Transcription factor CabHLH035 promotes cold resistance and homeostasis of reactive oxygen species in pepper

Plant basic helix-loop-helix(bHLH)transcription factors(TFs)play central roles in various abiotic stresses.However,its role in plant cold resistance is largely unknown.Previously,we characterised CaNAC035 in pepper,which positively regulates tolerance to cold,salt and drought stresses tolerance.Here,we identified CabHLH035,a CaNAC035-interacting protein in pepper.To explore its functions in cold stress tolerance,we silenced the gene in pepper via virus-induced gene silencing(VIGS)and overexpressed the gene in Arabidopsis.The results showed that CabHLH035 expression was induced by cold treatment,and silencing of CabHLH035 decreased cold stress tolerance.Conversely,overexpression of CabHLH035 in Arabidopsis increased cold stress tolerance.To investigate homologs genes of C-repeat binding factor(CBF)pathway proteins and reactive oxygen species(ROS)marker gene expression blocking by CabHLH035,we performed yeast one-hybrid(Y1H),dual luciferase and electrophoretic mobility shift assay experiments.The results showed that CabHLH035 bound to the region upstream of the CaCBF1A and CaAPX promoters.Additionally,CaCBF1A bound to the CaDHN4 promoter.Taken together,our results showed that CabHLH035 plays a crucial role in cold stress tolerance and its potential as a target for breeding cold-resistant crops.The findings provide a basis for studying the functions and regulatory network of cold stress tolerance in pepper.

Melatonin mitigates cold-induced damage to pepper seedlings by promoting redox homeostasis and regulating antioxidant profiling

This study assessed the influence of exogenous ME in the mitigation of cold damage in pepper seedlings. Melatonin(ME) is a dynamic molecule that helps plants cope with stress in several ways. Cold stress(CS) is one of the most important environmental factors that restrict plant growth and yield. Pepper(Capsicum annuum L.) is a valuable commercial crop, highly sensitive to CS. Thus, identifying an efficient strategy to mitigate cold damage is critical for long-term pepper production. For this purpose, the roots of pepper seedlings were pretreated with ME(5 μmol · L^(-1)) and exposed to CS for 7 d. The results indicated that CS suppressed pepper growth, hampered photosynthetic capacity, and damaged root architecture in pepper plants. In contrast, the production of reactive oxygen species(ROS), malondialdehyde(MDA), electrolyte leakage(EL), proline, and soluble sugars were enhanced in plants under CS. ME(5 μmol · L^(-1)) pretreatment reduced the negative effects of CS by recovering plant growth, root traits, gas exchange elements, and pigment molecules compared to CS control treatment. Furthermore, ME application efficiently reduced oxidative stress markers [hydrogen peroxide(H_(2)O_(2)), superoxide ion(O_(2)^(·-)), EL, and MDA] while increasing proline and soluble sugar content in pepper leaves. ME application combined with CS further increased antioxidant enzymes and related gene expression. Collectively, our results confirmed the mitigating potential of ME supplementation for CS by maintaining pepper seedling growth,improving the photosynthesis apparatus, regulating pigments, and osmolyte content.

Hydrolytic and Antimicrobial Activities of Bacteria Isolated from Fermented Peppers Sold in Markets at Brazzaville, Republic of the Congo

Microorganisms are omnipresent in all environments and play mainly the role of transformers, thanks to the multiple enzymes they are able to produce. In order to valorize fermented foods in the Republic of the Congo, this work aimed to characterize and study some properties of microorganisms isolated from samples of peppers sold in three markets of Brazzaville. A numeration of the total aerobic mesophilic flora (TAMF) was made in a solid medium, allowing the evaluation of each sample’s microbial concentration. The microbial mass varied from 2.8 × 105 CFU/g for the Ouénzé sample to 1.8 × 104 CFU/g for the Total sample and 2 × 104 CFU/g for the Moungali market sample. The evaluation of the enzymatic properties of the Bacillus isolates showed that 68.42% were capable of producing cellulases and 78.94% were capable of producing amylases and proteases. Antimicrobial activities revealed that 63.15% of the isolates were able to secrete inhibitory substances against E. coli and Staphylococcus aureus. Molecular analysis by PCR amplification, sequencing of the 16S rRNA gene and BLAST bioinformatics analysis provides newly identified bacteria strains with new accession numbers in GenBank: Bacillus thuringiensis MBCBR322 (OP474008), Bacillus megaterium MBCBJ1822 (OP476493), Bacillus thuringiensis MBCBR222 (OP476494), Priestia megaterium MBCBJ2022 (OP476495) and Lactobacillus paraplantarum MBCBR1522 (OP476496). Multiple sequences alignment of identified sequences with their homologs of GenBank has shown high similarities. The phylogenetic inference assay has provided the two groups of strains observed in this study, and the two groups are very coherent with the phylogeny of the reference.

Variety Selection and Green Cultivation Technology of Courtyard Ornamental and Edible Peppers

In order to comply with the development trend of the multifunctional use of peppers,we conducted an investigation into the characteristics and features of varieties,potting management techniques,and the methods of extending the fruit ornamental period and other aspects of courtyard ornamental and edible peppers.A set of cultivation techniques suitable for courtyard ornamental and edible peppers has been developed,including timely sowing and seedling,nutrient soil preparation,water and fertilizer management,trimming and pruning,preservation of flowers and fruits,green prevention and control of diseases and pests,harvesting,and so on.

Fine-mapping and transcriptome analysis of the photosensitive leaf-yellowing gene CaLY1 in pepper(Capsicum annuum L.)

Leaf color is directly related to altered photosynthesis.Hence,leaf yellowing mutants can be widely used for the researching plant physiology and functional genomes,for cultivating new varieties of popular horticultural plants,and for identifying hybrid purity(as markers).Here,we constructed a^(60)Co-γF_(2)population from the leaf-yellowing mutant R24 via radiation mutation with the inbred line WT21 of pepper.Genetic analysis showed that the leaf-yellowing of the mutant was controlled by a single recessive gene.By applying the Bulk Segregation Analysis and Kompetitive Allele Specific PCR markers,the leaf-yellowing gene CaLY1(Capsicum annuum Leaf yellow 1)was mapped on chromosome 9,SNP5791587-SNP6011215,with a size of 214.5 kb.One non-synonymous mutated gene Capana09g000166 was found in the interval.The gene encoded a Psb X,which is the core complex of PSⅡ.Transcriptome analysis further showed that 2301 differentially expressed genes were identified under shading treatment for 24 h in R24.The Gene Ontology enrichment pathways were related to photosynthesis light harvesting,cell wall,activity of quercetin 3-O-glucosyltransferase and flavonoid metabolic process,which likely regulate the response of pepper leaves to different light levels.Functional enrichment analysis indicated that the most abundant pathways were photosynthesis antenna proteins and metabolic.

A novel single-base mutation in CaSGR1 confers the stay-green phenotype in pepper

The stay-green trait is of considerable importance in extending the shelf life of green pepper fruit(Capsicum annuum L.)and in enhancing the appearance of ornamental plants.The study revealed the genetic and regulatory mechanisms of the stay-green trait in pepper,which will aid in the selection of ornamental pepper varieties.In this study,a pepper mutant with stay-green fruit named TNX348 was identified from a germplasm resource bank.Two segregating populations were constructed using the stay-green mutant TNX348 and then used in bulked segregant analysis combined with RNA sequencing and linkage analyses.The causal gene of the stay-green trait was mapped to an approximately 131-kb region,and a senescence-induced chloroplast protein gene,CaSGR1(Capana01g000359),was identified as a candidate gene.Sequencing analysis revealed a G→A single-base mutation of CaSGR1 in TNX348 that led to early termination of translation.Based on the single-base mutation,a single nucleotide polymorphism(SNP)marker co-segregating with the stay-green trait was developed.Furthermore,in transcriptome analysis,expression patterns of 11 hormone transduction-related transcription factors,such as abscisic acid-insensitive(ABI),abscisic acidresponsive element-binding factor(ABF),and NAC transcription factor,were similar or opposite to that of CaSGR1.The results indicated that the transcription factors might mediate chlorophyll degradation by regulating the expression of CaSGR1.

Red light induces salicylic acid accumulation by activating CaHY5 to enhance pepper resistance against Phytophthora capsici

Pepper(Capsicum annuum L.)is frequently challenged by various pathogens,among which Phytophthora capsici is the most devastating to pepper production.Red light signal acts as a positive induction of plant resistance against multiple pathogens.However,little is known about how the red light signal affects pepper resistance to P.capsici infection(PCI).Here,we report that red light regulates salicylic acid(SA)accumulation by activating elongated hypocotyl5(CaHY5),a basic leucine zipper(bZIP)transcription factor,thereby decreasing pepper susceptibility to PCI.Exogenous SA treatment reduced pepper susceptibility to PCI,while silencing of CaPHYB(a red light photoreceptor)increased its susceptibility.PCI significantly induced CaHY5 expression,and silencing of CaHY5 reduced SA accumulation,accompanied by decreases in the expression levels of phenylalanine ammonia-lyase 3(CaPAL3),CaPAL7,pathogenesis-related 1(CaPR1),and CaPR1L,which finally resulted in higher susceptibility of pepper to PCI.Moreover,CaHY5 was found to activate the expression of CaPAL3 and CaPAL7,which are essential for SA biosynthesis,by directly binding to their promoters.Further analysis revealed that exogenous SA treatment could restore the resistance of CaHY5-silenced pepper plants to PCI.Collectively,this study reveals a critical mechanism through which red light induces SA accumulation by regulating CaHY5-mediated CaPAL3 and CaPAL7 expression,leading to enhanced resistance to PCI.Moreover,red light-induced CaHY5 regulates pepper resistance to PCI,which may have implications for PCI control in protected vegetable production.

Microscopic and metabolic investigations disclose the factors that lead to skin cracking in chili-type pepper fruit varieties

The hydrophobic cuticle encasing the fruit skin surface plays critical roles during fruit development and post-harvest.Skin failure often results in the fruit surface cracking and forming a wound-periderm tissue made of suberin and lignin.The factors that make the fruit skin susceptible to cracking have yet to be fully understood.Herein,we investigated two varieties of chili peppers(Capsicum annuum L.),Numex Garnet,whose fruit has intact skin,and Vezena Slatka,whose fruit has cracked skin.Microscopical observations,gas chromatography-mass spectrometry,biochemical and gene expression assays revealed that Vezena Slatka fruit form a thicker cuticle with greater levels of cutin monomers and hydroxycinnamic acids,and highly express key cutin-related genes.The skin of these fruit also had a lower epidermal cell density due to cells with very large perimeters,and highly express genes involved in epidermal cell differentiation.We demonstrate that skin cracking in the Vezena Slatka fruit is accompanied by a spatial accumulation of lignin-like polyphenolic compounds,without the formation of a typical wound-periderm tissues made of suberized cells.Lastly,we establish that skin cracking in chili-type pepper significantly affects fruit quality during post-harvest storage in a temperature-dependent manner.In conclusion,our data highlight cuticle thickness and epidermal cell density as two critical factors determining fruit skin susceptibility to cracking in chili-type pepper fruit.

Effect of Dandelion(Taraxacum mongolicum Hand.-Mazz.)Intercropping with Different Plant Spacing on Blight and Growth of Pepper(Capsicum annuum L.)

Intercropping of crops that can secrete bacteriostatic active substances can not only inhibit the occurrence of disease but also have an important effect on plant growth.However,the effects of dandelion intercropping on pepper blight control and pepper growth remain unclear.In this study,the control effect of dandelion on pepper blight was studied by inoculating the pepper leaves with Phytophthora infestans,and it also discusses the correlation of the occurrence of pepper epidemic disease with the pepper canopy environment,soil environment,pepper photo-synthesis,and yield index.The results showed that best plant distance for dandelion intercropping was 20 cm(P20),and the control effect reached 43.31%.As compared to the CK,SOD enzyme,POD enzyme,and PAL enzyme were significantly up-regulated during the growth of pepper;chlorophyll content in pepper leaves was significantly increased;photosynthetic characteristics were significantly increased;stem diameter and yield of crop pepper were effectively improved;and the quality of the pepper product was better,but intercropping dandelion resulted in a significant decrease of nutrients in the soil environment of pepper,so a reasonable intercropping distance was needed.The correlation analysis shows that the incidence of pepper blight(A)was significantly positively correlated with soil temperature(Q),intercellular carbon dioxide(L),and canopy air temperature(O).The incidence of capsicum blight(A)was significantly negatively correlated with chlorophyll content(F),net photo-synthetic rate(K),stomatal conductance(M),ww rate(N),soil sucrase activity(W),vitamin C(AB),and leaf PAL enzyme(J).Finally,it was deduced that intercropping dandelion could effectively control the occurrence of pepper blight while also demonstrating a complex interaction with the pepper growing environment.

Effects of Polygonatum odoratum Polysaccharides on the Quality of White Chili Peppers in Jars

[Objectives]This study was conducted to investigate the effects of Polygonatum odoratum polysaccharides on the quality of white chili peppers in jars.[Methods]White chili peppers were pickled by adding different concentrations of P.odoratum polysaccharides with traditional pickling technology,and its nitrite content,pH and sensory quality were analyzed and detected.[Results]The results showed that,compared with the control group without adding P.odoratum polysaccharides,the nitrite content in white chili peppers in jars decreased after adding P.odoratum polysaccharides.When the addition amount of P.odoratum polysaccharides was 0.002%,the inhibition rate of nitrite reached the maximum.Under the same fermentation time,P.odoratum polysaccharides could significantly reduce the pH value of pickles.When the addition amount of P.odoratum polysaccharide was 0.002%and the pickling time was 3 months,the sensory quality of white chili peppers in jars was the best.At this point,white chili peppers in jars had rich aroma and tasted soft and sour,and the sour and salinity were suitable.As the fermentation time continued to increase,the chili aroma of white chili peppers in jars gradually decreased,and the sour taste became more pronounced.[Conclusions]This study can provide reference for the safety and quality control of white chili peppers in jars.

Effect of Slow and Controlled Release Fertilizers on the Yield and Nutrient Use Efficiency of Hot Pepper in Qiu-bei

[Objective] The aim of this study was to investigate the impacts of slow and controlled release fertilizers（SCRF）on the yield of qiubei hot pepper,its nutrient use efficiency and environment.[Method] Using Qiubei hot pepper（Capsicum frutescens L.）as the experimental material,we studied the fertilization effect and environment-protecting effect of SCRF.[Result] The result showed that SCRF could improve the agronomic characteristics of hot pepper.Compared to singly applied common fertilizers,SCRF increased economic yield by 20.90% and economic benefit by 13 234.35 Yuan/hm2,and the ratio of output to input was improved by 47.93%.In comparison with common straight fertilizers at same NPK proportion and rate,SCRF increased economic yield by 5.26% and economic benefit by 5 554.80 Yuan/hm2,and the ratio of output to input was improved by 9.91%.Under the reduced use of SCRF by 20%,SCRF increased economic yield by 12.38% and economic benefit by 9595.20 Yuan/hm2 compared with singly applied common fertilizers,and the ratio of output to input was improved by 65.95%.SCRF improved nitrogen,phosphorus and potassium use efficiencies by 12.42-17.53,3.35-5.24 and 5.37-14.02 percents respectively.[Conclusion] As the result of much reduced N and P application rates,SCRF would significantly economize fertilizer resources and minimize the pollution caused by the loss of fertilizer nutrients,which is of practical importance for environment protection.

AFLP Analysis of Cytoplasmic Male-sterile Line and Its Maintainer Line of Pepper

[Objective] The aim of this study was to analyze the cytoplasmic male sterile line 21A and its maintainer line 21B of peppers by AFLP,and lay the foundation for further studies on molecular mechanism of the cytoplasmic male sterility in peppers.[Method] Cytoplasmic male sterility(CMS)line 21A and its maintainer line 21B were analyzed by AFLP to obtain the specific amplified fragments of cytoplasmic male sterile line 21A,while the specific amplified fragments were recovered or sequenced,and analyzed by BLAST...

Screening of SSR Core Primers for Purity Identification of Pepper(Capsicum) Hybrids

[Objective] This study aimed to screen a set of SSR core primers suitable for purity identification of pepper （Capsicum） hybrids. [Method] DNA fingerprint of 100 pepper hybrids was analyzed using 17 SSR primers. [Result] According to the polymorphism and heterozygosity, Hpms1-214, Es395 and Hpmsl-5 were determined as three preferred core primers for purity identification of pepper hybrids. By using these three preferred core primers, 97 pepper hybrids （accounting for 97%） had heterozygous band pattern with at least one primer. Es330, Es363, Epms923, Es120 and Es64 were determined as candidate core primers for purity identification of pepper hybrids. Specific primers of 14 varieties were obtained, which could be used to further screen parent-complementary primers of each pepper hybrid. [Con- clusion] This study laid the foundation for constructing standard DNA fingerprints for purity identification of pepper hybrids.

Screening of Low Temperature Resistance Germplasm Resources of Pigment Pepper

Objective The aim of this study was to explore the method and standard for rapidly screening low temperature-resistant pepper germplasm resources and provide a theoretical basis for the breeding of low temperature-resistant pepper. [ Method ] With 110 pigment pepper seeds as the materials, their germination vigor under optimum temperature and suboptimal temperature were determined by means of roll rapid germination, and seeds with different genetic types were evaluated from aspects of germination vigor and its interval division. [ Result ] 37 pepper seeds with stronger low temperature resistance were screened. [ Conclusion]This study provides an important basis for screening low temperature-resistant pepper germplasm resources.

Current Situation and Countermeasures of Breeding for Processing Pepper in China

Research progress on the heterosis, cytoplasmic male sterility and biotechnology breeding of processing pepper （Capsicum annuum L.） was introduced, and the problems of limited germplasm resources and varieties of processing pep- per, large gap with the foreign companies were pointed out. Some suggestions were proposed on strengthening innovation and collection of germplasm resources, breed- ing processing pepper varieties, enhancing breeding level of processing pepper and its market competitiveness.

Discussion on AFLP Molecular Markers in Piper methysticum and Pepper

The aim of the research was to discuss the genetic relationships between Piper methysticum, Pepper and other wild species in Pepper genus. DNA was extracted from leaves which belonged to 28 germplasms including 6 materials of P. methysticum, 21 maerials of cultivated and wild Pepper, 1 material of Peperomia pellucida belonged to different genus. Premiers with good band-type and high polymorphism and resolution were selected from 64 pairs of primers for AFLP amplification and the clustering analysis was conducted with MVSP3.13f software. 191 bands were amplified by 4 pairs of premiers, 189 of which had polymorphism, being 98.6%. 28 germplasms were classified into 6 different groups at the genetic similarity coefficient of 0.52 by silver staining AFLP, in which 6 materials of Piper methysticum were clustered into a single group, indicating that P. methysticum belonged to Pepper family of Pepper genus but were distantly related to the others. The research provided the basis for selecting rootstocks for P. methysticum graft, molecular identification of P. methysticum and the fingerprint construction of P. methysticum.

Effects of Slow-release Fertilizer and Balanced Fertilization on Nitrogen, Phosphorus and Potassium Uptake in Peppers

Field plot experiment was conducted to study the effects of two slow-re- lease fertilizers and balanced fertilization on dry matter accumulation, yield, fertilizer use efficiency, nitrogen, phosphorus and potassium uptake of peppers at Jiangna Town, Yanshan County, Yunnan Province in 2011. The results showed that the dry matter accumulation in dried pepper plant, pepper yield, nitrogen, phosphorus, potassium uptake in peppers were significantly increased in all the fertilizer treat- ments, compared with those in control （no fertilizer）. Compared with conventional fertilization, balanced fertilization, slow-release compound fertilizer and slow-release urea fertilizer significantly increased dried pepper economic output by 20.94%, 17.5% and 14.54%, nitrogen uptake in dried peppers by 21.53%,18.46% and 13.19%, phosphorus uptake in dried peppers by 14.08%, 15.76% and 10.44%, potassium uptake in dried peppers by 22.66%, 15.73% and 16.28%; they also in- creased nitrogen and potassium use efficiency, but reduced potassium use efficiency due to the increased potassium addition. In treatments with balanced fertilization, slow-release compound fertilizer and slow-release urea fertilizer, the nitrogen utiliza- tion was 5.84%, 7.14% and 8.33% higher and the phosphorus utilization was 3.32%, 3.27% and 2.47% higher than those in treatment with conventional fertiliza- tion. In addition, the nitrogen application could be reduced by 20%-50% by bal- anced fertilization and the two slow-release fertilizers, thereby reducing environmen- tal pollution. Slow-release fertilizers could also reduce the frequency of fertilization and labor costs.

In vitro Regeneration Culture of Pepper and Its Application in Breeding

This paper summarized the technology of haploid production, protoplast culture, organ regeneration culture of pepper and the key factors affecting in vitro regeneration culture of pepper, including explants, seedling age, medium,genotype and plant growth regulator, then pointed out several main problems, in order to provide the reference for building an efficient in vitro regeneration culture system of pepper and its application in breeding.

Influences of Carbon Sources and Plant Growth Regulators on Anther Culture Efficiency of Pepper

[Objective]In order to increase anther culture efficiency of pepper.[Method]MS culture media and Bolajiaohong were used in this experiment to study the influences of carbon sources and concentrations on anther callus induction of pepper.Jiayu was taken as a material to study influences of plant growth regulators and concentrations on anther callus induction of pepper according to L16（4^5） orthogonal design.[Result]The average callus and embryoid induction rates of maltose at all concentrations were higher than these of sucrose but the difference was not significant.Taking maltose or sucrose as a carbon source,3% to 6% concentration was good for increasing induction frequencies of calli and embryoids.However,If the concentration was over 6%,the induction rates were declined dramatically with the increase of sugar concentration.The influences of growth regulators on induction rate of calli were listed as 2,4-D﹥ZT﹥NAA﹥KT﹥6-BA;the influences on induction rates of embryoids were listed as 2,4-D﹥NAA﹥ZT﹥KT﹥6-BA.The 2,4-D,ZT,NAA and KT had signficant or extremely significant influences on induction rates of calli and embryoids.2,4-D,ZT at 1.0 mg/L and NNA,KT at 0.5 mg/L had the best effects.The influences of ZT on calli and embryoids were better than those of KT and 6-BA.1.0 mg/L 2,4-D ＋1.0 mg/L ZT ＋0.5 mg/L KT ＋0.5 mg/L 6-BA was the best regulator combination for induction culture of Jiayu anther.[Conclusion]The experiment provided research basis for anther culture of pepper.

Studies on Thermo-sensitivity of Ningxia Hot Pepper Male Sterile Line

[Objective] This study was conducted to identify the thermo-sensitivity of the male sterile line （NYB） of Ningxia hot pepper. [Method] The pol en abortion rates of pepper male sterile line under artificial y control ed temperature condition and nature temperature condition were measured and compared. [Result] The pol en abortion was the sign of NYB male sterility. The pol en abortion rate changed little under different temperature conditions, while pol en amount greatly reduced at low temperature, indicating that NYB was genetical y stable in field. [Conclusion] The re-sults can provide some theoretical references for the application of NYB pepper male sterile line in breeding work.