifications of p53,the mechanisms of mutp53 accumulation,and its gain-of-function,based on previous findings.Additionally,this review discusses its impact on metabolic homeostasis,ferroptosis,genomic instability,the tumor microenvironment,and cancer stem cells,and highlights recent advancements in mutp53 research.

搭载实践十号卫星的不同小麦品种突变体鉴定与分析

为探讨卫星搭载对小麦农艺性状、高分子量麦谷蛋白亚基(HMW-GS)和分子水平变异的影响,本研究利用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)技术、简单重复序列(SSR)标记技术对实践十号返回式科学实验卫星搭载周麦18、周麦22和汶农14小麦品种干种子的SP3代农艺性状变异和SP5代突变系的HMW-GS和SSR分子标记多态性进行鉴定与分析。结果发现,航天诱变SP3代农艺性状与其野生型存在明显差异,且不同小麦品种对太空环境的敏感性不同。航天搭载能够诱发高分子量麦谷蛋白亚基和基因组DNA发生变异,发现3个小麦品种高分子量麦谷蛋白亚基的变异频率分别为2.15%、3.66%和5.21%,其中汶农14突变体HMW-GS亚基变异频率最高。21个SSR分子标记检测结果显示,周麦18和周麦22的航天诱变突变体与其野生型差异标记数量均不超过2个,而汶农14航天诱变的部分突变体与其野生型间差异标记数量较多,且株高和穗型等性状发生了遗传分离。综上,航天诱变使小麦基因组和蛋白组发生变异,所创制的优异特色突变体可为小麦育种的遗传改良和功能基因的研究利用提供丰富的材料基础。

Antioxidase System of a Rice Mutant with Low Chlorophyll b

[Objective] The mitigative effect of antioxidase system of a rice mutant with low chlorophyll b on photooxidative damage was studied.[Method] A rice mutant with low chlorophyll b and its wild type were taken as experimental materials to comparatively research their peroxide (H2O2) contents, the activity and isozymes of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) in chloroplast.[Result] Compared with the wild type, there were many kinds of SOD, POD and CAT isozymes in leaf cells and chloroplast cell of mutant, and the activity of SOD, POD and CAT isozymes in leaf cells and chloroplast cell of mutant was also correspondingly higher. Under intense light condition, the H2O2 content of chloroplast in mutant was less than that in the wild type. [Conclusion] The higher activity of scavenging active oxygen can relieve the photooxidative damage made by excessive light energy of intense light on photosynthetic membrane, which is an important reason for higher photosystem Ⅱ (PS II) stability of this mutant.

Genetic Analysis of a Small Grain Mutant Induced by T-DNA Insertion in Rice

[Objective] The aim of this study is to understand the genetic characteristics of a grain shape mutant and its possible role in genetic improvement of grain yield in rice. [Method] On the basis of the collection of T-DNA tag lines, the progeny of homozygous plants carrying T-DNA insertion were screened for mutants with mutated phenotypes. The genetic analysis of the mutant and test for the linkage between the mutated phenotype and the T-DNA insertion were carried out to determine its genetic characteristics. [Result] In the present study, a grain shape mutant induced by T-DNA insertion in rice was identified, which showed small grain. Genetic analysis of the mutant showed that the two types of phenotype, normal and small grain in the segregating populations derived from the T-DNA heterozygotes, fit the ratio of 3∶1. Test for Basta resistance showed that all the mutants were resistant while the normal plants segregated for resistant and susceptible by the ratio of 2∶1. The results indicated that the mutant phenotype cosegregated with Bar gene. The small grain mutant caused by T-DNA insertion was confirmed by PCR amplification aiming at T-DNA. [Conclusion] The grain shape mutant is useful for isolation of the tagged gene and genetic improvement in rice.